台灣位處於亞熱帶，夏季常遭受颱風、豪雨的侵襲，引起淹水或土壤過濕，造成農作物品質受損、產量銳減，也使農業經濟蒙受慘重損失，物價飆漲、民生不安。番茄是重要的世界性果菜，目前雖有耐熱品種，但在淹水耐受性上，仍有改善的空間；而台灣夏季生產番茄時以茄子作為根砧，以抵抗高溫淹水所產生的逆境。
本研究以不耐淹水之番茄‘台南亞蔬6號’(Lycopersicon esculentum cv. TNVEG 6)與耐淹水茄子(Solanum melongena cv. EG203)為試驗材料，苗齡45~50天時給予室溫23 ℃及高溫38 ℃淹水與不淹水處理，淹水處理的時間分別為0、6 hr、12 hr、1天、2天和4天，處理後再以全自動二維蛋白快速層析分離系統(ProteomeLabTM PF-2D)分析高溫淹水處理前後葉片之蛋白質，再以MALDI-TOF MS與MS/MS分析，利用生物資訊網站Mascot比對分析蛋白質的序列，了解其功能，探討高溫淹水期間蛋白質的差異表現及高溫淹水之相關生理。
根據蛋白質第一維分析結果，番茄高溫淹水四天後，pI 4~8蛋白質總量明顯減少，可能因為高溫淹水逆境破壞蛋白質，而茄子於淹水兩天後，其pI 4~8蛋白質量總量明顯增加，可能是抵抗高溫淹水逆境之蛋白質產生。
蛋白質第二維結果利用DeltaVue分析，比對番茄常溫未淹水和高溫淹水4天，發現有22個有差異的peaks；番茄常溫未淹水和高溫逆境4天，發現有11個有差異的peaks；茄子常溫未淹水和高溫淹水2天，發現14個有差異的peaks；茄子常溫未淹水和高溫逆境2天，發現有11個有差異的peaks，經MALDI-TOF MS與MS/MS分析後，得到20個與高溫淹水逆境有關之蛋白質，分別為：Acyl carrier protein、Fructose-1.6-bisphosphatase、Dehydrogenase、Mitogen activated protein kinase、Ni/Fe hydrogenase、GTP-binding protein、Oxygen-evolving enhancer protein、Deoxyribose-phosphate aldolase、Methionine sulfoxide reductase、Aldehyde dehydrogenase、Flavoprotein、Ribulose bisphosphate carboxylase/oxygenase (RuBisCO)、Myosin、Glutathione peroxidase、Hydrogenase、calcium-binding protein、Deformylase、Adenylyltransferase、sucrose-phosphate synthase、NADH dehydrogenase。
Taiwan is located within the subtropical zone. Typhoons or heavy rains in the summer which result in flooding and over wetted soil, not only lead to damage in quality and yield of the crops, but also impact the agricultural economy and the prices of crops. Tomato is one of the most important fruit crops in the world. Nowadays, even though there are heat tolerant cultivars of tomato, improvement in its flooding tolerance is still needed. Eggplant is one of the flooding tolerant species. It is used as the rootstock of tomato in summer in Taiwan.
In this research tomato Lycopersicon esculentum cv. TNVEG 6 and eggplant Solanum melongena cv. EG203 were needas the experimental materials. Forty five to fifty days plants were treated separately at 23℃ and at 38℃ with flooding and non-flooding stress for 0、6、12、24、48、96 hours. We investigated growth of the plants before and after each treatment. ProteomeLabTM PF-2D was utilized to analyze protein extracts from the leaves, and PF-2D MALDI-TOF MS or MS/MS were also used to analyze the sample. We also combined bioinformation mascot web site and ProteoVue DeltaVue software were combined for the sequence alignment for our protein spots to comprehend their functions.
According to the results of our PF-2D first dimension, after exposure to heat and flooding stress for four days, the quantity of tomato total protein from isoelectric point (pI) 4 to 8 were obviously decreased. A possible interpretation of this result may be that the heat and the flooding stress could damage the proteins. On the other hand, the quantity of total proteins from isoelectric point 4 to 8 are significantly increased in eggplant under heat and flooding stress after two days. This result shows that eggplants might produce some specific proteins to resist heat and flooding stresses.
DeltaVue software was used to analyze the second dimension results acquired from PF-2D. When comparing tomato at 23℃ without flooding to 38℃ with flooding stress after four days, there are 22 significantly different peaks in tomato. And there are 11 significantly different peaks in tomato comparing 23℃ without flooding to 38℃ without flooding stress after four days. While comparing eggplant at 23℃ without flooding to 38℃ with flooding stress after two days, there are 14 significantly different peaks. And there are 11 significantly different peaks in eggplant comparing 23℃ without flooding to 38℃ without flooding stress after two days. After MALDI-TOF MS and MS/MS analysis, there are 20 meaningful proteins in MS results: Acyl carrier protein、Fructose-1.6-bisphosphatase、Dehydrogenase、Mitogen activated protein kinase、Ni/Fe hydrogenase、GTP-binding protein、Oxygen-evolving enhancer protein、Deoxyribose-phosphate aldolase、Methionine sulfoxide reductase、Aldehyde dehydrogenase、Flavoprotein 、Ribulose bisphosphate carboxylase/oxygenase、Myosin、Glutathione peroxidase、Hydrogenase、calcium-binding protein、Deformylase、Adenylyltransferase、sucrose-phosphate synthase、NADH dehydrogenase.
