Cloning And Expression Analysis Of CHI And F3'H Genes In Impatiens Uliginosa

The Impatiens uliginosa is an annual herb of the Impatiens genus,which is mainly distributed in the southwest and south China.The flower pattern is peculiar,the color is beautiful,the tolerance is strong,and it has certain application value in the garden.As a more important trait of ornamental plants,flower color has always been one of the most important breeding objectives of gardening and gardening workers.In this paper,we mainly studied the CHI and F3'H genes in the synthetic pathway of phosin anthocyanin,and cloned the full-length cDNA,and performed real-time fluorescence quantitative expression,focusing on the sequence characteristics of CHI and F3'H genes and their The role in the formation of Impatiens uliginosa.The results obtained through experiments are as follows:(1)Two copies of the CHI gene of Impatiens uliginosa were obtained from the transcriptome sequencing results.The full-length cDNA and intron of the CHI gene of Impatiens uliginosa were obtained by RACE technique and DNA whole genome amplification using the RACE technique and DNA whole genome amplification.The following results were obtained: the cDNA sequence of IuCHI1 gene was 940 bp in length and 741 bp in exon.Containing three introns;the IuCHI2 gene has a full-length cDNA of 949 bp and an exon of 747 bp,containing three introns.The sequence homology analysis of the CHI gene by nucleotide blast showed that the mRNA sequences of CHI gene showed high homology with many species in the Gennebank database,such as golden tea,pomegranate,purple bell Suzuki,pear,Olives,etc.(2)Three copies of the Impatiens uliginosa F3'H gene were obtained from the transcriptome sequencing results.The full-length cDNA and intron sequences of F3'H gene of Impatiens uliginosa were obtained by RACE and DNA whole genome amplification using the RACE technique and DNA whole genome amplification.The following results were obtained: the full-length cDNA sequence of IuF3'H1 gene was1683 bp.The exon is 1560 bp and contains two introns.The cDNA sequence of IuF3'H2gene is 1683 bp in length and 1560 bp in exon.It contains two introns.The full-length cDNA sequence of IuF3'H3 gene is 1586 bp,exon.1560 bp,containing two introns.Sequence homology analysis of the F3'H gene by nucleotide blast showed that the mRNA sequence of the F3'H gene showed high homology with many species in the Gennebank database,such as grapes,rhododendrons,sesame seeds,rose,and morning glory.Flowers,etc.(3)The phylogenetic tree constructed by MEGA7,together with the other genes in the NCBI,was sequenced by MEGA7.The results showed that IuCHI1 and IuCHI2 of Impatiens uliginosa were clustered,indicating that the CHI gene of Impatiens uliginosa is It is more conservative during evolution.The closest relationship with the CHI gene of Impatiens uliginosa is white celery,which is an aquatic herb,and the phoenixphoenix is also often found in streams or shallow waters,indicating that the phylogenetic tree results are partially in accordance with the botanical tradition.This indicates that CHI gene has certain reference value in the research and application of its own phylogeny;the system constructed by MEGA7 together with the amino acid sequence encoded by the open reading frame of the F3'H gene mRNA sequence of Impatiens uliginosa and other species in NCBI The developmental tree showed that IuF3'H2 and IuF3'H3 of Impatiens uliginosa were clustered together,while IuF3'H1was in another branch,indicating that IuF3'H1 may be the collateral of IuF3'H2 and IuF3'H3 source.(4)By quantitative analysis of the fluorescence of CHI gene and F3'H gene in the four developmental stages of red and white squid,the CHI gene can be found in the four developmental stages of the colored flower red scorpion phoenix(flower bud)The expression of CHI gene may have an intrinsic correlation with the accumulation of red anthocyanin in the period of initial period,initial bloom,full bloom,and Xiehua period.During the four developmental stages of phoenix,the expression of CHI gene was the highest in the third period,and the expression of CHI gene was relatively small in other periods.It is speculated that it may be related to the accumulation of flavonoids.The expression level of F3'H gene in the colored flower red Impatiens uliginosa is higher than that in the colorless flower white Impatiens uliginosa,and the expression of F3'H and the pigment formed by the phoenix water phoenix color are presumed.Accumulation has a certain relationship.