| 摘要: |
| ‘东农冬麦1号（Dn1）’是首个能在严寒地带大面积种植、返青率超过85%的强抗寒冬小麦品种，为探索冬小麦中REVEILLE 7-like基因的功能，以及低温胁迫下外源油菜素内酯（BR）对该基因表达的影响。本研究利用RT-PCR法克隆了冬小麦（Dn1）REVEILLE 7-like基因的cDNA序列，并对其进行了生物信息学分析，冬小麦三叶期时叶面喷施BR，并于4 ℃、0 ℃、-10 ℃、-25 ℃对冬小麦叶片和分蘖节进行取样，用qRT-PCR技术分析该基因在以上不同处理组冬小麦叶片和分蘖节中的表达模式。结果表明，（1）REVEILLES属于MYB类转录因子家族，REVEILLE 7-like基因全长1251 bp，共编码416 个氨基酸，编码蛋白为不稳定亲水性蛋白，主要定位于细胞核；（2）REVEILLE 7-like与节节麦、二粒小麦亲缘关系最近，蛋白序列总相似度为78.62%；（3）REVEILLE 7-like基因启动子包含参与逆境响应、激素响应及MYB结合位点的顺式作用元件；（4）表达模式分析发现，对照组中REVEILLE 7-like基因的表达量在0 ℃、-10 ℃、-25 ℃时与4 ℃时相比较均有显著提高。处理组与对照组相比，该基因的表达量在-10 ℃、-25 ℃时均显著提高，推测该基因在冬小麦受到低温胁迫时起重要作用，且BR可以提高该基因的表达，进一步提高冬小麦的耐寒性。 |
| 关键词: 寒地冬小麦，油菜素内酯，REVEILLE 7-like，基因克隆，低温胁迫 |
| DOI：10.11931/guihaia.gxzw202407003 |
| 分类号:Q945.3；S512.1 ???????????? |
| 基金项目:黑龙江省省属本科高校基本科研业务费(145309512)； 齐齐哈尔大学研究生创新科研项目(QUZLTS_CX2023022) |
|
| Effects of exogenous Brassinosteroids on REVEILLE 7-like gene expression in winter wheat under low temperature stress |
|
CHEN Yushu1, ZHANG Junbao1, WANG Xuesong1, SHAO Qingyi1, YANG Sen1, CAO Jiaang1, LIU Lijie1,2*
|
|
1. Collegel of Life Sciences and Agriculture and Forestry, Qiqihar University, Qiqihar, Heilongjiang 161006, China; 2. Heilongjiang Provincial Collaborative Innovation Center of Agrobiological Preparation Industrialization, Qiqihar, Heilongjiang 161006, China
|
| Abstract: |
| Dongnong Dongmai No.1 (Dn1) is the first strong cold-resistant variety that can be planted in a large area in the cold zone, and the greening rate is over 85%. To explore the function of the REVEILLE 7-like gene in winter wheat and the effect of exogenous brassinolide (BR) on the expression of this gene under low temperature stress. In this study, the cDNA sequence of REVEILLE 7-like gene of winter wheat (Dn1) was cloned by RT-PCR, and its bioinformatics analysis was carried out. At the three-leaf stage of winter wheat, BR was sprayed on the leaves, and the leaves and tillers of winter wheat were sampled at 4℃, 0℃, -10℃ and -25℃. The expression patterns of this gene in the leaves and tillers of winter wheat in the above different treatment groups were analyzed by qRT-PCR. The results showed that, (1)REVEILLES belongs to MYB transcription factor family, and the full length of REVEILLE 7-like gene is 1251bp, which encodes 416 amino acids. The encoded protein is an unstable hydrophilic protein, mainly located in the nucleus. (2)REVEILLE 7-like has the closest genetic relationship with Aegilops tauschii and Eremmer wheat, and the total similarity of protein sequence is 78.62%. (3)REVEILLE 7-like gene promoter contains cis-acting elements involved in adversity response, hormone response and MYB binding site. (4)The expression pattern analysis showed that the expression of REVEILLE 7-like gene in the control group was significantly higher at 0℃, -10℃ and -25℃ than at 4℃. At -10℃ and -25℃, the expression of this gene in BR treatment group was significantly higher than that in control group, which suggested that this gene played an important role in winter wheat under low temperature stress, and BR could improve the expression of this gene and further improve the cold tolerance of winter wheat. |
| Key words: winter wheat, brassinolide, REVEILLE 7-like, gene cloning, low temperature stress |
