菜心<i>BcCIPK23</i>基因克隆及表达特性分析

朱云娜; 杨景辉; 王斌; 王玉昆; 周裕荣; 卢威宇; 刘建国

doi:10.16768/j.issn.1004-874X.2023.09.009

菜心BcCIPK23基因克隆及表达特性分析

Cloning and Expression Characteristics Analysis of BcCIPK23 in Flowering Chinese Cabbage

摘要

摘要:
目的 CIPK23基因在植物生长发育和矿质营养吸收中具有重要作用，但在菜心中关于BcCIPK23基因及其功能的研究尚无报道，该研究可为探讨BcCIPK23在菜心植株生长发育和氮代谢中的生物学功能提供理论参考。
方法以‘油绿501’菜心为材料，采用同源克隆法克隆到菜心BcCIPK23全长序列，对其编码蛋白进行生物信息学分析，探究其组织表达特性和对不同氮源的响应。
结果菜心BcCIPK23基因CDS全长为1 450 bp，编码482个氨基酸，分子量是53.41 kD，理论等电点为9.28，具有CIPK家族成员的STKc-SnRK3和CIPK-C保守结构域。系统进化树分析表明，菜心BcCIPK23和白菜BrCIPK23进化关系最近。BcCIPK23在菜心叶片、叶柄、薹茎、根系、花等组织中均有表达，但主要集中在叶片、叶柄、薹茎中表达；BcCIPK23表达也受氮源及其水平的影响，无论供NO₃^-还是供NH₄⁺，其表达水平均随氮浓度增加而降低。推测该基因在叶片、薹茎发育和氮素吸收利用中发挥关键作用。
结论菜心BcCIPK23为CIPK基因家族成员，主要在叶片等生长活跃的器官高表达，且可响应氮源及氮浓度变化，参与植物生长发育和氮代谢过程。

Abstract:
Objective CIPK23 plays an important role in the development and mineral absorption in many plants, however, there are no relevant reports on CIPK23 gene and its function in flowering Chinese cabbage. The research will provide some theoretical references for further study of the function of BcCIPK23 in the growth and development and nitrogen metabolism in flowering Chinese cabbage.
Method The full-length cDNA sequence of BcCIPK23 was obtained by homologous cloning method from flowering Chinese cabbage variety 'Youlv 501'. The encoded BcCIPK23 protein were analyzed by bioinformatics, and the expression characteristics of BcCIPK23 in different tissues and its response to different nitrogen sources of flowering Chinese cabbage were analyzed.
Result The CDS full length of BcCIPK23 gene in flowering Chinese cabbage was 1 450 bp, encoding 482 amino acids, with a molecular weight of 53.41 kD, and a theoretical isoelectric point of 9.28. The BcCIPK23 sequence contained the domains of STKc-SnRK3 and CIPK-C, which were the conserved domains of CIPK family. The phylogenetic analysis showed that BcCIPK23 from flowering Chinese cabbage was antigenically closely related to BrCIPK23 from Brassica rapa. The qRT-PCR results showed that BcCIPK23 was expressed in leaves, petioles, stalks, roots, flowers and other tissues of flowering Chinese cabbage, but was mainly expressed in leaves, petioles and stalks. The expression level of BcCIPK23 was significantly affected by different sources and concentrations of nitrogen, decreased significantly with the increase of NO₃^- or NH₄⁺ concentration. It was predicted that BcCIPK23 may play a key role in the growth and development of leaves and stalks, and the absorption and utilization of nitrogen in flowering Chinese cabbage.
Conclusion BcCIPK23 is a member of the CIPK gene family, which is highly expressed in leaves and other active organs. It can respond to changes in nitrogen sources and nitrogen concentrations, and is involved in plant growth and development and nitrogen metabolic processes.

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