摘要
为了解植物类固醇5α--还原酶基因（DET2）作为油菜素内酯（BR）生物合成途径中限速酶的作用。利用RACE技术克隆获得棉花的Gh DET2基因,并进行生物信息学分析,然后对棉花‘新陆早17号’幼苗使用PEG6000模拟干旱胁迫24 h,进行不同浓度油菜素内酯（brassinolide,BR）的喷施处理,分析Gh DET2基因在棉花幼苗根、茎、叶中的表达情况。序列分析表明Gh DET2基因的开放阅读框有774 bp,编码257个氨基酸。荧光定量分析结果显示,用BR处理棉苗3 h和6 h后Gh DET2基因表达量在叶中明显提高,而在根和茎中只略微提高,12 h和24 h时Gh DET2基因表达量较前期还有所降低。喷施BR在短时间内促进Gh DET2基因表达量的升高,推测BR喷施能够短期高效地增强棉花的抗旱能力。
This research aimed to understand the function of plant steroid 5α--reductase gene（DET2） as a rate-limiting enzyme in brassinolide（BR） biosynthetic pathway. The cotton ＇XLZ17＇ Gh DET2 gene was cloned by using RACE and bioinformatics analysis was carried out, then PEG6000 was used to simulate drought stress on cotton ＇XLZ17＇ seedlings for 24 h with different concentrations of BR spray treatment, and Gh DET2 gene expression in seedling roots, stems and leaves was analyzed. Sequence analysis showed that the open reading frame of Gh DET2 gene was 774 bp, encoding 257 amino acids. Fluorescent quantitative analysis showed that Gh DET2 gene expression in seedling leaves treated with BR for 3 h and 6 h increased significantly but only slightly increased in roots and stems. However, Gh DET2 gene expression with BR for 12 h and 24 h decreased compared with the previous treatment. Spraying BR could promote Gh DET2 gene expression in a short time, which might suggest that BR spraying could enhance drought resistance of cotton effectively in a short time.
作者
郑冰峰
马玉柱
卢华丹
孙森
安汶铠
张富春
Zheng Bingfeng ,Ma Yuzhu, Lu Huadan ,Sun Sen, An Wenkai ,Zhang Fuchun(Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi, 83004)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2018年第2期859-866,共8页
Genomics and Applied Biology
基金
新疆大学大学生创新训练项目
国家自然科学基金-新疆联合基金重点项目(U1303282)共同资助
关键词
棉花
类固醇5α-还原酶
干旱胁迫
油菜素内酯喷施
表达分析
Cotton, Steroid 5α-reductase gene （DET2）, Drought stress, Brassinolide spraying, Expression analysis
