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Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (07): 983-992.doi: 10.3724/SP.J.1006.2017.00983


Developing of Specific Transcription Sequences P21461 and P33259 on D. villosum 6VS and Their Application of Molecular Markers in Identifying Wheat-D. villosum Breeding Materials with Powdery Mildew Resistance

LIU Chang,LI Shi-Jin,WANG Ke,YE Xing-Guo,LIN Zhi-Shan*   

  1. National Key Facility for Crop Gene Resources and Genetic Improvement / Key Laboratory of Crop Genetics and Breeding, Ministry of Agriculture / Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2016-07-07 Revised:2017-03-01 Online:2017-07-12 Published:2017-03-25
  • Contact: Lin Zhishan, E-mail: linzhishan@caas.cn E-mail:15120067976@163.com
  • Supported by:

    This study was supported by the National Research and Development Program (2016YFD0102002) and the Agricultural Science and Technology Innovation Program of CAAS.


Dasypyrum villosum carries powdery mildew (PM) resistance gene Pm21 and PmV on its chromosome arms of 6V#2S and 6V#4S, respectively. The two resistant genes are co-segregated with exogenous chromosome arms in the offspring of hybrids between translocation lines and common wheat varieties. Developing polymorphic expression sequences to identify the two exogenous chromosomal arms is of great significance for their application in genetics and breeding, especially for 6V#4S chromosome, on which hereditary informations relatively lack. In this study, transcriptome of leaves of 6V#4S?6DL translocation line Pm97033 and common wheat line Wan7107 inoculated with Bgt was used as resources for sequences screening. As results, two unigenes of P21461 and P33259 from 6V#4S were identified by differential gene screening, synteny analysis, genomic DNA amplification and products sequencing of D. villosum accordingly. By using a pair of designed primer P461-5 based on P21461 sequence, it was found that there were 30 bp of InDel and 4 nt polymorphism between chromosomal arms of 6V#2S and 6V#4S. By the newly developed marker P461-5a, which retains the InDel polymorphism and truncates the product size amplified by P461-5, several wheat varieties and advanced lines with strong resistance to powdery mildew were identified to contain Pm21 or PmV. This marker showed potential application in discriminating D. villosum resources and assisted selection breeding for PM resistance in wheat. A marker P259-1 developed according to sequence of P33259 could specifically amplify a fragment from the wheat lines containing 6V#4S chromosome arm, but not amplify any fragment in the wheat lines containing 6V#2S?6AL translocation chromosome. Thereby, P259-1 can be used as a 6V#4S?6DL specific molecular marker in wheat backgrounds. The qRT-PCR assay showed that the expression of P21461 was not induced by PM. While transcriptional levels of P33259 increased about two times at 12 h and 24 h after inoculating the pathogens, suggesting that it might be involved in early interaction between Pm97033 and the pathogen.

Key words: Dasypyrum villosum–Triticum aestivum translocation line, Transcriptome, Powdery mildew resistance, Molecular markers

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