兰州大学机构库 >生命科学学院
BAS1转录因子筛选及SERK4四种剪切方式功能研究
Alternative TitleIdentification 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
KeywordBRs 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 AbstractWe 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
Identifierhttps://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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