| Research on the relationship between the content of gossypol and tannin in different cotton varieties and the resistance of resistance to verticillium wilt defoliating relevance,while it also clear the relationship between disease resistance and the content of gossypol and tannin in different cotton tissues. This article established a determination method to detect the content of gossypol and tannin. Using Verticillium dahliae spore suspension to infect 12 cotton varieties which is with different disease resistances, and determining the content of gossypol and tannin in the leaves and roots.at the same time, select the corresponding cotton varieties which is not inoculated as controls;When plants were abused by pathogenic bacteria,they would secrete a kind of protein compounds, which was a kind of active defense response inside plants.PevD1 is a secrete elicitor from Verticillium dahliae, it was very significant to detect disease resistance of cotton on Verticillium dahilae with investigate the induce resistance and mechanisms of PevD1; An ANR gene related to secondary metabolite was cloned by RT-PCR technologies. And using bioinformatics analysis science and differential gene expression analysis of biological information, explain its role in Gossypium barbadense tannin biosynthetic approach. Provide scientific theoretical basis for the future regulation of the use of gene technology to change the resistance of cotton verticillium wilt resistance increased. Conclusions are as following:1. The measurement method has been improved and indicate that the determination method established in this topic had high precision and good repeatability(gossypol RSD=1.06% and tannin RSD=1.64%) as well as high recovery rate and accuracy degree(Recovery of gossypol 98%~102%, tannin 98%~102.3%). After induced by Verticillium wilt of cotton,gossypol content was higher than that not induced, in the case of no inoculation,the average content of gossypol in the roots is 8.65% higher than that in the leaves, and after inoculation, the content is 4.56% higher. Moreover,the content of gossypol in disease-resistant cultivars was always higher than that in susceptible cultivars, and there was a significant positive correlation between the content of gossypol and the disease resistance of cotton varieties(leaves without inoculation R2=-0.877, leaves after inoculation R2=-0.886,roots without inoculation R2=-0.982 and roots after inoculation R2=-0.882). In the case of no inoculation, the content of tannin in leaves was 4.56% higher than that in roots, and after inoculation, the content is 1.90% higher. The content of tannin in the leaves and roots of the relative disease index varieties without inoculation was always higher than the susceptible cultivars. But the trend was contrary after inoculation. There was a significant negative correlation between the tannin content and the relative disease index of cotton varieties(leaves after inoculation R2=-0.985, roots after inoculation R2=-0.976), while there was asignificant positive correlation in the case of no inoculation(leaves without inoculation R2=0.961, roots without inoculation R2=0.763).2. PevD1 inducible can enhanced the expression of pathogenesis-related genes LAR、PAL 、 DFR 、 CCR 、 cadinene synthase,and it can also increase the activity of defense enzyme,such as PAL, CAT, SOD.Explanation PevD1 protein triggered the plant immune responses and provided a scientific basis for elucidating plant-elicitor interactions.3. Cloned a cotton Verticillium wilt resistant realated Gene,which is named GbANR gene,and the GenBank accession number is KR107510, GbANR gene contains an Open Reading Frame of 1011 bp,which was encoded a polypeptide of 336 amino acids. A comparison of the nucleotide sequences of homologous ANR genes from some other plants indicated that GbANR gene had a range of 72% to 99% identity in nucleotide sequence with homologues of other plants.the predict result shows that the molecular weight was 36.326 kD and an isoelectric point was 5.55. The result of Real-Time PCR indicated that the expression level of GbANR gene in leaves is higher than that in stem and root, and the expression level of the resistant cotton variety xinhai13 was higher than that in those of the susceptible cotton variety H.D-2. Verticillium wilt fungus stress can quickly induce the expression of GbANR in disease-resistant material leaves. And analyze of the expression pattern found that the gene may involved in the early response of Verticillium dahilae in roots. |
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