Identification of Zinc Finger Protein DnaJ-Like Gene Family in Capsicum annuum and Its Expression Analysis Responses to High Temperature Stress

doi:10.13560/j.cnki.biotech.bull.1985.2022-1033

Abstract

Abstract:

Plant DanJ-Like zine finger protein（DNAJE）is one of the important protein found in recent years, which plays an important role in photosynthesis, development and motility of plastids and the responses of plants to stresses. Here, the DNAJE whole gene family（CaDNAJE）was identified by the whole genome data of pepper（Capsicum annuum）, and the characteristic of gene and protein, comparative evolution, collinearity and transcriptional expression under high temperature stress were analyzed. Totally, 28 members of CaDNAJE genes were identified, and they were distributed unevenly on 10 chromosomes and 3 Scaffolds. The amino acid length, molecular mass and isoelectric point were between 99-470 aa, 10.05-52.26 kD and 4.75-9.64, respectively. Moreover, these encoded proteins were mainly located in the chloroplast and nucleus. The comparative evolution of pepper, tomato（Solanum lycopersicum）and Arabidopsis analysis can be divided into 2 subgroups: GroupⅠ and GroupⅡ, and the CaDNAJE protein in the same subgroup had similar conserved motifs and gene structures. Cis-acting element analysis in the promoter region showed that CaDNAJEs contained a large numbers of functional elements related to light response, hormone response and stress. There were 10 pairs of collinearity genes between pepper and Arabidopsis, but only one pair within pepper species. The transcriptome analysis of different pepper tissues indicated that most members of GroupⅠ were highly expressed in all tissues, while most members of GroupⅡ had low or no expression except for CaDNAJE2 and CaDNAJE9. The content of hydrogen peroxide in pepper leaves increased rapidly and then decreased. The content of malondialdehyde kept increasing and peroxidase activity also had been improved to different extent. High temperature stress induced the high expression of the heat shock factor/heat shock protein（Hsf/HSP）and CaDNAJE genes, which played a regulation role in different stress time. It is suggested that CaDNAJE family genes are involved in response to high temperature stress and then increased tolerance to reduce cell damage.

Key words: pepper, DnaJ-Like zinc finer protein, genome-wide identification, high temperature stress, gene expression, regulation

DUAN Min-jie, LI Yi-fei, YANG Xiao-miao, WANG Chun-ping, HUANG Qi-zhong, HUANG Ren-zhong, ZHANG Shi-cai. Identification of Zinc Finger Protein DnaJ-Like Gene Family in Capsicum annuum and Its Expression Analysis Responses to High Temperature Stress[J]. Biotechnology Bulletin, 2023, 39(1): 187-198.

Figures/Tables 9

Table 1 Primers used for quantitative real-time PCR

基因名称 Gene name	上游引物 Forward primer（5'-3'）	下游引物 Reverse primer（5'-3'）
CaDNAJE2	TACCCCGAAAAGTGTCCGC	TCGTCGAAAACCACCCTATGAG
CaDNAJE3	TTCTGCTCTATGGGGCTCTTAC	TTTTCCTCCACGGGTTCCT
CaDNAJE11	AACACTCAGCCCTGCTTCCC	GCTGAATGCCACTCCCTTGAC
CaDNAJE13	TGGTGGTGCCGATGAGATT	GCATTCGTCGCAGAGGATAA
CaDNAJE15	CACCAAAGTGAGGAGCATCG	AATCCACCAAGAAATCCAGCA
CaDNAJE17	ATTCCAATCCCAGCCACAA	CAACAGCAGCGATGACCAA
CaDNAJE19	GCTCAGCAAGCGGTAAAT	TGCCATCACCATCCCAGT
CaDNAJE21	CCCTTTCACTTCCAACCCC	TCAGTCCCATTTAGCCCATTTA
CaDNAJE23	CGGTGCCGCTCTGGTGAAATC	CGACAACCCACGTCCCCATCT
CaDNAJE24	TGGGAAGAGTCTATTGATGAACG	ATTGGTAGACACGCTTGGTTTT
CaDNAJE27	ACGCAAAGTAAGAAGAAAGCAGC	CGAGAACAGTATCCAAGCCCAC
CaHsfB5	AAAGTTCCAGAAAGGGTGC	TCCATACATTGAGTGAGGC
CaHSP22.0	CCCCACTCACCATCCCAAAA	CCCCACTGATTCTCAACACCCTA
CaHSP16.4	ATGTCAAAGATGATCAGCTTATTG	GGGTACTCAACACGGGACAC
CaUBI3	GGACCCAAACCCCGATGAT	CACCCCAAGCACAATAAGACA

Table 2 Conserved motif of the DNAJE proteins in pepper

基序 Motif	长度 Length/aa	氨基酸保守基序 Conserved sequence of amino acid
motif 1	20	CEGCGGSGFVPCSTCNGSGK
motif 2	33	EDKIVIYFTSLRGIRKTFEDCNTVRMILESFRV
motif 3	45	LQNVLEGKKVSLPRVFIKGRYIGGAEEIKQL- HEEGELKKLLEGJP
motif 4	15	CPECNENGLIRCPIC
motif 5	15	VDERDVSMDSGFREE

Fig. 1 Chromosomal location of DNAJE gene family members in pepper（Capsicum annuum） Chromosomal color ranging from green to red indicates the density of gene

Fig. 2 Phylogenetic tree of DNAJE gene family members in pepper, Arabidopsis and tomato（Solanum lyc-opersicum） The blue solid dot indicates pepper DNAJE protein, yellow triangle indicates tomato DNAJE protein, and red star indicates Arabidopsis DNAJE protein

Fig. 3 Analysis of conserved motifs, gene structure and cis-acting elements of DNAJE gene family in pepper A: CaDNAJE protein evolution. B: Conserved motif. C: Gene structure. D: cis-acting element distribution

Fig. 4 Collinearity analysis of DNAJE gene family between pepper and Arabidopsis Chr and the number 1-5 indicate the chromosomes of pepper and Arabidopsis, respectively. The red lines indicate DNAJE collinearity between interspecific. The purple lines indicate DNAJE collinearity between intraspecific. The Gray lines indicate represent collinearity of all gene membres between interspecific

Fig. 5 Expression profile analysis of pepper DNAJE genes in different tissues and fruit development A, B, C and D indicate the clustering of different expression patterns

Fig. 6 Expression analysis of genes related to high temperature stress in pepper * indicates significant differences at 0.05 level, and ** indicates significant differences at 0.01 level

Fig. 7 Physiological and biochemical responses of pepper seedlings to high temperature stress

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