Molecular basis of ergosterol-induced protection of grape against botrytis cinerea: induction of type I LTP promoter activity, WRKY, and stilbene synthase gene expression.
Type I lipid transfer proteins (LTPs) are basic, 9-kDa cystein-rich proteins believed to be involved in plant defense mechanisms. A 2,100-bp fragment containing the coding region of Vitis vinifera lipid transfer protein 1 (VvLTP1) and 1,420-bp of its promoter region was isolated by screening a grape genomic library. In silico analysis revealed several putative, defense-related, cis-regulatory elements such as W- and MYB-boxes, involved in the binding of WRKY and MYB transcription factors, respectively. The 5'-truncated versions of the VvLTP1 promoter were generated, cloned in front of the beta-glucuronidase (GUS) reporter gene, and introduced in tobacco plants and grapevine cell suspensions using Agrobacterium spp. Single MYB- and the W-boxes identified on the 0.250-kbp fragment were sufficient to induce GUS activity in transgenic tobacco plants after transient expression of MYB and WRKY. Ergosterol, a nonspecific fungal elicitor, induced GUS activity in transgenic grapevine cell suspensions transformed with the 1,420- and 750-bp promoter containing a palindromic arrangement of two W-boxes but not the 650- or 250-bp fragment, where only one W-box was present. Moreover, ergosterol triggered WRKY, VvLTP1, and stilbene synthase gene expression in grape plantlets and enhanced protection against Botrytis cinerea. The molecular basis of ergosterol-induced protection is discussed.
Laquitaine
Laurent
L
Laboratoire de Physiologie et Biochimie Végétales, UMR CNRS 6161, Université de Poitiers, Bâtiment Botanique, 40 Avenue du Recteur Pineau, 86022 Poitiers Cedex, France.
Gomès
Eric
E
François
Julie
J
Marchive
Chloé
C
Pascal
Stéphanie
S
Hamdi
Saïd
S
Atanassova
Rossitza
R
Delrot
Serge
S
Coutos-Thévenot
Pierre
P
D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001483:Base Sequence; D020171:Botrytis / Q000502:physiology*; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D019295:Computational Biology; D004875:Ergosterol / Q000494:pharmacology*; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D015698:Genomic Library; D005966:Glucuronidase / Q000032:analysis; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020598:Proto-Oncogene Proteins c-myb / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D027843:Vitis / Q000235:genetics* / Q000378:metabolism / Q000382:microbiology*
D052179:Antigens, Plant; D002352:Carrier Proteins; D010940:Plant Proteins; D020598:Proto-Oncogene Proteins c-myb; D014157:Transcription Factors; C511920:lipid transfer proteins, plant; D000217:Acyltransferases; C022674:stilbene synthase; D005966:Glucuronidase; D004875:Ergosterol