Effect of fruit maturity on UV-B-induced post-harvest anthocyanin accumulation in red Chinese sand pear

The effect of fruit maturity on UV-B-induced post-harvest anthocyanin accumulation in red Chinese sand pear (Pyrus pyrifolia Nakai) cultivar ‘Mantianhong’ was evaluated. During the irradiation, compared with the fruit harvested at 20 days before harvest (DBH) and 10 DBH, the mature fruit (harvested at commercial harvest date) had higher soluble solids content, soluble sugars concentration but lower firmness and starch content. In addition, higher content of anthocyanin has been detected in mature fruits than in immature fruits due to the significant increase in the expression of genes related to anthocyanin biosynthesis, especially PpCHS, PpF3H, PpANS, PpUFGT, PyMYB10 and PpbHLH in red Chinese sand pears. Hierarchical clustering analysis suggested that most genes related to anthocyanin biosynthesis showed a coordinate expression pattern. These findings are helpful in understanding the molecular mechanism of anthocyanin biosynthesis and regulation, which could lead to the development of new technologies for improving fruit color in Chinese sand pears and other fruits.     

Identification and functional analysis of anthocyanin biosynthesis genes in <Emphasis Type="Italic">Phalaenopsis</Emphasis> hybrids

Phalaenopsis species are among the most popular potted flowers for their fascinating flowers. When their whole-genome sequencing was completed, they have become useful for studying the molecular mechanism of anthocyanin biosynthesis. Here, we identified 49 candidate anthocyanin synthetic genes in the Phalaenopsis genome. Our results showed that duplication events might contribute to the expansion of some gene families, such as the genes encoding chalcone synthase (PeCHS), flavonoid 3′-hydroxylase (PeF3′H), and myeloblastosis (PeMYB). To elucidate their functions in anthocyanin biosynthesis, we conducted a global expression analysis. We found that anthocyanin synthesis occurred during the very early flower development stage and that the flavanone 3-hydroxylase (F3H), F3′H, and dihydroflavonol 4-reductase (DFR) genes played key roles in this process. Over-expression of Phalaenopsis flavonoid 3′,5′-hydroxylase (F3′5′H) in petunia showed that it had no function in anthocyanin production. Furthermore, global analysis of sequences and expression patterns show that the regulatory genes are relatively conserved and might be important in regulating anthocyanin synthesis through different combined expression patterns. To determine the functions of MYB2, 11, and 12, we over-expressed them in petunia and performed yeast two-hybrid analysis with anthocyanin (AN)1 and AN11. The MYB2 protein had strong activity in regulating anthocyanin biosynthesis and induced significant pigment accumulation in transgenic plant petals, whereas MYB11 and MYB12 had lower activities. Our work provided important improvement in the understanding of anthocyanin biosynthesis and established a foundation for floral colour breeding in Phalaenopsis through genetic engineering.     

Expression of genes involved in the anthocyanin biosynthesis pathway in white and red fruits of Fragaria pentaphylla and genetic variation in the dihydroflavonol-4-reductase gene

Structural and regulatory genes control fruit colors in plants. Real-time quantitative PCR results showed significantly higher expression levels of structural genes (FpCHS, FpDFR, FpANS, and FpUFGT) as well as of the regulatory gene MYB10 in red fruits of Fragaria pentaphylla compared to white fruits. These genes were strongly associated with anthocyanin accumulation within fruits. The full-length sequence of the FpDFR gene in red fruits of F. pentaphylla had a length of 2080 bp, was separated by five introns, and shared 95% homology with the F. vesca DFR sequence. Twenty-seven SNPs were detected in the FpDFR gDNA sequences between red and white fruits. Among these, transition substitutions were more frequent than transversions (66.7% vs. 33.3%), and a larger number of nucleotide variants existed in introns compared to exons (70.4% vs. 29.6%). A Chi-square test showed only three SNPs significantly associated with fruit color. Combined with structural analyses of the FpDFR protein and an expression analysis of the anthocyanin pathway genes, these results indicate that trans-regulation might contribute to color control in F. pentaphylla.     

RNAi-Mediated Silencing of the Flavanone 3-Hydroxylase Gene and Its Effect on Flavonoid Biosynthesis in Strawberry Fruit

Anthocyanin biosynthesis requires the activities of several enzymes in vivo. Flavanone 3-hydroxylase (F3H) converts flavanone into dihydroflavanol at an early step in the anthocyanin biosynthesis pathway. In this study we constructed an RNAi gene-silencing vector that encodes a hairpin F3H RNA. Agrobacterium strain GV3101 harboring the F3H RNAi vector was injected into strawberry fruits which were still attached to the plants 14 days after pollination. The phenotype was observed 10 days postinjection, and fruits were tested by RT-PCR and northern blot assays. The results showed that the F3H gene was downregulated by approximately 70 % in the agroinfiltrated fruits compared with the control. HPLC–MS analysis showed that anthocyanin content was greatly reduced, flavonol was also decreased, and the levels of p-coumaroyl glucoside and p-coumaroyl-1-acetate were markedly increased. We conclude that the precursors were shunted to the phenylpropanoid pathway, and that F3H is one of the key enzymes required for the biosynthesis of flavonoids in strawberry fruit. According to our results, reducing gene function via RNA interference is a rapid, simple, and effective way to identify gene function in strawberry fruit.