摘要
为研究铁皮石斛(Dendrobium catenatum Lindl.)应对盐胁迫的抗性机制,利用PCR技术从铁皮石斛cDNA中克隆丝氨酸/苏氨酸蛋白激酶基因CIPK24和类钙调素B亚基蛋白基因CBL1,用生物信息学技术对其编码蛋白的进化关系进行了分析;采用实时荧光定量PCR技术分析它们的表达模式;并构建酵母表达载体,利用酵母双杂交技术研究这两个蛋白的互作关系。结果表明,铁皮石斛CIPK24和CBL1分别与拟南芥CIPK24以及CBL1蛋白的亲缘关系最近;空间表达模式分析发现CIPK24和CBL1基因在花萼、合蕊柱等生殖器官中表达量较高,而在营养器官中表达量较低;盐处理铁皮石斛组培苗后,CIPK24在根中受盐胁迫诱导上调表达,在茎和叶中表达量变化不明显;而CBL1基因在根和叶中受盐胁迫诱导均呈现上调表达,且根中CIPK24和CBL1基因的表达模式相似。酵母双杂交结果表明铁皮石斛CIPK24和CBL1蛋白能够互作。本研究表明,根中CIPK24和CBL1通过互作可能在铁皮石斛响应盐胁迫中起到重要作用,为进一步研究铁皮石斛耐盐性提供理论参考。
In order to study the resistant mechanism of Dendrobium catenatum response to salt stress, a serine/threonine protein kinase gene CIPK24 and calcineurin B-like protein gene CBL1 were cloned from the D.catenatum cDNA by PCR method in this study. Phylogenetic relationships were assayed using bioinformation technique. The expression profiles of the two genes were analyzed by quantative real time PCR(RT-qPCR). The two genes were inserted into yeast expressing vectors, respectively. The interaction between CIPK24 and CBL1 was assayed through yeast two-hybrid system. Phylogenetic tree showed that DcCIPK24 and DcCBL1 were similar with Arabidopsis CIPK24 and CBL1, respectively. The spatial expression pattern of CIPK24 and CBL1 showed that they were highly expressed in reproductive organs such as sepals and gynostemium, and weakly expressed in vegetative organs. The CIPK24 gene was up-regulated by salinity in roots, but not obviously changed in stems and leaves of D. catenatum tissue culture seedlings. While the CBL1 gene was up-regulated both in roots and leaves when exposed to salt treatment. Meanwhile, the expression profiles of CIPK24 and CBL1 were similar in roots.Yeast two-hybrid test showed that CIPK24 could interact with CBL1. These results indicated that CIPK24 and CBL1 may play an important role in the roots in response to salt stress, and may provide theoretical foundation for further research on the salt tolerance of Dendrobium catenatum.
作者
张婷婷
罗琴
傅思毅
王健
宋希强
周扬
Zhang Tingting;Luo Qin;Fu Siyi;Wang Jian;Song Xiqiang;Zhou Yang(Key Laboratory for Quality Regulation of Tropical Horticultural Plants of Hainan Province,Key Laboratory of Germplasm Resources Biology of Tropical Special Ornamental Plants of Hainan Province,College of Horticulture,Hainan University,Haikou,570228;Key Laboratory of Ministry of Education for Genetics and Germplasm Innovation of Tropical Special Trees and Ornamental Plants,College of Forestry,Hainan University,Haikou,570228;Hainan Key Laboratory for Biotechnology of Salt Tolerant Crops,Hainan University,Haikou,570228)
出处
《分子植物育种》
CAS
北大核心
2021年第16期5326-5334,共9页
Molecular Plant Breeding
基金
海南省自然科学基金(319MS009,318QN189)
海南省自然科学基金创新团队项目(2018CXTD331)共同资助
海南大学科研启动项目(KYQD[ZR]1845)。
作者简介
同等贡献:张婷婷;同等贡献:罗琴;通信作者:周扬,zhouyang@hainanu.edu.cn。
