Cloning And Functional Validation Studies Related To Transcription Factors MYB46, A Hawthorn Lignin Synthesis Gene

Hawthorn (Crataegus spp.), Also called shanlihong, Shan Lihong, is a kind of important deciduous tree in the rose family (Rosaceae). Hawthorn fruit development, a most important process is kernel hardening, This is related to the deposition of lignin. The hawthorn is covered in a hard, woody, nuclear skin (inner peel), This affects its edible value, Is also a problem in the processing. In Chinese hawthorn resources, there is a very important resources named soft core Hawthorn,the nucleus is soft, Inner peel semi lignin, Can be eaten with the flesh. In order to explore the molecular mechanism of the formation of the core of hawthorn, in this paper, we choose hawthorn with hard core and hawthorn with soft core as test materials, through Illumina Genome Analyzer Ⅱx, screen out MYB46 gene. To explore the relationship between MYB46 gene and lignin biosynthesis. The main research is as follows:1.By analyzing the sequence data of the Hawthorn group, Two genes that were annotated as MYB46 transcription factors were screened out,8_Unigene_BMK.9503 and 8_Unigene_BMK.10344.Two varieties were selected from hawthorn in hard core and hawthorn in soft core, Take the flower after the 18d-30d of hawthorn fruit,Through the analysis of qRT-PCR experiment. It showed that the 8_Unigene_BMK.9503 gene expression higer in hawthorn with hard core than hawthorn with soft core. Further bioinformatics analysis showed that, Only 8_Unigene_BMK.9503 is the real MYB46 gene. So we choose 8_Unigene_BMK.9503 gene for the next experiment, and named it CpMYB46.2.Primers for 8_Unigene_BMK.9503 region design of CDS gene from Hawthorn, Amplification of CpMYB46 gene from the ’Kaiyuan soft seed’ and ’weeping shanlihong’. Gene cloning Analysis by sequencing results, The similarity of the results of the ’Kaiyuan soft seed’ and the transcriptional group was found to 98.6%,’The weeping shanlihong’ similarity and transcriptome sequencing results is 98.02%.3.CpMYB46 gene expression vector was constructed,8_Unigene_BMK.9503 gene expression vector, Agrobacterium mediated transformation of Arabidopsis thaliana and apple, To express it in Arabidopsis and apple, There were 27 transgenic Arabidopsis plants with 8_Unigene_BMK.9503 gene and 6 Apple transgenic plants with 8_Unigene_BMK.9503 gene, The lignin content and CpMYB46 expression levels of Arabidopsis thaliana and apple transgenic plants showed that the CpMYB46 gene had the function of regulating lignin synthesis.