Enzyme Activity Characteristics and Expression of Cu/Zn-SOD Gene in Tea Flowers

doi:10.3969/j.issn.1000-2561.2020.06.014

Abstract

Abstract:

In order to study the expression characteristics of Cu/Zn-SOD gene in the flowers within different developmental stages and various cultivars of tea plant flowers, and to discuss the correlationship between the expression of Cu/Zn-SOD gene and its enzyme activity, four developmental stages of cultivar ‘Longjing 43’ tea flower and 15 tea plant cultivars of fresh flowers were studied. The expression of three Cu/Zn-SOD genes was quantitatively analyzed by real-time fluorescent quantitative PCR techniques, and the enzyme activity of the three Cu/Zn-SOD was also determined simultaneously. The lowest Cu/Zn-SOD enzyme specific activity was found in the fourth stage, and the difference was significant when compared with the other developmental stages. The specific activity of Cu/Zn-SOD enzyme in different cultivars of tea flowers was significantly different. The Cu/Zn-SOD enzyme specific activity of ‘Zijuan’ tea flower was the highest, followed by ‘Huangjinya’ and ‘Baiye 1’. The activity of Cu/Zn-SOD enzyme of ‘Huangguanyin’ tea flower was the lowest. The expression level of Cu/Zn-SOD differed among the developmental stages, and the expression of cytoplasmic Cu/Zn-SOD1 gene was much higher than those of chloroplast type Cu/Zn-SOD2 and peroxisome Cu/Zn-SOD3. The expression of Cu/Zn-SOD in different cultivars was significantly different. Cu/Zn-SOD1 had the highest expression in cultivar ‘Huangmudan’ tea flower, Cu/Zn-SOD2 gene had the highest expression in cultivars ‘Baiye 1’ and ‘Huangguanyin’ tea flowers, and Cu/Zn-SOD3 gene had the highest expression in cultivar ‘Zijaun’ tea flowers. The result of correlation analysis showed that the specific activity of Cu/Zn-SOD had no correlation with Cu/Zn-SOD gene expression in different developmental stages of ‘Longjing 43’ tea flowers, but had considerable correlation with Cu/Zn-SOD3 gene expression in different tea cultivars. In this study, both tea cultivars and suitable plucking stage with high Cu/Zn-SOD activity were preliminarily expounded, which would provide references for the further research on SOD.

Key words: tea plant flowers, Cu/Zn-SOD gene, real time fluorescence quantitative PCR, Cu/Zn-SOD enzyme activity, cultivar ‘Longjing 43’

CLC Number:

S571.1

WANG Mengxin,WU Guohuo,CUI Lin,HAN Baoyu. Enzyme Activity Characteristics and Expression of Cu/Zn-SOD Gene in Tea Flowers[J]. Chinese Journal of Tropical Crops, 2020, 41(6): 1167-1173.

Figures/Tables 9

Fig. 1 Tea flowers in four developmental stages

Tab. 1 Primer sequences for expression analysis of target genes

基因Gene	NCBI登录号 Accession in NCBI	引物Primer	序列Sequences
Cu/Zn-SOD1	KR078346	CSD1-F	ATGCTGGTGGAAGGATTG
Cu/Zn-SOD1	KR078346	CSD1-R	ATCCCAAGTCTCAACCAAT
Cu/Zn-SOD2	KR078347	CSD2-F	ACGAACTTAGCCTTTCCACT
Cu/Zn-SOD2	KR078347	CSD2-R	CAAGCAAGTCACTAGTACACAG
Cu/Zn-SOD3	KR819175	CSD3-F	AGGTGCAAGAGTCGGATGT
Cu/Zn-SOD3	KR819175	CSD3-R	AGCGAGATGATATGAGGGTT
GAPDH	GE651107	GAPDH-F	TTGGCATCGTTGAGGGTCT
GAPDH	GE651107	GAPDH-R	CAGTGGGAACACGGAAAGC

Fig. 2 Specific activity of Cu/Zn-SOD in different developmental stages of ‘Longjing 43’ tea flower

Fig. 3 Difference in activity of Cu/Zn-SOD in tea flowers of fifteen tea plants cultivars Different lowercase letters represent significant difference among different cultivars at 0.05 level.

Fig. 4 Expression analysis of Cu/Zn-SOD genes in four developmental stages of ‘Longjing43’ tea flowers Different lowercase letters represent significant difference among stages at 0.05 level.

Fig. 5 Expression levels of three Cu/Zn-SOD genes in four developmental stages of ‘Longjing43’ tea flowers Different lowercase letters represent significant difference among different genes at the same stages at 0.05 level.

Fig. 6 Difference in expression levels of three kinds of Cu/Zn-SOD genes in tea flowers of fifteen cultivars of tea plants Different lowercase letters represent significant differences among different cultivars at 0.05 level.

Tab. 2 Correlation between the expression of each Cu/Zn- SOD gene and Cu/Zn-SOD activity within the third developmental stage in tea flowers of fifteen tea cultivars

基因 Genes	相关系数 Correlation coefficient	显著性水平 Significance level
Cu/Zn-SOD1	0.035	P>0.05
Cu/Zn-SOD2	0.185	P>0.05
Cu/Zn-SOD3	0.555	P<0.05

Tab. 3 Correlation between expression of every Cu/Zn- SOD gene with Cu/Zn-SOD activity in whole developmental stages of ‘Longjing 43’ tea flowers

基因 Genes	相关系数 Correlation coefficient	显著性水平 Significance level
Cu/Zn-SOD1	-0.789	P>0.05
Cu/Zn-SOD2	0.702	P>0.05
Cu/Zn-SOD3	-0.177	P>0.05

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