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作物学报 ›› 2021, Vol. 47 ›› Issue (8): 1511-1521.doi: 10.3724/SP.J.1006.2021.01082

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

普通小麦-六倍体中间偃麦草易位系的抗条锈鉴定及应用评估

王音1(), 冯志威3, 葛川3, 赵佳佳2, 乔玲2, 武棒棒2, 闫素仙2, 郑军2,3,*(), 郑兴卫2,3,*()   

  1. 1山西师范大学临汾学院自然科学系, 山西临汾041000
    2山西农业大学小麦研究所, 山西临汾041000
    3山西农业大学农学院/作物遗传与分子改良山西省重点实验室, 山西太原 030000
  • 收稿日期:2020-10-22 接受日期:2021-01-13 出版日期:2021-08-12 网络出版日期:2021-03-02
  • 通讯作者: 郑军,郑兴卫
  • 作者简介:王音, E-mail: wangyinstar@163.com
  • 基金资助:
    中国博士后科学基金项目(2020M670701);山西省重点研发计划项目(201903D221074);山西省重点研发计划项目(201903D221075);山西省重点实验室项目(201705D111008-22)

Identification of seedling resistance to stripe rust in wheat-Thinopyrum intermedium translocation line and its potential application in breeding

WANG Yin1(), FENG Zhi-Wei3, GE Chuan3, ZHAO Jia-Jia2, QIAO Ling2, WU Bang-Bang2, YAN Su-Xian2, ZHENG Jun2,3,*(), ZHENG Xing-Wei2,3,*()   

  1. 1Department of Natural Sciences, Shanxi Normal University, Linfen 041000, Shanxi, China
    2Institute of Wheat Research, Shanxi Agricultural University, Linfen 041000, Shanxi, China
    3College of Agriculture, Shanxi Agricultural University/Shanxi Key Laboratory of Crop Genetics and Molecular Improvement, Taiyuan 030000, Shanxi, China
  • Received:2020-10-22 Accepted:2021-01-13 Published:2021-08-12 Published online:2021-03-02
  • Contact: ZHENG Jun,ZHENG Xing-Wei
  • Supported by:
    China Postdoctoral Science Foundation(2020M670701);Key Research and Development Program in Shanxi Province(201903D221074);Key Research and Development Program in Shanxi Province(201903D221075);Shanxi Key Laboratory Program(201705D111008-22)

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摘要：

小麦近缘属种中含有丰富的抗病资源, 前期从普通小麦与六倍体中间偃麦草的杂交、回交后代中, 鉴定获得稳定的小麦易位系新种质ZH811。为发掘和利用该易位系的抗性基因, 对ZH811进行苗期分小种的条锈病鉴定、田间农艺性状考察、分子细胞学特性和主要品质性状分析; 利用随机扩增多态性DNA (Random Amplified Polymorphic DNA, RAPD)开发易位片段的特异标记。结果表明, ZH811苗期对条锈菌条中29、31、32、33、34以及水源4、5和7号的混合菌种具有较强的抗性, 其抗性源于5D染色体短臂含有来源于Ee基因组的小片段易位; SCAR标记(Sequence Characterized Amplified Region, SCAR)和黑芒可作为易位片段追踪的分子和形态标记; ZH811的主要农艺性状接近当前黄淮北片麦区的主推品种, HMW-GS组合类型是“1, 17+18, 5+10”, 3个位点均为优质亚基, 各项品质指标达到中强筋的标准; 易位片段具有增加穗粒数的效应, 不含降低品质的连锁累赘。ZH811可为培育高产、抗病和优质小麦新品种提供重要的中间材料。

关键词: 条锈, 易位系, 原位杂交, 分子标记

Abstract:

There are abundant disease resistance resources in relative genus of wheat. The translocation line ZH811 derived from progeny of wheat and Th. intermedium hybrids, was selected for evaluation of stripe rust resistance. A series of stripe rust races such as CYR29, CYR31, CYR32, CYR33, CYR34, Suwon-4, Suwon-5, and Suwon-7 were used to record stripe responses at seedling stage. Agronomical traits, quality traits, and molecular cytogenetic analysis were also performed. Moreover, specific molecular markers located on alien segment was developed by RAPD method. The results indicated that ZH811 was highly resistant to all tested races at seedling stage. And it further proved that the resistance was conferred by a small-fragment-translocation from the Ee genome on 5DS chromosome. The SCAR markers and black awn traits could be used to trace the translocation fragment. ZH811 possessed similar agronomic traits with the commercial cultivars in the Yellow-Huaihe-Haihe Rivers region of wheat. The translocation fragment may be associated with the increase of grain number. The components of the Glu-1 were good-quality subunits, including 1, 17+18, and 5+10, and each quality index met the standard of moderate gluten. The alien chromosomal fragment had no obvious linkage drag to grain quality performance. Based on these findings, ZH811 could be used as a potential material for wheat breeding in high yield, disease resistance, and high quality.

Key words: stripe rust, translocation line, in situ hybridization, molecular marker

表1

试验所需植物材料及信息"

材料名称
Plant materials		 材料信息
Information of plant materials
ZH811, ZH577 F5 (中国春/中间偃麦草)//晋麦33///临4133组合F3中选育
Deriving from F3 [F5(CS/Z1141)//Jinmai 33///Lin 4133]
ZH811×ZH577正反交F2群体
ZH811×ZH577 reciprocal F2 populations		 ZH811和ZH577进行杂交获得的正反交F2群体
F2 populations generated from reciprocal crosses between ZH811 and ZH577
ZH811×ZH577 BC1群体
ZH811×ZH577 BC1 populations		 ZH811×ZH577回交一代群体
The first backcross generations of ZH811 and ZH577
ZH811×ZH577 DH群体
ZH811×ZH577 DH populations		 以ZH811和ZH577为亲本构建的双单倍体群体
Double haploid population of a cross between ZH811 and ZH577
中国春、晋麦33、临4133
Chines Spring, Jinmai 33, Lin 4133		 易位系的亲本
Parent materials of the translocation line
铭贤169
Mingxian 169		 感病对照
Susceptible control variety
良星99
Liangxing 99		 品质性状对照
Quality traits control
师栾02-1、烟农19
Shiluan 02-1, Yannong 19		 高分子量麦谷蛋白亚基对照
High-Molecular-Weight Glutenin Subunits (HWM-GS) control varieties
Z1141 六倍体中间偃麦草
Th. intermedium (2n = 42, StStEeEeEbEb)
PI98526 二倍体长穗偃麦草
Th. elongatum (2n = 14, EeEe)
PI531712 百萨偃麦草
Th. bessarabicum (2n = 14, EbEb)
PI313960 拟鹅观草
Pseudoroegneria strigose (2n = 14, StSt)

图1

苗期抗病性鉴定"

表2

ZH811和ZH577后代群体中抗、感单株数和分离比适合性检验"

群体世代
Generation		 不同抗病类型的株系数量
No. of plants with different infection type		 理论比
Expected ratio		 χ2 P
抗病Resistant 感病Susceptible
正交F2 Direct cross F2 133 47 3:1 0.12 0.73
反交F2 Reciprocal cross F2 112 38 3:1 0.01 0.93
BC1 54 46 1:1 0.64 0.42

图2

ZH811 (A, B)和亲本晋麦33 (C, D)、临4133 (E, F)的GISH/FISH分析 A, C, E: 利用中国春基因组做封阻, 中间偃麦草基因组为探针的GISH分析; B, D, F: 利用探针pAs1 (绿色信号)和pSc119.2 (红色信号)进行FISH分析。白色箭头指示小麦-中间偃麦草易位片段(A)和5D染色体(B~F)。标尺为10 μm。"

图3

不同材料的标记鉴定结果 A: ZH811和ZH577的RAPD扩增(对应泳道从左到右分别是DL2000 marker、ZH811和ZH577退火温度46℃扩增、ZH811和ZH577退火温度50℃扩增、ZH811和ZH577退火温度54℃扩增); 白色箭头指示ZH811和ZH577扩增出的差异条带; B: SCAR标记不同材料中的扩增结果(对应泳道从左到右分别是DL2000 marker、ZH811、ZH577、中国春CS、临4133、晋麦33、中间偃麦草ZH1141、二倍体长穗偃麦草PI98526、百萨偃麦草PI531712、拟鹅观草PI313960、ddH2O);"

图4

ZH811×ZH577 DH部分株系的抗病表型及对应的SCAR标记和芒色鉴定"

图5

ZH811和ZH577植株形态"

图6

易位与非易位系农艺性状表现"

图7

高分子量麦谷蛋白亚基的SDS-PAGE分析 从左到右依次为师栾02-1 (SL02-1)、ZH811、烟农19 (YN19)、晋麦33号(JM33)、中国春(CS)。图中数字表示各条带代表的高分子量麦谷蛋白亚基类型。"

表3

不同材料的品质性状"

品种
Cultivar		 蛋白质含量
Protein content (%)		 沉降值
Zeleny
(mL)		 湿面筋含量
Wet gluten
content (%)		 形成时间
Formation time (min)		 稳定时间
Dough stability time stability time (min)		 最大拉伸阻力
Max resistance to drag (E.U.)		 拉伸面积
Stretch area (cm2)
ZH811
ZH577		 14.57±1.53 a 30.25±2.75 ab 32.87±2.32 a 5.82±0.41 a 7.88±0.23 a 368.00±12.37 a 88.00±3.12 a
14.96±1.87 a 32.84±2.11 a 32.26±2.88 a 5.75±0.33 a 8.15±0.17 a 342.00±13.59 a 89.00±4.69 a
良星99
Liangxing 99		 13.32±0.84 b 26.81±1.26 b 30.61±1.82 b 2.10±0.44 b 2.70±0.23 b 193.00±15.27 b 42.00±3.72 b
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