Journal of Integrative Agriculture ›› 2023, Vol. 22 ›› Issue (2): 642-649.DOI: 10.1016/j.jia.2022.08.068

• • 上一篇    

JIA-2021-1796 高粱SnRK1亚基编码基因的鉴定及其组合模式研究

  

  • 收稿日期:2021-10-12 接受日期:2022-01-27 出版日期:2023-02-20 发布日期:2022-01-27

Characterization of subunits encoded by SnRK1 and dissection of combinations among these subunits in sorghum (Sorghum bicolor L.)

XIAO Qian-lin*, HUANG Tian-hui*, ZHOU Chang, CHEN Wei-xi, CHA Jian-kui, WEI Xi-mei, XING Fang-yu, QIAN Meng-ya, MA Qian-nan, DUAN Hong, LIU Zhi-zhai   

  1. College of Agronomy and Biotechnology, Southwest University, Chongqing 400715, P.R.China

  • Received:2021-10-12 Accepted:2022-01-27 Online:2023-02-20 Published:2022-01-27
  • About author:XIAO Qian-lin, Tel: +86-23-68251410, E-mail: xiaoql1853@swu.edu.cn; HUANG Tian-hui, E-mail: 1772599750@qq.com; Correspondence LIU Zhi-zhai, Tel: +86-23-68251410, E-mail: liu003@swu.edu.cn * These authors contributed equally to this study.
  • Supported by:

    This work was supported by the National Natural Science Foundation of China (32001607) and the Fundamental Research Funds for the Central Universities of Southwest University, China (SWU118087).  

摘要:

蔗糖非发酵相关蛋白激酶1SnRK1)是一类重要的丝氨酸/苏氨酸蛋白激酶,广泛参与了植物的生长发育,并通过代谢及生理过程参与植物对生物与非生物胁迫的应答,同时在植物的碳水化合物分布及糖信号转导中也发挥着重要作用。本研究通过序列比对,从高粱基因组中鉴定到8SnRK1亚基编码基因,其中3个编码α亚基(SnRK1α1 - SnRK1α3)、3个编码β亚基(SnRK1β1 - SnRK1β3),以及编码γ (SnRK1γ)和βγ(SnRK1βγ)亚基的编码基因各1个。这8个高粱SnRK1基因分布于第1-3、第7、第95条染色体,与来自玉米、水稻的SnRK1基因存在共线性,并且这些基因的编码产物在相同亚基上表现出了高度的同源性。基于qRT-PCR的分析结果显示,在8个高粱SnRK1基因中,除SnRK1α3在籽粒中低表达以及SnRK1β2在穗中高表达外,其他基因在其他组织中均表现出相似的表达特性。基于高粱叶片原生质体的亚细胞定位结果表明,α1、α2、α3、γ4个亚基主要定位于细胞器,而在细胞膜和部分细胞器上则可以检测到β1、β2、β3、及βγ4个亚基的定位信号。此外,酵母双杂交分析发现,8个高粱SnRK1亚基存在α1-β2-βγ、α2-β3-γ、α3-β3-γ等3种不同的组合模式。这些研究结果,为后续高粱SnRK1亚基的功能解析等研究奠定了良好基础。

Abstract:

Sucrose nonfermenting-related protein kinase 1 (SnRK1) is one of the critical serine/threonine protein kinases.  It commonly mediates plant growth and development, cross-talks with metabolism processes and physiological responses to biotic or abiotic stresses.  It plays a key role in distributing carbohydrates and sugar signal transporting.  In the present study, eight SnRK1 coding genes were identified in sorghum (Sorghum bicolor L.) via sequences alignment, with three for α subunits (SnRK1α1 to SnRK1α3), three for β (SnRK1β1 to SnRK1β3), and one for both γ (SnRK1γ) and βγ (SnRK1βγ).  These eight corresponding genes located on five chromosomes (Chr) of Chr1–3, Chr7, and Chr9 and presented collinearities to SnRK1s from maize and rice, exhibiting highly conserved domains within the same subunits from the three kinds of cereals.  Expression results via qRT-PCR showed that different coding genes of SnRK1s in sorghum possessed similar expression patterns except for SnRK1α3 with a low expression level in grains and SnRK1β2 with a relatively high expression level in inflorescences.  Results of subcellular localization in sorghum leaf protoplast showed that SnRK1α1/α2/α3/γ mainly located on organelles, while the rest four of SnRK1β1/β2/β3/βγ located on both membranes and some organelles.  Besides, three combinations were discovered among eight SnRK1 subunits in sorghum through yeast two hybrid, including α1-β2-βγ, α2-β3-γ, and α3-β3-γ.  These results provide informative references for the following functional dissection of SnRK1 subunits in sorghum.

Key words: sorghum (Sorghum bicolor L.) , SnRK1 , expression analysis , combination pattern