甜瓜CDPK和CRK基因家族的鑒定及表達分析
發(fā)布時間：2018-10-16 14:41
【摘要】：甜瓜是重要的經(jīng)濟作物,在我國栽培面積廣泛,但它的生長發(fā)育易受逆境的影響。CDPK是一種植物中常見的絲氨酸/蘇氨酸蛋白激酶,在植物生長發(fā)育和抵御逆境方面發(fā)揮重要作用。本研究利用生物信息學方法,在全基因組范圍內(nèi),對甜瓜的CDPK和CRK基因家族進行鑒定,并分析其染色體分布、基因結(jié)構(gòu)、進化關系、基因復制方式及與擬南芥的共線性關系。同時,運用實時熒光定量PCR,檢測不同甜瓜組織及逆境脅迫(白粉病、鹽害、低溫及ABA)后CDPK和CRK基因的表達量,篩選到與逆境脅迫密切相關的基因。本研究取得的結(jié)果如下:1、在甜瓜全基因組數(shù)據(jù)庫中鑒定出18個CmCDPKs和7個CmCRKs,這25個基因分布在除3號染色體外的11條甜瓜染色體上,其中4號染色體上分布的CDPK基因最多。2、甜瓜CDPK和CRK基因的進化和結(jié)構(gòu)分析。通過進化關系分析,將甜瓜CDPKs分為四個亞族(CDPK I-IV),CDPK I-III亞族中的CDPK基因外顯子數(shù)目和內(nèi)含子相位較相似,含有7或8個外顯子。與其他三個亞族相比,CDPK IV亞族中的CmCDPK14有12個外顯子,其外顯子-內(nèi)含子結(jié)構(gòu)與CRK基因較相似。用甜瓜、擬南芥、水稻和番茄的CDPK和CRK蛋白序列構(gòu)建系統(tǒng)進化樹。結(jié)果顯示,110個CDPKs被分為四個亞族,每個物種的CDPK在四個亞族中均有分布。在系統(tǒng)發(fā)育進化樹上,CDPK IV和CRK I的進化關系較為緊密。3、甜瓜CDPK和CRK的復制方式及與擬南芥的共線性分析。通過分析,在甜瓜中鑒定出兩對片段復制的CDPK基因。同時在甜瓜和擬南芥中找出了20對共線性的CDPK基因和2對共線性的CRK基因。4、甜瓜CDPK和CRK基因的表達模式。運用實時熒光定量PCR,檢測了甜瓜CDPK和CRK基因在不同組織及不同逆境脅迫后的表達量。結(jié)果表明,除了CmCDPK3,其余的基因在根、莖、葉、花和卷須的至少一個器官中分布。大多數(shù)基因在白粉和ABA處理后下調(diào)表達,在鹽和低溫脅迫后上調(diào)表達。與其他處理相比,甜瓜CDPK和CRK基因?qū)浜γ{迫的響應較為強烈。以上結(jié)果說明甜瓜CDPK和CRK基因參與逆境信號的轉(zhuǎn)導。5、成功克隆CmCDPK7、CmCDPK10和CmCDPK15的CDS序列,連接pBI121構(gòu)建了表達載體。
[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.
【學位授予單位】：西北農(nóng)林科技大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S652;Q943.2
本文編號：2274680
[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.
【學位授予單位】：西北農(nóng)林科技大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S652;Q943.2
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