Cloning and Analysis of Protein Kinase OSK4 Gene of Kentucky Bluegrass and Its Response to Abiotic Stress

doi:10.11733/j.issn.1007-0435.2022.02.010

Acta Agrestia Sinica ›› 2022, Vol. 30 ›› Issue (2): 339-347.DOI: 10.11733/j.issn.1007-0435.2022.02.010

Cloning and Analysis of Protein Kinase OSK4 Gene of Kentucky Bluegrass and Its Response to Abiotic Stress

CHEN Yang^1,2, GAO Yan-song^1,2, LI Hong-ying³, ZHAO Qing-feng¹, CAO Ye-ping¹, DI Hao-yang¹, JIN Yi-feng^1,2

1. College of Life Science and Agro-Forestry, Qiqihar Univesity, Qiqihar, Heilongjiang Province 161006, China;
2. Heilongjiang Province Key Laboratory of Resistance Gene Engineering and Preservation of Biodiversity in Cold Areas, Qiqihar, Heilongjiang Province 161006, China;
3. School of science, Qiqihar University, Qiqihar, Heilongjiang Province 161006, China

Received:2021-07-15 Revised:2021-09-05 Published:2022-03-10

草地早熟禾蛋白激酶OSK4基因的克隆及对非生物胁迫响应分析

陈阳^1,2, 高岩松^1,2, 李洪影³, 赵清峰¹, 曹业萍¹, 邸浩洋¹, 金一锋^1,2

1. 齐齐哈尔大学生命科学与农林学院, 黑龙江齐齐哈尔 161006;
2. 抗性基因工程与寒地生物多样性保护黑龙江省重点实验室, 黑龙江齐齐哈尔 161006;
3. 齐齐哈尔大学理学院, 黑龙江齐齐哈尔 161006

通讯作者: 金一锋,E-mail:Jinyifeng8368215@126.com
作者简介:陈阳(1986-),女,黑龙江齐齐哈尔人,副教授,博士,主要从事草坪草逆境分子生物学研究,E-mail:Chenyang8368215@126.com
基金资助:
黑龙江省普通高等学校青年创新人才培养计划项目(UNPYSCT-2018102)；黑龙江省省属高等学校基本科研业务费科研项目(135309365)；齐齐哈尔市科学技术局科学技术计划项目(NYGG-201917)；齐齐哈尔大学大学生创新创业项目(202110232361、202110232371、202110232392)共同资助

Abstract

Abstract: In this study,the OSK4 gene of the sucrose non-fermenting-1-related protein kinase 1b (SnRK1b) subfamily of Poa pratensis L. was cloned and bioinformatically analyzed. The gene expression of different tissues was analyzed under drought,nitrogen and phosphorus stresses. The results showed that the OSK4 gene of Poa pratensis L. contained typical STKc_ AMPK_alpha,UBA_ SnRK1_plant and AMPKA_C domains,wiht a homologous similarity of 94.5% to Brachypodium distachyon. Real-time fluorescence quantitative PCR demonstrated a tissue-specificity expression of the OSK4 gene,with that of root>stem>spike>leaf. It was also found that drought stress significantly inhibited the expression of OSK4 gene in roots and leaves. The relative expression of OSK4 gene in Poa pratensis L was up-regulated with the increase of NaNO₃ concentration,while low-phosphorus stress (0.01 mM KH₂PO₄) could promote the expression of OSK4 gene. Overall,this study may offer valuable references for future research on the regulatory mechanisms of the SnRK1b subfamily in Poa pratensis L under abiotic stress as well as on the stress-resistant mechanisms of SnRK1 in turfgrass.

Key words: Kentucky Bluegrass, Protein Kinase, SnRK1, OSK4, Expression Analysis

摘要： 本研究以草地早熟禾(Poa pratensis L)为试材，对蔗糖非发酵1相关蛋白激酶SnRK1b亚家族OSK4基因进行克隆、生物信息学分析，并分析不同组织部位以及干旱、氮素、磷素胁迫下该基因表达情况。研究结果：草地早熟禾OSK4基因包含典型的STKc_ AMPK_alpha,UBA_SnRK1_plant，AMPKA_C结构域，与二穗短柄草(Brachypodium distachyum (L.) Beauv.)同源相似度为94.5%；实时荧光定量PCR结果表明，OSK4基因在组织部位中表达水平存在组织特异性，OSK4基因相对表达量高低顺序为根部>茎部>穗>叶部；干旱胁迫显著抑制根部、叶部OSK4基因的表达水平；草地早熟禾OSK4基因相对表达量随着NaNO₃浓度的增加而上调；低磷环境(0.01 mM KH₂PO₄)可促进OSK4基因的表达。研究结果为探究草地早熟禾SnRK1b亚家族在非生物胁迫下的调节机制，为丰富草坪草SnRK1抗逆机制提供研究基础。

关键词: 草地早熟禾, 蛋白激酶, SnRK1, OSK4, 表达分析

CLC Number:

Q945.78

CHEN Yang, GAO Yan-song, LI Hong-ying, ZHAO Qing-feng, CAO Ye-ping, DI Hao-yang, JIN Yi-feng. Cloning and Analysis of Protein Kinase OSK4 Gene of Kentucky Bluegrass and Its Response to Abiotic Stress[J]. Acta Agrestia Sinica, 2022, 30(2): 339-347.

陈阳, 高岩松, 李洪影, 赵清峰, 曹业萍, 邸浩洋, 金一锋. 草地早熟禾蛋白激酶OSK4基因的克隆及对非生物胁迫响应分析[J]. 草地学报, 2022, 30(2): 339-347.

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Cloning and Analysis of Protein Kinase OSK4 Gene of Kentucky Bluegrass and Its Response to Abiotic Stress

草地早熟禾蛋白激酶OSK4基因的克隆及对非生物胁迫响应分析

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