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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (10): 1442-1447.doi: 10.3724/SP.J.1006.2018.01442

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

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 Online:2018-10-10 Published:2018-07-17
  • Contact: Lin-Sheng WANG E-mail:964965931@qq.com
  • 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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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

Table 1

Primers used as specific markers for discriminating translocation chromosome"

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

Fig. 1

Fluorescence in situ hybridization (FISH) of translocation line T5AS-7LrL·7LrS (2n = 44) Sequential GISH-FISH of mitotic metaphase chromosomes. In panel A, L. racemosus genomic DNA was labeled with fluorescein-12-dUTP and visualized with green signals. In panel B, Oligo-pSc119.2-2 was labeled with TARAM and visualized with red signals ,and Oligo-pAs1-2 was labeled with 6-FAM and visualized with green signals. The arrows show translocation chromosomes."

Fig. 2

C-banding and FISH of translocation chromosome T5AS-7LrL·7LrS Chromosomes from left to right are C-banded 5A, C-banded T5AS-7LrL·7LrS, FISH T5AS-7LrL·7LrS, T5AS-7LrL·7LrS with red Oligo-pSc119.2-2, 5A with green Oligo-pSc119.2-2, FISH 7Lr, and C-banded 7Lr."

Fig. 3

Chromosome FISH at MI of PMC of translocation line T5AS-7LrL·7LrS L. racemosus genomic DNA was labeled with Fluorescein-l2-dUTP and visualized with green signals. The arrow shows the ring bivalent formed by a pair of translocation chromosomes T5AS-7LrL·7LrS."

Fig. 4

PCR profiles amplified with primers BE591127, BQ168298, and BE591737 1: Chinese Spring; 2: Leymus racemosus (Lr); 3: DA7Lr; 4: T5AS-7LrL·7LrS."

Table 1

Field evaluation of scab resistance (diseased spikelet rate) in different materials (%)"

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**

Fig. 5

Evaluation of scab resistance of translocation line T5AS-7LrL·7LrS A: T5AS-7LrL·7LrS; B: Chinese Spring; C: Mianyang 85-45; D: Sumai 3."

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