甜瓜CDPK和CRK基因家族的鉴定及表达分析

发布时间：2018-10-16 14:41

【摘要】：甜瓜是重要的经济作物,在我国栽培面积广泛,但它的生长发育易受逆境的影响。CDPK是一种植物中常见的丝氨酸/苏氨酸蛋白激酶,在植物生长发育和抵御逆境方面发挥重要作用。本研究利用生物信息学方法,在全基因组范围内,对甜瓜的CDPK和CRK基因家族进行鉴定,并分析其染色体分布、基因结构、进化关系、基因复制方式及与拟南芥的共线性关系。同时,运用实时荧光定量PCR,检测不同甜瓜组织及逆境胁迫(白粉病、盐害、低温及ABA)后CDPK和CRK基因的表达量,筛选到与逆境胁迫密切相关的基因。本研究取得的结果如下:1、在甜瓜全基因组数据库中鉴定出18个CmCDPKs和7个CmCRKs,这25个基因分布在除3号染色体外的11条甜瓜染色体上,其中4号染色体上分布的CDPK基因最多。2、甜瓜CDPK和CRK基因的进化和结构分析。通过进化关系分析,将甜瓜CDPKs分为四个亚族(CDPK I-IV),CDPK I-III亚族中的CDPK基因外显子数目和内含子相位较相似,含有7或8个外显子。与其他三个亚族相比,CDPK IV亚族中的CmCDPK14有12个外显子,其外显子-内含子结构与CRK基因较相似。用甜瓜、拟南芥、水稻和番茄的CDPK和CRK蛋白序列构建系统进化树。结果显示,110个CDPKs被分为四个亚族,每个物种的CDPK在四个亚族中均有分布。在系统发育进化树上,CDPK IV和CRK I的进化关系较为紧密。3、甜瓜CDPK和CRK的复制方式及与拟南芥的共线性分析。通过分析,在甜瓜中鉴定出两对片段复制的CDPK基因。同时在甜瓜和拟南芥中找出了20对共线性的CDPK基因和2对共线性的CRK基因。4、甜瓜CDPK和CRK基因的表达模式。运用实时荧光定量PCR,检测了甜瓜CDPK和CRK基因在不同组织及不同逆境胁迫后的表达量。结果表明,除了CmCDPK3,其余的基因在根、茎、叶、花和卷须的至少一个器官中分布。大多数基因在白粉和ABA处理后下调表达,在盐和低温胁迫后上调表达。与其他处理相比,甜瓜CDPK和CRK基因对冷害胁迫的响应较为强烈。以上结果说明甜瓜CDPK和CRK基因参与逆境信号的转导。5、成功克隆CmCDPK7、CmCDPK10和CmCDPK15的CDS序列,连接pBI121构建了表达载体。
[Abstract]:Muskmelon is an important cash crop, which is widely cultivated in China, but its growth and development are susceptible to stress. CDPK is a common serine / threonine protein kinase in plants. Play an important role in plant growth and development and resistance to adversity. In this study, the CDPK and CRK gene families of muskmelon were identified by bioinformatics, and their chromosomal distribution, gene structure, evolutionary relationship, gene replication and co-linear relationship with Arabidopsis were analyzed. Meanwhile, real-time fluorescence quantitative PCR, was used to detect the expression of CDPK and CRK genes in different muskmelon tissues and under stress (powdery mildew, salt injury, hypothermia and ABA). The results are as follows: 1. In the whole genome database of muskmelon, 18 CmCDPKs and 7 CmCRKs, genes were identified on 11 muskmelon chromosomes except chromosome 3. The distribution of CDPK gene on chromosome 4 was the most. 2. Evolution and structure analysis of CDPK and CRK genes in muskmelon. The CDPKs of melon was divided into four subfamilies (the number of exons of CDPK gene and the phase of intron were similar in CDPK I-IV), CDPK I-III subfamily), which contained 7 or 8 exons. Compared with the other three subfamilies, CmCDPK14 has 12 exons in the, CDPK IV subfamily, and its exon-intron structure is similar to that of the CRK gene. Phylogenetic tree was constructed using CDPK and CRK protein sequences of melon, Arabidopsis thaliana, rice and tomato. The results showed that 110 CDPKs were divided into four subfamilies, and the CDPK of each species was distributed in the four subfamilies. The relationship between, CDPK IV and CRK I was close in phylogenetic tree. 3. Replication of CDPK and CRK in muskmelon and collinear analysis with Arabidopsis thaliana. Two pairs of replicating CDPK genes were identified in muskmelon by analysis. At the same time, 20 pairs of co-linear CDPK genes and 2 pairs of co-linear CRK genes were found in muskmelon and Arabidopsis thaliana. 4. The expression patterns of CDPK and CRK genes in muskmelon were also found. The expression of CDPK and CRK genes in muskmelon was detected by real-time fluorescence quantitative PCR,. The results showed that the genes except CmCDPK3, were distributed in at least one organ of roots, stems, leaves, flowers and tendrils. Most genes were down-regulated after white powder and ABA treatment, and up-regulated after salt and low temperature stress. Compared with other treatments, the response of CDPK and CRK genes to chilling stress was stronger in muskmelon. These results indicated that the CDPK and CRK genes were involved in the stress signal transduction. 5. The CDS sequences of CmCDPK7,CmCDPK10 and CmCDPK15 were cloned and ligated into pBI121 to construct the expression vector.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S652;Q943.2

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