title: Analysis of rice genes induced by striped stemborer (Chilo suppressalis) attack identified a promoter fragment highly specifically responsive to insect feeding.
abstract: The objective of this study was to identify rice gene promoters that are specifically induced by feeding of the striped stemborer (Chilo suppressalis). Two PCR-selected cDNA subtractive libraries were constructed from the rice variety Minghui 63. Up- and down-regulated cDNAs induced by C. suppressalis feeding were arrayed on nylon membranes. After array hybridization and Northern blot analysis, a cDNA (B1-A04) encoding a putative subtilisin/chymotrypsin inhibitor was found to be rapidly and highly induced by C. suppressalis feeding, compared with mechanical wounding. The putative promoter region, spanning from -1,569 to +446 relative to the transcriptional initiation site was isolated, fused to the GUS gene (beta-glucuronidase reporter gene) and introduced by Agrobacterium-mediated transformation to rice. In non-infested plants, the GUS activity driven by this promoter fragment was detected in culms and panicles, but not in leaves and sheaths. At 6 h after insect feeding, GUS activity was significantly induced in sheaths and culms, but not in leaves. GUS activity and native B1-A04 gene were not induced by JA and ABA treatment. A serial deletion analysis revealed two regions (-1,569 to -1,166 and -1,166 to -582) that negatively regulate the gene expression in sheaths of non-infested plants but not in insect-infested plants. An electrophoretic mobility shift assay (EMSA) identified 7 DNA fragments with various binding activities with nuclear proteins from mechanically wounded, insect-infested and untreated plants, and their possible roles in gene regulation were speculated. This promoter fragment should have utility in development of insect resistant transgenic crops.
authors.affiliation: National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Hongxia
authors.initials: H
authors.lastname: Hua
authors.forename: Qing
authors.initials: Q
authors.lastname: Lu
authors.forename: Meng
authors.initials: M
authors.lastname: Cai
authors.forename: Caiguo
authors.initials: C
authors.lastname: Xu
authors.forename: Dao-Xiu
authors.initials: DX
authors.lastname: Zhou
authors.forename: Xianghua
authors.initials: X
authors.lastname: Li
authors.forename: Qifa
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D017931:DNA Primers; D018076:DNA, Complementary; D005247:Feeding Behavior; D017343:Genes, Plant; D007313:Insecta / Q000502:physiology*; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics* / Q000469:parasitology; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary