海雀稗PvCIPK9基因克隆及耐盐功能鉴定

doi:10.11733/j.issn.1007-0435.2023.10.005

草地学报 ›› 2023, Vol. 31 ›› Issue (10): 2938-2948.DOI: 10.11733/j.issn.1007-0435.2023.10.005

海雀稗PvCIPK9基因克隆及耐盐功能鉴定

宋志钢¹, 刘颖², 刘瑞¹, 卢少云¹

1. 华南农业大学生命科学学院, 广东广州 510642;
2. 青海大学农牧学院, 青海西宁 810016

收稿日期:2023-03-16 修回日期:2023-04-25 出版日期:2023-10-15 发布日期:2023-11-02
通讯作者: 卢少云,E-mail:turflab@scau.edu.cn
作者简介:宋志钢(1998-),男,汉族,河南许昌人,硕士研究生,主要从事草坪草种质资源与育种研究,E-mail:1375934198@qq.com
基金资助:
国家自然科学基金项目（32271765）资助

Cloning and Identification of Salt Tolerance Function of Paspalum vaginatum PvCIPK9

SONG Zhi-gang¹, LIU Ying², LIU Rui¹, LU Shao-yun¹

1. College of Life Science, South China Agricultural University, Guangzhou, Guangdong Province 510642, China;
2. College of Agriculture and Animal Husbandry, Qinghai University, Xining, Qinghai Province 810016, China

Received:2023-03-16 Revised:2023-04-25 Online:2023-10-15 Published:2023-11-02

摘要/Abstract

摘要： 盐生植物海雀稗（Paspalums vaginatium）是一种极其耐盐的多年生草本植物。本研究从海雀稗转录组数据中筛选对盐胁迫响应的PvCIPK9基因，克隆了PvCIPK9基因CDS序列全长，并对其表达模式、启动子元件等进行分析。为研究PvCIPK9基因的耐盐功能，构建了PvCIPK9过表达载体，采用农杆菌介导的花序侵染法转化拟南芥（Arabidopsis thaliana），获得过表达PvCIPK9的转基因拟南芥（Arabidopsis thaliana），对转基因拟南芥的耐盐性进行鉴定。结果表明，盐胁迫条件下，转基因拟南芥的生长表型显著优于野生型，转基因拟南芥在盐胁迫下的种子萌发率及相对生长量均显著高于野生型；此外，盐处理后转基因拟南芥的抗氧化酶活性也显著高于野生型，以上结果初步说明过表达PvCIPK9基因增强了拟南芥对盐胁迫的抵抗能力。本研究为深入解析海雀稗PvCIPK9在植物耐盐性中的作用奠定了基础。

关键词: 耐盐性, 海雀稗, 拟南芥, PvCIPK9

Abstract: The halophyte Paspalum vaginatum is a perennial herb with extremely salt tolerance. In this study,PvCIPK9 gene responsive to salt stress was screened from the transcriptome of Paspalum vaginatum,the full length of CDS sequence of PvCIPK9 gene was cloned,and its expression pattern and promoter elements were analyzed. In order to further study the salt stress resistance of PvCIPK9 gene,the overexpression vector of PvCIPK9 was constructed and transgenic Arabidopsis was obtained by agrobacterium-mediated inflorescence infection. Subsequently,salt tolerance of transgenic Arabidopsis was analyzed. The results showed that the growth phenotype of transgenic Arabidopsis was significantly better under salt stress compared to wild type,and the seed germination rate and relative growth yield of transgenic Arabidopsis were significantly higher under salt stress than that of wild type. In addition,the antioxidant enzyme activity of transgenic Arabidopsis was also significantly higher after salt treatment. The above results showed that overexpression of PvCIPK9 gene enhanced the resistance to salt stress in Arabidopsis. This study is helpful to further understanding the molecular function of PvCIPK9 in response to salt stress,and provides a theoretical basis for genetic improvement of crop salt tolerance.

Key words: Salt tolerance, Paspalum vaginatum, Arabidopsis thaliana, PvCIPK9

中图分类号:

Q945

宋志钢, 刘颖, 刘瑞, 卢少云. 海雀稗PvCIPK9基因克隆及耐盐功能鉴定[J]. 草地学报, 2023, 31(10): 2938-2948.

SONG Zhi-gang, LIU Ying, LIU Rui, LU Shao-yun. Cloning and Identification of Salt Tolerance Function of Paspalum vaginatum PvCIPK9[J]. Acta Agrestia Sinica, 2023, 31(10): 2938-2948.

0
/ / 推荐

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://manu40.magtech.com.cn/Jweb_cdxb/CN/10.11733/j.issn.1007-0435.2023.10.005

https://manu40.magtech.com.cn/Jweb_cdxb/CN/Y2023/V31/I10/2938

参考文献

[1] LUAN S,KUDLA J,RODRIGUEZ-CONCEPCION M,et al. Calmodulins and calcineurin B-like proteins[J]. The Plant Cell,2002,14(suppl 1):S389-S400
[2] KUDLA J,BATISTIC O,HASHIMOTO K. Calcium Signals:The lead currency of plant information processing[J]. The Plant Cell,2012,22(3):541-563
[3] DEFALCO T A,BENDER K W,SNEDDEN W A. Breaking the code:Ca2+ sensors in plant signalling[J]. Biochemical Journal,2010,425(1):27-40
[4] GONG D,GUO Y,SCHUMAKER K S,et al. The SOS3 family of calcium sensors and SOS2 family of protein kinases in Arabidopsis[J]. Plant Physiology,2004,134(3):919-926
[5] HASHIMOTO K,KUDLA J. Calcium decoding mechanisms in plants[J]. Biochimie,2011,93(12):2054-2059
[6] BATISTIC O,KUDLA J. Plant calcineurin B-like proteins and their interacting protein kinases[J]. Biochimica et Biophysica Acta (BBA)-Molecular Cell Research,2009,1793(6):985-992
[7] ALBRECHT V. The NAF domain defines a novel protein-protein interaction module conserved in Ca²⁺-regulated kinases[J]. The EMBO Journal,2001,20(5):1051-1063
[8] OHTA M,GUO Y,HALFTER U,et al. A novel domain in the protein kinase SOS2 mediates interaction with the protein phosphatase 2C ABI2[J]. The National Academy of Sciences of the USA,2003,100(20):11771-11776
[9] CHEN X,HUANG Q,ZHANG F,et al. ZmCIPK21,a maize CBL-interacting kinase,enhances salt stress tolerance in Arabidopsis thaliana[J]. International Journal of Molecular Sciences,2014,15(8):14819-14834
[10] SUN T,WANG Y,WANG M,et al. Identification and comprehensive analyses of the CBL and CIPK gene families in wheat (Triticum aestivum L.)[J]. BMC Plant Biology,2015,15(1):12870
[11] 李慧,路依萍,汪小凯,等. CBL互作蛋白激酶GmCIPK10增强大豆耐盐性[J]. 作物学报,2023,49(5):1272-1281
[12] HUANG C,DING S,ZHANG H,et al. CIPK7 is involved in cold response by interacting with CBL1 in Arabidopsis thaliana[J]. Plant Science,2011,181(1):57-64
[13] KIM K N,LEE J S,HAN H,et al. Isolation and characterization of a novel rice Ca²⁺-regulated protein kinase gene involved in responses to diverse signals including cold,light,cytokinins,sugars and salts[J]. Plant Mol Biol,2003,52(6):1191-1202
[14] DE LA TORRE F,GUTIERREZ-BELTRAN E,PAREJA-JAIME Y,et al. The tomato calcium sensor cbl10 and its interacting protein kinase cipk6 define a signaling pathway in plant immunity[J]. The Plant Cell,2013,25(7):2748-2764
[15] KURUSU T,HAMADA J,NOKAJIMA H,et al. Regulation of microbe-associated molecular pattern-induced hypersensitive cell death,phytoalexin production,and defense gene expression by calcineurin B-like protein-interacting protein kinases,OsCIPK14/15,in rice cultured cells[J]. Plant Physiology,2010,153(2):678-692
[16] RAO X,ZHANG X,LI R,et al. A calcium sensor-interacting protein kinase negatively regulates salt stress tolerance in rice (Oryza sativa)[J]. Functional Plant Biology,2011,38(6):441
[17] LEE G,CARROW R N,DUNCAN R R. Photosynthetic responses to salinity stress of halophytic seashore paspalum ecotypes[J]. Plant Science,2004,166(6):1417-1425
[18] 解新明,卢小良. 海雀稗种质资源的优良特性及其利用价值[J]. 华南农业大学学报,2004(S2):64-67
[19] 宗俊勤,高艳芝,陈静波,等. 淹水胁迫对4种暖季型草坪草光合特性的影响[J]. 热带作物学报,2021,42(1):130-139
[20] 刘一明,程凤枝,王齐,等. 四种暖季型草坪植物的盐胁迫反应及其耐盐阈值[J]. 草业学报,2009,18(3):192-199
[21] 吴雪莉,郭振飞,陈申秒,等. 海滨雀稗耐逆性研究进展[J]. 草地学报,2019,27(5):1117-1125
[22] ZHANG Y,SU J,DUAN S,et al. A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes[J]. Plant Methods,2011,7(1):30
[23] CHEN J,GUO Z,FANG J,et al. Physiological responses of a centipedegrass mutant to chilling stress[J]. Agronomy Journal,2013,105(6):1814-1820
[24] ZHUO C,LIANG L,ZHAO Y,et al. A cold responsive ethylene responsive factor from Medicago falcata confers cold tolerance by up-regulation of polyamine turnover,antioxidant protection,and proline accumulation[J]. Plant,Cell & Environment,2018,41(9):2021-2032
[25] BRADFORD M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Analytical Biochemistry,1976,1-2(72):248-254
[26] TANG R J,WANG C,LI K,et al. The CBL-CIPK calcium signaling network:Unified paradigm from 20 years of discoveries[J]. Trends in Plant Science,2020,25(6):604-617
[27] PANDEY G K,CHEONG Y H,KIM B G,et al. CIPK9:a calcium sensor-interacting protein kinase required for low-potassium tolerance in Arabidopsis[J]. Cell Research,2007,17(5):411-421
[28] 周三,韩军丽,赵可夫. 泌盐盐生植物研究进展[J]. 应用与环境生物学报,2001,7(5):496-501
[29] CHEN X,GU Z,FENG L I U,et al. Molecular analysis of rice CIPKs involved in both biotic and abiotic stress responses[J]. Rice Science,2011,18(1):1-9
[30] 贾会丽,王学敏,高涛,等. 转MsLEA4-4基因拟南芥株系的耐盐性分析[J]. 草地学报,2020,28(3):597-605
[31] 徐金龙,张文静,向凤宁. 植物盐胁迫诱导启动子及其顺式作用元件研究进展[J]. 植物生理学报,2021,57(4):759-766
[32] BATISTIC O,WAADT R,STEINHORST L,et al. CBL-mediated targeting of CIPKs facilitates the decoding of calcium signals emanating from distinct cellular stores[J]. The Plant Journal,2010,61(2):211-222
[33] WAADT R,SCHMIDT L K,LOHSE M,et al. Multicolor bimolecular fluorescence complementation reveals simultaneous formation of alternative CBL/CIPK complexesin planta[J]. The Plant Journal,2008,56(3):505-516
[34] LU L,CHEN X,ZHU L,et al. NtCIPK9:a calcineurin B-like protein-interacting protein kinase from the halophyte Nitraria tangutorum,enhances Arabidopsis salt tolerance[J]. Frontiers in Plant Science,2020(11):11-12
[35] KANWAR P,SANYAL S K,MAHIWAL S,et al. CIPK9 targets VDAC3 and modulates oxidative stress responses in Arabidopsis[J]. The Plant Journal,2022,(109):241-260
[36] WANG N,TAO B,MAI J,et al. Kinase CIPK9 integrates glucose and abscisic acid signaling to regulate seed oil metabolism in rapeseed[J]. Plant Physiology,2023,191(3):1836-1856
[37] GARCÍA G,CLEMENTE-MORENO M J,DÍAZ-VIVANCOS P,et al. The apoplastic and symplastic antioxidant system in onion:Response to long-term salt stress[J]. Antioxidants,2020,9(1):67
[38] HERNÁNDEZ-HERNÁNDEZ H,JUÁREZ-MALDONADO A,BENAVIDES-MENDOZA A,et al. Chitosan-PVA and copper nanoparticles improve growth and overexpress the SOD and JA genes in tomato plants under salt stress[J]. Agronomy,2018,8(9):175
[39] NALIWAJSKI M,SKLODOWSKA M. The relationship between the antioxidant system and proline metabolism in the leaves of cucumber plants acclimated to salt stress[J]. Cells,2021,10(3):609
[40] QIU Q S,GUO Y,DIETRICH M A,et al. Regulation of SOS1,a plasma membrane Na+/H+ exchanger in Arabidopsis thaliana,by SOS2 and SOS3[J]. The National Academy of Sciences of the USA,2002,99(12):8436-8441
[41] ZHU J. Regulation of ion homeostasis under salt stress[J]. Current Opinion in Plant Biology,2003,6:441-445
[42] LIU L,REN H,CHEN L,et al. A protein kinase,calcineurin B-like protein-interacting protein kinase 9,interacts with calcium sensor calcineurin B-like protein3 and regulates potassium homeostasis under low-potassium stress in Arabidopsis[J]. Plant Physiology,2013,161(1):266-277
[43] LARA A,RÓDENAS R,ANDRÉS Z,et al. Arabidopsis K⁺ transporter HAK5-mediated high-affinity root K+ uptake is regulated by protein kinases CIPK1 and CIPK9[J]. Journal of Experimental Botany,2020,71(16):5053-5060
[44] SINGH A,YADAV A K,KAUR K,et al. A protein phosphatase 2C,AP2C1,interacts with and negatively regulates the function of CIPK9 under potassium-deficient conditions in Arabidopsis[J]. Journal of Experimental Botany,2018,69(16):4003-4015

海雀稗PvCIPK9基因克隆及耐盐功能鉴定

Cloning and Identification of Salt Tolerance Function of Paspalum vaginatum PvCIPK9

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	叶文兴, 张胜男, 刘雪婷. 12个草地早熟禾品种幼苗耐盐性分析[J]. 草地学报, 2023, 31(9): 2740-2747.
[2]	张立霞, 李霄, 许蕾, 旦真措, 刘亚娇, 苏德毕力格, 丛丽丽, 杨青川, 龙瑞才. 蒺藜苜蓿miR167c调控生长和花器官发育的功能验证[J]. 草地学报, 2023, 31(10): 2925-2937.
[3]	任智新, 史建楠, 何佳星, 王晔, 范希峰, 李润枝, 于春欣, 彭真, 高月荣, 段留生. 盐胁迫对青绿苔草生长及生理特性的影响[J]. 草地学报, 2022, 30(8): 2045-2052.
[4]	余如刚, 王雪茄, 王国良, 徐智明, 高晴晴, 杜雪玲, 张宇龙. 紫花苜蓿品种苗期耐盐性分析及评价指标筛选[J]. 草地学报, 2022, 30(7): 1781-1789.
[5]	张磊, 王玉祥, 李瑞强, 楼明祝, 陈映霞, 张志杰, 张博. 盐水连续浇灌下107份黄花苜蓿苗期耐盐性评价[J]. 草地学报, 2022, 30(12): 3317-3325.
[6]	李娟霞, 白小明, 张翠, 刘娅娜, 冉福, 李萍, 陈辉. 7个野生一年生早熟禾种质萌发期耐盐性综合评价[J]. 草地学报, 2022, 30(11): 2937-2948.
[7]	王玉祥, 汪鹏, 李姣, 李倩, 张博. 盐水浇灌下六个三叶草的耐盐性评价[J]. 草地学报, 2022, 30(11): 2996-3008.
[8]	陈筱冉, 张铭笑, 严建萍, 刘燕蓉, 张万军. 紫花苜蓿MsCCD4基因克隆及耐盐功能鉴定[J]. 草地学报, 2022, 30(10): 2572-2580.
[9]	贾会丽, 王学敏, 高涛, 石永红, 王运琦, 吴欣明, 刘建宁, 方志红, 董宽虎. 转MsLEA4-4基因拟南芥株系的耐盐性分析[J]. 草地学报, 2020, 28(3): 597-605.
[10]	李雪, 沙栢平, 高雪芹, 王焱, 伏兵哲. 不同紫花苜蓿种质材料萌发期耐盐性鉴定与综合评价[J]. 草地学报, 2020, 28(2): 437-445.
[11]	张则宇, 李雪, 王焱, 常巍, 高雪芹, 伏兵哲. 59份苜蓿种质材料苗期耐盐性评价及耐盐指标筛选[J]. 草地学报, 2020, 28(1): 112-121.
[12]	吴雪莉, 郭振飞, 陈申秒, 庄黎丽. 海滨雀稗耐逆性研究进展[J]. 草地学报, 2019, 27(5): 1117-1125.
[13]	张燕, 朱慧森, 白永超, 黄志超, 胡婧, 董宽虎, 李存福. 3个居群野生草地早熟禾耐盐性比较研究[J]. 草地学报, 2018, 26(5): 1215-1222.
[14]	张寒, 潘香逾, 王秀华, 李家丽, 姜慧新, 赵岩. 苜蓿萌发期耐盐性综合评价与耐盐种质筛选[J]. 草地学报, 2018, 26(3): 666-672.
[15]	郭童天, 刘博文, 汤学友, 张雄, 孙娈姿. 干旱预处理对一年生黑麦草耐盐性的影响[J]. 草地学报, 2018, 26(2): 435-440.