Molecular cloning of allelopathy related genes and their relation to HHO in Eupatorium adenophorum.
In this study, conserved sequence regions of HMGR, DXR, and CHS (encoding 3-hydroxy-3-methylglutaryl-CoA reductase, 1-deoxyxylulose-5-phosphate reductoisomerase and chalcone synthase, respectively) were amplified by reverse transcriptase (RT)-PCR from Eupatorium adenophorum. Quantitative real-time PCR showed that the expression of CHS was related to the level of HHO, an allelochemical isolated from E. adenophorum. Semi-quantitative RT-PCR showed that there was no significant difference in expression of genes among three different tissues, except for CHS. Southern blotting indicated that at least three CHS genes are present in the E. adenophorum genome. A full-length cDNA from CHS genes (named EaCHS1, GenBank ID: FJ913888) was cloned. The 1,455 bp cDNA contained an open reading frame (1,206 bp) encoding a protein of 401 amino acids. Preliminary bioinformatics analysis of EaCHS1 revealed that EaCHS1 was a member of CHS family, the subcellular localization predicted that EaCHS1 was a cytoplasmic protein. To the best of our knowledge, this is the first report of conserved sequences of these genes and of a full-length EaCHS1 gene in E. adenophorum. The results indicated that CHS gene is related to allelopathy of E. adenophorum.
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
Huiming
H
Guo
Xixiang
X
Pei
Fanghao
F
Wan
Hongmei
H
Cheng
D000217:Acyltransferases / Q000235:genetics; D036603:Ageratina / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000502:physiology*; D019747:Aldose-Ketose Isomerases / Q000235:genetics / Q000378:metabolism; D015139:Blotting, Southern; D003001:Cloning, Molecular; D019295:Computational Biology; D017124:Conserved Sequence / Q000235:genetics; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant / Q000235:genetics*; D006903:Hydroxymethylglutaryl CoA Reductases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism; D005810:Multigene Family / Q000235:genetics; D009284:Naphthols / Q000737:chemistry / Q000494:pharmacology*; D009928:Organ Specificity / Q000187:drug effects / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D010675:Pheromones / Q000737:chemistry / Q000494:pharmacology*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction
D009097:Multienzyme Complexes; D009284:Naphthols; D010675:Pheromones; D010940:Plant Proteins; D010088:Oxidoreductases; D006903:Hydroxymethylglutaryl CoA Reductases; C114231:1-deoxy-D-xylulose 5-phosphate reductoisomerase; D000217:Acyltransferases; C020141:flavanone synthetase; D019747:Aldose-Ketose Isomerases