Response to Low Temperature Stress and Screening and Validation of Interaction Protein of CqCHP1 in Chieh-qua

doi:10.16420/j.issn.0513-353x.2022-0854

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (11): 2376-2386.doi: 10.16420/j.issn.0513-353x.2022-0854

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Response to Low Temperature Stress and Screening and Validation of Interaction Protein of CqCHP1 in Chieh-qua

WANG Min, YANG Songguang, LIU Wei, HE Xiaoming, SHI Shaoqi, LIU Wenrui, CHEN Lin, JIANG Biao, PENG Qingwu^*()

Guangdong Key Laboratory for New Technology Research of Vegetables，Vegetable Research Institute，Guangdong Academy of Agricultural Sciences，Guangzhou 510640，China

Received:2023-03-21 Revised:2023-09-18 Online:2023-11-25 Published:2023-11-28
Contact: PENG Qingwu

Abstract

Abstract:

In this study，CqCHP1 was cloned from‘Yueguang Chieh-qua’，which encoded cysteine/ histidine-rich C1 domain protein. The gene’s open reading frame was 816 bp with 271 amino acids. The secondary structure indicated that CqCHP1 contained main random curl and 23 serine phosphorylation sites. Phylogenetic tree and amino acid sequence alignment analysis showed that CqCHP1 had the highest homology with cucumber Csa_4G297435. CqCHP1 was expressed in different tissues and specifically induced by low temperature. And at seedling and fruiting stages，CqCHP1 showed higher expression in resistance materials than sensitive materials，while at vegetative growth and flowering periods，it’s the opposite. The protein was located in the nucleus. Analysis of interaction proteins obtained by yeast screening showed that CqCHP1 could interact with bHLH92 in nucleus. And the inteaction was verified by Bimolecular fluorescence complementation（BIFC）and Luciferase complementation imaging assay（LCI）.

Key words: chieh-qua, low temperature, CHP1 protein, protein-interation, bHLH92 transcription factor

WANG Min, YANG Songguang, LIU Wei, HE Xiaoming, SHI Shaoqi, LIU Wenrui, CHEN Lin, JIANG Biao, PENG Qingwu. Response to Low Temperature Stress and Screening and Validation of Interaction Protein of CqCHP1 in Chieh-qua[J]. Acta Horticulturae Sinica, 2023, 50(11): 2376-2386.

Figures/Tables 9

References 31

[1]	Bai L, Liu Y M, Mu Y, Anwar A, He C X, Yan Y, Li Y S, Yu X C. 2018. Heterotrimeric G-protein γ subunit CsGG3.2 positively regulates the expression of CBF genes and chilling tolerance in cucumber. Frontiers in Plant Science, 9:488. doi: 10.3389/fpls.2018.00488 URL
[2]	Chen Feng, Yan Jin-qiang, Li Mei-lan, Jiang Biao. 2021. Construction and identification of a homogenized Yeast Two-hybrid cDNA Library in Benincasa hispida. Guangdong Agricultural Sciences, 48 (8):58-64. (in Chinese)
	陈凤, 闫晋强, 李梅兰, 江彪. 2021. 冬瓜均一化酵母双杂交cDNA文库的构建及鉴定. 广东农业科学, 48 (8):58-64.
[3]	Chen Qing-jun, Wang Yong-jian, Zhang Hai-ying, Wang Shao-hui, Zhang Fu-man. 2005. Research progress on low temperature and weak light tolerance of cucumber. China Vegetables,(5):31-34. (in Chinese)
	陈青君, 王永健, 张海英, 王绍辉, 张福墁. 2005. 黄瓜低温弱光耐受性研究进展. 中国蔬菜,(5):31-34.
[4]	Chinnusamy V. 2003. ICE1:a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes & Development, 17 (8):1043-1054. doi: 10.1101/gad.1077503 URL
[5]	Chinnusamy V, Zhu J H, Zhu J K. 2007. Cold stress regulation of gene expression in plants. Trends Plant Sci, 12 (10):444-451. doi: 10.1016/j.tplants.2007.07.002 pmid: 17855156
[6]	Feng H L, Ma N N, Meng X, Zhang A, Wang J R, Chai S, Meng Q W. 2013. A novel tomato MYCtype ICE1-like transcription factor,SlICE1a,confers cold,osmotic and salt tolerance in transgenic tobacco. Plant Physiol. Biochem, 73:309-320.
[7]	Feng Xiao-ming. 2011. Cloning and functional characterization of an apple bHLH gene MdCIbHLH1[Ph. D. Dissertation]. Tai’an: Shandong Agricultural University. (in Chinese)
	冯晓明. 2011. 苹果bHLH基因MdCIbHLH1克隆及功能分析[博士论文]. 泰安: 山东农业大学.
[8]	Guy C, Kaplan F, Kopka J, Selbig J, Hincha D K. 2010. Metabolomics of temperature stress. Physiologia Plantarum, 132 (2):220-235. doi: 10.1111/ppl.2008.132.issue-2 URL
[9]	He X M, Xie D S, Chen Q H, Peng Q W. 2007. Chieh-qua biotechnology:progress and prospects. Asia and Aust J of Sci Biotech, 1:19-22.
[10]	Hwang I S, Choi D S, Kim N H, Kim D S. 2014. The pepper cysteine/histidine-rich DC 1 domain protein CaDC1 binds both RNA and DNA and is required for plant cell death and defense response. New Phytol, 201 (2):518-530. doi: 10.1111/nph.12521 pmid: 24117868
[11]	James H H, Alexandra C N, Peter J P, Peter M B, Yasutomi N. 1997. Taxonomy and function of C 1 protein kinase C homology domains. Protein Science, 6 (2):477-480. doi: 10.1002/pro.v6:2 URL
[12]	Jiang Y Q, Yang B, Deyholos M K. 2009. Functional characterization of the Arabidopsis bHLH92 transcription factor in abiotic stress. Mol Genet Genomics, 282:503-516. doi: 10.1007/s00438-009-0481-3 URL
[13]	Kidokoro S, Shinozaki K, Yamaguchi-Shinozaki K. 2022. Transcriptional regulatory network of plant cold-stress responses. Trends in Plant Sciences, 27 (9):922-935. doi: 10.1016/j.tplants.2022.01.008 URL
[14]	Kozik E U, Wehner T C. 2008. A single dominant gene Ch for chilling resistance in cucumber seedlings. Journal of the American Society for Horticultural Science, 133 (2):225-227. doi: 10.21273/JASHS.133.2.225 URL
[15]	Li Qiong, Li Li-li, Hou Juan, Luo Ren-ren, Wang Rui-dan, Hu Jian-bin, Huang Song. 2022. Advances on mechanism of cucurbit crops in response to low-temperature stress. Acta Horticulturae Sinica, 49 (6):1382-1394. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-0575 URL
	李琼, 李丽丽, 侯娟, 罗忍忍, 王瑞丹, 胡建斌, 黄松. 2022. 瓜类作物响应低温胁迫机制的研究进展. 园艺学报, 49 (6):1382-1394. doi: 10.16420/j.issn.0513-353x.2021-0575 URL
[16]	Li Shuang, Shi Guang-li, Chen Shan-shan, Zhou Yan-li, Zhao Chun-bo, Li Sheng-nan. 2021. Cloning and sequence analysis of cucumber cold tolerance gene CsRAV2. Molecular Plant Breeding, 19 (21):7012-7017. (in Chinese)
	李爽, 石广丽, 陈姗姗, 周艳丽, 赵春波, 李胜男. 2021. 黄瓜抗寒基因CsRAV2的克隆与序列分析. 分子植物育种, 19 (21):7012-7017.
[17]	Maruyama K, Sakuma Y, Kasuga M, Ito Y, Seki M, Goda H, Shimada Y, Yoshida S, Shinozaki K, Yamaguchi-Shinozaki K. 2010. Identification of cold-inducible downstream genes of the Arabidopsis DREB1A/CBF 3 transcriptional factor using two microarray systems. The Plant Journal, 38 (6):982-993. doi: 10.1111/tpj.2004.38.issue-6 URL
[18]	Megha S, Basu U, Kav Nat N V. 2018. Regulation of low temperature stress in plants by microRNAs. Plant Cell Environ, 41 (1):1-15. doi: 10.1111/pce.v41.1 URL
[19]	MiuraI K, Shiba H, Ohta M, Kang S W, Sato A, Yuasa T, Lwaya-inoue M, Kamada H, Ezura H. 2012. SlICE1 encoding a MYC-type transcription factor controls cold tolerance in tomato, Solanum lycopersicum. Plant Biotechnol J, 29 (3):253-260.
[20]	Miura K, Furumoto T. 2013. Cold Signaling and cold response in plants,International Journal of Molecular Sciences, 14:5312-5337.
[21]	Newton A C. 1995. Protein kinase C:structure,function,and regulation. Journal of Biological Chemistry, 270 (48):28495-28498. doi: 10.1074/jbc.270.48.28495 pmid: 7499357
[22]	Sakamoto H. 2004. Arabidopsis Cys2/His2-Type zinc-finger proteins function as transcription repressors under drought,cold,and high-salinity stress conditions. Plant Physiology, 136 (1):2734-2746. doi: 10.1104/pp.104.046599 pmid: 15333755
[23]	Song Y, Zhang X Y, Li M Z, Yang H, Fu D Y, Lv J, Ding Y L, Gong Z Z, Shi Y T, Yang S H. 2021. The direct targets of CBFs:in cold stress response and beyond. Journal of Integrative Plant Biology, 63 (11):1874-1887. doi: 10.1111/jipb.v63.11 URL
[24]	Talanova V V, Titov A F, Topchieva I V, Malysheva I E. 2008. Effect of stress factors on expression of the gene encoding a CBF transcription factor in cucumber plants. Doklady Biological Sciences. MAIK Nauka/Interperiodica, 423 (1):419-421.
[25]	Toledo-ortiz G, Huq E, Quail P H. 2003. The Arabidopsis basic/helix-loop-helix transcription factor family. Plant Cell, 15 (8):1749-1770. doi: 10.1105/tpc.013839 pmid: 12897250
[26]	Wang D, Guo Y, Wu C, Yang G, Li Y, Zheng C. 2008. Genome-wide analysis of CCCH zinc finger family in Arabidopsis and rice. BMC Genomics 9:44. doi: 10.1186/1471-2164-9-44
[27]	Wang Hong-fei. 2014. Germplasm identification of tolerance to low temperature and QTL mapping for low temperture adapbility in cucumber[M. D. Dissertation]. Chinese Academy of Agricultural Sciences. (in Chinese)
	王红飞. 2014. 黄瓜种质资源低温耐受性的鉴定评价及 QTL 初步定位研究[硕士论文]. 北京: 中国农业科学院.
[28]	Wang Y J, Zhang Z J, He X J, Zhou H L, Wen Y X, Dai J X, Hang J S, Chen S Y. 2003. A rice transcription factor OsbHLH1 is involved in cold stress response. Theor Appl Genet, 107:1402-1409. doi: 10.1007/s00122-003-1378-x URL
[29]	Wei Xin, Wang Han-tao, Wei Heng-ling, Fu Xiao-kang, Ma Liang, Lu Jian-hua, Wang Sheng-fen, Yu Shu-xun, Lu Di-mian. 2020. Cloning and drought resistance analysis of GhWRKY33 in upland cotton. Scientia Agricultura Sinica, 53 (22):4537-4549. (in Chinese) doi: 10.3864/j.issn.0578-1752.2020.22.002
	魏鑫, 王寒涛, 魏恒玲, 付小康, 马亮, 芦建华, 王省芬, 喻树迅. 陆地棉. 2020. GhWRKY33的克隆及抗旱功能分析. 中国农业科学, 53 (22):4537-4549. doi: 10.3864/j.issn.0578-1752.2020.22.002
[30]	Xue R, Liu Y, Zheng Y, Wu Y J, Li X J, Pei F K, Ni J Z. 2012. Three-dimensional structure and mimetic-membrane association of consensus 11-amino-acid motif from soybean LEA3 protein. Peptide Science, 98 (1):59-66.
[31]	Zhao Yanqing, Du Jianchang, Wang Panqiao, Qin Xiaodong, Chne Jinfeng. 2019. Identification and expression analysis of NAC transcription factor gene family under low temperature in Cucumis sativus var.hardwickii. Acta Horticulturae Sinica, 46 (7):1303-1319. (in Chinese)
	赵艳青, 杜建厂, 王盼乔, 秦晓东, 陈劲枫. 2019. 哈氏黄瓜NAC转录因子的鉴定及低温表达分. 园艺学报, 46 (7):1303-1319. doi: 10.16420/j.issn.0513-353x.2018-0717

用途	名称	引物序列（5′-3′）
Use	Name	Primer sequence
基因克隆Gene cloning	CqCHP1-F	ATGGCGCTGGTACCCGGAAA
	CqCHP1-R	CTAAGCACCAACAGAACCGA
实时荧光定量PCR	CqCHP1-QF	ACATACTGCTCCGATTCCCC
Quantitative real-time PCR	CqCHP1-QR	TTGGGGTGAATATTGACTCC
亚细胞定位 Sublocation	1305-GFP-CqCHP1-F	CGAGCTGTACAGATCTATGGCGCTGGTACCCGGAAA
	1305-GFP-CqCHP1-R	GGCCGCTTTAAGATCTCTACTAAGCACCAACAGAACCGA
酵母双杂 Yeast two-hybrid	BD-CqCHP1-F	GAATTCCCGGGGATCCGTATGGCGCTGGTACCCGGAAA
	BD-CqCHP1-R	GCAGGTCGACGGATCCCTAAGCACCAACAGAACCGA
	AD-bHLH92-F	CATCGATACGGGATCCATGGATGATAGCTTTTCTGT
	AD-bHLH92-R	CGAGCTCGATGGATCCTTAGCTTTCAAATTCTGGAC
双分子荧光互补	CqCHP1-BIFC-N-F	CCCTAGGAACTAGTATGGCGCTGGTACCCGGAAA
Bimolecular fluorescence	CqCHP1-BIFC-N-R	GATCCTCCACTAGTCTAAGCACCAACAGAACCGA
complementation	bHLH92-BIFC-C-F	TACGTAGTCACGTGATGGATGATAGCTTTTCTGT
	bHLH92-BIFC-C-R	CCGGACGTCACGTGTTAGCTTTCAAATTCTGGAC
荧光素酶互补	bHLH92-nLUC-F	ACGGGGGACGAGCTCGGTACCATGGATGATAGCTTTTCTGTG
Luciferase complementation	bHLH92-nLUC-R	CGCGTACGAGATCTGGTCGACGCTTTCAAATTCTGGACCCACA
imaging assay	CqCHP1-cLUC-F	TACGCGTCCCGGGGCGGTACCATGGCGCTGGTACCCGGAAA
	CqCHP1-cLUC-R	ACGAAAGCTCTGCAGGTCGACTCAAGCACCAACAGAACCGAAAAT

用途	名称	引物序列（5′-3′）
Use	Name	Primer sequence
基因克隆Gene cloning	CqCHP1-F	ATGGCGCTGGTACCCGGAAA
	CqCHP1-R	CTAAGCACCAACAGAACCGA
实时荧光定量PCR	CqCHP1-QF	ACATACTGCTCCGATTCCCC
Quantitative real-time PCR	CqCHP1-QR	TTGGGGTGAATATTGACTCC
亚细胞定位 Sublocation	1305-GFP-CqCHP1-F	CGAGCTGTACAGATCTATGGCGCTGGTACCCGGAAA
	1305-GFP-CqCHP1-R	GGCCGCTTTAAGATCTCTACTAAGCACCAACAGAACCGA
酵母双杂 Yeast two-hybrid	BD-CqCHP1-F	GAATTCCCGGGGATCCGTATGGCGCTGGTACCCGGAAA
	BD-CqCHP1-R	GCAGGTCGACGGATCCCTAAGCACCAACAGAACCGA
	AD-bHLH92-F	CATCGATACGGGATCCATGGATGATAGCTTTTCTGT
	AD-bHLH92-R	CGAGCTCGATGGATCCTTAGCTTTCAAATTCTGGAC
双分子荧光互补	CqCHP1-BIFC-N-F	CCCTAGGAACTAGTATGGCGCTGGTACCCGGAAA
Bimolecular fluorescence	CqCHP1-BIFC-N-R	GATCCTCCACTAGTCTAAGCACCAACAGAACCGA
complementation	bHLH92-BIFC-C-F	TACGTAGTCACGTGATGGATGATAGCTTTTCTGT
	bHLH92-BIFC-C-R	CCGGACGTCACGTGTTAGCTTTCAAATTCTGGAC
荧光素酶互补	bHLH92-nLUC-F	ACGGGGGACGAGCTCGGTACCATGGATGATAGCTTTTCTGTG
Luciferase complementation	bHLH92-nLUC-R	CGCGTACGAGATCTGGTCGACGCTTTCAAATTCTGGACCCACA
imaging assay	CqCHP1-cLUC-F	TACGCGTCCCGGGGCGGTACCATGGCGCTGGTACCCGGAAA
	CqCHP1-cLUC-R	ACGAAAGCTCTGCAGGTCGACTCAAGCACCAACAGAACCGAAAAT

Response to Low Temperature Stress and Screening and Validation of Interaction Protein of CqCHP1 in Chieh-qua

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