干旱胁迫下柠条锦鸡儿叶片SSH文库构建及CkWRKY1基因克隆

doi:10.11707/j.1001-7488.20130709

Abstract

Abstract:

Caragana korshinskii is an important cultivated shrub with high ecological and forage value, and widely distributes in Northwest China. C. korshinskii has a strong tolerance to drought, cold, heat, saline and poor soil. In order to study its resistance mechanisms to drought and explore for drought response related genes, a suppression subtractive hybridization (SSH) library of C. korshinskii under drought stress was constructed. Totally 1 286 ESTs from the SSH library were obtained and the average length was 475 bp. As many as 645 Unigenes were identified through the EST sequence assembly. Many drought-related genes were annotated by Blasting, such as the genes encoding dehydration related LEA and DHN proteins as well as transcription factors including MYB, bZIP, WRKY, NAC, and bHLH families. A candidate gene encoding a member of WRKY transcription factor family was cloned by rapid amplification of cDNA ends technique, and was named as CkWRKY 1 (GenBank accession No. JX987095). The protein deduced from the cDNA was composed from 330 amino acids with a conserved region "WRKYGQK". Real-time-quantitative PCR analysis showed that drought stress enhanced CkWRKY 1 expression at the transcriptional level, indicating that CkWRKY 1 might be involved in the drought resistance mechanisms of C. korshinskii at molecular level.

Key words: Caragana korshinskii, drought stress, suppression subtractive hybridization (SSH) library, WRKY, gene cloning

CLC Number:

S718.46

Yang Qi, Zhang Tao, Wang Ying, Li Gao, Yin Jiajia, Han Xiaomin, Qi Liwang, Li Guojing, Wang Ruigang. Construction of a Suppression Subtractive Hybridization Library of Caragana korshinskii Under Drought Stress and Cloning of CkWRKY 1 Gene[J]. Scientia Silvae Sinicae, 2013, 49(7): 62-68.

References

冯健,齐力旺,孙晓梅,等.2010. 落叶松扦插生根过程 SSH 文库构建及部分基因的表达分析.林业科学,46(6): 27-34.

郭九峰,孙国琴,乔慧蕾. 2012. 水分胁迫柠条锦鸡儿叶片均一化全长cDNA文库的构建及EST 分析. 华北农学报, 27(3): 67-71.

李冉,娄永根. 2011. 植物中逆境反应相关的 WRKY 转录因子研究进展. 生态学报,31(11): 3223-3231.

李彦瑾,赵忠,孙德祥,等. 2008.干旱胁迫下柠条锦鸡儿的水分生理特征.西北林学院学报, 23(3): 1-4.

牛西午. 2003. 柠条研究. 北京:科学出版社.

秦伟,赵光耀,曲志才,等. 2010. 小麦白粉病菌诱导的 TaWRKY 34 基因的鉴定与分析. 作物学报,36(2): 249-255.

王美珍, 王学敏, 王赞,等. 2009. 柠条GDP-甘露糖-3',5'-异构酶基因的克隆、序列分析及其植物表达载体的构建. 中国草地学报, 31(5): 75-83.

王文芳.2010. 冷诱导下柠条锦鸡儿SSH文库的构建. 呼和浩特: 内蒙古农业大学硕士学位论文.

王学敏, 高洪文. 2010. 柠条锦鸡儿 NCED4 基因的克隆及对逆境胁迫的响应. 植物分子生物学与现代农业——全国植物生物学研讨会论文摘要集.天津: 中国遗传学会.

王志会,夏新莉,尹伟伦. 2007. 不同种源的柠条锦鸡儿的生理特性与抗旱性.东北林业大学学报,35(9): 27-29.

徐当会,方向文,宾振钧. 2012. 柠条适应极端干旱的生理生态机制——叶片脱落和枝条中叶绿体保持完整性. 中国沙漠,32(3):691-697.

杨洪晓,王学全,卢琦,等.2010. 行带式柠条锦鸡儿林在内蒙古四子王旗退耕还草工程中的应用.林业科学, 46(11): 36-42.

杨文斌,任建民.1997.柠条抗旱的生理生态与土壤水分关系的研究.生态学报, 17(3): 239-244.

Guo Zejian, Kan Yunchao, Chen Xujun, et al. 2004. Characterization of a rice WRKY gene whose expression is induced upon pathogen attack and mechanical wounding. Acta Botanica Sinica, 46(8): 955-964.

Hirao T, Fukatsu E, Watanabe A. 2012. Characterization of resistance to pine wood nematode infection in Pinus thunbergii using suppression subtractive hybridization. BMC Plant Biology, 12(1):13

Huang T, Duman J G. 2002.Cloning and characterization of a thermal hysteresis (antifreeze) protein with DNA-binding activity from winter bittersweet nightshade, Solanum dulcamara. Plant Molecular Biology,48(4): 339-350.

Leelatanawit R, Klinbunga S, Aoki T, et al. 2008. Suppression subtractive hybridization (SSH) for isolation and characterization of genes related to testicular development in the giant tiger shrimp Penaeus monodon. BMB Rep, 41(11): 796-802.

Norelli J L, Farrell Jr R E, Bassett C L, et al. 2009. Rapid transcriptional response of apple to fire blight disease revealed by cDNA suppression subtractive hybridization analysis. Tree Genetics & Genomes, 5(1): 27-40.

Pnueli L, Hallak-Herr E, Rozenberg M, et al. 2002. Molecular and biochemical mechanisms associated with dormancy and drought tolerance in the desert legume Retama raetam. The Plant Journal, 31(3): 319-330.

Ramamoorthy R, Jiang S Y, Kumar N, et al. 2008. A comprehensive transcriptional profiling of the WRKY gene family in rice under various abiotic and phytohormone treatments. Plant and Cell Physiology, 49(6): 865-879.

Wang X, Chen X, Liu Y, et al. 2010. CkDREB gene in Caragana korshinskii is involved in the regulation of stress response to multiple abiotic stresses as an AP2/EREBP transcription factor. Molecular Biology Reports, 38(4): 2801-2811.

Zhou Q Y, Tian A G, Zou H F, et al. 2008. Soybean WRKY-type transcription factor genes, GmWRKY 13 , GmWRKY 21 , and GmWRKY 54 , confer differential tolerance to abiotic stresses in transgenic Arabidopsis plants. Plant Biotechnology Journal, 6(5): 486-503.

[1]	Wang Yilin, Wang Weifeng, Zhang Yunxiang, Chang Shujun, Guo Jinping. Responses of Leaf Morphological Structure and Physiological Characteristics of Populus euramericana cv. ‘BYu’ to Drought Stress [J]. Scientia Silvae Sinicae, 2019, 55(4): 42-50.
[2]	Jiang Cheng, Zhou Houjun, Zhao Yanqiu, He Hui, Chu Liwei, Song Xueqin, Lu Mengzhu. Effects of CDD Gene on the Growth and Development of Populus alba×P. glandulosa ‘84K’ in Response to Drought and Salt Stresses [J]. Scientia Silvae Sinicae, 2019, 55(2): 33-40.
[3]	Fu Bing, Ye Xia, Wang Huiyu, Chen Peng, Li Jidong, Zheng Xianbo, Tan Bin, Feng Jiancan. Identification of WRKY Transcription Factors in Jujube and Their Responses to ‘Candidatus Phytoplasma ziziphi’ and Salicylic Acid or Methyl Jasmonate Treatments [J]. Scientia Silvae Sinicae, 2018, 54(8): 65-78.
[4]	Ren Jian, Yin Yuqing, Zhang Huihui, Chen Dian, Wang Kexin, Wang Yong. Cloning and Analysis of AmCesA1 Gene in Acacia mangium [J]. Scientia Silvae Sinicae, 2018, 54(8): 79-87.
[5]	Zhang Jiangtao, Yang Shuhong, Zhu Di, Zhu Yanlin, Liu Youquan. Physiological Response of Annual Grafted Seedlings of Poplar 2025 and Its Two Bud Mutation Varieties to Drought Stress and Evaluation of Drought Resistance [J]. Scientia Silvae Sinicae, 2018, 54(6): 33-43.
[6]	Zhang Qin, Xu Zongda, Zhao Kai, Li Xiaowei, Zhang Luosha, Zhang Qixiang. Isolation and Biological Function Analysis of Anthocyanin Regulatory Gene PmMYB1 from Prunus mume [J]. Scientia Silvae Sinicae, 2018, 54(10): 64-72.
[7]	Du Mingfeng, Ding Guijie, Zhao Xizhou. Responses to Continuous Drought Stress and Drought Resistance of Different Masson Pine Families [J]. Scientia Silvae Sinicae, 2017, 53(6): 21-29.
[8]	Huang Juan, Chen Cun, Zhang Weixi, Ding Changjun, Su Xiaohua, Huang Qinjun. Effects of Drought Stress on Anatomical Structure and Photosynthetic Characteristics of Transgenic JERF36 Populus alba×P. berolinensis Seedling Leaves [J]. Scientia Silvae Sinicae, 2017, 53(5): 8-15.
[9]	You Hanli, Yuan Yihang, Li Changjiang, Zhang Lingyun. Cloning and Expression Analysis of MYB Homologous Gene PwMYB20 from Picea wilsonii [J]. Scientia Silvae Sinicae, 2017, 53(5): 23-32.
[10]	Quan Wenxuan, Ding Guijie. Dynamic of Volatiles and Endogenous Hormones in Pinus massoniana Needles under Drought Stress [J]. Scientia Silvae Sinicae, 2017, 53(4): 49-55.
[11]	Luo Qinghong, Ning Husen, He Miao, Ji Xiaomin, Lei Chunying. Ecophysiological Responses of Five Sandy Shrubs to Drought Stress [J]. Scientia Silvae Sinicae, 2017, 53(11): 29-42.
[12]	Chu Wenyuan, Wang Yujiao, Zhu Dongyue, Chen Zhu, Yan Hanwei, Xiang Yan. Selection of Novel Reference Genes in Poplar under Salt and Drought Stresses [J]. Scientia Silvae Sinicae, 2017, 53(10): 70-79.
[13]	Liu Fangchun, Ma Hailin, Ma Bingyao, Du Zhenyu, Jing Dawei, Xing Shangjun. Effect of Plant Growth-Promoting Rhizobacteria on Photosynthetic Characteristics in Walnut Seedlings under Drought Stress [J]. Scientia Silvae Sinicae, 2015, 51(7): 84-90.
[14]	Luo Qunfeng, Xu Meng, Feng Yuanheng, Xu Jiajuan, Li Huogen. Cloning and Bioinformatics of Chalcone Synthase Gene (CHS) in Liriodendron tulipifera and Characterization of Its Tissue Expression [J]. Scientia Silvae Sinicae, 2015, 51(5): 37-45.
[15]	Wang Jianyong, Tan Xiaofeng, Chen Hongpeng, Zeng Yanling, Long Hongxu, Mei Fangfang, Liu Kai. Cloning and Prokaryotic Expression of a Malonyl-CoA:ACP Transacylase Gene from Camellia oleifera [J]. Scientia Silvae Sinicae, 2015, 51(3): 148-154.

Construction of a Suppression Subtractive Hybridization Library of Caragana korshinskii Under Drought Stress and Cloning of CkWRKY 1 Gene

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments