biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 52:462-468, 2008 | DOI: 10.1007/s10535-008-0091-7

Cloning of differential expression fragments in cauliflower after Xanthomonas campestris inoculation

Y. Gu1,2, Y. W. Mao1, C. G. Wang1, Q. C. Zhao2, D. L. Sun2, W. Q. Song1,*
1 Department of Genetics and Cell Biology, College of Life Sciences, Nankai University, Tianjin, P.R. China
2 Tianjin Kernel Vegetable Research Institute, Tianjin, P.R. China

A near isogenic line (NIL) of Brassica oleracea var. botrytis with resistant and susceptible lines C712 and C731, was used in this study. More than 100 differentially expressed cDNA fragments were obtained from black rot resistant cauliflower plants obtained using cDNA-amplified fragment length polymorphism (AFLP) after infection with the pathogen. Thirteen of these fragments were cloned and subjected to reverse Northern blot analysis using both infected and control cDNA pools. Two positive clones, M2 and M6, were isolated. Northern dot blot and Northern blot analyses showed that M2 was constitutively expressed, whereas M6 contained a gene that was differentially expressed during pathogen infection. Moreover, M6 cDNA fragment was also highly expressed 16-24 h after H2O2 treatment. Southern blots showed that M6 is a single copy gene in the cauliflower genome, and encodes a protein with 84 % homology to gene on Arabidopsis chromosome 1. The deduced M6 protein has 91 % positive homology with the Arabidopsis 2A6 protein, which regulates ethylene synthesis; 76 % homology with a 1-aminocyclopropane-1-carboxylate oxidase (ACO), the last enzyme in ethylene synthesis; and 70 % homology with an ethylene induced DNA binding factor. These results suggest that M6 gene fragment is a new H2O2 downstream defense related gene fragment and can be induced by Xanthomonas campestris pv. campestris and H2O2.

Keywords: black rot resistance; Brassica oleracea var. botrytis; cDNA-AFLP; H2O2 treatment
Subjects: Arabidopsis thaliana; Brassica oleracea; chromosomes; genetic transformation; Xanthomonas campestris

Received: September 1, 2006; Accepted: February 14, 2007; Published: September 1, 2008  Show citation

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Gu, Y., Mao, Y.W., Wang, C.G., Zhao, Q.C., Sun, D.L., & Song, W.Q. (2008). Cloning of differential expression fragments in cauliflower after Xanthomonas campestris inoculation. Biologia plantarum, 52(3), 462-468. doi: 10.1007/s10535-008-0091-7
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