Identification，Expression Analysis and Interaction Validation of CsIDM in Tea Plants

doi:10.16420/j.issn.0513-353x.2021-1218

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (8): 1679-1696.doi: 10.16420/j.issn.0513-353x.2021-1218

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

Identification，Expression Analysis and Interaction Validation of CsIDM in Tea Plants

LI Yuteng¹, CHEN Yao², REN Hengze², LI Congcong¹, WANG Haoqian¹, CAO Hongli¹, YUE Chuan¹^,^*(), HAO Xinyuan²^,^*(), WANG Xinchao²

¹ College of Horticulture，Fujian Agriculture and Forestry University，Fuzhou 350002，China
² National Center for Tea Improvement，Tea Research Institute Chinese Academy of Agricultural Sciences，Hangzhou 310008，China

Received:2022-11-12 Revised:2023-05-05 Online:2023-08-25 Published:2023-08-23
Contact: YUE Chuan, HAO Xinyuan

Abstract

Abstract:

In this study，using different tea plant genomes as a reference，three CsIDM genes were identified and cloned from Camellia sinensis‘Longjing 43’. CsIDM1，belongs to the histone acetyltransferase family，has a typical HAT conserved domain，and is located in the nucleus. CsIDM2 and CsIDM3，which belong to the heat shock protein family，have typical ACD conserved domains，which are located in the nucleus，cell membrane and cytoplasm. The promoter region of CsIDMs contains a variety of cis-acting elements that respond to environmental signals，including light response，plant hormone response，stress response and plant growth-related elements. Expression analysis showed that CsIDMs expressed higher in flower buds，axillary buds and stems，but lower in fully open flowers，which suggests a certain tissue specificity. CsIDMs were differentially expressed under drought，high temperature and biotic stresses. During the formation and release of dormancy in overwintering buds，the expression of CsIDM1 showed a high-low-high expression pattern in the stages of paradormancy，endodormancy and ecodormancy. It was confirmed that CsIDM2 interacts with CsIDM3 to form a heterodimer by bimolecular fluorescence complementation. CsIDM2 and CsIDM3 interact with CsMBD5 and CsMBD16，respectively. In summary，CsIDMs play an important role in the stress response and bud dormancy removal of tea plants，and may participate in the regulation of methylation through the formation of protein complexes.

Key words: Camellia sinensis, demethylation, IDM complex, subcellular localization, expression analysis, BiFC assay

LI Yuteng, CHEN Yao, REN Hengze, LI Congcong, WANG Haoqian, CAO Hongli, YUE Chuan, HAO Xinyuan, WANG Xinchao. Identification，Expression Analysis and Interaction Validation of CsIDM in Tea Plants[J]. Acta Horticulturae Sinica, 2023, 50(8): 1679-1696.

Figures/Tables 13

References 45

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用途 Application	序列名称 Sequence name	引物序列（5′-3′） Primer sequence
基因克隆 Gene clone	CsIDM1	F：GGTACTGGTGTTCATTGATTAGAGCAAAGAAAG R：TTATTTTTATTTCTCATTATCTTGCAACATAGTGG
	CsIDM2	F：ATGGATGACTTGTCTCATGCT R：TTATATCCCTTTCATCACAATTCCTTC
	CsIDM3	F：ATTATTCTGGTTGATTTCTAGCCCCA R：TCATCTGCATAAAATATGAACACCATTC
qRT-PCR	Q-CsIDM1	F：ACTTTCGGATCAACTACCAAACGAT R：AAACATGGACCACTTCCCGTCA
	Q-CsIDM2	F：GATCTATTTCCTCCCCGGCTT R：TTCTAGCCCAGTTAGCCGCTTG
	Q-CsIDM3	F：GATGGGCCTTTCATGATGCC R：ATGCAGATTTGCCGACTCCA
内参基因 Reference gene	Q-CsPTB	F：TGACCAAGCACACTCCACACTATCG R：TGCCCCCTTATCATCATCCACAA
亚细胞定位 Subcellular localization	CsIDM1-GFP	F：CGAGAGTGTCGGAGCTATGTTATTCAACAAAGAAATCG R：GCCCTTGCTCACCATGTCGACTTTCTCATTATCTTGCAACA
	CsIDM2-GFP	F：GGTACCCGGGGATCCTCTAGAATGGATGACTTGTCTCATGC R：GCCCTTGCTCACCATGTCGACTATCCCTTTCATCACAATTC
	CsIDM3-GFP	F：GGTACCCGGGGATCCTCTAGAATGTGTACGTGCCCAAAGCA R：GCCCTTGCTCACCATGTCGACCTCATATTTCATGATGATAG
BIFC载体构建 BIFC vector construction	CsIDM1-BIFC	F：CGAACGATAGTTAATTAAATGTTATTCAACAAAGAAATC R：ACTGCCACCTCCTCCACTAGTTTTCTCATTATCTTGCAACA
	CsIDM2-BIFC	F：CGAACGATAGTTAATTAAATGGATGACTTGTCTCATGC R：CTGCCACCTCCTCCACTAGTTATCCCTTTCATCACAATTC
	CsIDM3-BIFC	F：CGAACGATAGTTAATTAAATGTGTACGTGCCCAAAGCAAAG R：CTGCCACCTCCTCCACTAGTCTCATATTTCATGATGATAG
	CsMBD5-BIFC	F：TACGAACGATAGTTAATTAAATGTCGACCTGCGAATCC R：CTGCCACCTCCTCCACTAGTGCTGCAAGAGAAATGAAATG
	CsMBD16-BIFC	F：TACGAACGATAGTTAATTAAATGGATGACAATGGCGCATC R：CACCACTGCCACCTCCTCCACTAGTCCATTCAAAATTTGG

用途 Application	序列名称 Sequence name	引物序列（5′-3′） Primer sequence
基因克隆 Gene clone	CsIDM1	F：GGTACTGGTGTTCATTGATTAGAGCAAAGAAAG R：TTATTTTTATTTCTCATTATCTTGCAACATAGTGG
	CsIDM2	F：ATGGATGACTTGTCTCATGCT R：TTATATCCCTTTCATCACAATTCCTTC
	CsIDM3	F：ATTATTCTGGTTGATTTCTAGCCCCA R：TCATCTGCATAAAATATGAACACCATTC
qRT-PCR	Q-CsIDM1	F：ACTTTCGGATCAACTACCAAACGAT R：AAACATGGACCACTTCCCGTCA
	Q-CsIDM2	F：GATCTATTTCCTCCCCGGCTT R：TTCTAGCCCAGTTAGCCGCTTG
	Q-CsIDM3	F：GATGGGCCTTTCATGATGCC R：ATGCAGATTTGCCGACTCCA
内参基因 Reference gene	Q-CsPTB	F：TGACCAAGCACACTCCACACTATCG R：TGCCCCCTTATCATCATCCACAA
亚细胞定位 Subcellular localization	CsIDM1-GFP	F：CGAGAGTGTCGGAGCTATGTTATTCAACAAAGAAATCG R：GCCCTTGCTCACCATGTCGACTTTCTCATTATCTTGCAACA
	CsIDM2-GFP	F：GGTACCCGGGGATCCTCTAGAATGGATGACTTGTCTCATGC R：GCCCTTGCTCACCATGTCGACTATCCCTTTCATCACAATTC
	CsIDM3-GFP	F：GGTACCCGGGGATCCTCTAGAATGTGTACGTGCCCAAAGCA R：GCCCTTGCTCACCATGTCGACCTCATATTTCATGATGATAG
BIFC载体构建 BIFC vector construction	CsIDM1-BIFC	F：CGAACGATAGTTAATTAAATGTTATTCAACAAAGAAATC R：ACTGCCACCTCCTCCACTAGTTTTCTCATTATCTTGCAACA
	CsIDM2-BIFC	F：CGAACGATAGTTAATTAAATGGATGACTTGTCTCATGC R：CTGCCACCTCCTCCACTAGTTATCCCTTTCATCACAATTC
	CsIDM3-BIFC	F：CGAACGATAGTTAATTAAATGTGTACGTGCCCAAAGCAAAG R：CTGCCACCTCCTCCACTAGTCTCATATTTCATGATGATAG
	CsMBD5-BIFC	F：TACGAACGATAGTTAATTAAATGTCGACCTGCGAATCC R：CTGCCACCTCCTCCACTAGTGCTGCAAGAGAAATGAAATG
	CsMBD16-BIFC	F：TACGAACGATAGTTAATTAAATGGATGACAATGGCGCATC R：CACCACTGCCACCTCCTCCACTAGTCCATTCAAAATTTGG

基因名称 Gene name	登录号 GenBank	氨基酸数 Amino acids number	分子质量/kD Molecular weight	pI	稳定系数 Instability index	亲水性 GRAVY	亚细胞定位 Subcellular localization
CsIDM1 AtIDM1	OK649843 AT3G14980	1 251 1 189	139.38 131.33	7.69 6.69	46.94 44.49	-0.451 -0.489	细胞核Nucleus 细胞核Nucleus
CsIDM2 AtIDM2	OK649845 AT1G54840	419 349	46.94 39.39	5.10 6.04	45.75 44.83	-0.251 -0.374	细胞核Nucleus 细胞核Nucleus
CsIDM3 AtIDM3	OK649846 AT1G20870	211 463	23.22 51.93	8.95 7.07	45.73 51.10	-0.470 -0.616	叶绿体Chloroplast 细胞核Nucleus

基因名称 Gene name	登录号 GenBank	氨基酸数 Amino acids number	分子质量/kD Molecular weight	pI	稳定系数 Instability index	亲水性 GRAVY	亚细胞定位 Subcellular localization
CsIDM1 AtIDM1	OK649843 AT3G14980	1 251 1 189	139.38 131.33	7.69 6.69	46.94 44.49	-0.451 -0.489	细胞核Nucleus 细胞核Nucleus
CsIDM2 AtIDM2	OK649845 AT1G54840	419 349	46.94 39.39	5.10 6.04	45.75 44.83	-0.251 -0.374	细胞核Nucleus 细胞核Nucleus
CsIDM3 AtIDM3	OK649846 AT1G20870	211 463	23.22 51.93	8.95 7.07	45.73 51.10	-0.470 -0.616	叶绿体Chloroplast 细胞核Nucleus

Identification，Expression Analysis and Interaction Validation of CsIDM in Tea Plants

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[2]	HOU Binghao, GAO Ting, WEI Yuede, GAO Shuilian, CAI Yinbi, CHEN Zhiming, JIN Shan, ZHANG Xingtan, WANG Pengjie, YE Naixing. Studies on Genetic Variation of‘Tieguanyin’Tea Trees Asexual Reproduction Based on High-depth Genome Resequencing [J]. Acta Horticulturae Sinica, 2023, 50(7): 1505-1517.
[3]	WANG Tonghuan, WU Yuxin, WU Yiyuan, LI Xinxin, LIU Mengyang, YANG Lianlian, LI Jiapeng, ZHANG Zhongshan, CAO Fang, ZHONG Xueting, WANG Zhanqi. Genome-wide Identification and Expression Analysis of the GRAS Gene Family in Response to Cold Stress in Chrysanthemum nankingense [J]. Acta Horticulturae Sinica, 2023, 50(4): 815-830.
[4]	WANG Zehan, YU Wentao, WANG Pengjie, LIU Caiguo, FAN Xiaojing, GU Mengya, CAI Chunping, WANG Pan, YE Naixing. WGCNA Analysis of Differentially Expressed Genes in Floral Organs of Tea Germplasms with Ovary-glabrous and Ovary-trichome [J]. Acta Horticulturae Sinica, 2023, 50(3): 620-634.
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[12]	LIANG Chen, SUN Ruyi, XIANG Rui, SUN Yimeng, SHI Xiaoxin, DU Guoqiang, WANG Li. Genome-wide Identification of Grape GRF Family and Expression Analysis [J]. Acta Horticulturae Sinica, 2022, 49(5): 995-1007.
[13]	XIAO Xuechen, LIU Mengyu, JIANG Mengqi, CHEN Yan, XUE Xiaodong, ZHOU Chengzhe, WU Xingjian, WU Junnan, GUO Yinsheng, YEH Kaiwen, LAI Zhongxiong, LIN Yuling. Whole-genome Identification and Expression Analysis of SNAT,ASMT and COMT Families of Melatonin Synthesis Pathway in Dimocarpus longan [J]. Acta Horticulturae Sinica, 2022, 49(5): 1031-1046.
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[15]	GAO Weilin, ZHANG Liman, XUE Chaoling, ZHANG Yao, LIU Mengjun, ZHAO Jin. Expression of E-type MADS-box Genes in Flower and Fruits and Protein Interaction Analysis in Chinese Jujube [J]. Acta Horticulturae Sinica, 2022, 49(4): 739-748.