Response of Ubiquitin-ligase Gene CsPUB21 to Different Abiotic Stress in Camellia sinensis

doi:10.16420/j.issn.0513-353x.2024-0378

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (3): 655-670.doi: 10.16420/j.issn.0513-353x.2024-0378

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

Response of Ubiquitin-ligase Gene CsPUB21 to Different Abiotic Stress in Camellia sinensis

DENG Shuqin, GAO Yingrui, LI Yutong, WANG Ying, GONG Chunmei, BAI Juan^*()

College of Horticulture，Northwest A & F University，Yangling，Shaanxi 712100，China

Received:2024-07-09 Revised:2025-01-12 Online:2025-03-25 Published:2025-03-25
Contact: BAI Juan

Abstract

Abstract:

The U-box ubiquitin E3 ligase gene CsPUB21 were cloned and functionally analyzed in Camellia sinensis‘Shaancha 1’. The length of the coding sequence of CsPUB21 is 1 242 bp，encoding a total of 413 amino acids with a conserved U-box domain at the N terminus and an ARM domain at the C terminus. The encoded protein，an unstable hydrophilic protein. The promoter region contains multiple cis-acting elements in response to drought，high salt and low temperature. The subcellular localization results indicated that CsPUB21 is localized on the cell membrane and cytoplasm. Gene expression pattern analysis showed that CsPUB21 was up-regulated by drought and salt stress treatments，and down-regulated by low temperature in different cultivars of C. sinensis. Overexpression of CsPUB21 gene in Arabidopsis significantly enhanced plants resistance to drought and salt stress. The qRT-PCR results showed that the expression level of CsPUB21 was up-regulated after drought and salt stress. Compared with the wild type，overexpression of CsPUB21 in Arabidopsis resulted in higher chlorophyll content and soluble sugar content under drought and salt stress，enhanced activities of antioxidant enzymes SOD，POD，and CAT，and decreased relative conductivity and MDA content. The research results indicated that the CsPUB21 plays a positive regulatory role in tea plant response to drought and salt stress.

Key words: Camellia sinensis, CsPUB21, gene cloning, abiotic stress, expression analysis, heterologous function verification

DENG Shuqin, GAO Yingrui, LI Yutong, WANG Ying, GONG Chunmei, BAI Juan. Response of Ubiquitin-ligase Gene CsPUB21 to Different Abiotic Stress in Camellia sinensis[J]. Acta Horticulturae Sinica, 2025, 52(3): 655-670.

Figures/Tables 12

References 46

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引物名称 Primer name	引物序列（5′-3′） Primer sequence	用途 Usage
1300-CsPUB21-F	GAGAACACGGGGGAC GAGCTCATGATTTCGTCTTGGCGGAG	植物过表达 Plant overexpression
1300-CsPUB21-R	CGCATAGCTAATCTG GGATCCTCAATTTTGAAGAGCTTCAACAAGA	植物过表达 Plant overexpression
qRT-CsPUB21-F	GAAGGGTTTGGCGAAGGAGA	荧光定量PCR Quantitative real time PCR
qRT-CsPUB21-R	AGCCCTCTCGCATATGCTTC
qRT-Csβ-actin-F	GGTAACATTGTGCTCAGTGGTGG
qRT-Csβ-actin-R	AACGACCTTAATCTTCATGCTGC

引物名称 Primer name	引物序列（5′-3′） Primer sequence	用途 Usage
1300-CsPUB21-F	GAGAACACGGGGGAC GAGCTCATGATTTCGTCTTGGCGGAG	植物过表达 Plant overexpression
1300-CsPUB21-R	CGCATAGCTAATCTG GGATCCTCAATTTTGAAGAGCTTCAACAAGA	植物过表达 Plant overexpression
qRT-CsPUB21-F	GAAGGGTTTGGCGAAGGAGA	荧光定量PCR Quantitative real time PCR
qRT-CsPUB21-R	AGCCCTCTCGCATATGCTTC
qRT-Csβ-actin-F	GGTAACATTGTGCTCAGTGGTGG
qRT-Csβ-actin-R	AACGACCTTAATCTTCATGCTGC

顺式作用元件 cis-Acting element	序列 Sequence	数量 Amount	功能 Function
MYB	CAACCA CAACTG	6 1	干旱、高盐、低温响应元件Drought，high salt，low temperature response element
MBS	CAACTG	1	干旱响应元件Drought response element
STRE	AGGGG	2	胁迫响应元件Stress response element
WUN-motif	AAATTACT	2	损伤诱导元件Damage inductive element
WRE3	CCACCT	1	损伤诱导元件Damage inductive element
W box	TTGACC	1	损伤与病原响应元件Damage and pathogen response element
ARE	AAACCA	1	厌氧诱导调控元件Anaerobic inducible regulatory element
Box 4	ATTAAT	5	光响应元件Photoresponsive element
ATC-motif	AGTAATCT	1	光响应元件Photoresponsive element
AAGAA-motif	GAAAGAA	1	脱落酸响应元件Abscisic acid response element
ABRE	TACGGTC	1	脱落酸响应元件Abscisic acid response element
MYC	CATGTG	4	抗脱落酸响应元件Abscisic acid resistant response element
F-box	CTATTCTCATT	1	赤霉素响应元件Gibberellin response element
CGTCA-motif	CGTCA	4	茉莉酸响应元件Jasmonic acid response element
TGACG-motif	TGACG	4	茉莉酸响应元件Jasmonic acid response element
ERE	ATTTTAAA	2	乙烯响应元件Ethylene responsive element
as-1	TGACG	4	根特异表达顺式作用调控元件Root specific expression of cis-acting regulatory elements

顺式作用元件 cis-Acting element	序列 Sequence	数量 Amount	功能 Function
MYB	CAACCA CAACTG	6 1	干旱、高盐、低温响应元件Drought，high salt，low temperature response element
MBS	CAACTG	1	干旱响应元件Drought response element
STRE	AGGGG	2	胁迫响应元件Stress response element
WUN-motif	AAATTACT	2	损伤诱导元件Damage inductive element
WRE3	CCACCT	1	损伤诱导元件Damage inductive element
W box	TTGACC	1	损伤与病原响应元件Damage and pathogen response element
ARE	AAACCA	1	厌氧诱导调控元件Anaerobic inducible regulatory element
Box 4	ATTAAT	5	光响应元件Photoresponsive element
ATC-motif	AGTAATCT	1	光响应元件Photoresponsive element
AAGAA-motif	GAAAGAA	1	脱落酸响应元件Abscisic acid response element
ABRE	TACGGTC	1	脱落酸响应元件Abscisic acid response element
MYC	CATGTG	4	抗脱落酸响应元件Abscisic acid resistant response element
F-box	CTATTCTCATT	1	赤霉素响应元件Gibberellin response element
CGTCA-motif	CGTCA	4	茉莉酸响应元件Jasmonic acid response element
TGACG-motif	TGACG	4	茉莉酸响应元件Jasmonic acid response element
ERE	ATTTTAAA	2	乙烯响应元件Ethylene responsive element
as-1	TGACG	4	根特异表达顺式作用调控元件Root specific expression of cis-acting regulatory elements

Response of Ubiquitin-ligase Gene CsPUB21 to Different Abiotic Stress in Camellia sinensis

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