作物杂志, 2018, 34(6): 10-16 doi: 10.16035/j.issn.1001-7283.2018.06.002

专题综述

马铃薯重要性状QTL定位及3个抗病性状分子标记辅助选育

王伟伟, 王洪洋, 刘晶, 梁静思, 李灿辉, 唐唯

云南师范大学马铃薯科学研究院,650500,云南昆明

Quantitative Trait Loci (QTL) Mapping and Three Resistance Traits Linkage Markers Selection in Potatoes

Wang Weiwei, Wang Hongyang, Liu Jing, Liang Jingsi, Li Canhui, Tang Wei

Joint Academy of Potato Science, Yunnan Normal University, Kunming 650500, Yunnan, China

通讯作者: 唐唯,讲师,研究方向为马铃薯晚疫病致病机制及防治

收稿日期: 2018-05-27   修回日期: 2018-08-19   网络出版日期: 2018-12-15

基金资助: 国家自然科学基金.  31660503
云南省应用基础研究计划青年项目.  2015FD015
全国大学生创新创业计划项目.  201710681007

Received: 2018-05-27   Revised: 2018-08-19   Online: 2018-12-15

作者简介 About authors

王伟伟,硕士研究生,研究方向为马铃薯对晚疫病的抗性机制 。

摘要

马铃薯是我国主要粮食作物之一,马铃薯分子育种研究具有重要意义。在二倍体马铃薯中,重要性状控制基因的QTL定位及克隆已经有大量报道。近年,随着同源四倍体分析软件的开发,四倍体马铃薯的遗传图谱构建和QTL定位也取得了突破性进展。分子标记是马铃薯育种的重要辅助手段,可快速准确筛选出多个优良性状。对马铃薯重要农艺性状的QTL定位和克隆,以及3个抗病性状分子标记辅助选育的研究进展进行概述,为加快马铃薯分子育种研究提供参考和实践依据。

关键词: 马铃薯 ; QTL ; 分子标记 ; 育种

Abstract

Potato is one of the main food crops in China, and research on potato molecular breeding research is of great significance. In the diploid potato, QTL mapping and cloning of important genes has been reported extensively; The development of tetraploid linked analysis software, genetic map construction in tetraploid potato and QTL mapping have also made breakthrough progress in recent years. Meanwhile, molecular markers are an important supplement method for potato breeding and it can quickly and accurately screen out multiple good traits. In this paper, the progress of the QTL mapping, cloning of important agronomic traits in potato and 3 resistance traits linkage markers breeding were summarized, to provid a reference and practical basis for accelerating potato molecular breeding.

Keywords: Potato ; QTL ; Molecular marker ; Breeding

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本文引用格式

王伟伟, 王洪洋, 刘晶, 梁静思, 李灿辉, 唐唯. 马铃薯重要性状QTL定位及3个抗病性状分子标记辅助选育[J]. 作物杂志, 2018, 34(6): 10-16 doi:10.16035/j.issn.1001-7283.2018.06.002

Wang Weiwei, Wang Hongyang, Liu Jing, Liang Jingsi, Li Canhui, Tang Wei. Quantitative Trait Loci (QTL) Mapping and Three Resistance Traits Linkage Markers Selection in Potatoes[J]. Crops, 2018, 34(6): 10-16 doi:10.16035/j.issn.1001-7283.2018.06.002

栽培种马铃薯(Solanum tuberosum L. 2n=4x=48)为茄科(Solanaceae)茄属(Solanum)一年生双子叶草本植物,1811年被引入欧洲大陆种植[1]。马铃薯引入我国的时间目前尚无定论,但在明代已有文字记载[2]。到了19世纪中叶,随着马铃薯传入我国并广泛种植,在粮食自给自足和农业经济中起着举足轻重的作用。2015年随着马铃薯主粮化战略的提出,我国对马铃薯的需求空前加大,但近年来,由于全球气候变暖、大气污染、种植方式的多样化,导致生物和非生物逆境胁迫增加,严重影响马铃薯产业的健康发展,因此急需新的育种资源和育种思路。高寒山区野生型地方品种的利用,优良性状的基因定位及在育种后代群体中的永久固定,以及提高育种后代的选择效率将成为解决上述逆境胁迫因素的重要手段。目前,全世界栽培的四倍体马铃薯中有99.5%是Solanum tuberosum[3],由于同源四倍体分离特点,传统马铃薯育种采用的杂交方式既耗费时间又耗费大量人力和物力。近年来,马铃薯全基因组测序拼接完成[4],使得育种家利用遗传学、分子生物学和基因组学来加快品种选育成为重要手段。马铃薯分子育种包括主要优良性状目标基因定位与克隆、分子标记辅助筛选和转基因3个方面。本文针对马铃薯优良性状的定位与克隆及分子标记辅助选育研究进展进行概述,为马铃薯育种研究提供理论和实践依据。

1 主要性状目标基因的QTL定位与克隆

1.1 四倍体马铃薯遗传图谱构建及重要性状QTL定位

目前,绝大部分马铃薯栽培种为普通栽培种(Solanum tuberosum ssp. tuberosum),为高度杂合的同源四倍体种,既有2/2式的均衡分离方式,即产生2个二价体;也存在3/1式的不均衡分离方式,即产生1个三价体和1个单价体(其中三价体和单价体配子是不育的);另外也可能出现4条同源染色体不分离,只产生1个四价体的情况。多数遗传作图和QTL定位软件均基于二倍体连锁分析模式,并不完全适用于四倍体,但也有相关研究报道。1998年利用二倍体分析软件JoinMap在四倍体马铃薯Ⅷ染色体发现1个复等位基因座对晚疫病抗性显著连锁,这在以往二倍体马铃薯的QTL定位中并没有发现[5];2014年利用JoinMap 4.0和MapQTL 4.0软件构建了长度593cM的遗传图谱,平均间距11.3cM,定位了11个和单株产量、结薯数、淀粉含量重要性状相关的QTLs[6]。2001年,同源四倍体遗传图谱构建方法被公布[7],基于上述方法,同源四倍体分析软件TetraploidMap第1版在2003年公布[8],但设计环境为LINUX,并且没有考虑四价体及双减数问题[9],2007年TetraploidMap公布Windows版本并修正上述问题[10],该软件为同源四倍体马铃薯的遗传图谱构建和QTLs定位提供了重要技术突破。如2014-2016年在四倍体水平上,利用AFLP、SNP和SSR的混合标记构建双亲本遗传图谱并利用区间作图(Interval Mapping)方法定位了试管薯块茎形成、花青素积累、抗晚疫病及抗病毒病等重要农艺性状的QTLs[11,12,13,14]

1.2 二倍体马铃薯重要农艺性状的QTL定位及相关基因的克隆

QTL(quantitative trait locus)是数量性状基因座,QTL定位是指确定控制数量性状的基因在基因组中的位置。定位的基础是遗传图谱的构建,利用遗传图谱可以找到与控制目的性状基因连锁的标记,图谱密度一般要小于10cM。在四倍体作图软件开发出来之前,国内外所报道的马铃薯遗传图谱大多数是基于二倍体野生种杂交分离群体完成的[15]。1988年,利用番茄的显性RFLP标记,报道了马铃薯第一张遗传图谱[16];1989年利用S. tubersum回交群体,构建了690cM的二倍体遗传图谱[17];以显性标记AFLP完成了马铃薯遗传图谱[18,19]。以基因座上的共显性标记SSR,2003年在二倍体种的3号和5号染色体上定位到了两个晚疫病抗性QTLs[20];2009年构建了遗传图谱并定位了30个抗晚疫病的QTLs[21];2010年研究构建了86个SSR标记、全长645cM的遗传图谱,并定位了3个重要性状QTLs[22];同时发现了晚疫病田间抗性和日照时间、生理小种显著相关QTL[23];2012年完成了BC31HP群体的810cM高密度遗传图谱,定位了6个晚疫病抗性的条件QTLs[24];2015年利用821对SSR引物构建了全长688cM、平均间距为4.38cM的高密度遗传图谱,并定位了1个与薯型相关的QTL LW04[25]。基于块茎形成,Schäfer-Pregl等[26]利用RFLP标记在二倍体群体中定位出8个有关产量的QTL和若干个控制块茎淀粉含量的QTL。花色素苷基因P编码类黄铜3′,5′-羟化酶合成紫色飞燕草色素,定位于第11染色体上[27];控制色素特异性表达基因定位于第10染色体上[28]。二氢黄酮醇4-还原酶(DFR)基因、类黄酮-3-O-葡萄糖基转移酶基因(3GT)、类黄酮-3′,5′-羟化酶(F3’,5’H)基因也被陆续克隆[29,30]。在抗虫方面,目前有17个马铃薯孢囊线虫抗性QTLs被定位到了7条染色体上[31]

2 DNA分子标记辅助育种

DNA分子标记技术利用PCR扩增,检测方法简单、快速、准确、可靠性高并且基因表达不受环境和季节影响。大多数分子标记都为共显性,能够很好鉴别出纯合基因与杂合基因来辅助育种,与通过表现型间接对基因型进行选择的传统育种方法比较,不受环境影响,不受等位基因显隐性干扰,结果可靠。此外,它对目标基因的检测可在育种早期进行选择,从而可以对起始材料进行筛选,大幅缩短了育种周期。

截至目前,国内外已有许多与马铃薯重要性状基因紧密连锁的标记被开发[32]。另外,分子标记在马铃薯种质资源鉴定及马铃薯遗传多样性研究中也发挥了巨大作用。

2.1 马铃薯重要性状基因及其标记的发掘

2.1.1 晚疫病 由致病疫霉(Phytophthora infestans)引起的马铃薯晚疫病是马铃薯生产上具有毁灭性的病害之一,是1845年爱尔兰大饥荒的罪魁祸首[33]。根据“基因对基因”假说[34],马铃薯栽培种的晚疫病抗性主要来源于主效抗病基因R1~R11以及其他一些广谱抗病基因。这些抗病基因主要来自S. demissum和S. bulbocastanum等野生种。如抗病基因R1的分子标记[32],R3的分子标记[35],R6、R7[36],主效抗病基因RB分子标记[37],R11[38]表1)。

表1   马铃薯主效抗晚疫病基因连锁标记

Table 1  Major resistance gene linkage markers to potato late blight

基因
Gene
染色体
Chromosome
来源Origin标记类型
Marker type
RB[37]8S. tuberosum×S. bulbocastanumPCR
Rpi-smiral[39]11S. tuberosumPCR
R1[40]5S. tuberosumAFLP
R2[41]4S. tuberosumPCR
R2[42]4S. tuberosumAFLP
R3[35]11S. tuberosumRFLP
R6, R7[36]11S. tuberosumRFLP
Rpi-ber[43]10S. berthaultiiESTs
R11[38]11S. tuberosumPCR
Rpi-blb1[44]8S. bulbocastanumPCR

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2.1.2 线虫 线虫种类较多,可以侵染马铃薯,最为常见的包括胞囊线虫和根结线虫。线虫侵染后,马铃薯块茎的大小及外观均发生畸变,引起减产。培育马铃薯线虫抗性品种,是目前防治线虫危害的较好方法之一。研究表明,抗马铃薯胞囊线虫的基因有17个已经被定位,其中4个高抗的垂直抗性基因:Gro1、Gpa2、H1和GroV1[31]。在马铃薯抗根结线虫的研究中,有7个连锁的分子标记被克隆或定位,分别是采用CAPs方法定位到的M39b和CT182[45],以及5个通过STS标记开发出的193I9、56F6、39E18、524K16、406L19[46]

2.1.3 马铃薯病毒 马铃薯病毒病是仅次于晚疫病的严重病害,目前发现的可以侵染马铃薯的病毒有近40种,最为严重的是马铃薯Y病毒和马铃薯卷叶病毒[47]。病毒是造成马铃薯品种资源退化的主要原因之一。目前抗病毒病基因及其连锁分子标记被陆续发现(表2)。

表2   马铃薯抗病毒病基因连锁标记

Table 2  Resistance gene linkage markers to potato virus

病毒Virus抗性基因Resistance gene来源Origin染色体Chromosome标记Mark
PVYRyadg[48]S. andigena11RYSC3/SCAR
PVYRyadg[48]S. andigena11RYSC4/SCAR
PVYRyadg[49]S. andigena11ADG2/CAPs
PVYRyadg[50]S. andigena11PYSC3/PCR based
PVYRysto[51]S. stoloniferum12YES3-3A/STS
PVYRysto[51]S. stoloniferum12YES3-3B/STS
PVYRysto[51]S. stoloniferum12SCARYSTO4/PCR based
PVYRy-fsto[52]S. stoloniferum12GP122718/CAPs
PVYRy-fsto[53]S. stoloniferum12GP122564/CAPs
PVYRychc[41]S. chacoense9Ry186/STS
PVYRychc[54]S. chacoense938-530/RAPD
PVYNytbr (HR)[55]S. tuberosum4TG506/ RFLP
PVYNy-1[56]S. tuberosum9Sldl1/CAPs
PVYNy-1[57]S. tuberosum9SC8951139/PCR based
PVYNy-2 (HR)[57]S. tuberosum11B11.6/CAPs
PLRVPLRV.1[58]S. chacoense11Nl271164/SCAR
PLRVPLRV.2[58]S. chacoense4GP76/SCAR
PLRVPLRV.3[58]S. chacoense5HM4-26/SCAR
PLRVPLRV.4[59]S. andigena11UB864600/SCAR
PLRVRlretb[60]S. tuberosum4、9DMB32-11/CAPs
PLRVRlretb[60]S. tuberosum4、91367-8a/CAPs
PLRVRlretb[60]S. tuberosum4、9C2-Atlg42990/CAPs
PLRVRladg[61]S. andigena5E35M48.192/AFLP
PLRVRladg[62]S. andigena5RGASC850/SCAR
PVANaadg (HR)[63]S. andigena11GP21/AFLP
PVSNs (HR)[53]S. andigena8SC811-260/CAPs
PVSNs[64]S. tuberosum8CP16/CAPs
PVMRm[65]S. megistacrolobum11GP283-320/CAPs
PVMRm[65]S. megistacrolobum11GP250-510/CAPs
PVMGm[65]S. gourlayi9SC878885/SCAR
PVXRx[66]S. tuberosum12GP34/CAPs
PVXRx1[67]S. andigena12RxSP/STS
PVXRx1[68]S. andigena12CP60/RFLP
PVXRx2[69]S. acaule5GP21/RFLP
PVXNbtbr (HR)[63]S. tuberosum5CT167/RFLP
PVXNb (HR)[63]S. tuberosum5SPUD237/CAPs
PVXNxphu (HR)[70]S. tuberosum9TG424/RELP

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2.2 胞质DNA作为分子标记用于马铃薯种质资源鉴定及亲缘关系分析

马铃薯叶绿体DNA(cpDNA)和线粒体DNA(mtDNA)结构简单、进化保守,其中cpDNA呈绝对的单亲母系遗传,mtDNA大多为单亲母系遗传。因此,cpDNA和mtDNA的多态性是揭示马铃薯遗传背景、追溯起源、研究马铃薯演化关系和育种亲本的重要分子标记。研究表明,马铃薯具有5种类型的cpDNA(T、W、A、C、S)和6种类型的mtDNA(α、β、γ、δ、ε、D),常用cpDNA/mtDNA的组合形式(如T/β、W/α、W/γ、A/ε等)表示不同的细胞质遗传背景[71,72]。其中T/β是普通栽培种(S. tuberosum ssp. tuberosum)细胞质的代表类型[73];D为墨西哥六倍体野生种(S. demissum)细胞质类型[72];W/α、W/γ在野生种中普遍存在,常用来表示野生种(除S. demissum外)细胞质的代表类型;同样,A/ε常用来表示安第斯栽培类群(Andigenum group)的细胞质类型[74]。Provan等[75]通过SSR分析比较了欧洲主栽马铃薯cpDNA和细胞核DNA(nDNA)的多态性,发现欧洲马铃薯遗传背景较为狭窄。Sukhotu等[76]利用RFLP标记分析了3 500份来源于马铃薯起源地安第斯山脉的品种中不同类型cpDNA的多态性,发现各品种间可能发生频繁的基因交流或具有相同的亲本来源。云南师范大学马铃薯科学研究院利用核基因组SSR标记和细胞质DNA标记,构建了上百份马铃薯种质资源的DNA分子指纹库[77],并制定了国家标准(马铃薯种薯真实性和纯度鉴定SSR分子标记,GB/T 28660-2012)。

3 小结和展望

目前,利用马铃薯的参考基因组在二倍体水平上已获得了多个重要农艺性状的QTLs,并完成了相关基因的克隆和功能验证。但同源四倍体马铃薯栽培种的基因组组装难度较大,尚处于起步阶段,且四倍体分析方法和软件不同于二倍体,目前还没有在多数马铃薯栽培种中运用到,因此,利用基因组注释研究、比较基因组研究开发基于同源四倍体的遗传作图软件是当前突破四倍体马铃薯遗传作图的几个主要解决手段。

马铃薯重要农艺性状分子标记是辅助育种的最有效手段之一,但目前开发的标记,首先并不完全是锚定性状决定基因的,很多标记都是和性状决定基因紧密连锁的,因此,对于多个数量性状决定基因,特别是具有上位效应的基因,标记和表型并不能一一对应;其次,很多重要性状标记是基于二倍体种开发的,在四倍体栽培种中的适应性还需要验证。总而言之,突破同源四倍体马铃薯重要性状的定位和克隆,以及开发高效的分子标记辅助育种,结合转基因和基因编辑技术,将在全基因组水平和传统遗传育种之间搭起一座桥梁,使得对现有马铃薯种质资源能够精准改造,大幅度提高马铃薯育种效率。

The authors have declared that no competing interests exist.
作者已声明无竞争性利益关系。

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Landrace potato cultivars are native to two areas in South America: the high Andes from eastern Venezuela to northern Argentina and the lowlands of south-central Chile. Potato first appeared outside of South America in Europe in 1567 and rapidly diffused worldwide. Two competing hypotheses suggested the origin of the "European" potato from the Andes or from lowland Chile, but the Andean origin has been widely accepted over the last 60 years. All modern potato cultivars predominantly have Chilean germplasm, explained as originating from breeding with Chilean landraces subsequent to the late blight epidemics beginning in 1845 in the UK. The Andean origin has been questioned recently through examination of landraces in India and the Canary Islands, but this evidence is inferential. Through a plastid DNA deletion marker from historical herbarium specimens, we report that the Andean potato predominated in the 1700s, but the Chilean potato was introduced into Europe as early as 1811 and became predominant long before the late blight epidemics in the UK. Our results provide the first direct evidence of these events and change the history of introduction of the European potato. They shed new light on the value of past breeding efforts to recreate the European potato from Andean forms and highlight the value of herbarium specimens in investigating origins of crop plants.

谷茂, 信乃俭 .

我国栽培马铃薯最早引进时间的辨析

中国农史, 1999(3):80-85.

[本文引用: 1]

Graham T, Guy H, Enrique C , et al.

Varietal change in potatoes in developing countries and the contribution of the International Potato Center:1972-2007.

International Potato Center (CIP), Social Sciences Working Paper, 2008.

[本文引用: 1]

Xu X, Pan S, Cheng S , et al.

Genome sequence and analysis of the tuber crop potato

Nature, 2011,475(7355):189-195.

DOI:10.1038/nature10158      URL     PMID:21743474      [本文引用: 1]

Abstract Potato (Solanum tuberosum L.) is the world's most important non-grain food crop and is central to global food security. It is clonally propagated, highly heterozygous, autotetraploid, and suffers acute inbreeding depression. Here we use a homozygous doubled-monoploid potato clone to sequence and assemble 86% of the 844-megabase genome. We predict 39,031 protein-coding genes and present evidence for at least two genome duplication events indicative of a palaeopolyploid origin. As the first genome sequence of an asterid, the potato genome reveals 2,642 genes specific to this large angiosperm clade. We also sequenced a heterozygous diploid clone and show that gene presence/absence variants and other potentially deleterious mutations occur frequently and are a likely cause of inbreeding depression. Gene family expansion, tissue-specific expression and recruitment of genes to new pathways contributed to the evolution of tuber development. The potato genome sequence provides a platform for genetic improvement of this vital crop. 2011 Macmillan Publishers Limited. All rights reserved

Meyer R C, Milbourne D, Hackett C A , et al.

Linkage analysis in tetraploid potato and association of markers with quantitative resistance to late blight (Phytophthora infestans)

Molecular and General Genetics, 1998,259(2):150-160.

DOI:10.1007/s004380050800      URL     PMID:9747706      [本文引用: 1]

We have constructed a partial linkage map in tetraploid potato which integrates simplex, duplex and double-simplex AFLP markers. The map consists of 231 maternal and 106 paternal markers with total map lengths of 990.965cM and 484.665cM. The longer of the two cumulative map lengths represents approximately 25% coverage of the genome. In tetraploids, much of the polymorphism between parental clones is masked by `dosage' which significantly reduces the number of individual markers that can be scored in a population. Consequently, the major advantage of using AFLPs – their high multiplex ratio – is reduced to the point where the use of alternative multi-allelic marker types would be significantly more efficient. The segregation data and map information have been used in a QTL analysis of late blight resistance, and a multi-allelic locus at the proximal end of chromosome VIII has been identified which contributes significantly to the expression of resistance. No late blight resistance genes or QTLs have previously been mapped to this location.

时启冬 .

四倍体马铃薯SSR遗传图谱的构建及若干性状的QTL定位分析

哈尔滨:东北农业大学, 2014.

URL     [本文引用: 1]

马铃薯(Solanum tuberosum L.)是世界第三大粮食作物。随着社会的发展和科技的进步,马铃薯育种已经从单一的高产育种到现在的多种用途专门培育的品质育种时代。马铃薯的工业加工对于马铃薯的品质性状的优质程度有着较高的要求,利用传统育种方法要消耗大量的时间,效率较低。当今分子生物学发展飞速,通过分子标记来构建遗传图谱对产量、品质性状进行QTL定位的辅助,将大大的提高育种效率,有效的加快对马铃薯优良品种的育成。 本研究以四倍体马铃薯87个F1代群体以及其父母本作为试验材料。利用SSR分子标记构建了一张遗传连锁图谱,对单株产量、淀粉含量等性状进行了QTL定位和分析。主要结论如下 1.利用Joinmap4.0对126对SSR标记进行遗传连锁分析,构建了一张包含有13个连锁群,覆盖11条染色体,并没有标记定位到第9号染色体上,56个SSR标记的四倍体马铃薯遗传连锁图谱。该图谱的总长度为593cm,覆盖率为80%左右,染色体覆盖基因组长度在2.4cm-92.1cm范围内,各个染色体上的标记数为2-10个,两标记之间的平均距离为11.3cm。本连锁遗传图谱间平均距离最大的为第7条染色体,平均距离为28.3cm;平均间距最小的为第1条染色体,平均距离为9.2cm。第二条染色体的标记数量最多,染色体上有10个标记,第3、第5和第8条染色体是最少的,分别有3个标记。 2.应用MapQTL4.0软件,利用区间作图法,对四倍体马铃薯的单株产量、单株结薯数量、商品薯率、干物质含量、淀粉含量、钾含量等性状进行QTL定位研究。控制马铃薯淀粉含量的QTL位点一共有3个:qSC-4、qSC-7和qSC-12分别位于第4条、第7条和第12条染色体上,LOD值为6.1、3.4和3.2,贡献度为36.1%、7%和12%。控制马铃薯单株结薯数量的QTL位点一共有2个:qTSET-2和qTSET-7分别位于第2和第7条染色体上,LOD值为2.7和3.2,贡献度为31%、36%。控制马铃薯单株产量的QTL位点一共有2个:qTY-2和qTY-11分别位于第2条和第4条染色体上,LOD值为2.7和9.1,贡献度为16%、47%。控制马铃薯商品薯率的QTL位点一共有1个:qCPR-2位于第2条染色体上,LOD值为5.7,贡献度为11%。控制马铃薯干物质含量的QTL位点一共有2个:qDMC-1-1和qDMC-1-2都位于第一条染色体上,LOD值为5.1和7.0,贡献度为36%、13%。控制马铃薯钾含量的QTL位点一共有1个:qPC-2。qPC-2位于第2跳染色体上,LOD值为4.0,贡献度为8.5% 3.本研究获得的全部11个QTL位点都为四倍体马铃薯的新QTL位点,部分前人二倍体马铃薯QTL定位有相同或邻近之处。本实验制作的图谱精细度只可用作初步的粗定位,但也给后续精细定位以及对以前定位工作的认证提供了可靠的证据

Luo Z W, Hackett C A, Bradshaw J E , et al.

Construction of a genetic linkage map in tetraploid species using molecular markers

Genetics, 2001,157(3):1369-1385.

DOI:10.1089/109065701750168806      URL     PMID:11238421      [本文引用: 1]

This article presents methodology for the construction of a linkage map in an autotetraploid species, using either codominant or dominant molecular markers scored on two parents and their full-sib progeny. The steps of the analysis are as follows: identification of parental genotypes from the parental and offspring phenotypes; testing for independent segregation of markers; partition of markers into linkage groups using cluster analysis; maximum-likelihood estimation of the phase, recombination frequency, and LOD score for all pairs of markers in the same linkage group using the EM algorithm; ordering the markers and estimating distances between them; and reconstructing their linkage phases. The information from different marker configurations about the recombination frequency is examined and found to vary considerably, depending on the number of different alleles, the number of alleles shared by the parents, and the phase of the markers. The methods are applied to a simulated data set and to a small set of SSR and AFLP markers scored in a full-sib population of tetraploid potato.

Hackett C A, Luo Z W .

Tetraploidmap:construction of a linkage map in autotetraploid species

Journal of Heredity, 2003,94(4):358-359.

DOI:10.1093/jhered/esg066      URL     PMID:12920109      [本文引用: 1]

TetraploidMap is a suite of Fortran 90 routines run from Microsoft Windows with a text-based input and output. TetraploidMap enables the user to assemble a linkage map from dominant and codominant (multiallelic) marker loci scored for the parents and full-sib progeny of a cross in an autotetraploid species. It includes routines for the inference of the parental genotypes, identification of linkage groups, two-point analysis to estimate the recombination frequency and LOD score between all pairs of marker in a linkage group, and locus ordering by simulated annealing.

Bourke P M, Voorrips R E, Visser R G , et al.

The double-reduction landscape in tetraploid potato as revealed by a high-density linkage map

Genetics, 2015,201(3):853-863.

DOI:10.1534/genetics.115.181008      URL     [本文引用: 1]

The creation of genetic linkage maps in polyploid species has been a long-standing problem for which various approaches have been proposed. In the case of autopolyploids, a commonly used simplification is that random bivalents form during meiosis. This leads to relatively straightforward estimation of recombination frequencies using maximum likelihood, from which a genetic map can be derived. However, autopolyploids such as tetraploid potato (Solanum tuberosum L.) may exhibit additional features, such as double reduction, not normally encountered in diploid or allopolyploid species. In this study, we produced a high-density linkage map of tetraploid potato and used it to identify regions of double reduction in a biparental mapping population. The frequency of multivalents required to produce this degree of double reduction was determined through simulation. We also determined the effect that multivalents or preferential pairing between homologous chromosomes has on linkage mapping. Low levels of multivalents or preferential pairing do not adversely affect map construction when highly informative marker types and phases are used. We reveal the double-reduction landscape in tetraploid potato, clearly showing that this phenomenon increases with distance from the centromeres.

Hackett C A, Milne I, Bradshaw J E , et al.

Tetraploid Map for Windows:linkage map construction and QTL mapping in autotetraploid species

Journal of Heredity, 2007,98(7):727-729.

DOI:10.1093/jhered/esm086      URL     PMID:17965198      [本文引用: 1]

An earlier program, TetraploidMap, enabled linkage analysis to be performed for autotetraploid species, with a text-based input and output. The current program, TetraploidMap for Windows, is considerably enhanced, and now beyond linkage analysis to perform quantitative trait locus (QTL) interval mapping, with a range of models and thresholds assessed by permutation tests. A Windows-based interface facilitates data entry and exploration. TetraploidMap for Windows is freely available from the Web site of Bioinformatics and Statistics Scotland at http://www.bioss.ac.uk/ (user-friendly software).

周俊 .

马铃薯(Solanum tuberosum L.)试管块茎形成的QTL定位及遗传分析

武汉:华中农业大学, 2014.

URL     [本文引用: 1]

马铃薯(Solanum tuberosum L.)在地球上的栽培历史超过8000年,广泛种植于158个国家和地区,是世界上最重要的粮食作物和经济作物之一,在保障世界粮食安全中发挥着重要作用。由于无性繁殖,病毒性退化是影响马铃薯产量的重要问题。脱毒种薯的产生解决了这一问题,而试管薯是脱毒种薯生产的首要环节。不同基因型马铃薯的试管薯形成能力差异较大,同时由于受到栽培马铃薯同源四倍体遗传复杂性的限制,关于马铃薯块茎形成的遗传研究多借助于降倍后的二倍体或者近缘二倍体野生种。即便如此,目前有关马铃薯块茎形成的遗传报道也十分有限,而且多是基于田间块茎的形成情况。而栽培马铃薯为同源四倍体,且试管薯的形成与田间块茎形成在环境条件的控制方面还有一定差异,而关于栽培马铃薯试管块茎形成的遗传研究目前还是空白。 本研究旨在四倍体水平上,构建马铃薯试管薯形成的遗传分离群体,在此基础上绘制遗传连锁图谱,并对试管薯形成相关QTL进行定位,以此分析马铃薯试管薯形成的遗传基础。主要研究结果如下: 1.基于组织培养条件下的试管薯形成表型,对来自4个杂交组合的119个马铃薯栽培种基因型材料在两种光周期条件下进行鉴定,发现8h/d光照时间处理比16h/d光照时间处理更有利于试管薯的形成;不同组合对光周期的敏感性不同,同一组合中不同基因型间亦存在较大差异。采用经典遗传理论对组合E中96个基因型材料的表型分离进行分析,x~2检测显示,试管薯形成与否可能受一对主效基因控制,其在两种光照条件下表现出不同的遗传效应,在短日照条件下表现为累加效应,长日照条件下则表现为互补效应。 2.从上述119个基因型材料中筛选得到极端表型材料E108(8h/d和16h/d光周期条件下均能形成试管薯)和E20(8h/d和16h/d光周期条件下均没有试管薯形成),构建了关于试管薯形成能力差异的F1分离群体MTI(237子代)。 3.对MT I群体237子代进行试管薯形成能力表型鉴定,发现该四倍体群体在两种光周期条件下的表型均呈偏性分布。在8h/d光周期条件下,结薯的基因型个数与不结薯的基因型个数分别为200和37,其比率(200:37)符合5:1(p=0.6629)。而16h/d光周期条件下,不结薯的基因型数与结薯的基因型数分别为163和74。长日照条件下能形成试管薯的这74个基因型在短日照条件下也形成试管薯。 4.利用来自128对AFLP引物组合、65对SSR引物和3对候选基因引物的711个标记,分别构建了双亲的遗传连锁图谱。父本图谱(E108)包含315个标记,全长948cM,由12个连锁群组成,所有12个连锁群都含有完整的4条同源染色体。依据其包含的64个SSR标记的定位信息,我们成功地将这12个连锁群与马铃薯12条染色体一一对应。母本图谱(E20)包含341个标记,由14个连锁群组成,全长1286cM,其中12个连锁群含有完整的4条同源染色体,其余2个连锁群分别覆盖到3条同源染色体。同样依据其包含的65个SSR标记的定位信息,我们将这14个连锁群与马铃薯染色体一一对应,其中第1和IV号染色体分别含有2个连锁群。 5.基于单标记分析方法,共检测到10个标记具有显著遗传效应,其中8个标记只在短日照条件下(8h/d光周期)具有显著效应,3个标记只在长日照条件下(16h/d光周期)具有显著效应,1个标记在两种光周期条件下均具有显著效应。这10个标记中的9个分别被定位在父本E108的第V号染色体和母本E20的第V、IX和XII号染色体上 6.利用双亲图谱进行区间做图(Interval Mapping),共定位到4个与马铃薯试管块茎形成相关的QTL。其中V号染色体上两个,分别为来自父本图谱的MT05和来自母本图谱的mt05;另外两个QTL均来自母本图谱,mt01-1位于I号染色体上,mt09位于IX号染色体。MT05符合加性效应,对表型变异的贡献率达到16.23%。mt01-1表现为双显性互补模型,mt05和mt09均符合单显性模型。mt05表现为正向遗传效应,mt09表现为负向遗传效应,mt01-1、mt05和mt09对表型变异的贡献率分别为6.60%、5.33%和4.81%。基于MT05的加性模型,父本E108产生的6种配子(Q12、Q13、Q14、Q23、Q24和Q34)中,显性基因型(Q-)与隐性基因型(qq)的比例为5:1,与MT I群体在短日照环境中试管薯形成与否的表型分布比例吻合,表明MT05位点中可能含有控制四倍体马铃薯试管薯形成的主效基因。 7.通过MT05与mt05、mt09之间的互作分析,我们认为MT I群体中试管薯形成表型是受主效QTL效应控制,并存在微效调节位点mt09。主效QTL MT05可,能是控制个体的试管薯形成与否,而mt09则可能影响个体的试管薯形成比例。 8.对3个块茎形成相关的候选基因(StSP6A, StCO和StCDF1)进行定位,结果显示,其中2个候选基因StSP6A和StCO在MT I群体中均不存在多态性,只有候选基因StCDFl被定位于父本图谱(E108)第V号染色体的短臂(16cM和18cM),且远离试管薯形成相关的主效QTL MT05的one-LOD置信区间(80-95cM)。上述结果表明,在MTI群体中定位到的与试管薯形成相关的QTLs与上述3个候选基因没有关联,即在该四倍体群体中定位的这些QTLs可能包含了新的与块茎形成相关的基因。 9.将MT05区段锚定到马铃薯DM基因组序列上发现,该区段内包含216个蛋白质编码基因。将这216个基因与本实验室高通量测序结果中可能与光周期调控试管薯形成以及蔗糖浓度影响试管薯形成的基因进行比对,结果显示上述216个基因中有54个在不同蔗糖浓度影响试管薯形成的过程中差异表达,表明本研究定位到的主效QTLMT05可能包含了涉及到蔗糖调节途径影响试管薯形成的基因。 本研究构建了近乎完整的马铃薯四倍体遗传连锁图谱,并首次定位了影响马铃薯试管块茎形成的QTL。基于表型分布、QTL的遗传效应模型以及QTL之间的互作分析,我们提出了该四倍体群体的试管薯形成是受到一对主效基因的调控,并存在微效基因的辅助修饰。而候选基因的定位结果显示本研究中定位到的QTLs可能包含了新的与块茎形成相关的基因。本研究的结果证实了前人报道的马铃薯块茎形成受到少数主效基因调控的遗传分析,揭示了马铃薯四倍体中可能存在新的块茎形成相关基因,提供了从试管块茎形成的角度寻找新的对于马铃薯块茎形成机理的认识和理解的可能性。

Massa A N, Manrique-Carpintero N C, Coombs J J ,et al.

Genetic linkage mapping of economically important traits in cultivated tetraploid potato (Solanum tuberosum L.)

G3 (Bethesda), 2015,5(11):2357-2364.

DOI:10.1534/g3.115.019646      URL     PMID:4632055      [本文引用: 1]

The objective of this study was to construct a single nucleotide polymorphism (SNP)-based genetic map at the cultivated tetraploid level to locate quantitative trait loci (QTL) contributing to economically important traits in potato (Solanum tuberosumL.). The 156 F1progeny and parents of a cross (MSL603) between “Jacqueline Lee” and “MSG227-2” were genotyped using the Infinium 8303 Potato Array. Furthermore, the progeny and parents were evaluated for foliar late blight reaction to isolates of the US-8 genotype ofPhytophthora infestans(Mont.) de Bary and vine maturity. Linkage analyses and QTL mapping were performed using a novel approach that incorporates allele dosage information. The resulting genetic maps contained 1972 SNP markers with an average density of 1.36 marker per cM. QTL mapping identified the major source of late blight resistance in “Jacqueline Lee.” The best SNP marker mapped 650.54 Mb from a resistance hotspot on the long arm of chromosome 9. For vine maturity, the major-effect QTL was located on chromosome 5 with allelic effects from both parents. A candidate SNP marker for this trait mapped 650.25 Mb from theStCDF1gene, which is a candidate gene for the maturity trait. The identification of markers forP. infestansresistance will enable the introgression of multiple sources of resistance through marker-assisted selection. Moreover, the discovery of a QTL for late blight resistance not linked to the QTL for vine maturity provides the opportunity to use marker-assisted selection for resistance independent of the selection for vine maturity classifications.

崔阔澍, 于肖夏, 于卓 , .

四倍体彩色马铃薯花青素含量及产量性状的QTL定位

草业学报, 2016,25(5):116-124.

DOI:10.11686/cyxb2015369      URL     Magsci     [本文引用: 1]

为确定彩色马铃薯薯块花青素含量、单株产量和商品薯率3个重要性状的QTL位点,以四倍体彩色马铃薯&#x02018;黑美人&#x02019;&#x000D7;&#x02018;MIN-021&#x02019;杂种F<sub>1</sub>代分离群体的210个单株无性株系及其亲本为材料,通过对这3个重要性状进行两年一点的观测试验,以及亲本间和杂种株系间的差异显著性分析,用TetraploidMap软件在已构建出的2张双亲的高密度彩色马铃薯分子遗传连锁图谱上分别定位其QTL。结果显示,这3个性状在亲本间和杂种株系间差异显著,且F<sub>1</sub>群体单株株系间各性状观测值均呈正态分布,适合QTL分析。在母本&#x02018;黑美人&#x02019;的遗传连锁图谱上检测到13个QTL,其中控制花青素含量的有5个、单株产量和商品薯率各有4个,遗传贡献率变幅为7.98%~19.62%。在父本&#x02018;MIN-021&#x02019;的遗传连锁图谱上检测到11个QTL,其中花青素含量和单株产量各有4个、商品薯率有3个,遗传贡献率范围在8.70%~21.62%之间。

刘龙超, 周云, 贺苗苗 , .

四倍体马铃薯SSR遗传图谱的构建及晚疫病抗性QTL初步定位

植物病理学报, 2016,46(1):84-90.

DOI:10.13926/j.cnki.apps.2016.01.010      URL     [本文引用: 1]

利用四倍体马铃薯栽培种‘大西洋’和‘陇薯6号’杂交得到的190个F,株系为作图群体,构建了四倍体马铃薯的分子遗传图谱,采用区间作图法对马铃薯晚疫病抗性进行了QTL初步定位。结果显示:通过对190个F1株系进行检测,共发现有7个与晚疫病抗性相关的QTL位点,分别分布在第5,6、7、10和11连锁群上;各位点的LOD值在2.70~10.32之间。其中主效QTL位点3个(LOD≥3.5),可解释17.37%~65.68%的表型变异。获得紧密连锁的特异标记(8183-210、8148-460)为进一步进行QTL精确定位提供了参考。

Felcher K J, Coombs J J, Massa A N , et al.

Integration of two diploid potato linkage maps with the potato genome sequence

PLoS ONE, 2012,7(4):e36347.

DOI:10.1371/journal.pone.0036347      URL     PMID:22558443      [本文引用: 1]

To facilitate genome-guided breeding in potato, we developed an 8303 Single Nucleotide Polymorphism (SNP) marker array using potato genome and transcriptome resources. To validate the Infinium 8303 Potato Array, we developed linkage maps from two diploid populations (DRH and D84) and compared these maps with the assembled potato genome sequence. Both populations used the doubled monoploid reference genotype DM1-3 516 R44 as the female parent but had different heterozygous diploid male parents (RH89-039-16 and 84SD22). Over 4,400 markers were mapped (1,960 in DRH and 2,454 in D84, 787 in common) resulting in map sizes of 965 (DRH) and 792 (D84) cM, covering 87% (DRH) and 88% (D84) of genome sequence length. Of the mapped markers, 33.5% were in candidate genes selected for the array, 4.5% were markers from existing genetic maps, and 61% were selected based on distribution across the genome. Markers with distorted segregation ratios occurred in blocks in both linkage maps, accounting for 4% (DRH) and 9% (D84) of mapped markers. Markers with distorted segregation ratios were unique to each population with blocks on chromosomes 9 and 12 in DRH and 3, 4, 6 and 8 in D84. Chromosome assignment of markers based on linkage mapping differed from sequence alignment with the Potato Genome Sequencing Consortium (PGSC) pseudomolecules for 1% of the mapped markers with some disconcordant markers attributable to paralogs. In total, 126 (DRH) and 226 (D84) mapped markers were not anchored to the pseudomolecules and provide new scaffold anchoring data to improve the potato genome assembly. The high degree of concordance between the linkage maps and the pseudomolecules demonstrates both the quality of the potato genome sequence and the functionality of the Infinium 8303 Potato Array. The broad genome coverage of the Infinium 8303 Potato Array compared to other marker sets will enable numerous downstream applications.

Bonierbale M W, Plaisted R L, Tanksley S D .

RFLP maps based on a common set of clones reveal modes of chromosomal evolution in potato and tomato

Genetics, 1988,120(4):1095-1103.

URL     PMID:17246486      [本文引用: 1]

Abstract Potato (Solanum tuberosum L.) and tomato (Lycopersicon esculentum) are members of the Solanaceae (nightshade family) and have the same basic chromosome number (x = 12). However, they cannot be cross-hybridized and, until now, it was unknown how conserved the gene order might be between these two species. We report herein the construction of a genetic linkage map of potato chromosomes based on genomic and cDNA clones from tomato. The potato map was drawn from segregation data derived from the interspecific cross S. phureja X (S. tuberosum X S. chacoense) (2n = 2x = 24), and consists of 135 markers defining 12 distinct linkage groups. Nearly all of the tomato probes tested hybridized to potato DNA, and in most cases, the copy number of the employed clones was the same in both species. Furthermore, all clones mapped to the same linkage group in both species. For nine chromosomes, the order of loci appears to be identical in the two species, while for the other three, intrachromosomal rearrangements are apparent, all of which appear to be paracentric inversions with one breakpoint at or near the centromere. These results are consistent with cytogenetic theory, previously untested in plants, which predicts that paracentric inversions will have the least negative effect on fitness and thus be the most likely form of chromosomal rearrangements to survive through evolutionary time. Linkage maps based on a common set of restriction fragment length polymorphism markers provide a basis for uniting the previously separate disciplines of tomato and potato genetics. Using these maps, it may now be possible to test theories about homologies or orthologies of other genes, including those coding for disease resistance and stress tolerances.

Gebhardt C, Ritter E, Debener T , et al.

RFLP analysis and linkage mapping in Solanum tuberosum

Theoretical & Applied Genetics, 1989,78(1):65-75.

DOI:10.1007/BF00299755      URL     PMID:24227032      [本文引用: 1]

Abstract A morphologically and agronomically heterogeneous collection of 38 diploid potato lines was analysed for restriction fragment length polymorphisms (RFLPs) with 168 potato probes, including random genomic and cDNA sequences as well as characterized potato genes of known function. The use of four cutter restriction enzymes and a fragment separation range from 250 to 2,000 bases on denaturing polyacrylamide gels allowed the detection of RFLPs of a few nucleotides. With this system, 90% of all probes tested showed useful polymorphism, and 95% of those were polymorphic with two or all three enzymes used. On the average, 80% of the probes were informative in all pairwise comparisons of the 38 lines with a minimum of 49% and a maximum of 95%. The percentage of heterozygosity was determined relative to each other for each line and indicated that direct segregation analysis in F1 populations should be feasible for most combinations. From a backcross involving one pair of the 38 lines, a RFLP linkage map with 141 loci was constructed, covering 690 cMorgan of the Solanum tuberosum genome.

Van Eck H J, Van der Voort J R, Draaistra J , et al.

The inheritance and chromosomal localization of AFLP markers in a non-inbred potato offspring

Molecular Breeding, 1995,1(4):397-410.

DOI:10.1007/BF01248417      URL     [本文引用: 1]

AFLP TM is a new technique to generate large numbers of molecular markers for genetic mapping. The method involves the selective amplification of a limited number of DNA restriction fragments out of complex plant genomic DNA digests using PCR. With six primer combinations 264 segregating AFLP amplification products were identified in a diploid backcross population from non-inbred potato parents. The identity of an AFLP marker was specified by the primer combination of the amplification product and its size estimated in bases. The segregating AFLP amplification products were mapped by using a mapping population with 217 already known RFLP , isozyme and morphological trait loci. In general, the AFLP markers were randomly distributed over the genome, although a few clusters were observed. No indications were found that AFLP markers are present in other parts of the genome than those already covered by RFLP markers. Locus specificity of AFLP markers was demonstrated because equally sized amplification products segregating from both parental clones generally mapped to indistinguishable maternal and paternal map positions. Locus specificity of AFLP amplification products will allow to establish the chromosomal identity of linkage groups in future mapping studies. Since AFLP technology is a multi-locus detection system, it was not possible to identify the AFLP alleles which belong to a single AFLP locus. The consequences of a genetic analysis based on single alleles, rather than on loci with two or more alleles on mapping studies using progenies of non-inbred parents are discussed.

金黎平, 刘杰, 方智远 .

二倍体马铃薯分子连锁图谱的构建

园艺学报, 2007,4(2):397-402.

DOI:10.3321/j.issn:0513-353X.2007.02.025      URL     [本文引用: 1]

Based on amp lified fragment length polymorphism (AFLP) markers, a molecular linkage map of potato was initially developed by using Fsegregating population from the cross between the diploid potato parents of 08675-21 and 09901-01. The genetic map included 24 main linkage group s, and the femalemaps covered 512 cM with an average distance of 6.83 cM between adjacentmarkerswhile the male2map s covered 578 cM with an average distance of 6.08 cM between adjacent markers.

Visker M, Keizer L, Eck H V , et al.

Can the QTL for late blight resistance on potato chromosome 5 be attributed to foliage maturity type?

Theoretical & Applied Genetics, 2003,106(2):317-325.

DOI:10.1007/s00122-002-1021-2      URL     PMID:12582858      [本文引用: 1]

We investigated the association between late blight resistance and foliage maturity type in potato by means of molecular markers. Two QTLs were detected for foliage resistance against Phytophthora infestans (on chromosomes 3 and 5) and one for foliage maturity type (on chromosome 5). The QTL for resistance to late blight and the QTL for foliage maturity type on chromosome 5 appeared to be mapped on indistinguishable positions. We were interested whether this genetic linkage was due to closely linked but different genes, or due to one (or more) gene(s) with pleiotropic effects. We therefore developed an approach to detect QTLs, in which resistance to late blight was adjusted for foliage maturity type. This analysis revealed the same two QTLs for resistance against P. infestans , but the effect of the locus on chromosome 5 was reduced to only half the original effect. This is a strong indication that the two indistinguishable QTLs for foliage maturity type and for late blight resistance on chromosome 5 may actually be one gene with a pleiotropic effect on both traits. However, there was still a significant effect on resistance against P. infestans on the locus on chromosome 5 after adjusting for foliage maturity type. Therefore we cannot rule out the presence of two closely linked QTLs on chromosome 5: one with a pleiotropic effect on both late blight resistance and foliage maturity type, and another with merely an effect on resistance. In addition, the two QTLs for resistance to late blight showed an important epistatic interaction, suggesting that QTLs for resistance affect each other's expression.

Danan S, Chauvin J E, Caromel B , et al.

Major-effect QTLs for stem and foliage resistance to late blight in the wild potato relatives Solanum sparsipilum and S. spegazzinii are mapped to chromosome X

Theoretical & Applied Genetics, 2009,119(4):705-719.

DOI:10.1007/s00122-009-1081-7      URL     PMID:19533081      [本文引用: 1]

To find out new resistance sources to late blight in the wild germplasm for potato breeding, we examined the polygenic resistance of Solanum sparsipilum and S. spegazzinii by a quantitative trait locus (QTL) analysis. We performed stem and foliage tests under controlled conditions in two diploid mapping progenies. Four traits were selected for QTL detection. A total of 30 QTLs were mapped, with a large-effect QTL region on chromosome X detected in both potato relatives. The mapping of literature-derived markers highlighted colinearities with published late blight QTLs or R-genes. Results showed (a) the resistance potential of S. sparsipilum and S. spegazzinii for late blight control, and (b) the efficacy of the stem test as a complement to the foliage test to break down the complex late blight resistance into elementary components. The relationships of late blight resistance QTLs with R-genes and maturity QTLs are discussed.

单友蛟, 刘杰, 卞春松 , .

马铃薯SSR遗传连锁图谱构建及3个重要农艺性状QTLs定位

中国蔬菜, 2010,1(18):10-14.

URL     Magsci     [本文引用: 1]

利用二倍体马铃薯杂交群体P1的167个基因型为作图群体,应用SSR标记构建了一张总长度为645 cM的遗传连锁图谱,该图谱包含86个SSR标记,标记间平均距离为7.5 cM。应用该图谱,利用复合区间作图法,对与马铃薯块茎相关的3个重要农艺性状进行QTL定位和遗传效应分析,结果在第5条染色体上检测到控制以上3个性状的QTL各1个,其中控制单株产量的QTL遗传贡献率为12.3 %,控制单薯质量的QTL遗传贡献率为16.1 %,控制块茎比重的QTL遗传贡献率为11.2 %。

Mihovilovich E, Munive S, Bonierbale M .

Influence of day-length and isolates of Phytophthora infestans on field resistance to late blight of potato

Theoretical & Applied Genetics, 2010,120(6):1265-1278.

DOI:10.1007/s00122-009-1254-4      URL     PMID:20063145      [本文引用: 1]

Main and interaction effects of day-length and pathogen isolate on the reaction and expression of field resistance to Phytophthora infestans were analyzed in a sample of standard clones for partial resistance to potato late blight, and in the BCT mapping population derived from a backcross of Solanum berthaultii to Solanum tuberosum . Detached leaves from plants grown in field plots exposed to short- and long day-length conditions were independently inoculated with two P. infestans isolates and incubated in chambers under short- and long photoperiods, respectively. Lesion growth rate (LGR) was used for resistance assessment. Analysis of variance revealed a significant contribution of genotype isolate day-length interaction to variation in LGR indicating that field resistance of genotypes to foliar late blight under a given day-length depended on the infecting isolate. An allele segregating from S. berthaultii with opposite effects on foliar resistance to late blight under long- and short day-lengths, respectively, was identified at a quantitative trait locus (QTL) that mapped on chromosome 1. This allele was associated with positive (decreased resistance) and negative (increased resistance) additive effects on LGR, under short- and long day-length conditions, respectively. Disease progress on whole plants inoculated with the same isolate under field conditions validated the direction of its effect in short day-length regimes. The present study suggests the occurrence of an isolate-specific QTL that displays interaction with isolate behavior under contrasting environments, such as those with different day-lengths. This study highlights the importance of exposing genotypes to a highly variable population of the pathogen under contrasting environments when stability to late blight resistance is to be assessed or marker-assisted selection is attempted for the manipulation of quantitative resistance to late blight.

李竟才 .

二倍体马铃薯遗传图谱构建及晚疫病抗性QTL定位

武汉:华中农业大学, 2012.

DOI:10.7666/d.Y2162708      URL     [本文引用: 1]

马铃薯(Solanum tuberosum L.)起源于南美洲安第斯山脉,是世界第四大粮食作物,第一大非谷类粮食作物。目前我国是世界上马铃薯生产第一大国,马铃薯产业已经成为我国农村经济发展的一个重要支柱产业。晚疫病(Phytophthora infestans)是世界范围内的马铃薯主要病害之一,是国际第一大作物病害。虽然化学防治是目前国内外最主要的晚疫病防治措施,但化学防治带来了更高的生产投入和环境污染。此外,晚疫病原菌的抗药性问题使化学防治的难度日益增加。因此,晚疫病持久抗性品种的选育越来越受到关注。但到目前为止,马铃薯生产中仍然缺乏优良的晚疫病持久抗性...

李梦台 .

二倍体马铃薯遗传图谱构建及薯形QTL分析

武汉:华中农业大学, 2015.

DOI:10.7666/d.Y2803558      URL     [本文引用: 1]

马铃薯(Solanum tuberosum L.)是世界上第一大非谷类粮食作物,被广泛种植于世界150多个国家和地区,每日有超过十亿人在消费食用。栽培马铃薯是一种高度杂合的同源四倍体作物,其遗传规律非常复杂,因此对于马铃薯遗传方面的研究多在二倍体水平上进行。马铃薯除了用于鲜食以外,还广泛用于食品加工。无论是用于鲜食市场还是加工行业,马铃薯薯形都是一个非常重要的性状。长薯形的马铃薯通常被加工成薯条,而圆薯形的马铃薯通常加工成薯片。尽管薯形是马铃薯一个非常重要的性状,但是目前关于马铃薯薯形遗传学方面的研究报道仍然十分有限。本研究旨在利用二倍体薯形分离群体,通过遗传图谱的构建,对马铃薯薯形相关的QTL进行定位,使我们对马铃薯薯形的遗传信息有更进一步的认识,为马铃薯薯形改良的分子标记辅助选择奠定基础。本实验室前期构建了一个二倍体群体,其母本是马铃薯测序种DM1-3(S.phureja),为纯合二倍体,长薯形,父本是40-3(S.chacoense),为杂合二倍体,圆薯形,本研究是在此基础上展开的。主要研究结果如下:1.分别在2014和2015年对两亲本及群体后代进行试管薯薯形鉴定,DM1-3为长形,40-3为圆形。两年薯形指数的相关性分析结果表明,相关系数r达到了0.92,达极显著水平。表明两年薯形表型数据的重复性很好,另一个方面也反映薯形性状比较稳定。另外,两年F1群体后代的平均薯形指数分别为2.79和2.85,明显小于两亲本薯形指数的中亲值4.515,表明圆薯形对长薯形是显性的。进一步用X2适合性测验群体后代薯形分布是否符合正态分布,结果表明群体后代的薯形指数分布不符合正态分布。2.遗传图谱构建:利用821对SSR引物在两亲本和部分后代单株中进行筛选,共获得116对特异性SSR引物。116个特异性引物扩增178个群体后代,共获得165个多态性标记,其中158个为父本的多态性标记,7个为母本的多态性标记。构建了父本40-3的遗传图谱,该图谱包含了157个标记,全长688.3 c M,平均间距为4.38 c M,覆盖了马铃薯的12条染色体。3.利用薯形与基因型标记的相关性分析结果表明,有13个标记与薯形显著相关,主要分布在马铃薯2号和4号染色体上。利用父本图谱进行MQM(多QTL作图),定位到一个与薯形相关的QTL位点(LW04)。而且在两年的薯形数据中结果一致,定位于马铃薯4号染色体的同一位置,对圆薯形有正向遗传效应,对表型变异的贡献率分别为6.1%和6.3%。

Schäfer-Pregl R, Ritter E, Hesselbach J , et al.

Analysis of quantitative trait loci (QTLs) and quantitative trait alleles (QTAs) for potato tuber yield and starch content

Theoretical & Applied Genetics, 1998,97(5/6):834-846.

DOI:10.1007/s001220050963      URL     [本文引用: 1]

Using RFLP markers, QTLs for tuber starch-content and tuber yield were mapped in two F 1 populations derived from crossing non-inbred di-haploid potato breeding lines. QTLs were identified and mapped, based on both single-marker tests and interval analyses. A model specifically developed for interval QTL analysis in non-inbred plant species was successfully applied for the first time to experimental data. Results of both methods of QTL analysis were similar but not identical. QTLs for tuber starch-content and tuber yield were analysed in segregating populations K31 and LH in five and two environments, respectively. Population K31 was fully genotyped whereas population LH was selectively genotyped according to high and low tuber-starch content. Eighteen putative QTLs for tuber starch-content were identified on all 12 potato linkage groups and eight putative QTLs for tuber yield were identified on eight linkage groups. Twenty of twenty six putative QTLs were reproducibly detected in at least two environments and/or mapping populations. Few major QTLs for tuber starch-content were highly stable across environments but were detected in only one of the two mapping populations analysed. Most QTLs for tuber yield were linked with QTLs for tuber starch-content suggesting that the effects on both traits are controlled by the same genetic factors. The results are discussed with respect to marker-assisted selection in potato.

Jung C S, Griffiths H M, De Jong D M , et al.

The potato P locus codes for flavonoid 3′,5′-hydroxylase

Theoretical & Applied Genetics, 2005,110(2):269-275.

DOI:10.1007/s00122-005-1987-7      URL     PMID:15565378      [本文引用: 1]

The potato P locus is required for the production of blue/purple anthocyanin pigments in any tissue of the potato plant such as tubers, flowers, or stems. We have previously reported, based on RFLP mapping in tomato, that the gene coding for the anthocyanin biosynthetic enzyme flavonoid 3′,5′-hydroxylase ( f3 ′ 5 ′ h ) maps to the same region of the tomato genome as P maps in potato. To further evaluate this association a Petunia f3 ′ 5 ′ h gene was used to screen a potato cDNA library prepared from purple-colored flowers and stems. Six positively hybridizing cDNA clones were sequenced and all appeared to be derived from a single gene that shares 85% sequence identity at the amino acid level with Petunia f3 ′ 5 ′ h . The potato gene cosegregated with purple tuber color in a diploid F 1 sub-population of 37 purple and 25 red individuals and was found to be expressed in tuber skin only in the presence of the anthocyanin regulatory locus I . A potato f3 ′ 5 ′ h cDNA clone was placed under the control of a doubled CaMV 35S promoter and introduced into the red-skinned cultivar ‘Désirée’. Tuber and stem tissues that are colored red in Désirée were purple in nine of 17 independently transformed lines.

De Jong W S, Eannetta N T, De Jong D M , et al.

Candidate gene analysis of anthocyanin pigmentation loci in the Solanaceae

Theoretical & Applied Genetics, 2004,108(3):423-432.

DOI:10.1007/s00122-003-1455-1      URL     PMID:14523517      [本文引用: 1]

Crop species in the Solanaceae , which includes tomato ( Lycopersicon esculentum ), potato ( Solanum tuberosum ), pepper ( Capsicum spp. ), and eggplant ( S. melongena ), exhibit natural variation in the types, levels, and tissue-specific expression patterns of anthocyanin pigments. While the identities of the genes underpinning natural variation in anthocyanin traits in these crops are largely unknown, many structural genes and regulators of anthocyanin biosynthesis have been isolated from the solanaceous ornamental species Petunia . To identify candidate genes that may correspond to loci controlling natural variation in the four crops, 13 anthocyanin-related genes were localized on a tomato F 2 genetic map. Gene map positions were then compared to mapped mutants in tomato and through comparative genetic maps to natural variants in potato, eggplant, and pepper. Similar map positions suggest that the tomato mutants anthocyaninless , entirely anthocyaninless , and anthocyanin gainer correspond to flavonoid 3′5′-hydroxylase ( f3′5′h ), anthocyanidin synthase, and the Petunia Myb domain trancriptional regulatory gene an2 , respectively. Similarly potato R , required for the production of red pelargonidin-based pigments, P , required for production of purple delphinidin-based pigments, and I , required for tissue-specific expression in tuber skin, appear to correspond to dihydroflavonol 4-reductase, f3′5′h and an2 , respectively. The map location of an2 also overlaps pepper A and eggplant fap 10.1, lla 10.1, lra 10.1, sa 10.1, pa 10.1 and ca 10.1, suggesting that a homologous regulatory locus has been subjected to parallel selection in the domestication of many solanaceous crops. To test the hypothesis that tomato anthocyaninless corresponds to f3′5′h , a portion of the gene was sequenced. A premature stop codon was observed in an anthocyaninless mutant, but not in wild-type.

肖继坪, 王琼, 郭华春 .

彩色马铃薯二氢黄酮醇4-还原酶(DFR)基因的克隆及生物信息学分析

分子植物育种, 2011,9(6):728-735.

[本文引用: 1]

肖继坪, 李俊, 郭华春 .

彩色马铃薯类黄酮-3-O-葡萄糖基转移酶基因(3GT)的生物信息学和表达分析

分子植物育种, 2015,13(5):1017-1026.

[本文引用: 1]

杨艳丽 . 云南马铃薯产业技术与经济研究. 北京: 科学出版社, 2016.

[本文引用: 2]

Ramakrishnan A P, Ritland C E, Blas Sevillano R H , et al.

Review of potato molecular markers to enhance trait selection

American Journal of Potato Research, 2015,92(4):455-472.

DOI:10.1007/s12230-015-9455-7      URL     [本文引用: 2]

Noncommercial varieties of potatoes ( Solanum tuberosum L.) harbor genetic potential for improvements of disease resistance and abiotic stress tolerance in commercial potato cultivars; however, introducing traits from noncommercial varieties to breeding stock can be extremely labor intensive. Molecular genetic markers closely associated with a trait can be used to decrease the time spent phenotyping varieties. Here we review genetic markers that have been used for marker-assisted selection (MAS) in potato. Most MAS markers have been used to detect disease resistance genes, and our review focuses on those markers. Complex traits such as cold, drought and viral tolerance can be studied by comparing expressed genes; next-generation sequencing technologies will help in the discovery of trait-specific molecular markers. This review aids in summarizing the potential of these molecular tools when breeding for complex traits in potato.

Fry W .

Phytophthora infestans:the plant (and R gene) destroyer

Molecular Plant Pathology, 2008,9(3):385-402.

DOI:10.1111/j.1364-3703.2007.00465.x      URL     PMID:18705878      [本文引用: 1]

Phytophthora infestans remains a problem to production agriculture. Historically there have been many controversies concerning its biology and pathogenicity, some of which remain today. Advances in molecular biology and genomics promise to reveal fascinating insight into its pathogenicity and biology. However, the plasticity of its genome as revealed in population diversity and in the abundance of putative effectors means that this oomycete remains a formidable foe.

Flor H H .

Current status of the gene-for-gene concept

Annual Review of Phytopathology, 1971,9(1):275-296.

DOI:10.1146/annurev.py.09.090171.001423      URL     [本文引用: 1]

EI-Kharbotly A, Leonards-Schippers C, Huigen D J , et al.

Segregation analysis and RFLP mapping of the R1 and R3 alleles conferring race-specific resistance to Phytophthora infestans in progeny of dihaploid potato parents

Molecular and General Genetics, 1994,242(6):749-754.

DOI:10.1007/BF00283432      URL     PMID:7908718      [本文引用: 2]

Phytophthora infestans (Mont.) de Bary is the most important fungal pathogen of the potato ( Solanum tuberosum ). The introduction of major genes for resistance from the wild species S. demissum into potato cultivars is the earliest example of breeding for resistance using wild germplasm in this crop. Eleven resistance alleles ( R genes ) are known, differing in the recognition of corresponding avirulence alleles of the fungus. The number of R loci, their positions on the genetic map and the allelic relationships between different R variants are not known, except that the R1 locus has been mapped to potato chromosome V The objective of this work was the further genetic analysis of different R alleles in potato. Tetraploid potato cultivars carrying R alleles were reduced to the diploid level by inducing haploid parthenogenetic development of 2 n female gametes. Of the 157 isolated primary dihaploids, 7 set seeds and carried the resistance alleles R1, R3 and R10 either individually or in combinations. Independent segregation of the dominant R1 and R3 alleles was demonstrated in two F 1 populations of crosses among a dihaploid clone carrying R1 plus R3 and susceptible pollinators. Distorted segregation in favour of susceptibility was found for the R3 allele in 15 of 18 F 1 populations analysed, whereas the RI allele segregated with a 1:1 ratio as expected in five F 1 populations. The mode of inheritance of the R10 allele could not be deduced as only very few F 1 hybrids bearing R10 were obtained. Linkage analysis in two F 1 populations between R1, R3 and RFLP markers of known position on the potato RFLP maps confirmed the position of the R1 locus on chromosome V and localized the second locus, R3 , to a distal position on chromdsome XI.

EI-Kharbotly A, Jacobs J M E, te Hekkert B T , et al.

Localization of Ds-transposon containing T-DNA inserts in the diploid transgenic potato:linkage to the R1 resistance gene against Phytophthora infestans (Mont.) de Bary

Genome, 1996,39(2):249-257.

DOI:10.1139/g96-034      URL     [本文引用: 2]

Colton L M, Groza H I, Wielgus S M , et al.

Marker-assisted selection for the broad-spectrum potato late blight resistance conferred by gene derived from a wild potato species

Crop Science, 2006,46(2):589-594.

DOI:10.2135/cropsci2005.0112      URL     [本文引用: 2]

ABSTRACT Potato(SolanumtuberosumL.)lateblight,causedbyPhytophthora infestans (Mont.) de Bary, is one of the most damaging diseases in any crop. Deployment of resistant varieties is the most effective way to control this disease. However, breeding for late blight resistance has been a challenge because the race-specific resistance genes intro- gressed from wild potato S. demissum Lindl. have been short lived and breeding for ''horizontal'' or durable resistance has achieved only moderate successes. We previously demonstrated that the high- level late blight resistance in a wild potato relative, S. bulbocastanum Dunal subsp. bulbocastanum, is mainly controlled by a single resis- tance gene RB. Transgenic potato lines containing the RB gene have showed strong late blight resistance, comparable to the backcrossed progenies derived from the somatic hybrids between potato and S. bulbocastanum. Here we report the development of a polymerase chain reaction-based DNA marker for tracking the RB gene in breed- ing populations derived from the potato 3 S. bulbocastanum somatic hybrids. Several marker-positive breeding lines showed the expected late blight resistance in greenhouse evaluations. Our results demon- strate that marker-based selection will allow us to effectively transfer the RB gene into potato using traditional breeding methods, an alter- native to deploying the RB gene through genetic transformation.

徐建飞, 黄三文, 金黎平 , .

马铃薯晚疫病抗性基因R11的遗传定位

作物学报, 2009,35(6):992-997.

DOI:10.3724/SP.J.1006.2009.00992      URL     Magsci     [本文引用: 2]

<p>由致病疫霉菌(<em>Phytophthora infestans</em>)引起的晩疫病是世界范围内最具毁灭性的马铃薯(<em>Solanum tuberosum</em>)病害。以含有晚疫病抗性基因<em>R11</em>的材料MaR11和不含已知抗性基因的品种Katahdin为亲本进行有性杂交,对获得的F<sub>1</sub>分离群体的83个基因型进行了晚疫病菌株接种鉴定和遗传分析。结果表明,<em>R11</em>为主效单基因,在MaR11中以单式形式(R<sub>11r11r11r11</sub>)存在。应用比较作图和分离群体分组分析(BSA),开发了6个与<em>R11</em>连锁的分子标记,并将<em>R11</em>定位于11号染色体长臂末端。<em>R11</em>距C2_At5g59960标记最近,约为2.4 cM。通过遗传图谱比较表明,<em>R11</em>较晚疫病抗性基因R3a和R10更靠近染色体端粒区。本研究所获得的遗传图谱为进一步构建<em>R11</em>高密度遗传图谱提供了基础。</p>

Tomczyńska I, Stefańczyk E, Chmielarz M , et al.

A locus conferring effective late blight resistance in potato cultivar Sárpo Mira maps to chromosome Ⅺ

Theoretical & Applied Genetics, 2014,127(3):647-657.

DOI:10.1007/s00122-013-2248-9      URL     PMID:3931936      [本文引用: 1]

Late blight of potato, caused by Phytophthora infestans , is one of the most economically important diseases worldwide, resulting in substantial yield losses when not adequately controlled by fungicides. Late blight was a contributory factor in The Great Irish Famine, and breeding for resistance to the disease began soon after. Several disease-resistant cultivars have subsequently been obtained, and amongst them Sárpo Mira is currently one of the most effective. The aim of this work was to extend the knowledge about the genetic basis of the late blight resistance in Sárpo Mira and to identify molecular markers linked to the resistance locus which would be useful for marker-assisted selection. A tetraploid mapping population from a Sárpo Mira02×02Maris Piper cross was phenotyped for foliar late blight resistance using detached leaflet tests. A locus with strong effect on late blight resistance was mapped at the end of chromosome XI in the vicinity of the R3 locus. Sárpo Mira’s genetic map of chromosome XI contained 11 markers. Marker 45/XI exhibited the strongest linkage to the resistance locus and accounted for between 55.8 and 67.902% of variance in the mean resistance scores noted in the detached leaflet assays. This marker was used in molecular marker-facilitated gene pyramiding. Ten breeding lines containing a late blight resistance locus from cultivar Sárpo Mira and the Rpi - phu1 gene originating from the late blight resistant accession of Solanum phureja were obtained. These lines have extended the spectrum of late blight resistance compared with Sárpo Mira and it is expected that resistance in plants containing this gene pyramid will have enhanced durability.

Kuhl J C, Bradeen J M, Kole C. Genetics,Genomics and Breeding of Sunflower. Boca Raton: CRC Press,USA, 2010: 111-113.

[本文引用: 1]

Mori K, Sakamoto Y, Mukojima N , et al.

Development of a multiplex PCR method for simultaneous detection of diagnostic DNA markers of five disease and pest resistance genes in potato

Euphytica, 2011,180(3):347-355.

DOI:10.1007/s10681-011-0381-6      URL     [本文引用: 2]

AbstractMultiplex PCR is practically a reasonable choice for molecular marker-assisted selection in potato breeding. We had developed and were using a multiplex PCR method for selection of resistance genes to cyst nematode (), Rx1 and ). Since then, more reliable and tightly linked markers for and , and a new marker for resistance to Ry

Li X, van Eck H J, Rouppe J , et al.

Autotetraploids and genetic mapping using common AFLP markers:the R2 allele conferring resistance to Phytophthora infestans mapped on potato chromosome 4

Theoretical & Applied Genetics, 1998,96(8):1121-1128.

DOI:10.1007/s001220050847      URL     [本文引用: 1]

Due to the complexity of tetrasomic inheritance, mapping studies in potato ( Solanum tuberosum L.) are generally conducted at the diploid level. In the present study we tested the feasibility of Bulked Segregant Analysis (BSA) using a tetraploid offspring for the identification of AFLP markers linked to the R2 allele, which confers race-specific resistance to Phytophthora infestans . Eleven bulk-specific AFLP markers, detected in fingerprints of 205 AFLP primer combinations, could be mapped in a linkage group encompassing the R2 locus. The efficiency of BSA at the tetraploid level, determined by the frequency of single-dose restriction fragments (SDRF), was much higher than expected on the basis of overall genetic dissimilarity between the parental clones. The fortuitous detection of AFLPs with linkage to the R2 allele is explained on the basis of specific genetic dissimilarity between cultivated potato and the chromosomal segment introgressed from S. demissum carrying the resistant R2 allele. AFLP markers common to those with linkage to R2 were visually recognized by their electrophoretic mobility in the AFLP fingerprint in a parental clone of a reference mapping population. Using these common AFLP markers we anchored the linkage group comprising the R2 allele to potato chromosome 4.

Rauscher G M, Smart C D, Simko I M , et al.

Characterization and mapping of Rpi-ber,a novel potato late blight resistance gene from Solanum berthaultii

Theoretical & Applied Genetics, 2006,112(4):674-687.

DOI:10.1007/s00122-005-0171-4      URL     PMID:16402191      [本文引用: 1]

Abstract Phytophthora infestans, the causal agent of late blight, threatens potato production worldwide. An important tool in the management of the disease is the use of resistant varieties. Eleven major resistance genes have been identified and introgressed from Solanum demissum. However, new sources of resistance are continually sought. Here, we report the characterization and refined genetic localization of a resistance gene previously identified as Rber in a backcross progeny of Solanum tuberosum and Solanum berthaultii. In order to further characterize Rber, we developed a set of P. infestans isolates capable of identifying each of the 11 R-genes known to confer resistance to late blight in potato. Our results indicate that Rber is a new resistance gene, different from those recognized in S. demissum, and therefore, it has been named RPi-ber according to the current system of nomenclature. In order to add new molecular markers around RPi-ber, we used a PCR-based mapping technique, named MASP-map, which located RPi-ber in a 3.9 cM interval between markers CT240 and TG63 on potato chromosome X. The location of RPi-ber coincides with an area involved in resistance to different pathogens of potato and tomato.

Wang M, Allefs S, van den Berg R G , et al.

Allele mining in Solanum:conserved homologues of Rpi-blb1 are identified in Solanum stoloniferum

Theoretical & Applied Genetics, 2008,116(7):933-943.

DOI:10.1007/s00122-008-0725-3      URL     PMID:18274723      [本文引用: 1]

Abstract Allele mining facilitates the discovery of novel resistance (R) genes that can be used in breeding programs and sheds light on the evolution of R genes. Here we focus on two R genes, Rpi-blb1 and Rpi-blb2, originally derived from Solanum bulbocastanum. The Rpi-blb1 gene is part of a cluster of four paralogues and is flanked by RGA1-blb and RGA3-blb. Highly conserved RGA1-blb homologues were discovered in all the tested tuber-bearing (TB) and non-tuber-bearing (NTB) Solanum species, suggesting RGA1-blb was present before the divergence of TB and NTB Solanum species. The frequency of the RGA3-blb gene was much lower. Interestingly, highly conserved Rpi-blb1 homologues were discovered not only in S. bulbocastanum but also in Solanum stoloniferum that is part of the series Longipedicellata. Resistance assays and genetic analyses in several F1 populations derived from the relevant late blight resistant parental genotypes harbouring the conserved Rpi-blb1 homologues, indicated the presence of four dominant R genes, designated as Rpi-sto1, Rpi-plt1, Rpi-pta1 and Rpi-pta2. Furthermore, Rpi-sto1 and Rpi-plt1 resided at the same position on chromosome VIII as Rpi-blb1 in S. bulbocastanum. Segregation data also indicated that an additional unknown late blight resistance gene was present in three populations. In contrast to Rpi-blb1, no homologues of Rpi-blb2 were detected in any material examined. Hypotheses are proposed to explain the presence of conserved Rpi-blb1 homologues in S. stoloniferum. The discovery of conserved homologues of Rpi-blb1 in EBN 2 tetraploid species offers the possibility to more easily transfer the late blight resistance genes to potato varieties by classical breeding.

Rouppe van der Voort J N A M, Janssen G J W, Overmars H , et al.

Development of a PCR-based selection assay for root-knot nematode resistance (Rmc1) by a comparative analysis of the Solanum bulbocastanum and S. tuberosum genome

Euphytica, 1999,106(2):187-195.

DOI:10.1023/A:1003587807399      URL     [本文引用: 1]

A PCR-based assay has been developed for marker assisted selection of root-knot nematode resistance ( Rmc1) in potato. To this end, a comparative genome analysis was carried out between Solanum bulbocastanum and S. tuberosum to identify PCR-based chromosome 11 alleles linked to Rmc1. The use of co-migrating AFLP markers, obtained by using primer combinations previously applied for AFLP analysis of the S. tuberosum genome, failed to align the AFLP map of the S. bulbocastanum genome with the S. tuberosum map. Apparently, the S. bulbocastanum genome is genetically too distantly related to the S. tuberosum genome for this type of analysis. Cleaved amplified polymorphic sequence (CAPS) markers were more readily applied for a comparative analysis within the region of interest. Rmc1 could be localized within a 4 cM interval between markers CT182 and M39b. It is demonstrated that the resistance spectrum of Rmc1 includes not only Meloidogyne chitwoodi and the related species M. fallax but also a genetically distinct population of M. hapla. The cost-efficiency of the CAPS markers applied for Rmc1 renders this approach as an attractive alternative for screening large segregating populations of potato for root-knot nematode resistance.

Zhang L H, Mojtahedi H, Kuang H , et al.

Marker-assisted selection of Columbia root-knot nematode resistance introgressed from Solanum bulbocastanum

Crop Science, 2007,47(5):2021-2026.

DOI:10.2135/cropsci2007.01.0003      URL     [本文引用: 1]

ABSTRACT The Columbia root-knot nematode (Meloidogyne chitwoodi Golden et al.) is a serious pest that reduces tuber quality of potato (Solanum tuberosum L.) in the U.S. Northwest and other parts of the world. A gene, RMc1(blb), derived from the Mexican wild species Solanum bulbocastanum Dunal, encodes resistance to this pest. An F1 mapping population with >250 individuals generated from an intraspecific cross between resistant and susceptible clones of S. bulbocastanum, SB22 and PT29, respectively, was used for marker screening and genetic linkage analysis. One amplified fragment length polymorphism marker and five sequence tagged site (STS) markers cosegregated with RMc1(blb). The five STS markers were developed from bacterial artificial chromosome (BAC) end sequences of BAC clones that were derived from another wild species, S. demissum Lindl, and contained homologs of resistance gene N against tobacco mosaic virus. These markers were tested on families that were part of the introgression of RMc1(blb) into advanced breeding lines in BC5. The utility of an efficient alternative to greenhouse and field phenotypic screening was demonstrated. The results of this study confirm that molecular markers closely linked to RMc1(blb) will assist in a selection program, reducing expense and time involved in root-knot nematode screening.

Brunt A A .

The main viruses infecting potato crops//

Loebenstein G,Berger P H,Brunt A A,et al. Virus and Virus-Like Diseases of Potatoes and Production of Seed-Potatoes, 2001: 65-67.

[本文引用: 1]

Kasai K, Morikawa Y, Sorri V A , et al.

Development of SCAR markers to the PVY resistance gene Ryadg based on a common feature of plant disease resistance genes

Genome, 2000,43(1):1-8.

DOI:10.1139/g99-092      URL     PMID:10701106      [本文引用: 2]

Sequence-characterized amplified regions (SCARs) were developed, based on nucleotide differences within resistance gene-like fragments isolated from a potato plant carrying the Ryadg gene, which confers extreme resistance to potato Y potyvirus (PVY). It originates from Solanum tuberosum subsp. andigena, and a susceptible potato plant. SCARs were tested using 103 potato breeding lines and cultivars with diverse genetic backgrounds derived from Europe, North America, and Japan. Two markers showed high accuracy for detection of the Ryadg gene. The SCAR marker RYSC3 was generated only in genotypes carrying Ryadg. The SCAR marker RYSC4 was detected in all genotypes carrying Ryadg but also in four PVY-susceptible genotypes. Neither marker was detected in genotypes carrying other Ry genes originating from different species than S. tuberosum subsp. andigena. Therefore, these SCAR markers should be powerful tools in marker-assisted selection for Ryadg in potato breeding programs, and should also be useful for cloning of the Ryadg gene.

Ottoman R, Hane D, Brown C , et al.

Validation and implementation of marker-assisted selection(MAS)for PVY resistance in a tetraploid potato breeding program

American Journal of Potato Research, 2009,86(4):304-314.

DOI:10.1007/s12230-009-9084-0      URL     [本文引用: 1]

The gene Ry adg from S. tuberosum ssp. andigena provides extreme resistance to PVY. This gene has been genetically mapped to chromosome XI and linked PCR-based DNA markers have been identified. Advanced tetraploid russeted potato clones developed by the U.S. Pacific Northwest Potato Breeding ( ri-State) Program with Ry adg PVY resistance were used in this study. The objective of this work was to assess the usefulness of molecular markers linked to Ry adg as a tool for selecting PVY resistance in a tetraploid potato breeding program. To achieve this, a full-sib tetraploid population segregating for Ry adg was screened with molecular markers linked to Ry adg , artificially inoculated with PVY O and evaluated in the greenhouse. A large percentage (96.4%) of the segregating lines showed coincidence between molecular markers and ELISA results at 40days after inoculation. This justifies the use of molecular markers as an alternative to artificial inoculation followed by ELISA . Segregation (resistant vs. susceptible) based on ELISA and molecular marker results in the full-sib population indicated the presence of Ry adg as a simplex in the PVY resistant parent OR00030-1. Additional full-sib populations segregating for the Ry adg gene coming from OR00030-1 and from a related clone, AOR00628-3, were evaluated under field conditions. MAS can be used as a fast and efficient tool to select for PVY resistance, reducing the number of PVY susceptible lines retained for succeeding field evaluations, and thereby increasing the odds of generating PVY resistant potato varieties.

Lopez-Pardo R, Barandalla L, Ritter E , et al.

Validation of molecular markers for pathogen resistance in potato

Plant Breeding, 2013,132:246-251.

DOI:10.1111/pbr.12062      URL     [本文引用: 1]

DNA markers have a large potential to improve efficiency and precision of conventional plant breeding programmes based on marker-assisted selection (MAS). In our study, we have evaluated the predictive abilities of the SCAR marker RYSC3 and the CAPS marker GP122564 with regard to the PVY resistance genes Ryadg and Rysto, respectively, and of marker TG689 linked to H1 conferring resistance to Globodera rostochiensis and marker HC associated with high levels of G. pallida resistance. The evaluations were made in 28 cultivars and accessions and in 219 progeny genotypes descending from ten different crosses. We observed in all evaluated cultivars and accessions the expected marker patterns according to their phenotypic classification into resistant and susceptible genotypes. However, in part considerable discrepancies were observed when analysing progeny of controlled crosses involving these resistance sources, particularly with respect to H1. Based on these results, practical aspects for the efficient implementation of marker-assisted selection are discussed, which consider the genetic origin of the material, costs aspects and methodology applied.

Song Y S, Schwarzfischer A .

Development of STS markers for selection of extreme resistance (Rysto) to PVY and maternal pedigree analysis of extremely resistant cultivars

American Journal of Potato Research, 2008,85(2):392-393.

DOI:10.1007/s12230-008-9012-8      URL     [本文引用: 3]

Two Ry sto STS markers, YES3-3A (34102bp) and YES3-3B (28602bp), were successfully developed from the Ry sto AFLP marker E+ACC/M+CTC-365. Both STS markers identified all 38 extremely resistant potato cultivars out of 188 European potato varieties from Germany, Hungary, Poland and The Netherlands, and also identified four extremely resistant MPI breeding lines out of five tested lines, in agreement with the phenotypic data of resistance level to PVY. In parallel, three different plastid-mitochondrial configurations (W/α, W/γ, and T/β) were analyzed by the PCR markers ALC_1,3 and ALM_4,5. Thirty-eight potato varieties were classified as the W/α cytoplasm type, 51 potato varieties were identified as carrying the W/γ cytoplasm type, and 99 potato varieties analyzed had the T/β cytoplasm type. All 38 marker-selected extremely resistant potato cultivars and four extremely resistant MPI breeding lines were identified as having the mitochondrial γ type in association with male sterility. Male sterility was validated by pollen germination tests in 38 potato cultivars carrying the mitochondrial α type or γ type. The male sterility of 17 extremely resistant potato cultivars and five potato cultivars carrying the W/γ cytoplasm type was confirmed by obtaining scarcely any or only few or wrinkled pollen grains and no subsequent pollen germination. On the other hand, pollen germination in all 16 potato cultivars with the W/α cytoplasm type was observed with different efficiency due to different genotypes. The Ry sto gene of extremely resistant potato cultivars and MPI breeding lines traced back to Solanum stoloniferum , and the maternal pedigree analysis of these cultivars clustered into three groups depending on three different primary crosses with S. stoloniferum -hybrids. This pedigree analysis confirmed the precision of Ry sto STS marker selection tagging the maternally inherited Ry sto gene derived from MPI breeding lines.

Flis B, Hennig J, Marczewski W , et al.

The Ry-fsto gene from Solanum stoloniferum for extreme resistant to potato virus Y maps to potato chromosome Ⅻ and is diagnosed by PCR marker GP122718 in PVY resistant potato cultivars

Molecular Breeding, 2005,15(1):95-101.

DOI:10.1007/s11032-004-2736-3      URL     [本文引用: 1]

A novel locus for extreme resistance to Potato virus Y (PVY), Ry-f sto , was identified on potato chromosome XII. The gene Ry-f sto has been introgressed from the wild potato species Solanum stoloniferum . Inheritance of Ry-f sto in the tetraploid potato population Rysto was consistent with the model of a single, dominant gene. Bulked segregant analysis identified an ISSR (inter-simple sequence repeat) marker UBC 857 980 linked to Ry-f sto . This marker mapped to linkage group XII of a reference potato RFLP (restriction fragment length polymorphism) map. Chromosome XII specific RFLP markers were converted into PCR-based STS and CAPS markers and tested for linkage with Ry-f sto in the population Rysto. CAPS marker GP122 718 was tightly linked to the resistance gene and was successfully used to identify Polish and German cultivars expressing extreme resistance to PVY. This indicates that the source of Ry-f sto has been widely utilized in various potato breeding programs and can be monitored by a diagnostic marker in marker-assisted selection.

Witek K, Strzelczyk-Zyta D, Hennig J , et al.

A multiplex PCR approach to simultaneously genotype potato towards the resistance alleles Ry-fsto and Ns

Molecular Breeding, 2006,18(3):273-275.

DOI:10.1007/s11032-006-9021-6      URL     [本文引用: 2]

A simple and robust multiplex PCR approach was developed for detection of the alleles Ry-f sto and Ns conferring resistance of potato to Potato Virus Y (PVY) and Potato Virus S (PVS), respectively. Cleaved amplified polymorphic sequence (CAPS) markers GP122 564 linked to Ry-f sto and SC811 260 linked to Ns were amplified in one PCR reaction and identified after simultaneous digestion of the amplicons with restriction enzymes Eco RV and Mbo I. Effectiveness of this procedure for marker-assisted selection was confirmed in 55 potato cultivars.

Hosaka K, Hosaka Y, Mori M , et al.

Detection of a simplex RAPD marker linked to resistance to potato virus Y in a tetraploid potato

American Journal of Potato Research, 2001,78(3):191-196.

DOI:10.1007/BF02883544      URL     [本文引用: 1]

Extreme resistance to potato virus Y, derived from a wild diploid species Solanum chacoense , was found in Japanese cultivar Konafubuki. The segregation ratio of resistant vs susceptible in the tetraploid population from Kita-akari (susceptible) x Konafubuki (resistant) indicated that the resistance gene followed a monogenic dominant fashion. Bulked DNA samples of resistant and of susceptible clones were screened with 306 decamer primers by PCR to find RAPD markers linked to the resistance. The RAPD marker 38-530 was reproducibly detected in the resistant clones with a recombination frequency of 16.3%. Except for Konafubuki the marker band was found only in a few limited parental lines and cultivars where the resistance is not involved. Thus, using Konafubuki as a resistance gene source, the RAPD marker 38-530 would be practically and widely useful in tetraploid breeding programs.

Celebi-Toprak F, Slack S A, Jahn M M .

A new gene,Nytbr,for hypersensitivity to potato virus Y from Solanum tuberosum maps to chromosome Ⅳ

Theoretical & Applied Genetics, 2002,104(4):669-674.

DOI:10.1007/s001220100749      URL     PMID:12582672      [本文引用: 1]

A diploid backcross population derived from a cross between Solanum tuberosum and Solanum berthaultii segregated for monogenic dominant hypersensitivity to Potato virus Y (PVY). We propose the symbol Ny tbr for this locus because plants carrying this gene develop necrosis after inoculation with PVY and the allele originated in S. tuberosum. The gene mapped to chromosome IV between TG316 and TG208 at LOD=2.72. This location does not correspond to any other mapped resistance genes in potato.

Szajko K, Strzelczyk-Zyta D, Marczewsli W .

Ny-1 and Ny-2 genes conferring hypersensitive response to potato virus Y (PVY) in cultivated potatoes:mapping and marker-assisted selection validation for PVY resistance in potato breeding

Molecular Breeding, 2014,34(1):267-271.

DOI:10.1007/s11032-014-0024-4      URL     PMID:4030098      [本文引用: 1]

Potato virus Y (PVY) is one of the most important viruses affecting potato ( Solanum tuberosum ) production. In this study, a novel hypersensitive response (HR) gene, Ny - 2 , conferring resistance to PVY was mapped on potato chromosome XI in cultivar Romula. In cultivars Albatros and Sekwana, the Ny - 1 gene was mapped on chromosome IX. In cv. Romula, the local lesions appeared in leaves inoculated with the PVY N-Wi isolate at 20 and 2802°C; PVY systemic infections were only occasionally observed at the higher temperature. In cvs. Albatros and Sekwana, expression of the necrotic reaction to virus infection was temperature-dependent. PVY N-Wi was localized at 2002°C; at 2802°C, the systemic, symptomless infection was observed. We developed the B11.6 1600 marker co-segregating with Ny - 2 and the S1d11 marker specific for the Ny - 1 gene. Fifty potato cultivars were tested with markers B11.6 and S1d11 and marker SC895 linked to the Ny - 1 gene in cv. Rywal. These results indicated the utility of these markers for marker-assisted selection of HR-like PVY resistance in potato breeding programs.

Szajko K, Chrzanowska M, Witek K , et al.

The novel gene Ny-1 on potato chromosome Ⅸ confers hypersensitive resistance to potato virus Y and is an alternative to Ry genes in potato breeding for PVY resistance

Theoretical & Applied Genetics, 2008,116(2):297-303.

[本文引用: 2]

Burkhart C R, Christ B J, Haynes K G .

Non-additive genetic variance governs resistance to Fusarium dry rot in a diploid hybrid potato population

American Journal of Potato Research, 2007,84(3):199-204.

DOI:10.1007/BF02986269      URL     [本文引用: 3]

Fusarium dry rot, caused by several Fusarium species , is a major storage disease of potatoes for which there is no fungicidal control. Levels of resistance in commercial potato germplasm are inadequate. The purpose of this study was to determine the inheritance of resistance to Fusarium dry rot in a diploid hybrid Solanum phureja-Solanum stenotomum population. Three tubers from each of four half-sibs from each of 38 diploid families were inoculated with a mixture of two isolates of Fusarium sambucinum and one isolate of Fusarium solani four times in both 2003-2004 and 2004-2005. Tubers were then incubated for 40 days at 15 C and 90% relative humidity. The surface diameter and depth of infected tissue were measured in two directions, at right angles to each other. Depth of infected tissue was determined by cutting the tuber in half through the inoculation point. Mean depth and diameter of infected tissue were analyzed. There were significant differences among clones. The experiment x clone interaction was also significant. Broad-sense heritabilities and their 95% confidence intervals for resistance to Fusarium dry rot in this population were estimated as 0.63 (0.50, 0.71) and 0.81 (0.76, 0.86) in 2003-2004 and 2004-2005, respectively, for mean diameter; and as 0.68 (0.57, 0.75) and 0.81 (0.75, 0.86) in 2003-2004 and 2004-2005, respectively, for mean depth. Narrow-sense heri-tabilities for mean diameter and depth were not significantly different from zero either year. Although there is genetic variation for resistance to Fusarium dry rot in this population, these results indicate that additive genetic variance is lacking or minimal, and therefore little or no genetic gain in resistance will be realized. A few highly resistant clones could, via 4x-2x crosses, theoretically transfer much of the dominance and epistatic variance governing resistance in this population to the tetraploid level.

Marczewski W, Flis B, Syller J , et al.

Two allelic or tightly linked genetic factors at the PLRV.4 locus on potato chromosome XI control resistance to potato leafroll virus accumulation

Theoretical & Applied Genetics, 2004,109(8):1604-1609.

DOI:10.1007/s00122-004-1780-z      URL     PMID:15448896      [本文引用: 1]

A novel locus for potato resistance to potato leafroll virus (PLRV) was characterized by inheritance studies and molecular mapping. The diploid parental clone DW 91-1187 was resistant to PLRV accumulation in both inoculated plants and their tuber progeny. The resistance to PLRV accumulation present in DW 91-1187 was not transmitted to any F 1 offspring when crossed with a PLRV susceptible clone. Instead, one half of the F 1 individuals exhibited undetectable amounts of PLRV as determined by ELISA during the primary infection assay, but accumulated PLRV in their tuber progeny plants. The other half was clearly infected both in the inoculated and tuber-born plants. The inheritance of resistance to PLRV accumulation may be explained by a model of two complementary alleles of a single gene ( PLRV.4 ) or by two complementary genes that are closely linked in repulsion phase. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers linked to the PLRV.4 locus were selected. The two complementary factors were closely linked in coupling phase to the alternative alleles UBC864 600 and UBC864 800 of DNA marker UBC864. These markers may be used for marker-assisted selection of genotypes having both factors for resistance to PLRV accumulation. The PLRV.4 locus was mapped to a central position on linkage group XI of the potato molecular map, where no resistance locus has been mapped previously.

Kuhl J C, Novy R G, Jonathan L W , et al.

Development of molecular markers closely linked to the potato leafroll virus resistance gene,Rlretb,for use in marker-assisted selection

American Journal of Potato Research, 2016,93(3):203-212.

DOI:10.1007/s12230-016-9496-6      URL     [本文引用: 3]

Potato leafroll virus (PLRV) is a major pathogen of potato with worldwide impact on seed and commercial production of potato. In North America, the primary varieties grown by industry are not resistant to PLRV and require the application of insecticides to control the aphid vector of PLRV, so as to prevent PLRV infection of the crop. The Solanum etuberosum- derived dominant gene Rlretb confers resistance to potato leafroll virus and has been mapped to chromosome 4. The previous closest marker to Rlretb , C2_At1g42990, was mapped 13.6 cM from the gene in a BC3 population. The development of additional molecular markers closely linked to Rlretb can significantly facilitate its use in breeding and its continued introgression into cultivated potato. Cleaved amplified polymorphic sequence markers were developed in the region surrounding C2_At1g42990 and mapped along with Rlretb in 102 BC4 progeny. Sixteen markers were identified surrounding Rlretb, with flanking markers at 2.1 and 9.3 cM. Two markers, 1367-8a and DMB32-11, both 2.1 cM from Rlretb, are shown to be well suited for marker assisted selection. Currently, breeding clones with the presence of Rlretb have been developed that comprise market classes characterized by having tubers that are round and white, or long and russet-skinned.

Velásquez A C, Mihovilvich E, Bonierbale M .

Genetic characterization and mapping of major gene resistance to potato leafroll virus in Solanum tuberosum ssp. andigena

Theoretical & Applied Genetics, 2007,114(6):1051-1058.

DOI:10.1007/s00122-006-0498-5      URL     PMID:17394033      [本文引用: 1]

Major gene inheritance of resistance to Potato leafroll virus (PLRV) was demonstrated in a parthenogenic population derived from the highly resistant tetraploid a ndigena landrace, LOP-868. This major gene or chromosome region seems to control a single mechanism for resistance to infection and virus accumulation in this source. About 149 dihaploid lines segregated in a ratio of 107 resistant to 32 susceptible, fitting the expected ratio for inheritance of a duplex gene under random chromatid segregation. A tetraploid AFLP map was constructed using as reference the ultra high density (UHD) map. All AFLP markers associated with PLRV resistance mapped to the same linkage group. Map position was confirmed by analysis of previously-mapped SSR markers. Rl adg is located on the upper arm of chromosome V , at 1 cM from its most closely linked AFLP marker, E35M48.192. This marker will be used to develop allele-specific primers or a pair of flanking PCR-based markers for their use in marker assisted selection.

Mihovilovich E, Aponte M, Lindqvist-Kreuze H , et al.

An RGA-derived SCAR marker linked to PLRV resistance from Solanum tuberosum ssp. andigena

Plant Molecular Biology Reporter, 2014,32(1):117-128.

DOI:10.1007/s11105-013-0629-5      URL     [本文引用: 1]

ABSTRACT A resistance gene analog (RGA)-derived sequence-characterized amplified region (SCAR) marker was successfully developed based on sequence homology with disease resistance genes of an AFLP molecular marker tightly linked to the Rl adg gene of Solanum tuberosum ssp. andigena. The new marker was designated as ‘RGASC850’ (RGA-derived SCAR) based on the size of the amplified fragment. ‘RGASC850’ could be efficiently used for monitoring introgression of Rl adg against backgrounds of improved gene pools with low likelihood of identifying false positives due to recombination. This SCAR proved to be highly predictive of Rl adg -based resistance, as it did not amplify potato leafroll virus (PLRV) resistance sources other than andigena, and thus would be useful in developing cultivars with complementary sources of resistance to PLRV. In addition, a cleaved amplified polymorphic sequence (CAPS) marker based on ‘RGASC850’ was developed capable of distinguishing genotypes carrying Rl adg . This CAPS marker would be useful for screening breeding populations derived from wide crosses, and confirming presence of the Rl adg gene in those parents amplifying the ‘RGASC850’ marker.

Jong W D, Forsyth A, Leister D , et al.

A potato hypersensitive resistance gene against potato virus X maps to a resistance gene cluster on chromosome 5

Theoretical & Applied Genetics, 1997,95(1/2):246-252.

DOI:10.1007/s001220050555      URL     [本文引用: 3]

The dominant Nb gene of potato confers strain-specific hypersensitive resistance against potato virus X (PVX). A population segregating for Nb was screened for resistance by inoculating with PVX strain CP2, which is sensitive to Nb . Through a combination of bulked segregant analysis and selective restriction fragment amplification, several amplified fragment length polymorphism (AFLP) markers linked to Nb were identified. These were cloned and converted into dominant cleaved amplified polymorphic sequence (CAPS) markers. The segregation of these markers in a Lycopersicon esculentum × L. pennellii mapping population suggested that Nb is located on chromosome 5. This was confirmed by examining resistant and susceptible potato individuals with several tomato and potato chromosome-5-specific markers. Nb maps to a region of chromosome 5 where several other resistance genes– including R1 , a resistance gene against Phytophthora infestans , Gpa , a locus that confers resistance against Globodera pallida , and Rx2 , a gene that confers extreme resistance against PVX–have previously been identified.

Marczewski W, Hennig J, Gebhardt C .

The potato virus S resistance gene Ns maps to potato chromosome Ⅷ

Theoretical & Applied Genetics, 2002,105(4):564-567.

DOI:10.1007/s00122-002-0976-3      URL     PMID:12582505      [本文引用: 1]

The dominant allele Ns confers in potato resistance to Potato virus S (PVS). To identify the chromosomal location of Ns , we mapped the Ns -linked marker SCG17 448 and the ISSR marker UBC811 600 to linkage group VIII of the RFLP map of a population that did not segregate for Ns . The map position of the Ns locus on chromosome VIII was confirmed with the detection of linkage between Ns and three RFLP markers, GP126, GP189 and CP16, known to be located in a corresponding region on potato chromosome VIII. PCR-based assays were developed for these RFLP markers. The PCR primers specific for GP126 generated polymorphic products (STS marker). In the case of markers GP189 and CP16, informative polymorphism was revealed in the Ns population after digestion with the restriction enzymes Hae III and Hin dIII, respectively. The genetic distance between Ns and the closest CP16 locus was 4.2 cM.

Marczewski W, Strzelczyk-Żyta D, Hennig J , et al.

Potato chromosomes Ⅸ and Ⅺ carry genes for resistance to potato virus M

Theoretical & Applied Genetics, 2006,112(7):1232-1238.

DOI:10.1007/s00122-006-0224-3      URL     PMID:16453130      [本文引用: 3]

Two new loci for resistance to potato virus M (PVM), Gm and Rm , have been mapped in potato. The gene Gm was derived from Solanum gourlayi , whereas, Solanum megistacrolobum is the source of the gene Rm . Gm confers resistance to PVM infection after mechanical inoculation. Rm induces a hypersensitive response in potato plants. Two diploid populations segregating for Gm and Rm , bulked segregant analysis (BSA) using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR), and available potato molecular maps were instrumental for mapping the resistance loci. The novel locus Gm was mapped to a central region on potato chromosome IX. The locus Rm was placed on the short arm of chromosome XI, close to the marker loci GP250 and GP283 , where a hotspot for monogenic and polygenic resistance to diverse pathogens is located in the potato and tomato genome.

Bebdahmane A, Kanyuka K, Baulcombe D C .

High-resolution genetical and physical mapping of the Rx gene for extreme resistance to potato virus X in tetraploid potato

Theoretical & Applied Genetics, 1997,95(1/2):153-162.

DOI:10.1007/s001220050543      URL     [本文引用: 1]

The Rx locus in potato confers extreme resistance to PVX. In the F 1 progeny of crosses between the PVX-susceptible cultivar Huinkel and the cultivar Cara ( Rx genotype) there was a 165:651 segregation of PVX resistance, indicating that Rx in Cara is present in the simplex condition. Using potato and tomato RFLP markers, we mapped Rx in Cara to the distal end of chromosome XII at a different position to the previously mapped Rx 1 locus. To generate a high-resolution linkage map in the vicinity of Rx a total 728 AFLP primer combinations were screened using DNA of bulked resistant and susceptible segregants. We also screened segregating populations for chromosomal recombination events linked to the Rx locus and identified 82 plants with recombination events close to Rx . Using these recombinant plants we have identified AFLPs that flank Rx and span an interval of 0.23 cM in a region of the genome where 1 cM corresponds to approximately 40061kb.

Wastie R L, Bradshaw J E ,

Mackay. Inheritance of resistance to fungal diseases of tubers

Wallingford:CAB International, 1994.

URL     [本文引用: 1]

The diseases considered in this chapter range from those caused by persistent soil borne fungi to those which are tuber-borne. The diseases are discussed in an order which reflects the extent to which the genetics of resistance is understood: Synchytrium endobioticum, dry rot (Fusarium solani, F. sulphureum [Gibberella cyanogena], F. sambucinum [G. pulicaris] and G. avenacea), Rhizoctonia solan...

Gebhardt C, Bellin D, Henselewski H , et al.

Marker-assisted combination of major genes for pathogen resistance in potato

Theoretical & Applied Genetics, 2006,112(8):1458-1464.

DOI:10.1007/s00122-006-0248-8      URL     PMID:16538512      [本文引用: 1]

Abstract Closely linked PCR-based markers facilitate the tracing and combining of resistance factors that have been introgressed previously into cultivated potato from different sources. Crosses were performed to combine the Ry ( adg ) gene for extreme resistance to Potato virus Y (PVY) with the Gro1 gene for resistance to the root cyst nematode Globodera rostochiensis and the Rx1 gene for extreme resistance to Potato virus X (PVX), or with resistance to potato wart (Synchytrium endobioticum). Marker-assisted selection (MAS) using four PCR-based diagnostic assays was applied to 110 F1 hybrids resulting from four 2x by 4x cross-combinations. Thirty tetraploid plants having the appropriate marker combinations were selected and tested for presence of the corresponding resistance traits. All plants tested showed the expected resistant phenotype. Unexpectedly, the plants segregated for additional resistance to pathotypes 1, 2 and 6 of S. endobioticum, which was subsequently shown to be inherited from the PVY resistant parents of the crosses. The selected plants can be used as sources of multiple resistance traits in pedigree breeding and are available from a potato germplasm bank.

Ritter E, Debener T, Barone A , et al.

RFLP mapping on potato chromosomes of two genes controlling extreme resistance to potato virus X (PVX)

Molecular and General Genetics, 1991,227(1):81-85.

DOI:10.1007/BF00260710      URL     PMID:1675423      [本文引用: 1]

Two different chromosomal locations of major genes controlling extreme resistance to potato virus X (PVX) were found by restriction fragment length polymorphism ( RFLP ) analysis of two populations segregating for the resistance. The resistance gene Rx1 mapped to the distal end of chromosome XII, whereas Rx2 was located at an intermediate position on linkage group V in a region where reduced recombination and segregation distortion have also been observed. These linkage anomalies were due to abnormal behaviour of the chromosome contributed by the resistant parent P34. The results presented were obtained using two different strategies for mapping genes of unknown location. One approach was the use of probes revealing polymorphic loci spread throughout the genome and resulted in the mapping of Rx1 . The second approach was based on the assumption of possible linkage between the resistance gene and clone-specific DNA fragments introduced from a wild potato species. Rx2 was mapped by adopting this strategy.

Tommiska J T, Watanabe N K, Valkonen T J P ,et al.

Mapping of the gene Nxphu that controls hypersensitive resistance to potato virus X in Solanum phureja IVP35

Theoretical & Applied Genetics, 1998,96(6):840-843.

DOI:10.1007/s001220050810      URL     [本文引用: 1]

The line IvP35 of the diploid (2n=2x=24) cultivated potato species Solanum phureja (family Solanaceae) expresses hypersensitive resistance (H) to potato X potexvirus (PVX). In this study, a diploid potato population was produced using IvP35 as the male parent and a diploid line of S. tuberosum (87HW13.7) as the female parent and tested for resistance to PVX. Data indicated that H to PVX in IvP35 is a dominant, monogenically inherited trait controlled by a single gene, named Nx phu , that is in a simplex condition ( Nxnx ). RFLP analysis carried out on the progeny lines revealed 4 markers (CT220, TG328, CT112 and TG424) from the long arm of chromosome IX that were linked to the hypersensitive phenotype; the closest linkage was observed with the marker TG424. Previous authors have shown that the same region of chromosome IX contains the gene Sw-5 for resistance to tomato spotted wilt tospovirus in Lycopersicon peruvianum (Solanaceae).

Lossl A, Adler N, Horn R , et al.

Chondriome-type characterization of potato:Mt α,β,γ,ε and novel plastid-mitochondrial configurations in somatic hybrids

Theoretical & Applied Genetics, 1999,99(1/2):1-10.

DOI:10.1007/s001220051202      URL     [本文引用: 1]

One hundred and eighty dihaploid clones used for protoplast fusions, and 144 tetraploid German potato cultivars were analysed for their cytoplasms using 11 homologous mt DNA-probes, and were classified as mitochondrial (mt) types α, β, γ, δ, and 07 according to their RFLP patterns. From the 4 x cultivars, 79 had the typical mt-type β of Solanum tuberosum being different from the 46 cvs which had the mt-α type and 19 others with mt-γ. A dendrogram shows their relationships to other Solanum species. The distantly related mt-07 was only found in di-haploids, and particularly in clones deriving from Solanum phureja and Solanum andigena . Accessory mt types will be actualized on website (http://www.edv.agrar.tu-muenchen.de/pbpz/ mm/mt/al1.htm). In order to evaluate the genetic potential of novel plastid-mitochondrial configurations we have analyzed four representative populations, which derive from different fusion-combination classes: [α (+) β], [α (+) γ], [α (+) δ] and [α (+) 07]. On the mitochondrial expression level, hybrids from an [α (+) 07] fusion could be distinguished by in-organello translation from [α (+) β] hybrids, and other di-haploids, by an additional translation product of 1561kDa. In fusion parents with mt-α and -γ an additional atp6 reading frame is detectable in sub-stoichiometric amounts by the use of specific PCR primers. The gene differs from the original 211 bp 3′ from the stop codon. Novel RFLP -patterns in 10% of the somatic hybrids were due to a high-rate replication of this pre-existing parental genome region. A second characteristic for somatic hybrids was the partial addition of parental mt sub-genomes. The major part of them revealed a new organization in their mt genomes at the mt-type characteristic loci rpl5 , rps14 , cob, rps10 , coxI and rpl2 , which contain recombination-specific repeats homologous to Petunia spp. and Nicotiana . A schematic model for the formation of novel mitochondrial genomes in potato somatic hybrids is provided.

Sanetomo R, Hosaka K .

A maternally inherited DNA marker,descended from Solanum demissum (2n=6x=72) to S. tuberosum (2n=4x=48)

Breeding Science, 2011,61(4):426-434.

DOI:10.1270/jsbbs.61.426      URL     PMID:3406774      [本文引用: 2]

A Mexican hexaploid wild potato species, Solanum demissum (dms), was only used as a female in previous breeding programs. The resulting clones with dms cytoplasm produced abundant, but non-functional pollen. A 170 bp DNA fragment, named Band 1, was originally detected in the F(1) hybrid between dms and S. tuberosum. In this study, the sequenced region was extended to 1,032 bp; nevertheless, it did not show any homology to known sequences. This extended region harboring Band 1 was, without introns, all transcribed to mRNA and was maternally inherited from dms to S. tuberosum through backcrosses. Three dms accessions, 168 accessions of 38 cultivated and closely related wild species, and 158 varieties and breeding lines were surveyed, which demonstrated that Band 1 was specific to dms and varieties and breeding lines with dms cytoplasm. Thus, Band 1 is a useful marker to distinguish dms cytoplasm, which enables us to design efficient mating combinations in breeding programs.

Powell W, Baird E, Duncan N , et al.

Chloroplast DNA variability in old and recently introduced potato cultivars

Annals of Applied Biology, 1993,123(2):403-410.

DOI:10.1111/j.1744-7348.1993.tb04102.x      URL     [本文引用: 1]

Chloroplast DNA (cpDNA) variability has been examined in a range of tetraploid European potato cultivars. The potato genotypes studied included primitive cultivars such as Lumpers (1806), Yam (1836), Myatts Ashleaf (1847) and more recently bred cultivars such as Brodick (1990). Three cpDNA pheno-types were detected and these probably represent original introductions from South America into Europe. The most common cp phenotype was the T type cpDNA (Hosaka & Hanneman, 1988) characteristic of cultivars descended from cv. Rough Purple Chili. The presence of the T type cpDNA in the cultivar Yam indicates that this genotype which is of Andigena origin shares a common cytoplasm with other 5. tuberosum ssp. tuberosum clones which have a Chilean type cytoplasm. The implications of these results are discussed in relation to the origin of the T type cpDNA. Methods for increasing the cytoplasmic diversity of the cultivated potato gene pool are proposed.

Hosaka K, Sanetomo R .

Development of a rapid identification method for potato cytoplasm and its use for evaluating Japanese collections

Theoretical & Applied Genetics, 2012,125(6):1237-1251.

DOI:10.1007/s00122-012-1909-4      URL     PMID:22696007      [本文引用: 1]

The cytoplasm of potatoes, characterized by the presence of T-type chloroplast DNA and β-type mitochondrial DNA, is sensitive to nuclear chromosomal genes that contribute to various types of male sterility. Past breeding efforts with various potato varieties have resulted in several different cytoplasms other than T/β. Varieties with Solanum stoloniferum -derived cytoplasm (W/γ) show complete male sterility, while those with S. demissum -derived cytoplasm (W/α) produce abundant, but non-functional pollen. Thus, identification of cytoplasmic types is important for designing efficient mating combinations. To date, only T-type chloroplast DNA can be accurately identified by a PCR marker. Here, we report a rapid identification technique by multiplex PCR, followed by restriction digestion with Bam HI in one reaction tube, and propose a new nomenclature for potato cytoplasm types (T, D, P, A, M, and W). Using this new technique, our collections of 748 genotypes, including 84 Japanese named varieties, 378 breeding lines and 26 landraces, and 260 foreign varieties and breeding lines, were grouped into cytoplasm types: T (73.902%), D (17.402%), P (4.502%), A (1.502%), M (0.302%), and W (2.402%). The utility of this marker system for breeding is discussed.

Provan J, Powell W, Dewar H , et al.

An extreme cytoplasmic bottleneck in the modern European cultivated potato (Solanum tuberosum) is not reflected in decreased levels of nuclear diversity. Proceedings

Biological Sciences, 1999,266(1419):633-639.

DOI:10.1098/rspb.1999.0683      URL     PMID:1689813      [本文引用: 1]

We have used the polymorphic chloroplast (cp) and nuclear simple sequence repeats (SSRs) to analyse levels of cytoplasmic and nuclear diversity in the gene pool of the European cultivated potato (Solanum tuberosum sap. tuberosum). Primers designed from the complete chloroplast sequence of tobacco (Nicotiana tabacum) were used to amplify polymorphic products in a range of potato cultivars. Combining the data from seven polymorphic cpSSR loci gave 26 haplotypes, one of which (haplotype A) accounted for 151 out of the 178 individuals studied and corresponded to the T-type cytoplasm previously identified in cultivated potatoes using chloroplast restriction fragment length polymorphism analysis. Phylogenetic and diversity analyses of the relationships between cpSSR haplotypes confirmed much higher levels of cytoplasmic diversity outwith the T-type group. Diversity levels at eight nuclear SSR loci, however, were not significantly different between cytoplasmic groups, suggesting a severe maternal bottleneck in the evolution of the modern cultivated potato. These results highlight the importance in quantifying levels of cytoplasmic as well as nuclear diversity and confirm the need for a change in breeding practices to increase levels of non-T-type cytoplasm in the cultivated gene pool, thus helping reduce problems associated with pollen sterility. This may be facilitated by germplasm analysis using cpSSRs, which will allow efficient selection of diverse cytoplasm donors.

Sukhotu T, Kamijima O, Hosaka K .

Genetic diversity of the Andean tetraploid cultivated potato (Solanum tuberosum L. subsp. andigena Hawkes) evaluated by chloroplast and nuclear DNA markers

Genome, 2005,48(1):55-64.

DOI:10.1139/g04-086      URL     [本文引用: 1]

易靖 .

马铃薯品种资源分子标记检测及其与重要生物学性状的关联分析

昆明:云南师范大学, 2016.

URL     [本文引用: 1]

我国马铃薯的育种研究先后经历了几个阶段,从最初的引种鉴定到现在的分子标记辅助育种,目的都是为了选育出更优质的马铃薯品种。对马铃薯种质资源的研究可以加速新品种选育的进程。本研究利用分子标记手段对云南师范大学保存的马铃薯种质资源的细胞核和细胞质基因组的遗传多样性进行了检测;对部分马铃薯品种的细胞质类型及其与重要生物学性状的关联进行了分析;同时利用形态学观察、细胞质和细胞核基因组多样性标记检测方法对不同来源的马铃薯合作88品种的真实性进行了检测。主要结果如下:1.利用4对叶绿体基因组标记引物和2对线粒体基因组标记引物对87份四倍体和9份二倍体马铃薯材料进行了检测。共检测出7种细胞质类型,分别是W/α[D]、W/γ[D]、W/α、W/γ、T/β、P和A/β型。国外引进品种的细胞质类型几乎全为T/β;云南省地方特色品种资源检测出A/β、W/α[D]和T/β三种类型;云南师范大学建立抗晚疫病育种群体B材料均为W/α[D],与国内外主栽品种细胞质类型不同。2.利用12对SSR标记引物对96份马铃薯种质资源进行了检测,获得96个马铃薯品种的分子指纹信息。按照细胞质遗传背景可将这些种质资源分为携带野生种(Wild,W)或普通栽培种(Tuberosum,T)细胞质基因组两个类群;比较而言,具有W细胞质类型的马铃薯种质资源的遗传多样性更丰富。3.对部分马铃薯品种的细胞质类型与某些重要性状的关联分析发现,具有W细胞质类型(包括安第斯栽培亚种细胞质类型【Andigena,A】)品种的抗性、成熟期和淀粉含量分别与T细胞质类型品种的抗性、成熟期和淀粉含量之间存在极显著差异;具有W细胞质类型品种的产量显著高于具有T细胞质类型的品种。4.利用形态学观察、细胞质和细胞核基因组多样性标记检测方法,对来自云南省12个主产区的143份―合作88‖样本进行了检测。形态学观察发现一份非合作88材料;细胞质基因组标记检测出一份非合作88材料;细胞核基因组SSR标记检测出23份样品与合作88标准品种不完全相同。上述结果表明,本文描述的细胞质基因组标记可以应用于马铃薯育种亲本选择和新品种辅助选育,核基因组SSR标记检测方法可揭示马铃薯种质资源的遗传多样性,但在品种真实性或纯度鉴定方面有待于进一步完善。

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