Identification of GRAS Gene Family and Expression Analysis Under Low Temperature Stress in Actinidia chinensis

doi:10.13560/j.cnki.biotech.bull.1985.2023-0324

Abstract

Abstract:

GRAS gene family is widely involved in plant growth and stress response. Low temperature is one of the important factors that restricts the production and distribution of kiwifruit（Actinidia chinensis）. The GRAS gene family of kiwifruit was identified, its expression under low temperature stress was analyzed, which may provide a theoretical basis for the research on cold resistance and variety breeding of kiwifruit. The GRAS gene family conserved domains were compared with the genome of A. chinensis ‘Hongyang’ reference, and the identified family members were analyzed by phylogenetic tree, protein physical, chemical properties, gene structure, protein tertiary structure, protein motif, cis-acting elements, collinearity, codon usage preference and gene expression mode. The results showed that there was a total of 79 GRAS family genes in kiwifruit genome, belonging to 8 subfamilies, and there were differences in gene and protein structures among each subfamily. Analysis of cis-acting elements showed the genes were involved in multiple plant hormone, growth and development, and stress response. Through codon preference analysis, it was found that the third base of GRAS family preferred to use pyrimidine bases G/T. Six genes were identified that may be involved in the low-temperature stress process in kiwifruit, and this hypothesis was validated by fluorescence quantitative PCR.

Key words: kiwifruit, GRAS gene family, codon analysis, low temperature stress

MAO Ke-xin, WANG Hai-rong, AN Miao, LIU Teng-fei, WANG Shi-jin, LI Jian, LI Guo-tian. Identification of GRAS Gene Family and Expression Analysis Under Low Temperature Stress in Actinidia chinensis[J]. Biotechnology Bulletin, 2023, 39(11): 297-307.

Figures/Tables 12

Table 1 Primers used for quantitative real-time PCR

基因Gene	正向引物Forward primer（5'-3'）	反向引物Reverse primer（5'-3'）
AcGRAS32	ACATTAGGCTCCCACCACTG	CTCCCGAGTCATTCAACCAT
AcGRAS3	AGTTCCGGATGTTGAGGTTG	TTTGAAGCGGGGTAAGAATG
AcGRAS53	TTCCAATCCTGCAATGAACA	AGGAATGATGCGGATCTGAC
AcGRAS79	GGTCGATGATCTTTCCGTGT	TCCCGAAGAGCTTCAGAAAA
AcGRAS6	GAACGAAGGTTTTCGCTCTG	ACTGGATCCCCTGAGCTTTT
AcGRAS42	AGTCGGCGATGAGCTTAAAA	GGAACCTTGTACAGCCGAAA
Actin	GTGCTCAGTGGTGGTTCAA	GACGCTGTATTTCCTCTCAG

Fig. 1 Phylogenetic evolutionary tree of kiwifruit GRAS gene family

Fig. 2 Physical and chemical properties of GRAS family

Fig. 3 Genetic structure of GRAS family

Fig. 4 Protein tertiary structure of some GRAS genes

Fig. 5 Domain of GRAS family

Fig. 6 Cis-acting element of GRAS gene family

Fig. 7 Collinearity analysis of GRAS family

Fig. 8 ENC-plot curve of kiwifruit GRAS family genes

Fig. 9 PR2-plot of kiwifruit MYB family genes

Fig. 10 Analysis of GRAS gene expression in kiwifruit during overwintering 1-3 represents kiwifruit branches in overwintering period on November 15, 2020, January 15, 2021 and March 15, 2021 respectively

Fig. 11 Expression analysis of 6 kiwifruit GRAS genes under low temperature treatment Different letters indicate that there are significant differences in genes at the level of 0.05 under different treatment times. *, ** indicated significant difference between treatment and control at 0.05 and 0.01 levels

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