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作物学报 ›› 2016, Vol. 42 ›› Issue (11): 1601-1608.doi: 10.3724/SP.J.1006.2016.01601

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

过表达TaPK-R1基因增强了小麦对纹枯病的抗性和耐冻性

罗美英1,2,荣玮2,魏学宁2,杨坤2,徐惠君2,禤维言1,*,张增艳2,*   

  1. 1广西大学农学院,广西南宁530004;2中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程/农业部麦类生物学与遗传育种重点实验室,北京100081
  • 收稿日期:2016-03-09 修回日期:2016-07-11 出版日期:2016-11-12 网络出版日期:2016-08-11
  • 通讯作者: 禤维言:E-mail:xuanweiyan_1@163.com;张增艳,E-mail:zhangzengyan@caas.cn
  • 基金资助:

    本研究由国家自然科学基金项目(31271799)资助。

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 Published:2016-11-12 Published online:2016-08-11
  • Contact: Xuan Weiyan:E-mail:xuanweiyan_1@163.com;Zhang Zengyan,E-mail:zhangzengyan@caas.cn
  • Supported by:

    ThisstudywassupportedbytheNationalNaturalScienceFoundationofChina(31271799).

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摘要：

小麦纹枯病菌(Rhizoctonia cerealis)和倒春寒是小麦生产中重要的生物和非生物胁迫因子，本研究目的是利用转基因技术改良小麦的纹枯病抗性和耐冷性。利用构建的转基因载体pAHC25-MYC-TaPK-R1，通过基因枪介导法，将小麦AGC蛋白激酶基因TaPK-R1导入春小麦品种扬麦20，获得TaPK-R1过表达的转基因小麦。对T1至T4代植株进行PCR、RT-PCR、qRT-PCR、Western blot分析，筛选到3个转基因小麦株系，外源TaPK-R1基因能够在其植物体内遗传和超量转录，并翻译成蛋白质。利用致病力不同的R. cerealis菌株R0301和WK207，分别对3个转基因株系T1至T2代和T3至T4代进行纹枯病抗性鉴定，发现TaPK-R1过量表达提高了转基因小麦的纹枯病抗性，3个转基因株系各世代的纹枯病平均病级为1.16~2.11, 病情指数为23.20~42.10，而对照扬麦20的纹枯病病级为2.55~3.60，病情指数为51.00~72.00。用?9℃低温处理三叶一心期小麦幼苗24 h，对照扬麦20的存活率为17%，3个转基因小麦株系的存活率分别为52%、79%和96%。上述结果表明，TaPK-R1过量表达能显著增强小麦对纹枯病的抗性及对冻害的耐受性, 所获得的3个转基因小麦株系可作为潜在的抗源材料。

关键词: 小麦, 纹枯病, 蛋白激酶, TaPK-R1, 冻害

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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