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作物学报 ›› 2018, Vol. 44 ›› Issue (10): 1442-1447.doi: 10.3724/SP.J.1006.2018.01442

• 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-大赖草易位系T5AS-7LrL·7LrS分子细胞遗传学鉴定

王林生(),张雅莉,南广慧   

  1. 河南科技大学农学院/洛阳市作物遗传改良与种质创新重点实验室, 河南洛阳 471023
  • 收稿日期:2018-03-07 接受日期:2018-06-12 出版日期:2018-10-10 网络出版日期:2018-07-17
  • 通讯作者: 王林生
  • 基金资助:
    本研究由国家自然科学基金项目(31501301);河南省国际合作项目()和河南省教育厅自然科学研究项目(2011A180011)资助(172102410052);河南省教育厅自然科学研究项目(2011A180011)

Molecular and Cytogenetic Identification of Triticum aestivum-Leymus racemosus Translocation Line T5AS-7LrL·7LrS

Lin-Sheng WANG(),Ya-Li ZHANG,Guang-Hui NAN   

  1. Key Laboratory of Crop Genetic Improvement and Germplasm Innovation / College of Agriculture, Henan University of Science and Technology, Luoyang 471023, Henan, China
  • Received:2018-03-07 Accepted:2018-06-12 Published:2018-10-10 Published online:2018-07-17
  • Contact: Lin-Sheng WANG
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31501301);the International Cooperation Program of Henan Province(172102410052);the Natural Science Research Program of Henan Education Department(2011A180011)

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摘要：

大赖草对赤霉病具有较好的抗性, 将大赖草赤霉病抗性基因转入普通小麦, 对拓宽小麦赤霉病抗性基础有重要意义。本研究在获得抗赤霉病普通小麦-大赖草异附加系基础上, 采用 60Co-γ射线(1200 Rad, 剂量率100 Rad min -1)处理小麦-大赖草二体附加系DA7Lr, 并用处理后的花粉授给去雄的普通小麦中国春, 对其M1代种子根尖细胞有丝分裂中期染色体进行GISH分析, 获得1株具有一条普通小麦-大赖草易位染色体的植株, 对其自交后代中具有2条易位染色体植株的花粉母细胞减数分裂中期I观察发现, 2条易位染色体形成了稳定的环状二价体, 表明该植株为纯合体。利用顺序GISH-双色FISH分析, 结合C-分带、小麦D组专化探针Oligo-pAs1-2和B组专化探针Oligo-pSc119.2-2, 进一步鉴定出该易位系为T5AS-7LrL·7LrS, 同时筛选出可追踪该易位系的3个EST-STS分子标记, 即BE591127、BQ168298和BE591737。该易位系的育成也为小麦赤霉病遗改良提供了新种质。

关键词: 普通小麦-大赖草易位系, 分子细胞遗传学, 赤霉病抗性, 60Co-γ射线

Abstract:

Leymus racemosus is highly resistant to wheat scab. The transfer of resistance genes from L. racemosus to common wheat (Triticum aestivum) is important for broadening the resistant sources against scab in common wheat. In this study, the pollen of DA7Lr, a T. aestivum-L. racemosus disomic addition line with scab resistance, was irradiated with 60Co-γ ray at 1200 Rad (100 Rad min -1) before pollinating to emasculated T. aestivum cv. Chinese Spring. One plant with one translocation chromosome was detected in the M1 generation by GISH. This plant was then self-pollinated and the pollen mother cells (PMCs) of the offspring plants with two translocation chromosomes were cytologically observed, and one ring bivalent was found at meiotic metaphase I, indicating that the plant with two translocation chromosomes was one translocation homozygote. The translocation line was proved to be T5AS-7LrL·7LrS by C-banding, and sequential GISH-FISH using Oligo-pAs1-2 and Oligo-pSc119.2-2 as probes. Three EST-STS markers (BE591127, BQ168298, and BE591737) were identified to be able to track the T5AS-7LrL·7LrS line. The translocation line also serves as an resistant source against wheat scab in wheat breeding programs.

Key words: Triticum aestivum-Leymus racemosus translocation line, molecular cytogenetics, scab resistance, 60Co-γ ray

表1

鉴定易位染色体的特异性引物"

引物名称
Primer name		 序列 Sequence (5′-3′) EST染色体
EST chromosome		 退火温度
Annealing temperature (°C)
正向 Froward 反向 Reverse
BE591127 GCAGCTCATCTTCATGGTCA CGTTGCAGCAATCAGTCCTA 7AS 7BS 7DS 60
BQ168298 GCTCTCGCTCATCATCAACA CTCGCAATGGTACCAAGGTT 7AS 7BS 7DS 60
BE591737 AGCAGCTAGGAGGGTGTCTG TAACCGCAGCTTTCTCATCC 7AS 7BS 7DS 60

图1

易位系T5AS-7LrL·7LrS荧光原位杂交(2n=44) 有丝分裂中期的顺次GISH-FISH, A图中绿色信号为Fluorescein-l2-dUTP标记的大赖草基因组DNA, B图中的红色信号为TARAM标记的Oligo-pSc119.2-2, 绿色信号为6-FAM标记的Oligo-pAs1-2, 箭头指示易位染色体。"

图2

易位染色体T5AS-7LrL·7LrS的C-分带及荧光原位杂交 从左至右依次为C-分带的5A、C-分带的T5AS-7LrL·7LrS、荧光原位杂交的T5AS-7LrL·7LrS、Oligo-pSc119.2-2 (红色)杂交的T5AS-7LrL·7LrS、Oligo-pSc119.2-2 (绿色)杂交的5A、荧光原位杂交的7Lr和C分带的7Lr。"

图3

易位系T5AS-7LrL·7LrS花粉母细胞减数分裂中期I染色体的荧光原位杂交 图中绿色信号为荧光素Fluorescein-l2-dUTP标记的大赖草基因组DNA, 箭头指示配对的二价体易位染色体T5AS-7LrL·7LrS。"

图4

引物BE591127、BQ168298和BE591737的PCR扩增结果 1: 中国春; 2: 大赖草; 3: DA7Lr; 4: T5AS-7LrL·7LrS。"

表1

不同材料赤霉病抗性(病小穗率)的田间鉴定"

材料
Material		 大田鉴定 Identified in field
2015 2016 2017
中国春 Chinese Spring 35.27 39.58±7.96 30.90±8.92
T5AS-7LrL·7LrS 8.70 11.22±5.12** 9.67±7.51**
苏麦3号Sumai 3 3.90 6.35±3.12** 5.69±2.14**
绵阳85-45 Mianyang 85-45 49.23 56.32±11.26** 47.83±13.25**

图5

易位系T5AS-7LrL·7LrS的赤霉病抗性鉴定 A: T5AS-7LrL·7LrS; B: 中国春; C: 绵阳85-45; D: 苏麦3号。"

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