| Cotton Verticillium wilt, a vascular disease of cotton caused by fungal soil-borned pathogen Verticillium dahliae, is a kind of worldwide spread diseases that is the most difficult to be controlled in cotton production. It is strongly considered that Developing resistant and tolerant cultivar is the most economic and effective ways to resolve this problem. Therefore, recently research has been focused on deeply understanding of the molecular mechanism about interactions between V. dahliae and the host, and also it does become the research hotpoint to clone the responsing genes which expressed during the affection stage.The objectives of this research were attempted to obtain the resistant-related candidate genes from upland cotton to offer new gene sources for transgenic breeding by identifying the response cDNA induced by the toxin of V. dahliae. Total RNA was isolated from the seedling roots of upland cotton (G. hirsutum L.var. zhongmiansuo12) induced by toxin of the pathogen V. dahliae stain V991 after 24 hours. Double strand cDNA was synthesized followed the protocols of the Smart?cDNA Systhesis Kit (CLONTECH Co.). 180 clones were acquired by the method of Supression Subtractive Hybridization, and 15 cDNA clones were identified positively using Reverse Northern Blot, which was named as Response Gene induced by toxin of Verticillium dahliae, for short as Vdrg. Sequences of Vdrg were analyzed online in the NCB1 Website and molecular mapping were also carried out by using two populations derived from G. hirsutum and G. barbadense hybrid combination of Pima S6 + Acala Maxxa and Pima S6 X Tamcot GCNH. The results showed as follows:Vdrgl has 501bp contained NBS-LRR conservative motifs, it seems like the NBS-LRR type resistance protein gene and locates on acme of Linkage Group D02, 5.6cM apart from marker pAR286E4C and Unig22D03bE6D. Vdrg2 (1048bp) is Serine/threonine phosphatases like protein gene and locates on Chr. 18(D), 166.0 cM apart from acme and 0.9 cM apart from marker Unig06G03aE6R, and also located on L.G..A01, 141.8cM apart from acme, 0.5cM from marker Coau2L21aE4D and 1.1cM from marker Coau2I17E4C, respectively. Vdrg3(409 bp) is likely to Zn-depdent hydrolyzase gene, locates on L.G..D04, 69.4cM apart from acme, 0.1 cM from marker A1212+E5C, and 0.8cM from marker A1682E6C, respectively. Vdrg4 (1538 bp) is PTS_HPR_SER Serine/threonine phosphatases like gene and locates on Chr.23(D), 38.1cM apart from acme, 0.9cM from GatelBD04cE6R (A01A05) and 4.6cM from Gafb23A18E4C. Vdrg5 (856bp) and Vdrg11 (757bp) are phosphofructokinase like genes. Vdrg5 locates on L.G.D02,165.4cM apart from acme, 3.7cM from pAR922E4D and 5.2cM from marker pAR570bE3C. Vdrg 11 was hybridized badly. Vdrg6(599bp, Vdrg7 (986) and Vdrgl 5 (231bp) are protein kinases like genes; Vdrg6 locates on L.G.A06, 77.7cM apart from acme, 1.0cM from marker Unig23D03bE3C and 0.7cM from marker pAR071E3C, and also locates on Chr.14(D), 102.1cM apart from acme, 0.2cM from marker A1449bE2D, 3.3cM A1222.09, respectively. Vdrg7 (986) and Vdrgl 5 show badly hybridizatioaVdrg8(409 bp) and Vdrg9 (409 bp) are chytochrome oxidize and deoxidize enzyme like genes, locate on L.G.D07,43.4cM apart from acme, 0.3cM from marker Gate4CA09cE6C and 1.6cM from marker pAR078cE3C, respectively. Vdrg10(632 bp) is glycosylationate protein like gene and locates on the L.G.A01, 56.9cM apart from acme, 0.9cM from marker pAR058E4C, 2.2 cM from marker pAR413aE3C, and also locates on the L.G.A02, 46.7cM apart from acme, on the same site with marker pVNC244E3C, 2.6cM apart from marker Gate2BF03E4, 5.4cM from marker Coau2A22bE3R, respectively. Vdrg12 (708bp) and Vdrg13(707) are hydroyase like genes and locate on the Chr.l8(D), 95.0cM apart from acme, 0.4cM from MSAT1721C and 2.2cM from Gate3AH03cE3C, respectively. Vdrg14 (606 bp) is predicted as cytochrome C like protein gene and located on the Chr.04(A), 37.1cM apart from acme, 1.4cM from Gate3BEllbE3D, 5.3cM apart from marker Unig28E02E5C.
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