2007-01-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/660213摘要：根據前人的研究，花粉母細胞形成發育期或番茄開花前1至2星期，對高溫最為敏感並導致高落果率。為了鑑定出耐熱品種較高之結實率之相關蛋白質體，本年度我們已經獲得數個蛋白質（均為下游之目標基因）與其對應基因，它們可能參與了耐熱與不耐熱品系間的差異。本計劃內我們擬生產重組蛋白並研究其生化性質以評估它們是否與熱的穩定性有關。過去５年來多個數量性狀基因座之分子基礎已被揭開，大部份都是屬於調節性基因而非下游之目標基因。調節性基因與蛋白通常表現量低而且不易用電泳偵測得到。為了找出造成耐熱與不耐熱品系不同之調節性基因，本研究擬進行二种試驗。假設這些調節性基因是來自耐熱品系內的特有基因，或是相同的表現基因但有不同的表現量。為尋找獨特的基因我們擬使用抑制性扣除式雜合，而研究不同的表現量我們將採用微陣技術使用Affymatrix公司出品含有個10000基因之晶片。執行此計畫，我們選用CLN2468D耐熱番茄與L04783對熱敏感番茄同時在人工氣候室內栽培，其溫控是25℃/20℃。我們得自微陣與抑制性扣除式雜合的調節性基因對改善亞熱帶地區之栽培番茄將有重要之貢獻。<br> Abstract: Developing anther one week before anthesis in tomatoes was reported as the most sensitive period to high temperature which caused a low fruit set. We have obtained several proteins and their corresponding genes (all downstream target genes) which are probably involved in the difference between heat-tolerant variety and heat-sensitive one. In this project, we will generate fusion proteins and study the biochemical properties to evaluate whether they are correlated to heat stability. In the past five years molecular basis of several of quantitative trait loci were revealed and most of them were regulatory genes rather than downstream target genes. Regulatory transcript and protein are relatively low in quantity and not easy to be detected in electrophoresis. To identify the regulatory genes responsible for difference of fruit set between heat-sensitive and heat-tolerant lines, in this project we will run two different experiments supposing that regulatory genes are either derived from unique genes only present in heat-tolerant line and/or different in expression level. To search for the unique gene, we will apply suppression subtractive hybridization method, and for expression level of transcripts we will use microarray with the chips from Affymatrix containing 10,000 genes. In doing so, two selected clones, CLN2468D, the heat-tolerant, and L04783, the heat-sensitive, will be grown at the same time in a phytotron at (natural photoperiod) and 25 oC /20 oC. The regulatory genes obtained from both microarray and suppression subtractive hybridization methods will have a great potential to improve the fruit set in growing tomato in subtropical areas.番茄微陣抑制性扣除式雜合調節性基因tomatomicroarraysuppression subtractive hybridizationRegulatory transcript