新麦草细胞分裂素调控基因CKX4克隆及表达分析

doi:10.11733/j.issn.1007-0435.2023.11.008

草地学报 ›› 2023, Vol. 31 ›› Issue (11): 3289-3298.DOI: 10.11733/j.issn.1007-0435.2023.11.008

新麦草细胞分裂素调控基因CKX4克隆及表达分析

任晓敏¹, 云岚^1,2, 李珍¹, 艾芊¹, 赵乔³, 石凤翎¹

1. 内蒙古农业大学草原与资源环境学院, 内蒙古呼和浩特 010018;
2. 草地资源教育部重点实验室, 内蒙古呼和浩特 010011;
3. 中国科学院深圳先进技术研究院, 广东深圳 518055

收稿日期:2023-05-24 修回日期:2023-07-03 出版日期:2023-11-15 发布日期:2023-12-01
通讯作者: 云岚,E-mail:yunlan@imau.edu.cn;赵乔,E-mail:iao.zhao@siat.ac.cn
作者简介:任晓敏(1998-),女,汉族,内蒙古乌兰察布人,硕士研究生,主要从事草种质资源与育种研究,E-mail:2937923090@qq.com
基金资助:
内蒙古自治区自然科学基金重点项目(2023ZD07)；内蒙古自治区种业创新重大示范工程“揭榜挂帅”项目(2022 JBGS0040)资助

Cloning and Expression Analysis of Cytokinin Regulatory Gene CKX4 in Psathyrostachys juncea

REN Xiao-min¹, YUN Lan^1,2, LI Zhen¹, AI Qian¹, ZHAO Qiao³, SHI Feng-ling¹

1. College of Grassland Resource and Environment, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia 010018, China;
2. Key Laboratory of Grassland Resources of Education Ministry, Hohhot, Inner Mongolia 010011, China;
3.ShenzhenInstitute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, Guangdong Province 518055, China

Received:2023-05-24 Revised:2023-07-03 Online:2023-11-15 Published:2023-12-01

摘要/Abstract

摘要： 分蘖数是影响新麦草饲草和种子产量的关键因素，其关键基因CKX4的克隆及表达分析可为新麦草产量的提高奠定基础。研究克隆了新麦草(Psathyrostachys juncea (Fisch.) Nevski)CKX4基因全长序列，对新麦草及其近缘物种的CKX4进行生物信息学分析；以多分蘖和少分蘖新麦草不同组织为材料，采用qRT-PCR及RNA-Seq方法研究CKX4的相对表达量；利用烟草瞬时转染和蛋白质印迹法分析CKX4-YFP的表达情况及CKX4蛋白的亚细胞定位。结果表明：克隆的新麦草CKX4基因全长为1 596 bp，11个近缘物种的CKX4均有相同的结构域；系统发育树显示新麦草与小麦等麦类作物的亲缘关系最近；CKX4蛋白存在GHS组氨酸残基和Leu(亮氨酸)残基的功能位点；CKX4基因在少分蘖材料中表达量显著高于多分蘖材料；亚细胞定位显示CKX4定位于细胞内。新麦草CKX4蛋白在调控新麦草生长发育中起关键作用，本研究将为新麦草产量性状精准改良提供理论依据和参考。

关键词: 新麦草, CKX4, 基因克隆, 表达分析

Abstract: Number of tillers is a key factor affecting forage and seed yield of Psathyrostachys juncea. Cloning and expression analysis of the key gene CKX4 can lay a foundation for improving the yield of P. juncea. In this study the full-length sequence of CKX4 gene of P. juncea was cloned,and the bioinformatics analysis of CKX4 in both P. juncea and its relative species was carried out. The relative expression of CKX4 in different tissues of P. juncea with more and less tillers was studied by qRT-PCR and RNA-Seq. The expression of CKX4-YFP and the subcellular localization of CKX4 protein were analyzed by transient transfection and Western blot. The results showed that the cloned CKX4 gene of P. juncea was 1 596 bp in size. CKX4s of 11 relative species had the same domain. The phylogenetic tree showed that P. juncea had the closest relationship with Triticum aestivum L. and other wheat crops. CKX4 protein situated in the functional sites of GHS histidine residue and Leu (leucine) residue. The expression of CKX4 gene in sparse tillering materials was significantly higher than that in densely tillering materials. The subcellular localization revealed that CKX4 located inside the cell. The CKX4 protein of P. juncea plays a key role in regulating the growth and development of P. juncea. This research would provide a theoretical basis and references for the precise improvement of yield traits of P. juncea.

Key words: Psathyrostachys juncea(Fisch.) Nevski, CKX4, Gene clone, Expression analysis

中图分类号:

S543

任晓敏, 云岚, 李珍, 艾芊, 赵乔, 石凤翎. 新麦草细胞分裂素调控基因CKX4克隆及表达分析[J]. 草地学报, 2023, 31(11): 3289-3298.

REN Xiao-min, YUN Lan, LI Zhen, AI Qian, ZHAO Qiao, SHI Feng-ling. Cloning and Expression Analysis of Cytokinin Regulatory Gene CKX4 in Psathyrostachys juncea[J]. Acta Agrestia Sinica, 2023, 31(11): 3289-3298.

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新麦草细胞分裂素调控基因CKX4克隆及表达分析

Cloning and Expression Analysis of Cytokinin Regulatory Gene CKX4 in Psathyrostachys juncea

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