播娘蒿耐寒基因DsKIN1的克

发布时间：2018-05-01 09:15

本文选题：播娘蒿 + DsKIN　；参考：《应用与环境生物学报》2017年03期

【摘要】：播娘蒿是极端耐寒的冷诱导植物,研究其低温胁迫下的冷响应基因对于揭示其抗寒性具有重要意义.根据拟南芥At KIN1和At KIN2同源比对,设计同源引物进行播娘蒿KIN基因克隆,通过RACE技术克隆得到Ds KIN1基因,利用Vector NTI11.5软件对其进行序列分析,采用Realtime-PCR测定Ds CBF1、Ds CBF2、Ds COR和Ds KIN1基因在不同处理方式[整株低温处理、同一植株局部低温处理和局部未受低温处理(两部分植株)]下各时间段(0.2 h、0.5 h、1 h、2 h、6 h、12 h、24 h)的表达,采用String数据库预测分析与Ds KIN1相互作用的蛋白.结果显示,Ds KIN1基因序列全长为462 bp,开放阅读框ORF长度为198 bp,编码66个氨基酸,分子量(Mr)为6.61×10~3,理论等电点为9.10,亲水性好.播娘蒿Ds CBF1和Ds CBF2在0.2 h开始表达,在2 h达到最大,随后下降.在冷诱导0.2 h后,Ds CBF调控的耐寒基因Ds COR和Ds KIN1基因开始表达,均呈现上调的趋势.整株、局部冷诱导植株与同一植株上未受低温胁迫部位的耐寒基因Ds COR和Ds KIN1表达模式类似.此外发现与Ds KIN1相互作用的蛋白大部分为低温诱导蛋白、盐和渗透压胁迫响应的信号蛋白、与渗透调节有关的家族蛋白、RING-H2锌指蛋白2B以及未知蛋白等.本研究说明播娘蒿局部冷诱导时,冷信号可进行传递,使未受冷诱导部位获得耐寒性能.
[Abstract]:Artemisia vulgaris is an extremely cold-tolerant cold-induced plant. It is important to study its cold-response gene under low temperature stress for revealing its cold resistance. According to the homologous comparison of at KIN1 and at KIN2 in Arabidopsis thaliana, homologous primers were designed to clone KIN gene of Artemisia vulgaris. Ds KIN1 gene was cloned by RACE technique and sequenced by Vector NTI11.5 software. Realtime-PCR was used to detect the expression of COR and Ds KIN1 genes in different treatments (whole low temperature treatment, local low temperature treatment and local non-cryogenic treatment (two parts of plants), and the expression of COR and Ds KIN1 gene in different treatments (0. 2 h ~ 0. 5 h ~ 1 h ~ 2 h ~ 6 h ~ (12) h ~ (24 h) in different treatments (whole plant low temperature treatment, same plant local low temperature treatment and local low temperature treatment (two parts plants). The proteins interacting with Ds KIN1 were predicted by String database. The results showed that the length of KIN1 gene was 462 BP, the length of open reading frame ORF was 198 BP, the molecular weight was 6.61 脳 10 ~ (-3), the theoretical isoelectric point was 9.10, and the hydrophilicity was good. The expression of Ds CBF1 and Ds CBF2 began at 0. 2 h, reached the maximum at 2 h, and then decreased. After 0. 2 h of cold induction, Ds COR and Ds KIN1 genes regulated by Ds CBF began to express, both of which were up-regulated. The expression patterns of Ds COR and Ds KIN1 were similar in whole and local cold induced plants. In addition, it was found that most of the proteins interacting with Ds KIN1 were hypothermia inducible proteins, signal proteins responsive to salt and osmotic stress, RING-H2 zinc finger proteins 2B and unknown proteins related to osmotic regulation. This study shows that the cold signal can be transmitted during local cold induction, so that the cold tolerance can be obtained.
【作者单位】：四川大学生命科学学院微生物与代谢工程四川省重点实验室;
【基金】：国家自然科学基金项目(31171447) “十二五”国家科技支撑计划项目(2014BAD02B02) 国家微生物资源平台项目(NIMR-2016-8) 四川省科技厅项目(2014GZX0005,2017JCPT0001)资助~~
【分类号】：Q943.2

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