Cloning of Phytochrome Interaction Factor CsPIF3a and Its Response to Light and Temperature Stress in Camellia sinensis

doi:10.13560/j.cnki.biotech.bull.1985.2024-0794

Abstract

Abstract:

Objective PIFs (Phytochrome interacting factors) are the core components in the photopigment-mediated photothermal signaling pathway. Studying the function of PIF3 in tea plant (Camellia sinensis (L.) O. Kuntze) provides important theoretical guidance for using light signaling pathways to avoid or mitigate the harm caused by adverse conditions. Method The CsPIF3a gene was cloned from tea plants through homology cloning and subsequently analyzed using bioinformatics tools. Expression patterns of CsPIF3a across various tissues and under different light and temperature conditions were assessed using real-time quantitative PCR (RT-qPCR). Additionally, tobacco transient expression assays and yeast two-hybrid assays were conducted to determine the protein localization and transcriptional activity of CsPIF3a. Result The open reading frame (ORF) of CsPIF3a was 2 163 bp in length, encoding a protein of 721 amino acids. The predicted molecular mass of the encoded protein was 76.97 kD. The theoretical isoelectric point was 5.70, having hydrophilic properties. The phylogenetic tree analysis showed that CsPIF3a was closely related to Actinidia eriantha Bentham. and contained a basic helix-loop-helix (bHLH) domain, an active phytochrome binding (APB) domain, and an APA domain. The secondary structure analysis of CsPIF3a revealed that it comprised 18.17% α-helix, 6.80% β-sheet, 1.53% β-turn, and 73.51% random coil, with 46.87% of the regions being disordered. Subcellular localization assays demonstrated that CsPIF3a was localized in the nucleus and presented transcriptional activity. Reverse transcription quantitative PCR (RT-qPCR) analysis indicated that CsPIF3a was predominantly expressed in the old leaves and stems of tea plants. Additionally, its expression was found to be responsive to temperature stress and correlated with light intensity. Conclusion CsPIF3a is localized in the nucleus and exhibits transcriptional activity. In response to temperature stress, the expression level of CsPIF3a gene was related to light intensity.

Key words: CsPIF3a, gene cloning, bioinformatics analysis, subcellular localization, transcriptional activity, expression analysis

BAN Qiu-yan, ZHAO Xin-yue, CHI Wen-jing, LI Jun-sheng, WANG Qiong, XIA Yao, LIANG Li-yun, HE Wei, LI Ye-yun, ZHAO Guang-shan. Cloning of Phytochrome Interaction Factor CsPIF3a and Its Response to Light and Temperature Stress in Camellia sinensis[J]. Biotechnology Bulletin, 2025, 41(4): 256-265.

Figures/Tables 10

References 31

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植物 Plant	GenBank登录号 GenBank accession	氨基酸数 Number of amino acids	相对分子量 Relative molecular weight/kD	理论等电点 Theoretical isoelectric point	碱性氨基酸 Basic amino acids/%	酸性氨基酸 Acidic amino acids/%	芳香族氨基酸 Aromatic amino acid/%	脂肪族氨基酸 Aliphatic amino acids/%	总平均疏水性 Total average hydrophobicity
茶树C. sinensis	XP_028108728.1	721	76.96	5.70	11	11	3	16	-0.547
杨梅M. rubra	KAB1199165.1	739	78.80	6.46	13	11	4	16	-0.506
胡桃J. microcarpa	XP_041007950.1	738	78.93	5.98	12	11	4	15	-0.551
欧榛C. avellana	XP_059456869.1	744	79.75	6.05	12	11	3	16	-0.502
夏栎Q. robur	XP_050273354.1	749	80.23	6.05	11	10	4	15	-0.546
酸枣Z. jujuba	XP_015874808.1	729	77.78	5.64	11	11	4	15	-0.553
水蜜桃P. persica	XP_020422550.1	729	78.19	5.67	11	11	4	15	-0.579
欧洲甜樱桃P. avium	XP_021830447.1	728	78.16	5.77	11	11	4	15	-0.584
橡胶树H. brasiliensis	XP_058000598.1	745	79.20	5.73	11	10	4	15	-0.555
葡萄V. vinifera	XP_010659764.1	709	75.86	6.30	12	10	4	15	-0.598
君迁子D. lotus	XP_052204847.1	721	76.31	5.97	11	10	4	16	-0.511
狭叶油菜C. lanceoleosa	KAI8022635.1	627	68.01	5.64	11	11	4	15	-0.636
红马银花R. vialii	XP_058191728.1	700	75.50	5.94	12	11	5	16	-0.471
拟南芥Arabidopsis thaliana	NP_001318964.1	524	56.99	6.11	13	12	5	15	-0.728
毛花猕猴桃A. eriantha	XP_057494369.1	723	77.24	6.56	12	10	4	16	-0.584

植物 Plant	GenBank登录号 GenBank accession	氨基酸数 Number of amino acids	相对分子量 Relative molecular weight/kD	理论等电点 Theoretical isoelectric point	碱性氨基酸 Basic amino acids/%	酸性氨基酸 Acidic amino acids/%	芳香族氨基酸 Aromatic amino acid/%	脂肪族氨基酸 Aliphatic amino acids/%	总平均疏水性 Total average hydrophobicity
茶树C. sinensis	XP_028108728.1	721	76.96	5.70	11	11	3	16	-0.547
杨梅M. rubra	KAB1199165.1	739	78.80	6.46	13	11	4	16	-0.506
胡桃J. microcarpa	XP_041007950.1	738	78.93	5.98	12	11	4	15	-0.551
欧榛C. avellana	XP_059456869.1	744	79.75	6.05	12	11	3	16	-0.502
夏栎Q. robur	XP_050273354.1	749	80.23	6.05	11	10	4	15	-0.546
酸枣Z. jujuba	XP_015874808.1	729	77.78	5.64	11	11	4	15	-0.553
水蜜桃P. persica	XP_020422550.1	729	78.19	5.67	11	11	4	15	-0.579
欧洲甜樱桃P. avium	XP_021830447.1	728	78.16	5.77	11	11	4	15	-0.584
橡胶树H. brasiliensis	XP_058000598.1	745	79.20	5.73	11	10	4	15	-0.555
葡萄V. vinifera	XP_010659764.1	709	75.86	6.30	12	10	4	15	-0.598
君迁子D. lotus	XP_052204847.1	721	76.31	5.97	11	10	4	16	-0.511
狭叶油菜C. lanceoleosa	KAI8022635.1	627	68.01	5.64	11	11	4	15	-0.636
红马银花R. vialii	XP_058191728.1	700	75.50	5.94	12	11	5	16	-0.471
拟南芥Arabidopsis thaliana	NP_001318964.1	524	56.99	6.11	13	12	5	15	-0.728
毛花猕猴桃A. eriantha	XP_057494369.1	723	77.24	6.56	12	10	4	16	-0.584

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