过量表达RdreB1BI对草莓果实品质及相关基因的影响

doi:10.3864/j.issn.0578-1752.2018.07.013

Abstract

Abstract: 【Objective】 The effect of exogenous RdreB1BI transformation to the fruit quality and related gene expression in ‘Benihoppe’ strawberry were analyzed to reveal the function and molecular mechanism of quality regulation in transgenic strawberry fruit. 【Method】 The edible organs on five RdreB1BI transgenic strawberry lines and non-transgenic ‘Benihoppe’ strawberries (fruits in full red stage) were used as material to test and determine the vertical diameter, transverse diameter, weight, contents of flavor substance (soluble sugar, soluble protein, ascorbic acid) and coloring materials (anthocyanin, flavonoids and total phenol). The structure of the exogenous gene and seven secondary metabolites synthesis pathway-related genes (RdreB1BI, Fractin, FvC4H, FvCCR2, FvGST, FvF3H, FvDFR and FvMYB306) and the elements of gene promoter were analyzed and were predicted by bioinformatics methods such as BLASTn, GENEFINDER, and PlantCARE. The expression of related genes was detected by qRT-PCR. The data were analyzed by 7300 system software and 2^-△△Ct method. The variance and correlation analysis of physiological, biochemical and molecular data were analyzed. The effects of RdreB1BI on the fruit quality and related gene expression of ‘Benihoppe’ strawberry were discussed comprehensively. 【Result】 The fresh weight of transgenic and non-transgenic strawberries was ranging from 11.75 to 15.42 g. The vertical and transverse diameters were ranging from 35.12 to 40.42 mm and 28.73 to 32.6 mm, respectively. Only the volume diameter of transgenic line 8 was significantly higher than that of line 7, and there were no significant differences between the other samples. The contents of anthocyanin in line 1 and line 7 were significantly higher than those of the control. The contents of total phenol in transgenic line1 and line 7 were significantly higher than that in control. The contents of total phenol in each line were significantly higher than that in the control. The soluble sugar contents in the transgenic line 1, line 7 and line 8 were significantly higher than that of the control (21.70 mg?g^-1 FW), which were 2.87, 3.39 and 3.35 times of the control. There was a positive correlation between soluble solids contents and soluble sugar contents (r=0.811*), but there was no significant difference in the soluble solids contents of the samples. The contents of amino acids in the fruiting fruits of the transgenic lines were ranging from 0.2580 to 0.3950 g/100 g FW. The content of amino acids in the control was 0.5151 g/100 g FW, which was significantly higher than that of the transgenic lines. The contents of titratable acid in the transgenic line 1 and line 7 were significantly higher than that in the control. The content of ascorbic acid in line 1 was 168.35 mg/100 g FW, which was significantly higher than that of the wild-type of 92.50 mg/100 g FW. The contents of soluble protein in transgenic line 9 and line 10 were 25.97 and 25.86 mg?g^-1 FW, respectively, which was significantly higher than that of control (22.93 mg?g^-1 FW). The expression levels of RdreB1BI, FvCCR2 (cinnamoyl-CoA reductase 2-like) and FvMYB306 (myb-related protein 306) in the transgenic lines were significantly higher than that of the wild-type. It was found that the promoter region of differentially expressed genes contained a variety of higher plant cis-acting elements, mainly were CAAT-box and TATA-box, which had enhanced transcription efficiency of eukaryotes. It also included some the cis-acting elements such as G-Box, G-box, MBS, ARE, 5UTR Py-rich stretch, which can affect the response of each gene to light and play a regulatory role in the phenylpropane metabolic pathway. 【Conclusion】 RdreB1BI regulated the related genes involved in the development and maturation of transformants, improved the effectiveness of light, activated the key genes of flavonoid biosynthetic pathway. Differentially expressed genes contained a large quantity of light-induced response elements. The expression of FvCCR2 and FvMYB306 promoted the synthesis of secondary metabolites such as flavonoids, phenols, anthocyanins and total phenols. The transformation of RdreB1BI gene resulted in a significant increase in the nutrient and coloring material contents and improved the fruit quality in strawberry.

Key words: strawberry (Fragaria×ananassa Duch.), RdreB1BI, overexpression, fruit quality, promotor

YAN YiChao, WAN ChunYan, GU XianBin, GUO ChengBao, CHEN YueHong, GAO ZhiHong. Effect of RdreB1BI Gene Overexpression on Fruit Quality and Related Gene Expression in Strawberry[J].Scientia Agricultura Sinica, 2018, 51(7): 1353-1367.

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