Cloning And Functional Verification Of Genes Related To Fusarium Wilt Resistance In Flavonoid Metabolism Pathway Of Island Cotton(Gossypium Barbadense L.)

Fusarium wilt is one of the important factors that threaten the production of Gossypium barbadense.L.To study the molecular mechanism of resistance to Fusarium wilt will provide solid foundation for cultivating resistant island cotton varieties and solve the problem of wilt disease.A total of 6 materials were selected for transcriptome and DEG analysis.The real-time quantitative(qRT-PCR)method was used to correlate the flavonoid metabolic pathway Genes were analyzed by disease resistance materials(06-146,Egyptian cotton 2,DJ-07-136,5917)and susceptible materials(Xinhai 14,Hai 92-3,Egyptian cotton 424).The genes related to flavonoid metabolic pathway were cloned and analyzed by bioinformatics,and the function of gene was verified by virus-mediated gene silencing.The specific results were as follows:1.The results of sequencing and DEG(Differentially Expressed Gene)analysis showed that the expression level of flavonoid metabolic pathway in disease resistant material was significantly higher than that in susceptible material,and the gene The expression level was higher than that before inoculation.2.The expression of ANR,CHI,ANS,CHS,DFR,TT7,and C4 H and FLS were analyzed by qRT-PCR.The results showed that the expression of ANR,CHI,The expression level in the diseased material was significantly higher than that in the susceptible material.The expression level of CHI gene in disease resistant material was significantly higher than that in susceptible material except 4 h.The expression of DFR gene was significantly higher than that of susceptible material at different times after inoculation.The expression level of TT7 gene did not change significantly in the susceptible material,and the expression level of TT7 gene was significantly up-regulated.3.The results showed that the expression level of TT7 and FLS gene was negatively correlated with the disease index at 40 h after inoculation,and the expression of CHI and DFR gene was significantly correlated with the disease index.The expression level was negatively correlated with the disease index.The expression level of CHI and LDOX gene was negatively correlated with disease index at 28 h after inoculation.There was a significant negative correlation between DFR gene and disease index at 10 h after inoculation.4.The flavonoid metabolic pathways genes: GbCHI,GbDFR and GbTT7 were cloned and analyzed by bioinformatics.The following results were obtained: GbCHI and GbDFR were acidic proteins except GbTT7.In addition to GbCHI,GbTT7 and GbDFR stabilized protein.GbCHI,GbDFR and GbTT7 are both hydrophilic proteins.GbDFR and GbTT7 have two and one transmembrane regions at the N-terminus.GbCHI belongs to the chalcone synthase family.GbTT7 belongs to the cytochrome P450 protease family,and GbDFR belongs to the SDR family,which is close to the rice,But far away from Gossypium arboreum,Gossypium hirsutum and Gossypium raimondii.It is speculated that these two genes may be related to resistance to Fusarium wilt in Gossypium barbadense.L.5.The virus-mediated gene silencing technique was successfully applied to the island cotton,and the positive control TRV-CLA appeared white phenotype 12-13 days after injection.The degree of whitening of different leaves was the first Three pieces of the fourth film.The results showed that VIGS-GbCHI and VIGS-GbDFR were more serious than the control,and the VIGS-GbDFR and VIGS-GbCHI were compared with the control.The incidence of VIGS-GbCHI was more severe than that of VIGS-GbDFR.It is speculated that GbCHI and GbDFR play an important role in the process of resistance to Fusarium wilt.