农业生物技术学报2013，Vol.21Issue(4)：396-406,11.DOI:10.3969/j.issn.1674-7968.2013.04.003
小麦染色体组蛋白泛素化与生理型雄性不育相关性的研究
Study on the Relationship between the Ubiquitination of Histones on Chromosomes and the Physiological Male Sterility Induced by Chemical Hybridizing Agent SQ-1 in Wheat(Triticum aestivum L.)
摘要
Abstract
Wheat(Triticum aestivum L.) as the main food crop, studying its utilization of heterosis and male sterility is the matter that has been tried to overcome in the world. To deeply research the mechanism of the physiological male sterility of wheat, the relationship between the ubiquitination of histones and the physiological male sterility was studied, which based on the research front of histone modifications. Used physiologically male sterility induced with chemical hybridizing agent SQ-1 and corresponding normal lines of complete fertility as experimental materials in wheat, investigated the rate of the physiological male sterility as well as their pistil fertility. The histones were extracted from the two lines' flowers. The ubiquitination levels of the histones were monitored with Western blot, and then cloned the cDNA functional sequence of RAD6, which was identified as an ubiquitin-conjugating enzyme of histone H2B. Bioinformatics technique was applied to analyze RAD6 such as sequence multiplication, constructed a phylogenetic tree, forecasted secondary structure and 3D structure of RAD6. Lastly, qRT-PCR was performed to analyze the expression of the RAD6 gene. The results showed that the chemical hybridizing agent SQ-1 had a sound effect on mile-killing. The ubiquitination level of histone H3 was almost the same between the male sterile and fertile lines. During mononuclear period, the ubiquitination of histone H2A was seemed to be existing in fertility, but disappearing in the sterility ones; the ubiquitination level of histone H2B in physiological male sterile wheat was seemed to be significantly less than that in fertile ones, meanwhile, the expression of the RAD6 gene in sterility wheats was apparently suppressed. These data indicated that SQ- 1 distinctly restrained the ubiquitination levels of histones H2A, H2B as well as the expression of the RAD6 gene in the mononuclear period, which may be influencing the translation of some key genes and evently result in physiological male sterility in wheat. Thus, it can be seen that there is a close relationship between the ubiquitination of the histones on chromosomes and the physiological male sterility induced by the chemical hybridizing agent SQ-1 in wheat. The research supplies scientific evidence for the ubiquitination of histones involve in the wheat physiological male sterility, meanwhile, it establishes the solid epigenetics study foundation for wheat physiological male sterility mechanism.关键词
小麦/杀雄剂SQ-1/免疫印迹/组蛋白泛素化/RAD6/荧光定量PCR/生物信息学Key words
Wheat/ Chemical hybridizing agent SQ-1/ Western blot/ Histone ubiquitination/ RAD6/ qRT- PCR/ Bioinformatics引用本文复制引用
桑青,刘红占,张改生,朱启迪,祝万万,张龙雨,车会学..小麦染色体组蛋白泛素化与生理型雄性不育相关性的研究[J].农业生物技术学报,2013,21(4):396-406,11.基金项目
国家高技术研究发展计划(863)重大专项(No.2011AA10A106)、国家自然科学基金项目(No.31071477,No.31171611)、高等学校博士学科点专项科研基金(No.20090204110024)和陕西省"13115"科技创新工程重大科技专项(No.2010ZDKG-68) （863）
