Expression and characterization of virus resistant gene in Solanum tuberosum L. cv. Golden valley
- Issued Date
- 2007
- Abstract
- Potato Virus Y (PVY), the type member of the genus potyvirus in the family Potyviridae is a widespread virus leading to severe damage in Solanaceae, which was loss crop yield up to 75% in Potato (Solanum tuberosum L.). PVY infected plants shows several symptoms like mosaic, yellowing, necrosis, mottle, vein clearing, vein bending and misshapen in tubers. Plants carrying the Ry gene are resistance to a number of potiviruses. Previously, we identified that the potato cv. Golden valley has rich in Ry gene overexpression. We isolated crude protein form cv. Golden valley and applied to the most virus-susceptible potato cv. Winter valley among the twelve Korean cultivars studied. Real-time PCR and ELISA results showed that in vivo application of Golden protein to cv. Winter valley could reduce the rate of PVY accumulation significantly. Analysis of Ry gene sequence was showed that there were about 91% sequence homologs between Solanum tuberosum subsp. andigena and Solanum tuberosum L. cv. Golden valley. Our second phase research work is going on to develop virus resistant potato cultivars by introducing, Ry gene and bio-pesticides for other legume crops simultaneously.
- Alternative Author(s)
- Youngmin Park
- Affiliation
- 조선대학교 일반대학원 생명공학과
- Department
- 일반대학원 환경생명공학과
- Awarded Date
- 2008-02
- Table Of Contents
- CONTENTS
LIST OF TABLES iii
LIST OF FIGURES iv
LIST OF ABBREVIATION v
ABSTRACT vi
INTRODUCTION 1
MATERIAL AND METHODS 3
1. Plant materials and Growth condition 3
2. Virus innoculation 3
3. ELISA analysis for detection in PVYO infected potato varieties 3
3-1. Total protein assay 3
3-2. ELISA test 4
4. Expression pattern of CP and Ry gene by RT-PCR 4
4-1. Total RNA extraction 4
4-2. cDNA synthesis 5
4-3. RT-PCR amplification 5
5. Application of Proteingolden 8
5-1. Proteingolden preparation. 8
5-2. Proteingolden treatment 8
6. Expression pattern of CP and Ry gene against PVYO using Quantitative (Real-time) PCR 8
7. Cloning of Rygolden gene 9
7-1. cDNA synthesis of Rygolden gene 9
7-2. Vector(pUC19) and Rygolden gene preparation 9
7-3. Vector(pUC19) and Rygolden gene ligation and transformation 12
7-4. Plant expression vector(pBI121) and Rygolden gene preparation 12
7-5. Vector(pBI121) and Rygolden gene ligation and transformation 15
RESULT AND DISCUSSION 17
1. The pattern of PVYO virus infection between potato lines 17
2. Selection of virus resistant potato using DAS-ELISA 21
3. Characterization of virus resistant and susceptible cultivars 24
3-1. Expression of CP gene 24
3-2. Expression of Ry gene 24
4. Ry gene expression in potato lines 27
5. The virus resistant effect of Proteingolden 27
6. Rygolden gene construction strategy and sequence differences 31
적요 36
REFERENCES 30
감사의글 42
- Degree
- Master
- Publisher
- 조선대학교 일반대학원 생명공학과
- Citation
- 박영민. (2007). Expression and characterization of virus resistant gene in Solanum tuberosum L. cv. Golden valley.
- Type
- Dissertation
- URI
- https://oak.chosun.ac.kr/handle/2020.oak/6985
http://chosun.dcollection.net/common/orgView/200000235872
- Appears in Collections:
- General Graduate School > 3. Theses(Master)
- Files in This Item:
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