Transcriptome Analysis And Anthocyanin Biosynthesis Genetic Expression In The Spathe Of Anthurium Andreanum

Anthurium andreanum is an important economic value as pot and cut flower. Its spathe is the main ornamental part, and the color is the most important factor to decide the commodity value.Traditional cross breeding needs long time and has a large variation in offspring, it’s difficult to selection.Mutation breeding gains mutant plants in a small variation rate, and it is hard to get the variation. Molecular breeding is a new method of cultivating new varieties. However, the lack of molecular study in A.andreanum background makes the molecular research lag behind. To improve the efficiency of A.andreanum breeding and selection, it is necessary to set up the basis for molecular genetic studies in A.andreanum. Thus establish the genome database for A.andreanum has a great significance. High-throughput sequencing, Illumina HiSeq/MiSeq method was adopted to do Transcriptome Sequence at 8 different development stages (S1-S8) of A.andreanum spathe. Meanwhile, S5 (Blooming phase:Red) and S8 (Senescence phase:Green) stages were measured by the Digital Gene Expression Profiling (DGE) in this study. The transcriptome database of A. Andreanum was enriched and the key genes in the pathway of anthocyanin biosynthesis related to spathe color were found out. This research provided a basic reference background for gene cloning, molecular marker selecting as well as genetic improvement breeding in A.andreanum. Main results are as follows:1. Through the whole stages of A.andreanum, a total of 56445573 clean reads (about 5.64 G) were obtained by Illumina HiSeq/MiSeq, including 64576 unigenes, and the length of unigenes was ranged 200 to 2000bp.Enriched the information of A.andreanum.2.12766374 (1.28G) and 18583390 (1.86G) trancriptome data were obtained by DGE at the stage of S5 and S8, respectively. There were 2316 unigenes had the different expression.3. Combines Transcriptome with Digital Gene Expession,10 different expression gene sequences were selected in the anthocyanin biosynthesis of A.andreanum spathe colour between S5 and S8. 10unigenes were annotated in 8 genes by blasting in NCBI, including Chalcone Synthase (CHS), Chalcone Isomerase(CHI), Flavanone-3p-hydroxylase(F3H), Flavonoid-3’-hydroxylase(F3’H), Dihydroflavonol-4-reductase(DFR), Anthocyanidin Synthase(ANS), Leucoanthocyanidin Reductase(LLAR) and Cinnamate-4-hydroxylase(C4H). Speculated that these 10 gene sequences play a key role in process of change in color in A.andreanum spathe development.4. Specific amplification products of 10 gene sequences were gained. qRT-PCR verification showed that 10 gene sequences expression quantity were down-regulated in S8, and it had the same tendency with DGE results. So,it proved the accuracy of the DGE sequencing results.5. By monitoring 10 gene sequences expressionin A.andreanum spathe development, S3 is a strong stage of anthocyanin synthesis. During S5, S6, S7 and S8 period,10 gene sequences expression quantity decreased gradually as spathe mature and were reduced at least in the S8 stage. The 10 gene sequences were the key gene during the color formation of A.andreanum spathe.