番茄植株感染番茄黃化捲葉泰國病毒種後之抗氧化生理及蛋白質體研究

The Study on Antioxidative Physiology and Proteomics of Tomato Plants Infected by Tomato Yellow Leaf Curl Thailand Virus

本研究探討番茄感染黃化捲葉泰國病毒（Tomato yellow leaf curl Tailand disease, TYLCTHD）後之抗氧化生理及蛋白質體，期有助於瞭解抗病機制。使用亞蔬-世界蔬菜中心選育出之番茄抗病品系CLN2777G-1-5及感病品種Tomatoll，病毒材料則為番茄黃化捲葉泰國病毒種（Tomato yellow leaf curl Thailand virus, TYLCTHV）。結果顯示，兩品種（系）之葉片超氧歧化酶（superoxide dismutase, SOD）與抗壞血酸過氧化酶（ascorbate peroxidase, APX）活性雖於接種TYLCTHV後差異不大，然抗病品系葉片過氧化物酶（peroxidase）活性於接種後第1天顯著高於感病品種，且在接種後第3天過氧化氫酶（catalase, CAT）活性亦顯著高於健康植株與接種無病毒粉蝨之植株。抗病品系在接種後葉片穀胱甘肽還原酶（glutathione reductase, GR）活性皆顯著較感病品種高。故推測抗病番茄在感染TYLCTHV後，植株為抵抗TYLCTHV，其葉片peroxidase活性先快速上升，於第3天CAT活性接續上升，GR活性則在感染後各階段皆呈現較高活性，而有助於抗病番茄品系CLN2777G-1-5植株抵抗TYLCTHV之感染。利用全自動二維蛋白快速層析分離系統（ProteomeLabTM PF-2D）及基質輔助雷射脫附游離飛行時間質譜儀（matrix-assisted laser desorption ionization time-of-flight, MALDI-TOF）分析，比對出兩品種（系）感染TYLCTHV後具差異表現之30個蛋白質，其參與光合作用、逆境防禦系統、氧化還原相關反應或蛋白質運輸、轉譯與裂解，或可作為番茄抗TYLCTHD育種策略及抗病篩選指標之參考。 This research aimed at the antioxidation and proteomics of tomato plants after infection of TYLCD to understand the resistance mechanism. The resistant tomato line CLN2777G-1-5 and susceptible cultivar Tomatoll which were selected and bred by AVRDC-World Vegetable Center, respectively. Tomato yellow leaf curl Thailand virus (TYLCTHV) was used as inoculum material. The results indicated that the activities of superoxide dismutase (SOD) and ascorbate peroxidase (APX) in leaves were not significantly different between 2 varieties (lines) after inoculation by TYLCTHV. However, peroxidase activity in leaves of the resistant line was significantly higher than susceptible cultivar on 1 day after inoculation. The catalase activity (CAT) in leaves of the resistant line was also significantly higher than the healthy resistant plant and the resistant plant inoculated by healthy-whitefly on 3 days after inoculation. The activity of glutathione reductase (GR) in leaves of the resistant line was significantly higher than that of susceptible variety after inoculation of TYLCTHV. Peroxidase activity in the leaves of the resistant line increased rapidly on the first day infected by TYLCTHV. CAT activity then increased on the third day. GR showed high activity at all stages of infection which contributed to the resistance of CLN2777G-1-5 plants to TYLCTHV infection. Proteomics of leaves were analyzed by ProteomeLabTM PF-2D and matrix-assisted laser desorption ionization time-of-flight. Thirty proteins with differential expression were identified from CLN2777G-1-5 and 'Tomatoll' after inoculation with TYLCTHV. These proteins involve photosynthesis, stress defense, oxidation-reduction reaction, and protein transport, translation, and lysis. The results may be helpful to plan strategy and select index for breeding TYLCTHV resistant tomato.