Effect Of AmiRNA-mediated FaCRYs Genes On Anthocyanin Biosynthesis In Strawberry

Strawberry(Fragaria�ananassa)is perennial herbaceous plants of Rosaceae Fragaria. The fruit contains a high concentration of flavonoids compounds, which has important health functions and significant economic value, so it is much loved by consumers and planters.Anthocyanin is one of the most widely distributed and widely studied in flavonoid compounds. It plays an important role in improving the quality of strawberry fruit.In recent years, the impact of environmental factors on the anthocyanin accumulation is gaining increasing attention, especially the effect of light the accumulation of anthocyanin. Many studies have shown that photoperiod, light intensity and light quality have influence on the anthocyanin synthesis, of which light quality plays a more important role, and short wavelengths light is more conducive to the accumulation of a large number of anthocyanin. However, light quality is how to control the anthocyanin accumulation, and its accumulation mechanism is not yet clear, needs to be further studied.In this paper, based on the materials of'Benihoppe'strawberry, using the amiRNAi technique and transgenic technique, silencing the FaCRYl and FaCRY2 genes in the genome at the same time, and final, we obtain the Facrys double mutant plants. Then treate mutant plants under white and 7:3 ratios of red light and blue light, by measuring anthocyanin content and the relative levels of expression of anthocyanin related structural genes and the relative levels of expression of anthocyanin related transcription factors, To study the function of FaCRYs in strawberry anthocyanin synthesis. The results obtained are as follows:First of all, we designed special primers according to the NCBI landing FaCRYl and FaCRY2 gene CDS sequences, and cloned the fragment of the strawberry FaCRY1 and FaCRY2 structure sequence. Sequence analysis showed that this sequence contains a FAD structure domain, it provides molecular sequence information for designing amiRNA interference.Second, use the methods of gene synthesis, successfully constructed the pre-amiRNA silence carrier by using MIR390a, and it provides carrier for transgene operation. Based on the materials of'Benihoppe'strawberry leaves, by using of agrobacterium mediated gene delivery technology and tissue culture technology, successfully obtain the double mutant plants.Third, the mutant plants and the negative control plants are treated under white light and 7:3 ratios of red light and blue light, then measure the anthocyanin contents and the expression of anthocyanin synthesis related genes. Analysis found that FaCRYl and FaCRY2 positive regulation of synthesis of anthocyanin, and its main function with anthocyanin synthetic regulation related transcription factors, to manage synthesis of anthocyanin through inhibiting or activing the transcription factors; speculated That FaCRY2 can act directly on the upstream regulatory factors of synthesis of anthocyanin gene, FaCRYl may control synthesis of anthocyanin through regulate upstream of CHS gene.