Study on the Cold Resistance Function of VaJAZ9 and VaCOI1 Genes in Vitis amurensis

doi:10.16420/j.issn.0513-353x.2023-0910

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (10): 2243-2254.doi: 10.16420/j.issn.0513-353x.2023-0910

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

Study on the Cold Resistance Function of VaJAZ9 and VaCOI1 Genes in Vitis amurensis

SUN Yutong, LIU Deshuai, HUANG Xuzheng, CHI Jingnan, QI Xun, YAO Wenkong^*()

College of Enology and Horticulture，Ningxia University，Yinchuan 750021，China

Received:2024-03-14 Revised:2024-07-16 Online:2024-10-25 Published:2024-10-21
Contact: YAO Wenkong

Abstract

Abstract:

In this study，the VaCOI1 gene from Vitis amurensis Rupr.‘Zuoshan-1’were cloned，and the gene function was analysed. The results show that the VaCOI1 gene is located on chromosome 13，and its ORF has a full-length of 1 797 bp，encoding 598 amino acids，and belongs to the F-box family. Clustering analysis indicates that the VaCOI1 protein performed a high homology with HuCOI1（Herrania umbratica）and TcCOI1（Theobroma cacao）. Bimolecular fluorescence complementation（BiFC）assays indicate that VaJAZ9 can interact with the VaCOI1 protein. The heterologously overexpressing of VaJAZ9 and VaCOI1 in Arabidopsis thaliana shows that at the low temperature，the VaCOI1 overexpressing plants had a higher survival rate and a lower conductivity than the wild type plants，whereas the VaJAZ9 overexpressing plants had a lower survival rate and a higher conductivity than the wild type plants. Quantitative real-time PCR（qRT-PCR）analysis show that the low temperature related genes including AtKIN1，AtCOR15A and AtRD29A were up-regulated in the VaCOI1 overexpressing A. thaliana，whereas these genes were down-regulated in overexpressing VaJAZ9 plants. In conclusion，the results show that the VaCOI1 acts as a positive regulator in response to low temperature stress，whereas the VaJAZ9 acts as a negative regulator.

Key words: Vitis amurensis, JA signaling pathway, JAZ9, COI1, cold resistance

SUN Yutong, LIU Deshuai, HUANG Xuzheng, CHI Jingnan, QI Xun, YAO Wenkong. Study on the Cold Resistance Function of VaJAZ9 and VaCOI1 Genes in Vitis amurensis[J]. Acta Horticulturae Sinica, 2024, 51(10): 2243-2254.

Figures/Tables 8

References 34

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用途 Application	引物 Primer	引物序列（5′-3′） Primer sequence
构建载体	COI1-2300-F	AGAACACGGGGGACGAGCTCATGGAGGATGGAAACGAGAGAAAG
Vector construction	COI1-2300-R	ACCATGGTGTCGACTCTAGACAACGTAAGAAAAGATGCAGGGTC
	JAZ9-2300-F	ACGAGCTCGGTACCCGGGGATCCATGTCGAGCTCCTCGGATATTGC
	JAZ9-2300-R	CCCTTGCTCACCATGGTGTCGACCTTTGGAGATTTGGCAGCCAAGC
BiFC验证	221-CE-JAZ9-XbaI-F	GAGAACACGGGGGACTCTAGAATGTCGAGCTCCTCGGATATTGC
BiFC verification	221-CE-JAZ9-XhoI-R	GTACATGGTACCCCGCTCGAGCTTTGGAGATTTGGCAGCCAAGC
	221-NE-COI1-BamHI-F	GGCCCAGGCCTACTAGTGGATCCATGGAGGATGGAAACGAGAGAAAG
	221-NE-COI1-XhoI-R	TACCCGGGAGCGGTACCCTCGAGCTACAACGTAAGAAAAGATGCAGGGTC
过表达植株检测	COI1-F	ATGGAGGATGGAAACGAGAGAAAG
Detection of	COI1-R	CTGCTCATCAGCACCTCTCTC
overexpressed plant	JAZ9--F	ATGTCGAGCTCCTCGGATATTGC
	JAZ9-R	CACGATCACTTGGCCACCGTAG
实时荧光定量PCR	RT-COI1-F	GCTTGCTTGACACAGAGGACCA
Real-time	RT-COI1-R	CCAATCACGTTCCTGGCCTCAA
Quantitative PCR	RT-AtActin-F	CTTGCACCAAGCAGCATGAA
	RT-AtActin-R	CCGATCCAGACACTGTACTTCCTT
	AtRT-CBF1-F	ACTTCGCTGACTCGGCTTGG
	AtRT-CBF1-R	ACGCACCTTCGCTCTGTTCC
	AtRT-CBF2-F	GGAATCAACCTGTGCCAAGGAA
	AtRT-CBF2-R	CCAACATCGCCTCTTCATCCATAT
	AtRT-CBF3-F	TTCCTCAGGCGGTGATTATATTCC
	AtRT-CBF3-R	CTCCGACGAACTCCTCTGTATATTG
	RT-RD29A-F	CAGAGGAACCACCACTCAACACA
	RT-RD29A-R	CTCTAGGTTTACCTGTTACGCCTG
	RT-RD29B-F	ATGGAGTCACAGTTGACACGTCCT
	RT-RD29B-R	CTTCTGGGTCTTGCTCGTCATACT
	RT-KIN1-F	ATGTCAGAGACCAACAAGAATGCC
	RT-KIN1-R	CTACTTGTTCAGGCCGGTCTTG
	RT-AtCOR15A-F	AGTCGTTGATCTACGCCGCTAA
	RT-AtCOR15A-R	TCTCACCATCTGCTAATGCCTCTT

用途 Application	引物 Primer	引物序列（5′-3′） Primer sequence
构建载体	COI1-2300-F	AGAACACGGGGGACGAGCTCATGGAGGATGGAAACGAGAGAAAG
Vector construction	COI1-2300-R	ACCATGGTGTCGACTCTAGACAACGTAAGAAAAGATGCAGGGTC
	JAZ9-2300-F	ACGAGCTCGGTACCCGGGGATCCATGTCGAGCTCCTCGGATATTGC
	JAZ9-2300-R	CCCTTGCTCACCATGGTGTCGACCTTTGGAGATTTGGCAGCCAAGC
BiFC验证	221-CE-JAZ9-XbaI-F	GAGAACACGGGGGACTCTAGAATGTCGAGCTCCTCGGATATTGC
BiFC verification	221-CE-JAZ9-XhoI-R	GTACATGGTACCCCGCTCGAGCTTTGGAGATTTGGCAGCCAAGC
	221-NE-COI1-BamHI-F	GGCCCAGGCCTACTAGTGGATCCATGGAGGATGGAAACGAGAGAAAG
	221-NE-COI1-XhoI-R	TACCCGGGAGCGGTACCCTCGAGCTACAACGTAAGAAAAGATGCAGGGTC
过表达植株检测	COI1-F	ATGGAGGATGGAAACGAGAGAAAG
Detection of	COI1-R	CTGCTCATCAGCACCTCTCTC
overexpressed plant	JAZ9--F	ATGTCGAGCTCCTCGGATATTGC
	JAZ9-R	CACGATCACTTGGCCACCGTAG
实时荧光定量PCR	RT-COI1-F	GCTTGCTTGACACAGAGGACCA
Real-time	RT-COI1-R	CCAATCACGTTCCTGGCCTCAA
Quantitative PCR	RT-AtActin-F	CTTGCACCAAGCAGCATGAA
	RT-AtActin-R	CCGATCCAGACACTGTACTTCCTT
	AtRT-CBF1-F	ACTTCGCTGACTCGGCTTGG
	AtRT-CBF1-R	ACGCACCTTCGCTCTGTTCC
	AtRT-CBF2-F	GGAATCAACCTGTGCCAAGGAA
	AtRT-CBF2-R	CCAACATCGCCTCTTCATCCATAT
	AtRT-CBF3-F	TTCCTCAGGCGGTGATTATATTCC
	AtRT-CBF3-R	CTCCGACGAACTCCTCTGTATATTG
	RT-RD29A-F	CAGAGGAACCACCACTCAACACA
	RT-RD29A-R	CTCTAGGTTTACCTGTTACGCCTG
	RT-RD29B-F	ATGGAGTCACAGTTGACACGTCCT
	RT-RD29B-R	CTTCTGGGTCTTGCTCGTCATACT
	RT-KIN1-F	ATGTCAGAGACCAACAAGAATGCC
	RT-KIN1-R	CTACTTGTTCAGGCCGGTCTTG
	RT-AtCOR15A-F	AGTCGTTGATCTACGCCGCTAA
	RT-AtCOR15A-R	TCTCACCATCTGCTAATGCCTCTT

Study on the Cold Resistance Function of VaJAZ9 and VaCOI1 Genes in Vitis amurensis

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