The Transcriptome Analysis Of Fruit Development And Verification Of Genes Related To Anthocyanidin Biosynthesis About ’Sanhuali’ (Japanese Plum)

Japanese plum(Prunus salicina Lindl.), which originates from Yangtze River Basin in China and then became a particular south subtropical cultivated population by artificial culture in South China, is the largest deciduous fruit trees in Guangdong Province.And the cultivated population mainly comprize ‘Sanhuali’ with red exo- and mesocarp,‘Zhusili’ and ‘Hongxianli’ with green exocarp and white mesocarp, ‘Sanyueli’ with red exocarp and white mesocarp et three categories. And the cultivated area of ‘Sanhuali’accounts for over 80% of the total area of plum in Guangdong. In this study, we sampled’Baicui Jimali’ and ’Huami Damili’ and by RNA-Seq, we mined the differentially expressed genes of fruit development, maturation and ripening and presented a reference gene set,mainly analysed differentially expressed genes of anthocyanin biosynthesis of fruit development and the cell wall metabolism related to fruit ripening softening. which can contribute to breeding varieties for lovely color and resistance to storage and transportation in future. The main findings are as follows:1. Based on transcriptome sequencing data of buds, flowers, embryo and different stage(75, 100, 130 days after flowers) fruits of ’Huami Damili’, we established a whole transcriptome database for ’Sanhuali’, and obtained 68,484 Unigenes. 33,630 Unigenes were annotated to GO, KEGG, NR, Pln TFDB etc databases. At the same time, 7,984 SSR were developed from Unigenes whose length are 1 kb or more.2. Based on four fruit transcriptome sequencing data of ’Huami Damili’, we screened and obtained 5,394 differentially expressed genes(DEGs). For those DEGs, with the transcription factor prediction analysis, KEGG analysis and expression patterns GO enrichment analysis, some highly expressed peroid genes module were found. With the protein to protein interaction network prediction analysis, some highly interaction modules were found in fruit developmaturation and maturation process. And we found that the most of predicted hormone-related genes were down-regulated and finally obtained 30 genes involved in hormone biosynthesis.3. By making an analysis to anthocyanin-related genes, we found that 41 structural genes, 26 transporter-related genes, 39 transcription factor and 3 down-regulated genes in pigment accumulation may be related to anthocyanin biosynthesis and transpoter.Based on successfully screened HMBS2 as quantitative reference gene, 10 anthocyanin structral genes and 9 MYB genes were verified with q RT-PCR. And the results show that compared to buds and flowers of ’Huami Damili’, the red color of fruit(130 days after flowers) may be affected by c23975.graph_c0(ANS) and c19863.graph_c0(UFGT),compared to ’Baicui Jimali’ ripe fruit(130 days after flowers), the deeper color of ’Huami Damili’ ripe fruit(130 days after flowers) may be affected by c21951.graph_c0(CHS2) and c19863.graph_c0(UFGT). And we found c6572.graph_c0 may be the one of effective MYB transcription factor that may have effect on fruit anthocyanins formation.we also have successfully cloned the CDS, full-length of c6572.graph_c0, and made predicted and found it contains R2R3 protein domain that is conservative in MYB transcription factor family.