小桐子抗逆相关基因的克隆和表达分析

发布时间：2018-04-25 16:35

  本文选题：小桐子 + 非生物胁迫　； 参考：《昆明理工大学》2017年硕士论文

【摘要】：小桐子(Jatrophacurcas L.)属于大戟科麻风树属,是近年来受到广泛关注的一种能源植物。为了不与粮食作物争地和改善生态环境的需要,小桐子一般被种植在贫瘠、干旱的土地上,面临多种逆境胁迫,极大地影响了它的产量与油质,限制了它的地域分布,故对其抗逆性的研究具有重要意义。本研究从小桐子成熟的叶片中克隆获得5个与其抗逆相关基因的cDNA全长,进行了生物信息学分析,并结合qRT-PCR方法研究了以上5个基因在小桐子不同组织、种子不同发育阶段和不同非生物胁迫处理下的表达模式,为有效挖掘和利用小桐子的抗性优良基因提供参考。本研究获得的主要结果如下:1、克隆得到JcNF-YB3和JcNF-YB10基因的CDS序列,分别包含549bp和528bp完全阅读框(ORF),编码182和175个氨基酸,都与油桐NF-YB家族基因序列一致性最高且亲缘关系最近。定量分析表明,JcNF-YB3和JcNF-YB10在小桐子根、茎、叶、花、果皮和种子中都有表达,JcNF-YB3在根中表达量最高,JcNF-YB10在种子中表达量最高。两个基因在种子发育期的表达量变化趋势一致,但是JcNF-YB10在种子整个生长发育期的表达量都要高于JcNF-YB3。JcNF-YB3和JcNF-YB10都在NaCl、PEG、4℃低温、机械损伤和ABA处理后表达量不同程度上调。2、克隆得到JcGSK基因的CDS序列,包含1230bpORF,编码409个氨基酸,与巴西橡胶树GSK蛋白的氨基酸序列一致性最高(94%)且亲缘关系最近。组织表达结果显示,JcGSK在小桐子根、茎、叶、花、果皮和种子中都有表达,在根中表达量最高,种子中的表达量次之,在种子发育过程中表达量的变化与种子生长发育趋势基本一致。小桐子幼苗在NaCl、PEG、4℃低温、机械损伤和ABA处理后JcGSK表达量不同程度上调。3、克隆得到JcYkt6基因的CDS序列,包含600bpORF,编码199个氨基酸,与香瓜和黄瓜的Ykt6蛋白序列一致性最高(91%)且亲缘关系最近。定量分析结果显示,JcYkt6基因在小桐子根、茎、叶、花、果皮和种子中都有表达,在种子中表达量最高。在种子发育期表达量变化过程与种子中储藏蛋白质合成趋势基本一致。NaCl、PEG、4℃低温、机械损伤和ABA处理下JcYkt6表达水平都不同程度上调。4、克隆得到Jc5β-StR基因的CDS序列,包含1173 bp ORF,编码390个氨基酸,与蓖麻5β-StR蛋白序列一致性最高(87%)且亲缘关系最近。Jc5β-StR是一个短链还原酶,属于SDR超家族。组织表达结果显示Jc5β-StR在小桐子根、茎、叶、花、果皮和种子中都有表达,在种子中表达量最高,在种子生长发育期表达量变化与种子生长发育的趋势基本一致。NaCl、PEG、4℃低温、机械损伤和ABA处理下,Jc5β-StR的表达量都不同程度上调。总之,JcNF-YB3、JcNF-YB10、JcGSK、JcYkt6和Jc5β-StR都具有组织表达特异性,可能参与调控小桐子器官生长发育,虽然在不同非生物胁迫条件下的表达模式不同,但是它们都能够参与小桐子对于非生物胁迫的应答,表明它们在小桐子的抗逆机制上也起到一定作用。
[Abstract]:Jatrophacurcas L. It belongs to the genus Euphorbiaceae, and is a kind of energy plant which has received extensive attention in recent years. In order not to compete with food crops for land and to improve the ecological environment, jatropha is generally planted on barren, arid land and is subject to various stresses, which greatly affect its yield and oil quality and limit its geographical distribution. Therefore, the study of its resistance is of great significance. In this study, the full-length cDNA of five genes related to stress resistance were cloned from mature leaves of Paulownia chinensis. The bioinformatics analysis was carried out, and the above five genes were studied in different tissues of Paulownia cuspidata with qRT-PCR method. The expression patterns of seeds at different developmental stages and under different abiotic stress provide a reference for the effective mining and utilization of the fine genes of resistance of Paulownia chinensis. The main results of this study were as follows: 1. The CDS sequences of JcNF-YB3 and JcNF-YB10 genes were cloned, including 549bp and 528bp complete reading frame, encoding 182 and 175 amino acids respectively. Quantitative analysis showed that JcNF-YB3 and JcNF-YB10 were expressed in the roots, stems, leaves, flowers, pericarp and seeds of Paulownia cuspidata. JcNF-YB3 expressed the highest amount in the root and JcNF-YB10 was the highest in the seeds. The expression of JcNF-YB10 was higher than that of JcNF-YB3.JcNF-YB3 and JcNF-YB10 at 4 鈩，

本文编号：1802122