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作物杂志,2021, 第5期: 20–27 doi: 10.16035/j.issn.1001-7283.2021.05.004

• 遗传育种·种质资源·生物技术 • 上一篇    下一篇

12个主产区历史小麦品种抗叶锈病基因分析

段振盈1(), 徐新玉1, 李星2, 李在峰2, 马骏3, 姚占军1()   

  1. 1河北农业大学农学院/华北作物种质资源研究与利用教育部重点实验室,071001,河北保定
    2河北农业大学植物保护学院/河北省病虫害生物防治工程技术研究中心,071001,河北保定
    3中国农业大学农学院,100193,北京
  • 收稿日期:2020-10-18 修回日期:2021-04-13 出版日期:2021-10-15 发布日期:2021-10-14
  • 通讯作者: 姚占军
  • 作者简介:段振盈,主要从事小麦抗病遗传育种研究,E-mail： dzy950910@163.com
  • 基金资助:
    “十三五”国家重点研发计划(2018YFD0300501)

Leaf Rust Resistance Gene Analysis of 12 Wheat Cultivars in Main Producing Areas

Duan Zhenying1(), Xu Xinyu1, Li Xing2, Li Zaifeng2, Ma Jun3, Yao Zhanjun1()   

  1. 1College of Agronomy, Hebei Agricultural University/North China Key Laboratory for Germplasm Resources of Education Ministry, Baoding 071001, Hebei, China
    2College of Plant Protection, Hebei Agricultural University/Control Center of Plant Disease and Plant Pests of Hebei Province, Baoding 071001, Hebei, China
    3College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China
  • Received:2020-10-18 Revised:2021-04-13 Online:2021-10-15 Published:2021-10-14
  • Contact: Yao Zhanjun

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摘要：

近年来,小麦叶锈病发生有加重趋势,培育抗病品种是减轻小麦叶锈病危害的环保有效途径。用12个小麦品种及35个含已知抗叶锈病基因的载体品系在苗期接种19个不同毒性的叶锈菌生理小种,通过基因推导和系谱分析发掘待测品种中的抗叶锈病基因,并通过分子标记检测进一步验证;在田间接种强毒性混合生理小种,进行成株期病情严重度与普遍率调查,筛选慢锈性品种。结果表明,在石新828、百农3217、济南2号、泰山1号、石特14、晋麦2148、烟农15、小偃6号、温麦6号共9个品种中检测到Lr1、Lr26、Lr34、Lr37和Lr46共5个抗叶锈病基因,其中部分品种中发现多个抗性基因。成株期筛选出百农3217、平阳27、济南2号、泰山1号、石特14、晋麦2148、碧蚂4号、烟农15、小偃6号、温麦6号共10个慢叶锈性品种,其中碧蚂4号和小偃6号等品种是我国小麦育种的骨干亲本,探究这些品种中的抗病基因对培育小麦抗叶锈病品种具有重要意义。

关键词: 小麦, 基因推导, 分子标记, 抗性基因, 慢锈品种

Abstract:

In recent years, wheat leaf rust has a tendency of aggravation, breeding resistant varieties is the most environmental friendly and effective way to reduce the harm of wheat leaf rust. Twelve wheat varieties and 35 donor lines were inoculated with 19 Chinese pathotypes of Puccinia triticina for leaf rust at seedling stage, and the rust resistance genes in the varieties to be tested were explored through gene derivation and genealogy analysis, and further verified by molecular marker detection. The severity and prevalence rates of wheat were investigated by inoculation of virulent rusts mixed with physiological small species at the stage of plant growth, and selected the slow rust varieties. The results showed that Lr1, Lr26, Lr34, Lr37 and Lr46 were detected in nine varieties of Shixin 828, Bainong 3217, Ji’nan 2, Taishan 1, Shite 14, Jinmai 2148, Yannong 15, Xiaoyan 6, Wenmai 6, among which several resistance genes were found in some varieties. Some of these varieties also contained multiple resistance genes and ten slow rust resistant varieties were selected from the identification of wheat rust-resistant leaves at the mature stage, including Bainong 3217, Pingyang 27, Ji’nan 2, Taishan 1, Shite 14, Jinmai 2148, Bima 4, Yannong 15, Xiaoyan 6, Wenmai 6, a total of ten slow rust peasant wheat varieties and Bima 4 and Xiaoyan 6 were the backbone parents of wheat breeding. It is still important to explore the resistance genes of these varieties for breeding new wheat varieties resistant to leaf rust.

Key words: Wheat, Gene postulation, Molecular marker, Resistance gene, Slow rust varieties

表1

分子标记的引物序列及PCR扩增程序

基因
Gene		 引物
Primer		 引物序列（5´→3´）
Sequence of primer (5´→3´)		 反应程序
Cycle condition		 连锁标记
Linked marker		 参考文献
Reference
Lr1 WR003 F GGGACAGAGACCTTGGTGGA 94℃ 2min;35个循环（94℃ 30s;65℃ 30s;72℃ 30s）;72℃ 2min;4℃保温 STS [24]
WR003 R GACGATGATGATTTGCTGCTGG
Lr9 J13/1 TCCTTTTATTCCGCACGCCGG 94℃ 2min;35个循环（94℃ 30s;68.5℃ 30s;72℃ 30s）;72℃ 2min;4℃保温 STS [25]
J13/2 CCACACTACCCCAAAGAGACG
Lr10 Fl.2245 GTGTAATGCATGCAGGTTCC 94℃ 2min;35个循环（94℃ 45s;60℃ 45s;72℃ 30s）;72℃ 3 min;4℃保温 STS [26]
Lr10-6/r2 AGGTGTGAGTGAGTTATGTT
Lr19 SCS265 F GGCGGATAAGCAGAGCAGAG 94℃ 2min;35个循环（94℃ 30s;65℃ 30s;72℃ 30s）;72℃ 2 min;10℃保温 SCAR [26]
SCS265 R GGCGGATAAGTGGGTTATGG
Lr19 SCS253 F GCTGGTTCCACAAAGCAAA 94℃ 2min;35个循环（94℃ 30s;60℃ 30s;72℃ 30s）;72℃ 2min;4℃保温 SCAR [27]
SCS253 R GGCTGGTTCCTTAGATAGGTG
Lr20 STS638 L ACAGCGATGAAGCAATGAAA 94℃ 2min;35个循环（94℃ 30s;60℃ 30s;72℃ 30s）;72℃ 2min;4℃保温 STS [28]
STS638 R GTCCAGTTGGTTGATGGAAT
Lr24 J9/1 TCTAGTCTGTACATGGGGGC 94℃ 2min;35个循环（94℃ 30s;60℃ 30s;72℃ 30s）;72℃ 2 min;4℃保温 STS [29]
J9/2 TGGCACATGAACTCCATACG
Lr26 ω-secalin F ACCTTCCTCATCTTTGTCCT 94℃ 2min;35个循环（94℃ 30s;65℃ 30s;72℃ 30s）;72℃ 2min;4℃保温 STS [30]
ω-secalin R CCGATGCCTATACCACTACT
Lr26 Glu-B3 F GGTACCAACAACAACAACCC 94℃ 2min;35个循环（94℃ 30s;65℃ 30s;72℃ 30s）;72℃ 2min;4℃保温 STS [31]
Glu-B3 R GTTGCTGCTGAGGTTGGTTC
Lr34 csLV34 F GTTGGTTAAGACTGGTGATGG 94℃ 2min;35个循环（94℃ 30s;55℃ 30s;72℃ 30s）;72℃ 2min;4℃保温 STS [32]
csLV34 R TGCTTGCTATTGCTGAATAGT
Lr37 VENTRIUP AGGGGCTACTGACCAAGGCT 94℃ 2min;35个循环（94℃ 30s;65℃ 30s;72℃ 30s）;72℃ 2min;4℃保温 STS [33]
LN2 TGCAGCTACAGCAGTATGTACACAAAA
Lr46 csLV46G22F F TCGACTTTGGAATGGAGTTGC 94℃ 2min;35个循环（94℃ 30s;60℃ 30s;72℃ 30s）;72℃ 2min;4℃保温 STS [34]
csLV46G22 R GGCGAAGATGCCATCATCCACCG

表2

35个载体品系、12个供试品种及感病对照品种郑州5389对19个菌系苗期抗性鉴定结果

品种（系）
Cultivar (line)		 侵染型Infection types to P. triticina pathotypes
FH
DR		 FH
JS①		 FH
GQ①		 PH
GN		 FH
JR		 FH
SQ		 FG
KQ		 PH
DQ		 PH
GL		 FH
JQ		 FH
BR		 FG
BQ		 FH
DQ①		 FH
GQ②		 FH
JT		 TG
KS		 FH
JS②		 PH
TS		 FH
DQ②
RL6003 (Lr1) 0 ; 1 3+ 1 ; 1 4 3+ ; ; ; 1 ; 1 4 1 3+ ;
RL6016 (Lr2a) ; 1 1 1 1 ; ; ; ; ; ; ; 1 1 ; 3+ ; 1 1
RL6047 (Lr2c) 3+ 4 3 3 4 3 3+ 3+ 3 3+ 3 3 3+ 4 3+ 3+ 4 3+ 3+
RL6002 (Lr3) 4 3+ 3+ 3 3+ 3+ 4 4 3+ 3+ 3+ 3+ 3+ 3+ 4 3+ 3+ 4 4
RL6010 (Lr9) 0 0 0 0 0 0 0 ; 0 0 0 0 0 0 0 0 0 ; 0
RL6005 (Lr16) 4 4 3+ 3+ 3+ 3+ 4 4 3+ 3+ 3+ 4 3+ 3 3+ 4 3+ 3+ 3+
RL6064 (Lr24) ; ; 0 ; ; ; ; ; ; ; ; ; ; ; ; ; ; 1 ;
RL6078 (Lr26) 3+ 3 4 3 4 3+ 1 3+ 3 3+ 3+ 1 3+ 3 3+ 2 3+ 4 3+
RL6007 (Lr3ka) 2 1 2 1 1 3 2 2 1 1 1 2 1 2 2 2 1 3+ 2
RL6053 (Lr11) 2 3+ 3 3+ 3+ 3 3 2 3+ 3 1 1 1 3 3 3 3 3+ 2
RL6008 (Lr17) 3+ 3+ 1+ 2 3 3 3 3 2 3+ 2 2+ 3 1 3+ 3+ 3 3+ 3
RL6049 (Lr30) 1 2 2 1 1 2 3 1 1 1 1 1 1 1 1 3+ 1 3+ 1
RL6051 (LrB) 3+ 3+ 3+ 3 3+ 3+ 4 3+ 4 3+ 4 3+ 3+ 4 4 3 4 3+ 4
RL6004 (Lr10) 4 3+ 3+ 2 4 3+ 3+ 3 1 3 4 4 3+ 4 4 4 4 4 3+
RL6013 (Lr14a) X 3 X 3+ X X 1 2 1 1 X 1+ 1 2 3 3+ 3 4 1
RL6009 (Lr18) 3 1 1 2 3 1 1 1 1 1+ 3 1 1 1 3 1 1 2 1
RL6019 (Lr2b) 2 3 1+ 1 3 3 2 2 1 3 1 2 2 2 3 3+ 2 1 2
RL6042 (Lr3bg) 3+ 4 3 3 3+ 3+ 3+ 4 3+ 4 3+ 3+ 3+ 3+ 3+ 4 3+ 4 4
RL4031 (Lr13) 3+ 3+ 2 2 2 4 3 3+ 3 3+ 2 3+ 3+ 3 3+ 3+ 3 4 3
RL6006 (Lr14b) 4 3+ 3+ 3 3+ 3+ 4 3+ 2 3+ 4 3+ 4 4 4 2 3+ 4 3+
品种（系）
Cultivar (line)		 侵染型Infection types to P. triticina pathotypes
FH
DR		 FH
JS①		 FH
GQ①		 PH
GN		 FH
JR		 FH
SQ		 FG
KQ		 PH
DQ		 PH
GL		 FH
JQ		 FH
BR		 FG
BQ		 FH
DQ①		 FH
GQ②		 FH
JT		 TG
KS		 FH
JS②		 PH
TS		 FH
DQ②
RL6052 (Lr15) 1 1 ; 1 0 ; 0 4 4 3 ; 0 ; ; 1 3+ 3+ 0 0
RL6040 (Lr19) ; 0 0 0 0 ; 0 0 0 ; 0 ; 0 0 0 0 ; ; ;
RL6092 (Lr20) ; 1 ; 3 1 3+ 1 3 3+ 1 1 ; 1 ; 1 1 3 1 1
RL6043 (Lr21) 3 2 1+ 1 3 3+ 2 1 1 2 1 1 1 1 2 1 1 1 2
RL6012 (Lr23) 3+ 3+ 3+ 3+ 3+ 4 3 3+ 3 3+ 3+ 3+ 4 3+ 3+ 3+ 3 3+ 3+
RL6079 (Lr28) 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 ; 0
RL6080 (Lr29) 1 1 1+ 1+ 1 1 1 2 1 1 1+ ; 2 ; 1 1 1 1 1
RL6057 (Lr33) 3+ 3 3+ 3 3+ 3+ 3 3+ 3+ 3+ 3+ 4 3+ 3+ 3+ 3 3 3+ 3+
E84018 (Lr36) 1 1+ 1+ 1+ 1 2 1+ 2+ 2 2 1 1 1 1 3 1 1 2 1
KS86NGRC02 (Lr39) 3 2 2 2 2 2 3 3 2 2 3+ 3+ 3 3 2 3+ 1 2+ 3+
KS91WGRC11 (Lr42) 2 2 1+ 1 1 0 1 1+ 1 2 1 1 1 1+ 1 1+ 1 1 1
RL6147 (Lr44) ; 3+ 3 3 3 3 3 2 1 3 3 3+ 3 2 3+ 2 3 3 4
RL6144 (Lr45) 2 3+ 2 4 1 4 2 3+ 3+ 3 1 2 2+ 2 3 2 1 2+ 3
PAVON76 (Lr47) 0 0 0 0 ; 0 0 1 0 0 0 0 0 0 0 0 0 ; 0
C78.5 (Lr51) ; ; ; ; ; ; ; ; ; ; 1 ; 1 ; ; ; ; 0 ;
石新828 Shixin 828 3 3 3 3 3+ 3+ 3 3 3 3+ 3 3+ 3 3+ 3+ 2 3+ 3 3+
百农3217 Bainong 3217 1 1 2 1+ 1 2 1 3 2 ; ; 1 ; 1 ; 3 1 2+ ;
平阳27 Pingyang 27 1 1 ; ; 1 1 1 2 2 1 1 1 ; ; ; 2 1 2 ;
济南2号Ji’nan 2 3 3 3+ 3 3+ 3+ 3+ 3+ 3+ 3+ 3 3 ; 2 3 2 1 4 3
泰山1号Taishan 1 3+ 3 3+ 3+ 3+ 3+ ; 3+ 3+ 3 3 1 3 2+ 3+ 1+ 3+ 3+ 3
石特14 Shite 14 1 1 1 2 2 2 ; 1 2 1 1 1 1 ; ; ; 1 2 2
晋麦2148 Jinmai 2148 3+ 3 3+ 3+ 3+ 3+ 3+ 4 3+ 3+ 4 3 3 3+ 3+ 3+ 3+ 4 3
碧蚂4号Bima 4 3 3+ 3+ 3 2 3 3+ 4 3+ 3+ 4 2 2+ 2+ 2 2+ 1 3+ 1
碧蚂1号Bima 1 3+ 3+ 3+ 3 3+ 3+ 1 4 3 3+ 4 ; 3 3 3+ 3 3+ 4 3+
烟农15 Yannong 15 2 1 1 ; 1 1 ; 2+ 2 ; ; ; 1 ; ; 2+ 1 2 1
小偃6号Xiaoyan 6 3+ 3+ 3+ 3+ 3+ 3+ 3+ 4 4 3+ 3+ 3 3 3 3+ 3+ 3 3+ 3
温麦6号Wenmai 6 3+ 3+ 3+ 3+ 3+ 3+ 3+ 4 3+ 3+ 3 3 3+ 3 3 3+ 3+ 3+ 3
郑州5389
Zhengzhou 5389		 3 4 3+ 4 3 4 3+ 3+ 4 3+ 4 4 4 3+ 4 3+ 3+ 4 3

图1

12个小麦品种中含有的抗叶锈病基因分子标记检测结果 M: DM2000标记或pBR322 DNA标记;Z：郑州5389;1~12：石新828、百农3217、平阳27、济南2号、泰山1号、石特14、晋麦2148、碧蚂4号、碧蚂1号、烟农15、小偃6号、温麦6号

表3

12个供试品种、慢锈对照SAAR和感病对照郑州5389成株期对混合小种侵染型、严重度、普遍率和病情指数结果

品种
Cultivar		 混合小种苗期侵染型
Mixed small species infection
type at seedling stage		 严重度Final disease severity 普遍率
General rate
(%)		 病情指数
Disease index
(%)
2017-2018 2018-2019 平均Average
SAAR 3 1 1 1 100 1
石新828 Shixin 828 4 40 30 35 100 35
百农3217 Bainong 3217 3 1 5 3 98 2.94
平阳27 Pingyang 27 3 1 5 3 100 3
济南2号Ji’nan 2 3+ 10 25 17.5 100 17.5
泰山1号Taishan 1 3 1 5 3 100 3
石特14 Shite 14 3 1 5 3 100 3
晋麦2148 Jinmai 2148 4 15 15 15 100 15
碧蚂4号Bima 4 3+ 5 15 10 100 10
碧蚂1号Bima 1 4 40 60 50 98 49
烟农15 Yannong 15 3 1 1 1 100 1
小偃6号Xiaoyan 6 3 5 10 7.5 100 7.5
温麦6号Wenmai 6 3 5 5 5 100 5
郑州5389 Zhengzhou 5389 4 80 90 85 100 85
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