Molecular characterization of Plutella xylostella (Lepidoptera: Plutellidae) phosphoglycerate mutase and its response to parasitoid Cotesia plutellae (Hymenoptera: Braconidae)
- 주제(키워드) Plutella xylotella , Cotesia plutellae , Parasitism , Phosphoglycerate mutase , Hypotrophy
- 발행기관 고려대학교 대학원
- 지도교수 한성식
- 발행년도 2013
- 학위수여년월 2013. 2
- 학위구분 석사
- 학과 일반대학원 생명공학과
- 세부전공 분자생물학
- 원문페이지 38 p
- 실제URI http://www.dcollection.net/handler/korea/000000037713
- 본문언어 영어
- 제출원본 000045746061
초록/요약
Abstract Diamondback moth, Plutella xylostella larvae damage leaves, buds, flowers, and seed-buds of cultivated cruciferous plants. The parasitized Plutella xylostella by Cotesia plutellae exhibits hypotrophied structures in the fat body compared with non-parasitized larvae. Insect fat body is a principal organ for metabolic processing of digestive products following absorption, and is an organ responsible for the storage and syntheses of carbohydrates, proteins and lipids. Phosphoglycerate mutases (PGMs) catalyze the reversible conversion of 3-phosphoglycerate and 2-phosphoglycerate as part of glycolysis and gluconeogenesis. A full-length cDNA encoding a PGM was cloned and characterized in the diamondback moth, Plutella xylostella (PxPGM, 768 bp). The PxPGM is 255 amino acids with a molecular mass of approximately 28 kDa. The deduced amino acid sequence showed significant homology with other insect PGMs from Papilio xuthus (91%), Papilio polytes (91%), Bombyx mori (91%) and Danaus plexippus (93%). The molecular phylogeny of insect PGM was examined using amino acid sequences from a number of insects. Quantitative real-time PCR showed that PxPGM mRNA was expressed during all larval developmental stages examined. Analysis of parasitized larvae by the endoparasitoid C. plutellae suggests that the PxPGM expression decrease following parasitoid challenge. Spatial expression analysis showed that PxPGM is found predominantly in the fat body. These results indicate that the parasitization by C. plutellae induce several changes of host physiology such as depression of feeding consumption.
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List of Table i
List of Figures ii
Abstract iii
Ⅰ. Introduction 1
Ⅱ. Materials and Methods 3
2.1. Insect rearing and parasitization 3
2.2. RNA isolation, cDNA synthesis and degenerate PCR 3
2.3. Rapid amplification of cDNA ends (RACE) 5
2.4. Sequencing and phylogenetic analysis 6
2.5. Quantitative real time PCR expression developmental stages 8
2.6. Quantitative real time PCR tissue-specific expression 9
Ⅲ. Results 10
3.1. Identification and characterization of the PxPGM 10
3.2. Phylogenetic relationship of PxPGM with other insect PGMs 10
3.3. Developmental stage and Tissue-specific expression of PxPGM 11
Ⅳ. Discussion 20
Ⅴ. References 23
