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 antioxidation and proteomics of tomato plants infected by Tomato yellow leaf curl Thailand virus

林煜恆(Yu-Heng Lin)

指導教授 ： 羅筱鳳

國立臺灣大學/生物資源暨農學院/園藝學研究所/碩士(2011年)

https://doi.org/10.6342/NTU.2011.01495

摘要

番茄黃化捲葉病毒(Tomato yellow leaf curl virus, TYLCV)所誘發之番茄黃化捲葉病(Tomato yellow leaf curl disease, TYLCD)為近年來影響世界番茄生產的嚴重病害，常造成番茄生產上100％的經濟損失。防治TYLCD目前除了抗病育種，尚無有效之防治方法。由於番茄植株感染TYLCD後之生理尚不明瞭，故本研究針對植株染病後之抗氧化生理及蛋白質體兩方面深入探討，期有助於瞭解番茄TYLCD之抗病機制。植物材料使用由亞洲蔬菜研究發展中心選育出之番茄抗病品系CLN2777G-1-5及感病品種Tomatoll，病毒材料為番茄黃化捲葉泰國病毒種(Tomato yellow leaf curl Thailand virus, TYLCTHV)。試驗結果顯示TYLCTHV不能藉由機械法接種至番茄植株，以粉蝨為媒介接種TYLCTHV至感病品種植株後，葉片數、株高、SPAD值及葉綠素螢光Fv/Fm值皆顯著下降，於第3天所有植株皆檢測出病毒，第7天植株表現出病徵；抗病品系則於接種後第28天方有83%植株檢測出病毒，且仍無病徵表現。雖然抗病品系於接種TYLCTHV後 1及3天超氧歧化酶(superoxide dismutase, SOD)活性顯著低於感病品種，但是抗病品系之過氧化氫酶(catalase, CAT)、抗壞血酸過氧化酶(ascorbate peroxidase, APX)、穀胱甘肽還原酶(glutathione reductase, GR)、過氧化物酶(peroxidase)活性皆可持續升高，而有助於植株抗病毒侵害之逆境。兩番茄品種(系)於接種TYLCTHV後葉片蛋白質總量皆增加。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，比對出番茄兩品種(系)感染TYLCTHV後具差異表現之30個蛋白質，這些蛋白質參與植物光合作用、逆境防禦系統、蛋白質運輸、蛋白質轉譯、蛋白質裂解及氧化還原相關反應。本研究結果或可作為番茄抗TYLCD育種策略及抗病篩選指標之參考。

關鍵字

番茄黃化捲葉泰國病毒種 ； 超氧歧化酶過氧化氫酶抗壞血酸過氧化酶穀胱甘肽還原酶過氧化物酶蛋白質體

並列摘要

Tomato yellow leaf curl disease (TYLCD) induced by Tomato yellow leaf curl virus is one of the most severe diseases of tomato. TYLCD defeats tomato production worldwide and often causes 100％ economic loss. There is no efficient method to control TYLCD, except for breeding resistant cultivars. However, the plant physiology concerning TYLCD infection remains totally unknown. This research studied 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 Asian Vegetable Research and Development Center, and Tomato yellow leaf curl Thailand virus (TYLCTHV) were used as experimental materials. The results indicated that TYLCTHV is transmitted by whitefly that not mechanical. The leaf number, plant height, relative chlorophyll content (SPAD value) and chlorophyll fluorescence Fv/Fm of the infected ‘Tomatoll’ plant, were significantly decreased. All plants of susceptible cultivar were detected the occurrence of TYLCTHV DNA and first symptom appeared on 3 and 7 days after inoculation. While eighty three percent plants of resistant line were not detected the occurrence of virus DNA until 28 days and did not show symptom. Activity of superoxide dismutase (SOD) in leaves of resistant line was significantly lower than susceptible cultivar on 1 and 7 days after inoculation. However, the activities of catalase (CAT)、ascorbate peroxidase (APX)、glutathione reductase (GR) and peroxidase of resistant line raised successively after inoculation. The raised activities of antioxidant enzymes may induce the resistance of CLN2777G-1-5 plants to TYLCTHV. Proteomics of leaves 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, protein transport, translation, protein lysis, and oxidation-reduction reaction. The results may be helpful to plan strategy and select index for breeding TYLCTHV resistant tomato.

並列關鍵字

Tomato yellow leaf curl Thailand virus ； superoxide dismutase ； catalase ； ascorbate peroxidase ； glutathione reductase ； peroxidase ； proteomics

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