作物杂志, 2019, 35(6): 14-19 doi: 10.16035/j.issn.1001-7283.2019.06.003

遗传育种·种质资源·生物技术

黄淮南片麦区新育成品种(系)中3个矮秆基因分子标记检测及其与农艺性状的关系

曹廷杰1, 张玉娥1, 胡卫国1, 杨剑1, 赵虹1, 王西成1, 周艳杰1, 赵群友2, 李会群3

1河南省农业科学院小麦研究所,450002,河南郑州

2南阳市种子管理站,473000,河南南阳

3濮阳市种子管理站,457000,河南濮阳

Detection of Three Dwarfing Genes in the New Wheat Cultivars (Lines) Developed in South Huang-Huai Valley and Its Association with Agronomic Traits

Cao Tingjie1, Zhang Yu’e1, Hu Weiguo1, Yang Jian1, Zhao Hong1, Wang Xicheng1, Zhou Yanjie1, Zhao Qunyou2, Li Huiqun3

1Wheat Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan, China

2Seed Management Department of Nanyang, Nanyang 473000, Henan, China

3Seed Management Department of Puyang, Puyang 457000, Henan, China

收稿日期: 2019-05-28   修回日期: 2019-08-21   网络出版日期: 2019-12-15

基金资助: 河南省农业科学院自主创新基金,河南省小麦产业技术体系(S2010-01-G03)

Received: 2019-05-28   Revised: 2019-08-21   Online: 2019-12-15

作者简介 About authors

曹廷杰,副研究员,主要从事小麦育种及重要性状遗传分析研究 。

摘要

利用矮秆基因RhtB1-b、RhtD1-b和Rht8特异分子标记对郑麦583和2015-2016年度参加河南省区域试验、河南省品种比较试验、国家黄淮南片区域试验及国家黄淮麦区品种比较试验的共630份小麦材料的基因型进行检测。结果表明,供试材料中检测到549份材料含有RhtB1-b基因;592份材料含有RhtD1-b基因;513份材料含有Rht8基因;422份材料同时含有3个矮秆基因,169份材料仅含有2个矮秆基因,说明3个主要的矮秆基因在河南小麦育种过程中被聚合使用。此外,分析发现,矮秆基因Rht8与株高和每公顷穗数,以及千粒重具有显著相关性。郑麦583等小麦品种聚合了这3个矮秆基因,具有优良的丰产性,通过选择和利用矮秆基因对于培育具有丰产性优点的小麦品种具有一定价值。

关键词: 小麦 ; 株高 ; 矮秆基因 ; 农艺性状

Abstract

In this study, Zhengmai 583 and 630 materials derived from variety comparison test and regional trials of Yellow-Huai wheat area and Henan Province at 2015-2016 were screened by specific molecular marker of wheat dwarf genes RhtB1-b, RhtD1-b and Rht8. The results showed that 549 wheat materials carried RhtB1-b gene; 592 wheat materials carried RhtD1-b gene; 513 wheat materials carried Rht8 gene; 422 materials carried all 3 dwarfing genes, and additional 169 lines carried 2 dwarfing genes which indicated that the 3 dwarf genes had been fixed in Henan wheat breeding progress. Moreover, Rht8 showed significantly correlation with reducing plant height, spike number per hectare and increasing 1000-kernel weight. Zhengmai 583 carried 3 dwarf genes, showed excellent fertility. The selection and utilization of dwarf genes are of certain value for the cultivation of wheat varieties with high yield.

Keywords: Wheat ; Plant height ; Dwarfing gene ; Agronomic traits

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曹廷杰, 张玉娥, 胡卫国, 杨剑, 赵虹, 王西成, 周艳杰, 赵群友, 李会群. 黄淮南片麦区新育成品种(系)中3个矮秆基因分子标记检测及其与农艺性状的关系[J]. 作物杂志, 2019, 35(6): 14-19 doi:10.16035/j.issn.1001-7283.2019.06.003

Cao Tingjie, Zhang Yu’e, Hu Weiguo, Yang Jian, Zhao Hong, Wang Xicheng, Zhou Yanjie, Zhao Qunyou, Li Huiqun. Detection of Three Dwarfing Genes in the New Wheat Cultivars (Lines) Developed in South Huang-Huai Valley and Its Association with Agronomic Traits[J]. Crops, 2019, 35(6): 14-19 doi:10.16035/j.issn.1001-7283.2019.06.003

半矮秆基因的应用是小麦获得高产抗倒的重要途径,可以降低株高从而达到减少倒伏风险,增加收获指数及提高产量的目的[1]

目前已命名的小麦矮秆基因有25个(Rht1-Rht25)[2,3,4,5],但在我国小麦生产上应用较为广泛的有RhtB1-b、RhtD1-b及Rht8。世界上超过70%的小麦品种均含有矮秆基因RhtB1-b和RhtD1-b中的1~2个[6]。其中RhtB1-b和RhtD1-b分别被定位在小麦4B和4D染色体上,为异源染色体上的同源基因[7]。RhtB1-b和RhtD1-b位点间具有加性效应,编码DELLA蛋白,主要功能为抑制植物对赤霉素(GA)的响应,但该蛋白并不结合GA受体蛋白GAINSENSITIVE DWARF1,因此,表现为对赤霉素不敏感,且由于RhtB1-b和RhtD1-b基因突变导致植株矮小。目前在小麦中鉴定出多个RhtB1-b和RhtD1-b基因的等位变异,其中Rht-B1c是在RhtB1-b基因编码区插入一个90bp的片段,造成DELLA蛋白保守区插入了30个氨基酸,相反,极度矮小植株的Rht-D1c位点的产生是由于Rht-D1b 基因发生了拷贝数变异,另一矮化位点RhtB1-b基因的提前终止,产生了Rht-B1d和Rht-B1e位点[8]

Rht8来自日本小麦品种Akakomugi,为赤霉素敏感类型,被定位于小麦2D染色体上,与SSR标记Xgwm261连锁,该分子标记目前用来检测Rht8基因,且使用较为广泛,在含有Rht8的小麦品种中可扩增出192bp的DNA片段[6,9]。Rht8对株高的降低效应为8%~12%,并且对小麦穗粒数和产量具有正效应[10],同时,该基因还具有抑制胚芽鞘和根生长的效应[11]

郑麦583是2012年河南省审定的小麦品种,株高适中,品质优良,丰产稳产性好,是目前河南省小麦生产主导品种之一。本研究以631份小麦品种(系)作为试验材料,利用3个重要矮秆基因RhtB1-b、RhtD1-b和Rht8特异的分子标记进行检测,同时对株高及产量三要素进行调查,旨在了解矮秆基因在小麦品种郑麦583和黄淮南片麦区新育成品种(系)中的存在情况及其对重要农艺性状的影响,为矮秆基因的进一步利用提供理论依据。

1 材料与方法

1.1 试验材料

供试材料共计631份,其中,郑麦583是河南省农业科学院小麦研究所选育的高产优质强筋小麦新品种,于2012年10月通过河南省审定(豫审麦2012003),其他630份为2015-2016年参加河南省区域试验、河南省品种比较试验、国家黄淮南片区域试验及国家黄淮麦区品种比较试验的小麦品种(系)。试验材料于2015年在河南省新乡市河南省农业科学院现代农业科技试验示范基地种植,采用随机区组试验,按照常规田间操作管理。每个品种种植6行,行长8.0m,行距20cm。

1.2 DNA提取

在小麦三叶期进行取样,采用CTAB法提取DNA,利用1%的琼脂糖凝胶电泳对DNA完整性进行检测,检测合格后利用紫外分光光度计检测波长260nm处的OD值并测定DNA浓度,随后将DNA稀释至50ng/μL,用于PCR反应。

1.3 RhtB1-b、RhtD1-b和Rht8分子标记检测

试验所用的RhtB1-b和RhtD1-b的分子标记参照Ellis等[12]提供的引物序列,根据携带RhtB1-b和RhtD1-b基因的农林10号作为阳性对照,Rht8分子标记参照Ellis等[6]发表的SSR引物Xgwm261,其阳性对照为Akakomugi,在该材料中可扩增出192bp的DNA片段。引物由北京擎科新业生物技术有限公司合成,根据其分子量将引物稀释至10μmol/L,PCR的反应体系为10μL:2× TSINGKE Master Mix 5μL,引物各0.5μL,DNA模板1.0μL,ddH2O 3.0μL,PCR反应条件为:95℃预变性4min;95℃30s,退火30s(退火温度参照表1),72℃延伸1min,共35个循环;72℃延伸7min。反应完成后利用8%聚丙烯酰胺凝胶电泳进行检测,经银染脱色后在日光灯下进行观察读带,使用标准分子量Marker Ⅰ[天根生化科技(北京)有限公司,MD101]进行对照。

表1   用于检测RhtB1-b、RhtD1-b及Rht8的分子标记

Table 1  Molecular markers for detection of RhtB1-b、RhtD1-b and Rht8

检测基因名称
ID of detection gene
正向引物序列(5′-3′)
Sequence of forward primer (5′-3′)
反向引物序列(5′-3′)
Sequence of reverse primer (5′-3′)
退火温度(℃)
Annealing temperature
RhtB1-bTCTCCTCCCTCCCCACCCAACCATCCCCATGGCCATCTCGAGCTA63
RhtD1-bCGCGCAATTATTGGCCAGAGATAGCCCCATGGCCATCTCGAGCTGCTA63
Rht8CTCCCTGTACGCCTAAGGCCTCGCGCTACTAGCCATTG55

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1.4 小麦农艺性状的调查

小麦抽穗后,每个品种选取1m均匀一致,有代表性的样段,在收获前随机选取20株测量株高,成熟后将样段内的植株全部拔取,进行室内考种,调查穗粒数和千粒重,计算每公顷穗数。

1.5 统计分析

对基因型和农艺性状表型进行整理,利用R(3.5.1版本,https://www.r-project.org/)对农艺性状进行描述性统计并绘制散点矩阵图,使用SPSS v22软件进行单因素方差分析。

2 结果与分析

2.1 矮秆基因RhtB1-b、RhtD1-b及Rht8的检测结果

利用引物对RhtB1-b、RhtD1-b及Rht8标记片段进行扩增,并在非变性PAGE胶中对其进行基因型检测。

根据RhtB1-b-F/RhtB1-b-R和RhtD1-b-F/RhtD1-b-R对631份小麦品种(系)进行检测,以对照农林10号作为前2个矮秆基因的阳性对照,分别检测到549份和592份小麦品种(系)材料含有RhtB1-b基因和RhtD1-b基因,条带大小分别约为400bp和254bp(图1)。

图1

图1   RhtB1-b (a)和RhtD1-b (b)引物在部分参试品种(系)中的扩增结果

M.Marker;1.农林10号;2.郑麦583;3.圣麦197;4.新良3号;5.光泰68;6.徐麦0054;7.泉麦890;8.西农501。下同

Fig.1   Amplification results of RhtB1-b (a) and RhtD1-b (b) primers in some tested cultivars (lines)

M. Marker; 1. Norin 10; 2. Zhengmai 583; 3. Shengmai 197; 4. Xinliang 3; 5. Guangtai 68; 6. Xumai 0054; 7. Quanmai 890; 8. Xinong 501. The same below


利用SSR分子标记Xgwm261检测Rht8基因,共检测出4种变异类型(图2),以阳性材料Akakomugi作为参照,共检测到513份小麦材料含有Rht8基因(192bp)。

图2

图2   Rht8 4种变异在测试品种(系)中的PCR扩增结果

9.丰德存麦12号;10.龙科1221。a.192bp;b.202bp;c.165bp;d.174bp

Fig.2   PCR amplification result of 4 varieties of Rht8 in test cultivars (lines)

9. Fengdecunmai 12; 10. Longke 1221. a. 192bp; b. 202bp; c. 165bp; d. 174bp


对631份试验材料的3个矮秆基因单倍型进行分析,5份小麦材料不含任何被检测的矮秆基因,占0.82%;422份小麦材料同时含有RhtB1-b、RhtD1-b及Rht8基因,其中郑麦583属于单倍型;169份小麦材料仅含有2个矮秆基因,占27.61%,其中98份小麦材料的单倍型为RhtB1-b/RhtD1-b/-(占16.01%),17份小麦材料的单倍型为RhtB1-b/-/Rht8(占2.78%),54份小麦材料的单倍型为-/RhtD1-b/Rht8(占8.82%);此外,16份小麦材料仅含1个矮秆基因,占2.61%,其中仅含有RhtB1-b基因的小麦品种(系)有3份(占0.49%),仅含有RhtD1-b基因的小麦品种(系)有10份(占1.63%),仅含有Rht8基因的小麦品种(系)有3份(占0.49%)(图3)。

图3

图3   631份小麦品种(系)中RhtB1-b、RhtD1-b、Rht8单倍型的分布频率

Fig.3   Distribution frequency of RhtB1-b, RhtD1-b and Rht8 haplotypes in 631 wheat cultivars (lines)


2.2 主要农艺性状描述性统计及相关性分析

631份小麦材料平均株高为76.99cm,分布范围为59.30~91.00cm,变异系数为0.06,郑麦583平均株高为76.00cm,较平均值略低。此外,对产量三要素进行调查,每公顷平均穗数为5.57×106,分布范围为3.80×106~6.63×106个,变异系数为0.08,其中郑麦583每公顷穗数为6.26×106个,高于平均值;千粒重平均为45.00g,分布范围为37.00~53.40g,变异系数为0.06,郑麦583千粒重为45.00g,与平均值相当;穗粒数平均值为34.80,分布范围为29.10~43.80,变异系数为0.06,郑麦583穗粒数为33.00,略低于平均值(表2)。

表2   631份小麦品种(系)材料农艺性状的描述性统计

Table 2  Descriptive statistics of agronomic traits in 631 wheat cultivars (lines)

性状Trait最小值
Minimum
最大值
Maximum
平均值
Mean
标准差
Standard deviation
变异系数(%)
Coefficient of variation
株高Plant height (cm)59.3091.0076.994.510.06
每公顷穗数Spike number per hectare (×106)3.806.635.570.450.08
千粒重1000-kernel weight(g)37.0053.4045.002.710.06
穗粒数Kernel number per spike29.1043.8034.802.190.06

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图4可知,4个农艺性状均表现为正态分布,其中,株高、每公顷穗数、千粒重和穗粒数在供试材料中分布最多的区间范围分别为70~80cm、3.80×106~6.63×106、43~47g、33~37。株高与每公顷穗数呈显著正相关,相关系数为0.28,与穗粒数表现为显著负相关,相关系数为-0.26。株高与千粒重之间不相关,每公顷穗数与千粒重、穗粒数之间呈现显著负相关,相关系数分别为-0.38与-0.58。千粒重与穗粒数之间也无显著相关关系。

图4

图4   4个农艺性状的散点矩阵图

柱状图表示4个性状的频率分布;左下角元件显示两两性状间的二维散点图(直线拟合线),右上角表示两两性状间的相关系数和显著性(“*” P<0.05)

Fig.4   Scatter plot matrix of 4 agronomic traits

Histogram represents the frequency distribution of 4 traits; Lower left quarter plot elements display the pairwise two-dimensional scatter plot (the straight fitting line); Upper right corner display the pairwise correlation coefficient ("*" P<0.05)


2.3 矮秆基因Rht8对主要农艺性状的影响

分别对Rht8等位基因间的株高、每公顷穗数、千粒重和穗粒数进行单因素方差分析,结果表明,Rht8对4个农艺性状的效应均达到显著水平,含有Rht8的小麦品种(系)株高降低、每公顷穗数减少、千粒重上升、穗粒数上升(表3)。不含有Rht8基因的小麦品种(系)平均株高较含有Rht8基因的品种(系)平均株高高1.56cm,每公顷穗数多0.12×106个,含有Rht8基因的小麦品种(系)平均千粒重和平均穗粒数比不含有Rht8基因的品种(系)分别高0.56g、0.87。

表3   Rht8对4个农艺性状的影响

Table 3  Effects of Rht8 on 4 agronomic traits

性状
Trait
位点
Locus
样本容量
Sample size
平均数
Mean
标准差
Standard error
最小值
Minimum
最大值
Maximum
F显著性
Significance
株高Plant height (cm)Rht8(+)51376.704.4459.3089.0011.530.001**
Rht8(-)11778.264.6163.9091.00
每公顷穗数Spike number per hectare (×106)Rht8(+)5135.540.363.806.6310.120.002**
Rht8(-)1175.660.484.406.63
千粒重1000-kernel weight (g)Rht8(+)51345.102.6637.8053.404.080.044*
Rht8(-)11744.542.9037.0050.90
穗粒数Kernel number per spikeRht8(+)51334.932.5829.1043.8015.190.000**
Rht8(-)11734.062.4129.4040.61

Note: "*" P<0.05; "**" P<0.01

注:“*” P<0.05;“**” P<0.01

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3 讨论

自20世纪60年代以来,矮秆基因在世界范围内得到广泛利用,但其在不同地理范围分布存在差异,美国小麦品种主要以RhtB1-b和RhtD1-b基因作为矮秆来源[13,14],而对Rht8基因的利用非常少,此外,保加利亚小麦品种则主要利用Rht8基因作为矮秆来源[15],但在德国小麦品种中没有检测到Rht8基因[16]。在中国,RhtB1-b、RhtD1-b和Rht8基因在小麦育种中的利用率不断增加[17],其中RhtD1-b基因的利用率最高,这与本研究的结果一致。RhtD1-b在不同地区小麦品种中分布不均匀,在山东小麦品种(系)中含有RhtD1-b基因的小麦品种(系)约54%,比RhtB1-b多20.67%,而很少有品种(系)同时含有两个基因[18]。对246份黄淮麦区当前主栽小麦品种、历史品种和农家品种中矮秆基因的检测发现,Rht1D-b的利用率最高[19],在安徽和四川小麦品种中则可能主要利用了Rht8作为矮秆来源[20,21]。在河南新育成的129份小麦品种中RhtD1-b和Rht8的分布均超过了50%[22]。本研究发现,携带矮秆基因RhtB1-b和RhtD1-b的小麦品种(系)占供试材料的比例超过90%或接近90%,说明RhtB1-b和RhtD1-b基因已在黄淮麦区小麦品种(系)中被固定,而Rht8基因的使用频率略低,这可能是由于携带Rht8基因的植株存在对赤霉素敏感、早期生长活力较低、面粉蛋白含量降低等不良效应[23,24]。同时研究发现3个矮秆基因已被聚合使用,这说明使用多个矮秆基因累加以降低株高,在小麦抗倒性育种中具有重要作用。

降低小麦株高可减轻倒伏所带来减产的风险,而降低株高对重要农艺性状具有不同的效应,有研究表明矮秆基因对有效分蘖数、小穗数、穗长影响不大,对穗粒数有一定的促进作用,基因Rht-D1b 和Rht-B1b对千粒重有一定的负效应,而Rht8有利于提高千粒重[25]。在本研究中,Rht8基因在河南省小麦新品种(系)中具有增加千粒重和穗粒数的效应,这与之前的研究结果相似[26,27],但同时却降低了每公顷穗数,表现了明显的三要素之间的协同,因而在进行大穗大粒的小麦育种工作中可以对该基因加以应用[19]。此外,Rht8基因与其他优异基因(如Ppd-D1)可以有效地进行聚合[28],在生产上具有很好的应用前景。郑麦583等小麦品种为河南省已审定的小麦品种,丰产性好(每公顷穗数表现突出),尽管有报道矮秆基因对部分品质参数具有不良的效应[29],但郑麦583表现出了优质的特性,说明矮秆基因和一些品质相关的优异位点可以聚合,郑麦583中聚合了3个矮秆基因,并且具有丰产性等优异表现,因此在以提升每公顷穗数作为育种目标时可使用郑麦583作为亲本。

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The gibberellin-responsive dwarfing gene Rht12 can significantly reduce plant height without changing seedling vigor and substantially increase ear fertility in bread wheat (Triticum aestivum. L). However, Rht12 delays heading date and anthesis date, hindering the use of Rht12 in wheat improvement. To promote early flowering of the Rht12 dwarf plants, the photoperiod-insensitive allele Ppd-D1a was introduced through a cross between Jinmai47 (Ppd-D1a) and Karcagi (Rht12). The results showed that Ppd-D1a can rescue the delaying effect of Rht12 on flowering time and promote earlier flowering by 9.0 days (163.2°Cd) in the Rht12 dwarf plants by shortening the late reproduction phase. Plant height was reduced by Rht12 (43.2%) and Ppd-D1a (10.9%), achieving dwarf plants with higher lodging resistance. Ear fertility, like the grain number per spike, was significantly increased by Rht12 (21.3%), while it was reduced by Ppd-D1a (6.5%). However, thousand kernel weight was significantly reduced by Rht12 (12.9%) but significantly increased by Ppd-D1a (16.9%). Finally, plant yield was increased by 16.4 and 8.2%, and harvest index was increased by 24.9 and 15.4% in the Rht12 dwarf lines and tall lines with Ppd-D1a, respectively. Clearly, there was an additive interaction between Rht12 and Ppd-D1 and the introduction of Ppd-D1a advanced the flowering time and improved the yield traits of Rht12 dwarf plants, suggesting that the combination of Rht12 and Ppd-D1a would be conducive to the successful use of Rht12 in wheat breeding programs.

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World wheat grain yields increased substantially in the 1960s and 1970s because farmers rapidly adopted the new varieties and cultivation methods of the so-called 'green revolution'. The new varieties are shorter, increase grain yield at the expense of straw biomass, and are more resistant to damage by wind and rain. These wheats are short because they respond abnormally to the plant growth hormone gibberellin. This reduced response to gibberellin is conferred by mutant dwarfing alleles at one of two Reduced height-1 (Rht-B1 and Rht-D1) loci. Here we show that Rht-B1/Rht-D1 and maize dwarf-8 (d8) are orthologues of the Arabidopsis Gibberellin Insensitive (GAI) gene. These genes encode proteins that resemble nuclear transcription factors and contain an SH2-like domain, indicating that phosphotyrosine may participate in gibberellin signalling. Six different orthologous dwarfing mutant alleles encode proteins that are altered in a conserved amino-terminal gibberellin signalling domain. Transgenic rice plants containing a mutant GAI allele give reduced responses to gibberellin and are dwarfed, indicating that mutant GAI orthologues could be used to increase yield in a wide range of crop species.

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The introduction of the Reduced height (Rht)-B1b and Rht-D1b semidwarfing genes led to impressive increases in wheat (Triticum aestivum) yields during the Green Revolution. The reduction in stem elongation in varieties containing these alleles is caused by a limited response to the phytohormone gibberellin (GA), resulting in improved resistance to stem lodging and yield benefits through an increase in grain number. Rht-B1 and Rht-D1 encode DELLA proteins, which act to repress GA-responsive growth, and their mutant alleles Rht-B1b and Rht-D1b are thought to confer dwarfism by producing more active forms of these growth repressors. While no semidwarfing alleles of Rht-A1 have been identified, we show that this gene is expressed at comparable levels to the other homeologs and represents a potential target for producing novel dwarfing alleles. In this study, we have characterized additional dwarfing mutations in Rht-B1 and Rht-D1. We show that the severe dwarfism conferred by Rht-B1c is caused by an intragenic insertion, which results in an in-frame 90-bp insertion in the transcript and a predicted 30-amino acid insertion within the highly conserved amino-terminal DELLA domain. In contrast, the extreme dwarfism of Rht-D1c is due to overexpression of the semidwarfing Rht-D1b allele, caused by an increase in gene copy number. We show also that the semidwarfing alleles Rht-B1d and Rht-B1e introduce premature stop codons within the amino-terminal coding region. Yeast two-hybrid assays indicate that these newly characterized mutations in Rht-B1 and Rht-D1 confer "GA-insensitive" dwarfism by producing DELLA proteins that do not bind the GA receptor GA INSENSITIVE DWARF1, potentially compromising their targeted degradation.

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Optimizing wheat height to maximize yield has been an important aspect which is evident from a successful example of green revolution. Dwarfing genes (Rht) are known for yield gains due to lodging resistance and partitioning of assimilates into ear. The available and commercially exploited sources of dwarfism in Indian spring wheat are Rht1 and Rht2 genes inspite of availability of over 20 dwarfing genes. Rht8 a Gibberellic acid sensitive dwarfing gene is another reduced height gene commercially exploited in some Mediterranean countries. Two F2 populations segregating for Rht1 and Rht8 genes with each comprising 398 and 379 plants were developed by crossing European winter wheat cultivars Beauchamp and Capitole with Indian spring wheat cultivar PBW 621. Different genotypic combinations for Rht1 and Rht8 genes were selected from these populations through linked molecular markers and selected F3:4 lines were evaluated for various agronomic traits in a replicated trial. Reduction in plant height with Rht8 and Rht1 averaged 2.86% and 13.3% respectively as compared to the group of lines lacking dwarfing gene. Reduction was spread along all the internodes of wheat culm and reduction was lower as progress towards the lower internode. Grain number per spike and highest yield was observed in lines carrying only Rht1 gene. Reduction in plant biomass was observed with either of the dwarfing gene. Longest coleoptile length and seedling shoot length averaged 4.4 ± 0.09 cm and 19.5 ± 0.48, respectively was observed in lines lacking any of the dwarfing gene. Negligible reduction of 6.75% and 2.84% in coleoptile length and seedling shoot length, respectively was observed in lines carrying only Rht8 gene whereas F3:4 lines with Rht1 gene showed 21.64% and 23.35% reduction in coleoptile length and seedling shoot length, respectively. Additive effect of genes was observed as double dwarfs showed 43.31% and 43.34% reduction in coleoptile length and seedling shoot length.

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"Perfect" markers for the Rht-B1b and Rht-D1b dwarfing genes in wheat

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PCR-based markers were developed to detect the point mutations responsible for the two major semi-dwarfing genes Rht-B1b ( Rht1) and Rht-D1b ( Rht2) in wheat. These markers were validated by testing 19 wheat varieties of known Rht genotype. They included Rht-B1b and Rht-D1b dwarfs, double-mutant varieties and tall wheats. These were correctly genotyped with the Rht-B1b and Rht-D1b-specific primers, as well as markers specific for the tall alleles Rht-B1a and Rht-D1a. Using a family of doubled-haploid lines segregating for Rht-B1b and Rht-D1b, the markers were mapped to the expected homoeologous regions of chromosomes 4B and 4D, respectively. Both markers were strongly correlated with a reduction in height, accounting for 23% ( Rht-B1b) and 44% ( Rht-D1b) of the phenotypic variance in the population. These markers will have utility in marker-assisted selection of the Rht-B1b and Rht-D1b genes in wheat breeding programs.

孙树贵, 李艳丽, 鲁敏 , .

67份美国小麦品种矮秆基因的分子标记检测

麦类作物学报, 2013,33(6):1087-1092.

DOI:10.7606/j.issn.1009-1041.2013.06.004      URL     [本文引用: 1]

为了明确矮秆基因在美国小麦品种中的分布特点并发掘较少利用的矮秆基因,本研究利用8个小麦矮秆基因的特异分子标记对67份美国小麦品种中的矮秆基因进行了检测。结果表明,67份美国小麦品种中,超过80%的品种(56个)含有矮秆基因,其中 Rht-B1bRht8基因频率较高,分别占62.7%和43.3%。仅有5个品种含有 Rht-D1b基因,同时发现3个品种含有 Rht5基因,2个品种含有 Rht12基因,未发现含有 Rht4Rht9Rht13基因的品种。在含有矮秆基因的品种中,35.8%的品种含有2个或2个以上的矮秆基因。其中有1个品种同时含有3个矮秆基因,有20个品种同时含有 Rht-B1bRht8基因,有3个品种同时含有 Rht-D1bRht8基因,有1个品种同时含有 Rht-D1bRht12基因,其余32个品种各含有1个矮秆基因。本研究未发现同时含有 Rht-B1bRht-D1bRht8以及同时含有 Rht-B1bRht-D1b的品种。

Guedira M, Brown-Guedira G, Sanford D V , et al.

Distribution of Rht genes in modern and historic winter wheat cultivars from the eastern and central USA

Crop Science, 2010,50(5):1811-1822.

DOI:10.2135/cropsci2009.10.0626      URL     [本文引用: 1]

Ganeva G, Korzun V, Landjeva S , et al.

Identification,distribution and effects on agronomic traits of the semi-dwarfing Rht alleles in Bulgarian common wheat cultivars

Euphytica, 2005,145(3):305-315.

DOI:10.1007/s10681-005-1742-9      URL     [本文引用: 1]

Bulgarian common wheat cultivars released in the period 1925–2003 were studied using the gibberellic acid (GA) test and microsatellite analysis of the Xgwm261 locus on chromosome 2DS to identify the semi-dwarfing (Rht) genes. The old cultivars, isolated through selection from landraces, carried rare alleles (211- and 215-bp) at Xgwm261 locus, and those developed by hybridisation to foreign cultivars, carried the 165- and 174-bp alleles. Forty-two (55.3%) of 76 modern cultivars were GA-responsive. The 192-bp allele, diagnostic for Rht8, was observed in 64 (84.2%) modern cultivars, of which 37 carry Rht8 alone, and 27 possess a combination of Rht8 and a GA-insensitive allele viz. Rht-B1d (17); Rht-D1b (6) and Rht-B1b (4). The 174-bp allele is present in seven cultivars, only one of which is photoperiod-sensitive, and the rest are day-length insensitive. The 203-bp allele was found in six modern cultivars. Cultivars carrying the Rht8 allele are the most widespread and some of them have been cultivated for a long period. Cultivars with the `Saitama 27' allele (Rht-B1d) are the most productive and are second in distribution in the country. The recently observed trend for increasing the proportion of cultivars with GA-insensitive Rht genes is probably due to their combination with the 192-bp allele of Xgwm261 locus tightly linked to the Ppd-D1, to the break of the link between the 174-bp allele and ppd-D1, and to the introduction of other genes influencing flowering time.

Knopf C, Becker H, Ebmeyer E , et al.

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Zhang X, Yang S, Zhou Y , et al.

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Euphytica, 2006,152(1):109-116.

DOI:10.1007/s10681-006-9184-6      URL     [本文引用: 1]

Reduction of plant height has played a significant role in improving wheat production and knowledge of dwarfing genes in Chinese wheat will be very important for developing high yielding cultivars. Molecular markers were used to detect the presence of genes Rht-B1b (Rht1), Rht-D1b (Rht2) and Rht8 in 220 wheat genotypes from autumn-sown wheat regions in China. They include landmark landraces, leading cultivars and core parents involved in wheat breeding from the 1950s to the present. Results indicated that Rht-D1b and Rht8 dominate with frequencies of 45.5% and 46.8%, respectively, followed by Rht-B1b with a frequency of 24.5%. The frequencies of Rht-B1b and Rht-D1b increased, from 8.6 to 32.2% and 36.2 to 53.4%, respectively, whereas the frequency of Rht8 has remained constant over time, when compared with cultivars released before and after 1990. This indicates that both the Rht-B1b and Rht-D1b were successfully used in wheat production in Chinese environments. Our study shows that Rht-B1b and Rht-D1b can be used in the post-anthesis heat stressed environments. Rht-B1b in Chinese wheats is derived from two sources, viz., Norin 10 and the Italian introduction St2422/464 (Rht-B1b and Rht8). The identity of Rht-B1b in these two sources still needs to be confirmed. Suwon 86 carrying both Rht-B1b and Rht-D1b, and Chinese cultivars, Huixianhong and Yaobaomai, are the primary sources of Rht-D1b in Chinese wheats. It is likely that Rht-D1b in Youbaomai derives from an unknown introduction. Italian introductions such as Funo and Abbondanza, and Lovrin 10 with the 1B/1R translocation, and Chinese landraces are the three major sources of Rht8. This information will be very valuable for wheat breeding in China, and internationally.

Mu M, Liu Y, Guo X , et al.

Distribution of Rht-Blb and Rht-Dlb in wheat cultivars in Shandong detected by molecular markers

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RFLP markers have proven to be a reliable and highly informative tool for characterizing genetic diversity in maize. Joint analysis of inbred lines and populations should provide valuable information with respect to (1) a better understanding of the genetic basis of present elite germplasm and (2) the identification of populations that may prove to be useful sources of genetic diversity for breeding programs. Sixty-two inbred lines of known heterotic groups and ten maize populations, some of them significant contributors to the genetic basis of the heterotic groups, were assayed at 28 RFLP loci. Joint data analyses first underlined that the populations displayed a large number of alleles that were absent in the set of inbred lines. Associations among inbreds and populations further proved consistent with pedigree data of the inbreds and provided new information on the genetical basis of heterotic groups. In particular, European flint inbreds were revealed to be as close to the Northeastern U.S. flint population studied as to the typical European populations. These results advocate the analysis of larger sets of populations by means of molecular markers in order to (1) gain insight into the history of maize germplasm and (2) set up appropriate strategies for the use of genetic resources in breeding programs.

周晓变, 赵磊, 陈建辉 , .

黄淮麦区小麦种质资源矮秆基因分布及其与农艺性状的关系

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257份小麦品种资源中矮秆基因的分子检测

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[本文引用: 1]

张德强, 宋晓朋, 冯洁 , .

黄淮麦区小麦品种矮秆基因Rht-B1b、Rht-D1b和Rht8的检测及其对农艺性状的影响

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Molecular mapping of gibberellin-responsive dwarfing genes in bread wheat

Theoretical and Applied Genetics, 2005,111(3):423-430.

DOI:10.1007/s00122-005-2008-6      URL     [本文引用: 1]

Opportunities exist for replacing reduced height (Rht) genes Rht-B1b and Rht-D1b with alternative dwarfing genes for bread wheat improvement. In this study, the chromosomal locations of several height-reducing genes were determined by screening populations of recombinant inbred lines or doubled haploid lines varying for plant height with microsatellite markers. Linked markers were found for Rht5 (on chromosome 3BS), Rht12 (5AL) and Rht13 (7BS), which accounted for most of the phenotypic variance in height in the respective populations. Large height differences between genotypes (up to 43 cm) indicated linkage to major height-reducing genes. Rht4 was associated with molecular markers on chromosome 2BL, accounting for up to 30% of the variance in height. Confirming previous studies, Rht8 was linked to markers on chromosome 2DS, whereas a population varying for Rht9 revealed a region with a small but significant height effect on chromosome 5AL. The height-reducing effect of these dwarfing genes was repeatable across a range of environments. The molecular markers developed in this study will be useful for marker-assisted selection of alternative height-reducing genes, and to better understand the effects of different Rht genes on wheat growth and agronomic performance.

Botwright T L, Rebetzke G J, Condon A G , et al.

Influence of the gibberellin-sensitive Rht8 dwarfing gene on leaf epidermal cell dimensions and early vigour in wheat (Triticum aestivum L.)

Annals of Botany, 2005,95(4):631-639.

DOI:10.1093/aob/mci069      URL     PMID:15655105      [本文引用: 1]

The gibberellin-insensitive Rht-B1b and Rht-D1b dwarfing genes are known to reduce the size of cells in culms, leaves and coleoptiles of wheat. Resulting leaf area development of gibberellin-insensitive wheats is poor compared to standard height (Rht-B1a and Rht-D1a) genotypes. Alternative dwarfing genes to Rht-B1b and Rht-D1b are available that reduce plant height, such as the gibberellin-responsive Rht8 gene. This study aims to investigate if Rht8 has a similar dwarfing effect on the size of leaf cells to reduce leaf area.

李杏普, 兰素缺, 冯延茹 .

Rht-B1b,Rht-D1b,Rht-B1c小麦矮秆基因及其互作对小麦农艺性状的影响

河北农业科学,2006(1):14-18.

URL     [本文引用: 1]

利用2套近等基因系,在人工模拟气候室,研究了不同Rht近等基因系营养性状的遗传差异.结果证明:Rht-B1b和Rht-D1b矮秆基因系不但具有很好的降秆作用,而且可以在生长前期形成较多的干物质和较坚硬的秆子,为后期籽粒产量的形成奠定基础;Rht-D1b半矮秆基因根系长度明显长于其它基因系,可能较其它基因系具有较强的耐旱能力;Rht-B1c基因系的分蘖数量及次生根或总根发生量多于其它基因系,无论苗高或株高均显著低于其它基因系,可在单纯的提高单株分蘖或次生根生长量及降秆为目的的育种中发挥一定作用.

Du Y, Chen L, Wang Y , et al.

The combination of dwarfing genes Rht4 and Rht8 reduced plant height,improved yield traits of rainfed bread wheat (Triticum aestivum L.)

Field Crops Research, 2018,215:149-155.

DOI:10.1016/j.fcr.2017.10.015      URL     [本文引用: 1]

Wang Y, Du Y, Yang Z , et al.

Comparing the effects of GA-responsive dwarfing genes Rht13 and Rht8 on plant height and some agronomic traits in common wheat

Field Crops Research, 2015,179:35-43.

DOI:10.1016/j.fcr.2015.04.010      URL     [本文引用: 1]

Zhang K, Wang J, Qin H , et al.

Assessment of the individual and combined effects of Rht8 and Ppd-D1a on plant height,time to heading and yield traits in common wheat

The Crop Journal, 2019.

DOI:10.1080/00028899908984503      URL     PMID:10635545      [本文引用: 1]

As part of a European Concerted Action on Male Reproduction Capability an exposure assessment survey was conducted among seasonal workers in the fruit growing sector in the Netherlands. Dermal exposure to the fungicides captan and tolylfluanid was measured using cotton gloves (12 persons) and skin pads on several body parts (12 persons). In addition, a set of exposure data was used from a study conducted recently among Dutch fruit growers. For harvesting activities, re-entry time appeared to be an important determinant of dermal exposure to captan and tolyfluanid. Explained variance of regression models was moderate to high (range 0.30-0.87). For captan, calculated half-life times from the most recent exposure survey were lower (glove data: 5 days; pad data: 6 days) compared with half-life times based on the previously conducted study (11 days). Possible explanations for the discrepancy are discussed. For tolylfluanid, estimated half-life times during harvesting were 2 and 3 days, based on pad and glove data, respectively. Prediction of captan exposure during other crop activities appeared to be far more difficult (explained variance equal to 0.06), although the estimated half-life time was comparable with that for harvesting. The data suggest that re-entry time gives useful information to group workers in broad exposure categories. Nonetheless, it was concluded that large studies are needed to evaluate the importance of re-entry time in more detail.

Jobson E M, Martin J M, Schneider T M , et al.

The impact of the Rht-B1b,Rht-D1b,and Rht-8 wheat semi-dwarfing genes on flour milling,baking,and micronutrients

Cereal Chemistry, 2018,95(6):770-778.

DOI:10.1002/cche.2018.95.issue-6      URL     [本文引用: 1]

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