BAS1转录因子筛选及SERK4四种剪切方式功能研究

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	BAS1转录因子筛选及SERK4四种剪切方式功能研究
Alternative Title	Identification of transcription factors controlling the expression of BAS1 and functional analysis of four alternative spliced isoforms of SERK4
	熊敏
Thesis Advisor	黎家
	2016-05-23
Degree Grantor	兰州大学
Place of Conferral	兰州
Degree Name	硕士
Keyword	BRs BAS1 转录因子 SERK4 剪切方式
Abstract	利用生物信息学方法对拟南芥BR代谢途径中的几个重要的酶BAS1，SOB7，UGT73C5，UGT73C6的转录因子进行预测，再构建pBAS1::GUS的纯合背景，将可能的转录因子超表达到该背景植株中，35S::HAT1-GFP，35S::HAT3-GFP植株表现为叶片偏下性生长，叶边缘略微向内卷曲，叶片颜色加深，35S::HAT3-GFP的表型较35S::HAT1-GFP相对严重一些，RT-PCR检测发现BAS1的表达被下调。将HAT1和HAT3超表达到pBAS1::GUS背景植株中，通过对其幼苗，莲座叶，花序，果荚的GUS染色与背景植株作比较发现，35S::HAT1-GFP和35S::HAT3-GFP的植株中GUS显色相对较浅，BAS1的表达量被下调，这与表型反应出的结果一致，也就说明HAT1和HAT3可能调控BAS1的表达。 SERKs家族是RLK家族的重要分支，SERK4与其他SERKs存在功能冗余，不管是缺失突变体还是超表达转基因植株都没有明显的表型。在拟南芥中，许多基因都存在不同的剪切方式，通过RNA-seq数据库序列查找比对发现SERK4存在四种的剪切方式，并通过PCR得到验证。在拟南芥各个组织器官中，这几种剪切方式的表达量并无明显差异；用不同激素、不同温度和胁迫条件处理，各种剪切方式的表达量变化趋势都基本一致；在bri1-5和serk3 serk4背景下超表达这四种剪切方式，只有第二种剪切方式能部分恢复bri1-5的表型，也能恢复serk3 serk4的子叶细胞死亡表型。上述结果表明SERK4的第二种剪切方式在BR信号途径以及SERK3和 SERK4控制细胞死亡过程中发挥功能。
Other Abstract	We identified several transcription factors of BR inactivating enzymes BAS1, SOB7, UGT73C5 and UGT73C6.To reveal thei role in regulating BAS1 expression, we generated a pBAS1::GUS line and analyzed the expression of GUS. We overexpressed transcription factors in Col-0 and found HAT1 and HAT3, can regulate BAS1. 35S::HAT1-GFP and 35S::HAT3-GFP plants displayed a phenotype with dark green leaves. When overexpressing HAT1 and HAT3 in pBAS1::GUS, the expression of BAS1 was down regulated. SERK4 plays a redundant role with BAK1. In Arabidopsis, many genes have several isoforms. Searching sequences from RNA-seq data, we found SERK4 have four isoforms, and the presence of four different isoforms was verified by RT-PCRs. We analyzed the expression levels of the four isoforms in different tissues and under different treatments,the results showed no difference. When overexpressing isoforms in bri1-5 or serk3 serk4, only the second isoform could suppress the deficient phenotypes of bri1-5 and the cell death phenotype of serk3 serk4. These results indicate that only the second isoform of SERK4 is functional in BR signaling and cell-death control pathways.
URL	查看原文
Language	中文
Document Type	学位论文
Identifier	https://ir.lzu.edu.cn/handle/262010/222057
Collection	生命科学学院
Recommended Citation GB/T 7714	熊敏. BAS1转录因子筛选及SERK4四种剪切方式功能研究[D]. 兰州. 兰州大学,2016.

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