Identification and expression analysis of NPR gene family members and cloning of cold-induced CsNPR3 gene in tea plants (Camellia sinensis)

doi:10.3969/j.issn.1004-1524.20220895

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (7): 1511-1522.DOI: 10.3969/j.issn.1004-1524.20220895

• Crop Science • Previous Articles Next Articles

Identification and expression analysis of NPR gene family members and cloning of cold-induced CsNPR3 gene in tea plants (Camellia sinensis)

XUE Chengjin¹(), ZHAO Lanxin¹, ZHAO Degang¹^,², HUANG Xiaozhen¹^,³^,^*()

1. The Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region, Ministry of Education, College of Life Sciences/Agricultural Engineering Research Institute, Guizhou University, Guiyang 550025, China
2. Guizhou Academy of Agricultural Sciences, Guiyang 550006, China
3. College of Tea Sciences, Guizhou University, Guiyang 550025, China

Received:2022-06-16 Online:2023-07-25 Published:2023-08-17
Contact: HUANG Xiaozhen

Abstract

Abstract:

Cold stress is the main abiotic stress that affects the yield and quality of tea plants (Camellia sinensis). The non-expresser of pathogenesis-related genes, NPRs are major immune regulatory proteins which have been implicated in plant disease resistance pathway. However, the function of NPR genes has not been well investigated in tea plants. In this study, we identified three members of NPR gene family which located on different tea plant chromosomes by using bioinformatics technology. They were named as CsNPR1, CsNPR2 and CsNPR3 (GenBank No. ON794747) respectively. These genes all contain highly conserved BTB/POZ domain, ankyrin repeat domain and multiple cysteine residues. Although the phylogenetic analysis showed that the number of NPR family members in tea plants was different from that of other species, it still showed highly conservation in evolution, which was the same as that of Arabidopsis and other species. Gene structure analysis showed that tea NPR family and Arabidopsis had similar exons-intron structure. Real-time fluorescent quantitative PCR (qRT-PCR) technique was used to identify the different tissue expression patterns of CsNPR1, CsNPR2 and CsNPR3, as well as their responses to cold stimulation and hormone salicylic acid (SA). Furthermore, CsNPR3, which was induced both by cold signal and SA was further cloned. It was found that there were variety differences in the amino acid sequence of the protein. This study can provide important scientific and theoretical basis for further clarifying the involvement of NPR gene family in cold stress and salicylic acid regulation in tea plants.

Key words: Camellia sinensis, cold stress, NPR, gene family

CLC Number:

S571.1

XUE Chengjin, ZHAO Lanxin, ZHAO Degang, HUANG Xiaozhen. Identification and expression analysis of NPR gene family members and cloning of cold-induced CsNPR3 gene in tea plants (Camellia sinensis)[J]. Acta Agriculturae Zhejiangensis, 2023, 35(7): 1511-1522.

Figures/Tables 10

References 27

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基因名称 Gene name	基因座 Gene locus	引物序列 Primer sequence(5'→3')	用途 Usage	产物大小 Product size/bp
CsNPR1	CSS0034352	F: TTGGTTGCTCTGTATCAGAG R: ATGATGTCAACATCGGACCT	qRT-PCR	161
CsNPR2	CSS0007599	F: ATGAGGAGTCGCTTGAGCCAT R: GCTGTTCTAGATGACAGCTAAG	qRT-PCR	160
CsNPR3	CSS0050405	F: TGTTGGCACACACATTGCA R: TCCTCAATTGATGCCTTCTC	qRT-PCR	140
CsNPR3	CSS0050405	F:TCTAGAATGACCCTTGATGATTCCT R:CCCGGGATATTGATGAGGGTGGTGGT	基因克隆 Cloning of gene	1 443
GAPDH		F: TTGGCATCGTTGAGGGTCT R: AGTGGGAACACGGAAAGC	内参基因 Reference gene	200

基因名称 Gene name	基因座 Gene locus	引物序列 Primer sequence(5'→3')	用途 Usage	产物大小 Product size/bp
CsNPR1	CSS0034352	F: TTGGTTGCTCTGTATCAGAG R: ATGATGTCAACATCGGACCT	qRT-PCR	161
CsNPR2	CSS0007599	F: ATGAGGAGTCGCTTGAGCCAT R: GCTGTTCTAGATGACAGCTAAG	qRT-PCR	160
CsNPR3	CSS0050405	F: TGTTGGCACACACATTGCA R: TCCTCAATTGATGCCTTCTC	qRT-PCR	140
CsNPR3	CSS0050405	F:TCTAGAATGACCCTTGATGATTCCT R:CCCGGGATATTGATGAGGGTGGTGGT	基因克隆 Cloning of gene	1 443
GAPDH		F: TTGGCATCGTTGAGGGTCT R: AGTGGGAACACGGAAAGC	内参基因 Reference gene	200

基本特征Basic characteristics	CsNPR1	CsNPR2	CsNPR3
基因登录号Accession number	CSS0034352	CSS0007599	CSS0050405
染色体Chromosome	Chr14	Chr8	Chr10
位置Location	73 483 419~73 488 814	162 894 169~162 902 685	154 564 692~154 567 524
编码蛋白质长度CDS/bp	1 764	1 761	1 443
蛋白质Protein/aa	587	586	480
分子量Molecular weight/ku	65 085.66	65 520.01	52 658.93
等电点pI	5.59	5.68	6.06
亚细胞定位Subcellular localization	胞浆Cytoplasmic	胞浆Cytoplasmic	胞浆Cytoplasmic

基本特征Basic characteristics	CsNPR1	CsNPR2	CsNPR3
基因登录号Accession number	CSS0034352	CSS0007599	CSS0050405
染色体Chromosome	Chr14	Chr8	Chr10
位置Location	73 483 419~73 488 814	162 894 169~162 902 685	154 564 692~154 567 524
编码蛋白质长度CDS/bp	1 764	1 761	1 443
蛋白质Protein/aa	587	586	480
分子量Molecular weight/ku	65 085.66	65 520.01	52 658.93
等电点pI	5.59	5.68	6.06
亚细胞定位Subcellular localization	胞浆Cytoplasmic	胞浆Cytoplasmic	胞浆Cytoplasmic

NPR homologues	CsNPR2	CsNPR3	AtNPR1	AtNPR2	AtNPR3	AtNPR4	AtNPR5	AtNPR6
CsNPR1	45.69	32.43	53.06	53.47	44.88	41.98	31.47	32.61
CsNPR2		34.29	39.33	40.29	62.48	57.63	34.28	35.17
CsNPR3			29.12	30.79	29.58	29.93	75.98	73.22
AtNPR1				62.29	39.07	37.12	28.65	29.72
AtNPR2					39.77	38.48	28.22	30.14
AtNPR3						68.22	31.96	31.50
AtNPR4							30.10	31.28
AtNPR5								80.70

Identification and expression analysis of NPR gene family members and cloning of cold-induced CsNPR3 gene in tea plants (Camellia sinensis)

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