NAC转录因子与乙烯合成及信号转导基因间的相互调控作用
发布时间：2018-08-15 16:51
【摘要】：NAC转录因子是植物特有的具有多种生物调控功能的转录因子。研究表明,NAC转录因子参与调控植物生长发育、激素信号转导途径及胁迫应答反应。本文利用生物信息学的方法对番茄NAC4-9转录因子进行结构分析和生物功能预测。利用病毒诱导的基因沉默技术(VIGS)对NAC转录因子编码基因及乙烯合成和信号转导相关基因进行沉默,以此研究NAC转录因子在番茄成熟及乙烯信号转导途径中的调控作用。主要研究结果如下:(1)生物信息学分析:对SNAC4-9转录因子蛋白的高级结构和生物调控功能预测结果显示,SNAC4和SNAC6与拟南芥ANAC056最相近;SNAC5和SNAC8具有相似的高级结构且与拟南芥ANAC072最相近;SNAC7和SNAC9与拟南芥ANAC029最相近且三者具有相似的三级结构。生物调控功能预测结果显示,SNAC4-9转录因子还可能参与调控生长素、脱落酸、茉莉酸等植物激素的信号转导途经。分析乙烯合成和信号转导相关基因LeACS4、LeACO1、NR、LeETR4、LeCTR1、LeERF2的启动子序列进行转录因子结合位点预测,发现乙烯相关因子可能参与调控茉莉酸、脱落酸和水杨酸等激素响应及高温、干旱等逆境胁迫反应。(2)VIGS技术对转录因子的生物功能研究:构建pTRV2-LeACS4/LeACO1/NR/LeETR4/LeCTR1/LeERF2载体,应用注射法侵染活体番茄植株,沉默番茄果实中的SNAC4/5/6/7/9和LeACS4/LeACO1/NR/LeETR4/LeCTR1/LeERF2/LeRIN基因。在农杆菌侵染10天后,沉默SNACs基因的番茄植株株高低于对照组。沉默SNACs及乙烯相关基因的植株,番茄果实表现出成熟延迟表型,而只注射含空TRV2载体的农杆菌菌液的对照组果实照常成熟。(3)基因沉默的番茄果实转录水平检测结果显示SNAC基因的沉默影响了番茄果实中乙烯合成和信号转导相关基因的表达情况。SNAC基因的沉默会导致乙烯合成基因LeACS2/LeACS4/LeACO1及乙烯受体基因NR/LeETR4表达水平降低。同时,SNAC基因的表达水平也受到乙烯合成及信号转导基因表达情况的影响,乙烯合成基因LeACS4/LeACO1和乙烯信号转导相关因子LeCTR1/LeERF2的编码基因的沉默抑制SNAC基因表达,乙烯受体基因NR/LeETR4的沉默诱导SNAC基因表达水平升高。(4)沉默SNAC4基因后,在成熟过程中,番茄果实的乙烯释放速率下降,并且延迟了乙烯释放高峰的出现时间。
[Abstract]:NAC transcription factors are plant specific transcription factors with various biological regulatory functions. The results showed that NAC transcription factors were involved in plant growth and development, hormone signal transduction pathway and stress response. In this paper, bioinformatics was used to analyze the structure and biofunction of tomato NAC4-9 transcription factors. Viral induced gene silencing technique (VIGS) was used to silence NAC transcription factor encoding genes and ethylene synthesis and signal transduction related genes in order to study the regulatory role of NAC transcription factors in tomato maturation and ethylene signal transduction pathway. The main results are as follows: (1) Bioinformatics analysis: the prediction of the advanced structure and bioregulatory function of SNAC4-9 transcription factor protein shows that SNAC4 and SNAC6 have similar high-level structures to Arabidopsis ANAC056 and SNAC5 and SNAC8. SNAC7 and SNAC9 are the most similar in ANAC072 and ANAC029 in Arabidopsis thaliana, and the three have similar tertiary structures. The prediction of biological regulation function indicates that SNAC4-9 transcription factor may also be involved in the regulation of auxin, abscisic acid, jasmonic acid and other plant hormones signal transduction pathway. By analyzing the promoter sequence of LeACS4, LeACO1, LeETR4, LeCTR1, LeERF2, and predicting transcription factor binding sites, we found that ethylene related factors may be involved in regulating the response of hormones such as jasmonic acid, abscisic acid and salicylic acid and high temperature. (2) study on the biological function of transcription factors by VIGS: construct pTRV2-LeACS4/LeACO1/NR/LeETR4/LeCTR1/LeERF2 vector, infect tomato plants in vivo by injection method, silence the SNAC4/5/6/7/9 and LeACS4/LeACO1/NR/LeETR4/LeCTR1/LeERF2/LeRIN genes in tomato fruit. After infection with Agrobacterium tumefaciens for 10 days, the plant height of tomato with SNACs gene silencing was lower than that of control group. Tomato fruits with silencing SNACs and ethylene-related genes showed delayed maturation phenotype. The fruit of control group treated with Agrobacterium tumefaciens containing empty TRV2 vector was mature as usual. (3) the transcriptional level of tomato fruit with gene silencing showed that the silencing of SNAC gene affected ethylene synthesis and signal transduction in tomato fruit. The silencing of SNAC gene resulted in the decrease of LeACS2/LeACS4/LeACO1 and NR/LeETR4 expression of ethylene biosynthesis gene. At the same time, the expression level of SNAC gene was also affected by ethylene synthesis and signal transduction gene expression. The silencing of ethylene synthesis gene LeACS4/LeACO1 and the encoding gene of ethylene signal transduction related factor LeCTR1/LeERF2 inhibited the expression of SNAC gene. The silencing of ethylene receptor gene NR/LeETR4 induced the increase of SNAC gene expression. (4) after silencing the SNAC4 gene, the ethylene release rate of tomato fruit decreased during maturation, and the emergence time of ethylene release peak was delayed.
【学位授予单位】：天津大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2
本文编号：2184825
[Abstract]:NAC transcription factors are plant specific transcription factors with various biological regulatory functions. The results showed that NAC transcription factors were involved in plant growth and development, hormone signal transduction pathway and stress response. In this paper, bioinformatics was used to analyze the structure and biofunction of tomato NAC4-9 transcription factors. Viral induced gene silencing technique (VIGS) was used to silence NAC transcription factor encoding genes and ethylene synthesis and signal transduction related genes in order to study the regulatory role of NAC transcription factors in tomato maturation and ethylene signal transduction pathway. The main results are as follows: (1) Bioinformatics analysis: the prediction of the advanced structure and bioregulatory function of SNAC4-9 transcription factor protein shows that SNAC4 and SNAC6 have similar high-level structures to Arabidopsis ANAC056 and SNAC5 and SNAC8. SNAC7 and SNAC9 are the most similar in ANAC072 and ANAC029 in Arabidopsis thaliana, and the three have similar tertiary structures. The prediction of biological regulation function indicates that SNAC4-9 transcription factor may also be involved in the regulation of auxin, abscisic acid, jasmonic acid and other plant hormones signal transduction pathway. By analyzing the promoter sequence of LeACS4, LeACO1, LeETR4, LeCTR1, LeERF2, and predicting transcription factor binding sites, we found that ethylene related factors may be involved in regulating the response of hormones such as jasmonic acid, abscisic acid and salicylic acid and high temperature. (2) study on the biological function of transcription factors by VIGS: construct pTRV2-LeACS4/LeACO1/NR/LeETR4/LeCTR1/LeERF2 vector, infect tomato plants in vivo by injection method, silence the SNAC4/5/6/7/9 and LeACS4/LeACO1/NR/LeETR4/LeCTR1/LeERF2/LeRIN genes in tomato fruit. After infection with Agrobacterium tumefaciens for 10 days, the plant height of tomato with SNACs gene silencing was lower than that of control group. Tomato fruits with silencing SNACs and ethylene-related genes showed delayed maturation phenotype. The fruit of control group treated with Agrobacterium tumefaciens containing empty TRV2 vector was mature as usual. (3) the transcriptional level of tomato fruit with gene silencing showed that the silencing of SNAC gene affected ethylene synthesis and signal transduction in tomato fruit. The silencing of SNAC gene resulted in the decrease of LeACS2/LeACS4/LeACO1 and NR/LeETR4 expression of ethylene biosynthesis gene. At the same time, the expression level of SNAC gene was also affected by ethylene synthesis and signal transduction gene expression. The silencing of ethylene synthesis gene LeACS4/LeACO1 and the encoding gene of ethylene signal transduction related factor LeCTR1/LeERF2 inhibited the expression of SNAC gene. The silencing of ethylene receptor gene NR/LeETR4 induced the increase of SNAC gene expression. (4) after silencing the SNAC4 gene, the ethylene release rate of tomato fruit decreased during maturation, and the emergence time of ethylene release peak was delayed.
【学位授予单位】：天津大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2
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