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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (11): 1601-1608.doi: 10.3724/SP.J.1006.2016.01601

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

TaPK-R1 Overexpressing Transgenic Wheat Lines Enhance ResistancetoSharp Eyespotand Freezing Stress

LUOMei-Ying1,2,RONGWei2,WEIXue-Ning2,YANGKun2,XUHui-Jun2,XUANWei-Yan1,*,ZHANGZeng-Yan2,*   

  1. 1 Agricultural College of Guangxi University, Nanning 530004, China; 2 The National Key Facility for Crop Gene Resources and Genetic Improvement / Key Laboratory of Biology and Genetic Improvement of Triticeae Crops, Ministry of Agriculture / Institute of Crop Science, Chinese Aca?demy of Agricultural Sciences, Beijing 100081, China
  • Received:2016-03-09 Revised:2016-07-11 Online:2016-11-12 Published:2016-08-11
  • Contact: Xuan Weiyan:E-mail:xuanweiyan_1@163.com;Zhang Zengyan,E-mail:zhangzengyan@caas.cn E-mail:luo_meiying@yeah.net
  • Supported by:

    ThisstudywassupportedbytheNationalNaturalScienceFoundationofChina(31271799).

Abstract:

Sharp eyespot caused by Rhizoctonia cerealis is a soil-borne disease and freezing stress is one of the major abiotic stresses in wheat production. The object of this study was to improve wheat resistance to sharp eyespot and freezing using transgenic technique. The transformation vector pAHC25-MYC-TaPK-R1 expressing the wheat AGC protein kinase gene TaPK-R1 was constructed and transformed into the spring wheat Chinese cultivar Yangmai 20 through particle bombardment. The transformed T1 to T4 plants were subjected to PCR, RT-PCR, qRT-PCR, and Western blot analyses. Three transgenic wheat lines were generated and screened, in which the introduced TaPK-R1 transgene was inherited and expressed in higher level. After inoculation with R. cerealis virulent-isolate R0301 or WK207, these three TaPK-R1-overexpressing transgenic wheat lines displayed significant improved resistance to sharp eyespot. The infection types of these 3 transgenic lines in T1 to T4 generations were 1.16–2.11, and their disease indices were 23.20–42.10. At the same time, the infection types and disease indexes of the non-transformed wheat Yangmai 20 were 2.55–3.60 and 51.00–72.00, respectively. The three-leaf wheat seedlings were treated with ?9 ?C for 24 hours. Freezing tolerances of the three transgenic lines were dramatically improved, whose survival rates were 52%, 79%, and 96%, respectively, and significantly higher (P < 0.01) than that of the non-transformed Yangmai 20 (survival rate of 17%). Our results indicate that resistance/tolerance to sharp eyespot and freezing stress could be significantly enhanced in TaPK-R1 overexpressing transgenic wheat. The three transgenic lines may serve as potential resource in wheat breeding aiming at resistance improvement to sharp eyespot and freezing stress.

Key words: Wheat, Sharpeyespot, Proteinkinase, TaPK-R1, Freezing, Enhancedresistance

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