Functional Analyses of Pepper Chitinase, Pathogenesis-Related Protein 1, Lipase and RING-Finger Protein Genes in Disease Resistance and Abiotic Stress Tolerance of Capsicum annuum and Arabidopsis thal
- 발행기관 고려대학교 대학원
- 발행년도 2004
- 학위명 박사
- 학과 대학원:농학과
- 식별자(기타) DL:000014914210
- 서지제어번호 000045212830
초록/요약
Transcriptional activation of specific pathogenesis-related (PR) genes and active defense by the PR proteins are required for the establishment of induced plant resistance in plants. This dissertation is aimed at the gene expression, promoter regulation and functional role of pepper PR genes, including basic class II chitinase (CAChi2), basic PR-1 (CABPR1), GDSL-type lipase (CALIP1) and C3-H-C4 type RING-finger protein (CARFP1) genes. The CALIP1 and CARFP1 proteins were identified as CABPR1-interacting proteins in the yeast two-hybrid system. These PR genes showed differential organ-specific expression in healthy pepper plants. The CAChi2, CABPR1, CALIP1 and CARFP1 genes were differentially expressed in the pepper leaves during the compatible and incompatible interactions with X. campestris pv. vesicatoria. The CAChi2, CABPR1 and CARFP1 gene expressions were not only locally induced by Colletotrichum coccodes infection, but were also involved in the establishment of systemic acquired resistance of pepper plants. The CAChi2, CABPR1, CALIP1 and CARFP1 genes were also differentially induced in pepper leaf tissues by a variety of environmental stress conditions, such as high salt, drought, low temperature, and ethylene, jasmonic acid and salicylic acid treatments. The CAChi2 transcripts were mainly localized in phloem cells of vascular tissues of pepper leaves infected with C. coccodes. The CALIP1-GFP and CARFP1-GFP proteins were targeted in the vascular tissues, and the cytoplasm and specific cellular organelles of Arabidopsis root cells, respectively. Immunogold labeling of PR-1 proteins occurred over cell walls and cytoplasm of the host and the oomycete pathogen, and at the interface between host and oomycete cell walls in tomato stem tissues inoculated with Phytophthora capsici. The CAChi2 and CABPR1 promoters were up-regulated locally and systemically by Pseudomonas syringae pv. tabaci infection. The CAChi2 and CABPR1 promoter activation was closely related to osmo
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TABLE OF CONTENTS
ABSTRACT ---------------------------------------------------------------------- i
TABLE OF CONTENTS -------------------------------------------------------- vi
LIST OF FIGURES ---------------------------------------------------------- xvii
CHAPTER
1. Introduction ------------------------------------------------------------- 1
1.1. Plant disease resistance ---------------------------------------------- 2
1.2. Protein-protein interactions in plant disease resistance ------------ 3
1.3. Pathogenesis-related (PR) gene ------------------------------------- 5
1.3.1. PR-1 family ------------------------------------------------------ 6
1.3.2. Chitinase family ------------------------------------------------- 9
1.3.3. Lipase ----------------------------------------------------------- 12
1.3.4. RING-finger protein -------------------------------------------- 14
1.4. Objectives of the study -------------------------------------------- 15
2. Induction by pathogen, salt and drought of a basic class II chitinase mRNA and its in situ localization in pepper
(Capsicum annuum) ---------------------------------------------------17
2.1. Abstract -------------------------------------------------------------- 18
2.2. Introduction ---------------------------------------------------------- 18
2.3. Materials and methods ---------------------------------------------- 21
2.3.1. Plants, fungus, and inoculation ------------------------------ 21
2.3.2. ABA, NaCl, and drought treatments ------------------------- 22
2.3.3. RNA isolation and Northern blot analysis -------------------- 22
2.3.4. In situ hybridization ------------------------------------------ 23
2.4. Results --------------------------------------------------------------- 25
2.4.1. Organ-specific regulation and tissue-specific localization
of pepper chitinase mRNA ----------------------------------- 25
2.4.2.
