Polyamine biosynthesis regulated by StARD expression plays an important role in potato wounded periderm formation = 상처난 감자의 주피 형성에 중요한 기능을 가진 StARD 발현에 의한 폴리아민 생합성 조절

다국어 초록 (Multilingual Abstract)

An Acireductone dioxygenase(ARD) gene of patatoes was isolated from the ESTs of potato post-suberization cDNA libraries.The highest expression levels of the StARD gene and the protein appeared 36h after suberization. An approximate nine-fold increase in ARD activity was detected at 36h after wounding. Real-time RT PCR analysis and immunolocalization studies revealed that StARD transcripts increase at the wound surface of potato tubers. The polyamine(PA) content increased significantly after wounding at the wound surface. The increased PA content and ARD activity may play an important role in wound periderm formation.

국문 초록 (Abstract)

감자에서 Acireductone dioxygenase(ARD)유전자는 potato postsuberization cDNA 라이브러리의 ESTs 에서 동정되었다. 감자 StARD 유전자와 단백질의 발현량은 슈베린층의 생성되는 36 시간 후이다. 상처난 후 36 시간 후에 ARD 의 활성은 아홉배에 달할 정도로 증가하엿다. 리얼타임 RT-PCR 분석과 Immunolocalization 실험결과 StARD 전사가 감자 괴경의 상처난 표면에서 증가함을 보여주었다. 감자의 상처 표면에서 상처난이후에 폴리아민의 농도가 증가하였다. 증가한 폴리아민과 ARD 활성은 상처난주피의 형성에 증요한 역할을 하고 있다.

목차 (Table of Contents)

• I. Introduction = 1
• II. Material and Methods = 16
• A. Plant material and treatment = 16
• B. Gene cloning and overexpression of StARD protein = 17
• C. Preparation of tuber protein extraction. = 18
• D. DNA and RNA gel blot analysis = 19
• E. Preparation of antiserum to ARD and western blot analysis. = 21
• F. Enzyme activity assays. = 23
• 1. α-keto acid assay = 23
• 2. Formate Assays. = 24
• 3. CO Assay. = 26
• G. Quantification of PAs = 26
• H. Histological anaylisis of wound periderm = 27
• I. Immunolocalization = 28
• J. Real-time RT-PCR = 28
• III. Result and Discussion = 30
• A. Cloning and characterization of StARD gene = 30
• B. Overexpression of StARD protein and its activity = 34
• C. Increased contents and biosynthesis of polyamines = 43
• D. Polyamine biosynthesis is more closely related to ARD expression than ethylene in potato suberization process = 47
• IV. Reference = 56
• Abstract = 61