用於茄科作物上具交互保護能力的辣椒葉脈斑駁病毒疫苗之開發

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Title:	用於茄科作物上具交互保護能力的辣椒葉脈斑駁病毒疫苗之開發
Other Titles:	Vaccine development for cross protection against pepper veinal mottle virus in Solanaceae crops
Authors:	王冠達 WANG, GUAN-DA
Contributors:	陳宗祺 CHEN, TSUNG-CHI 醫學檢驗暨生物技術學系
Keywords:	植物病毒疫苗;交互保護;茄科作物 plant virus vaccine;cross-protection;solanaceous crops
Date:	2023
Issue Date:	2023-11-22
Abstract:	番茄 (Solanum lycopersicum L.) 及番椒 (Capsicum annuum L.) (包括甜椒及辣椒) 為全球最重要之蔬果類茄科作物，全球種植總面積約3880萬公頃，台灣種植總面積逾1萬公頃。在臺灣，種植番茄及番椒時經常遭受蚜蟲傳播之Potyvirus屬病毒危害，如馬鈴薯病毒Y (potato virus Y，PVY)、辣椒斑駁病毒 (pepper mottle virus，PepMoV) 及辣椒葉脈斑駁病毒 (pepper veinal mottle virus，PVMV) 等，其中以PVMV最為常見。為了獲得對PVMV具交互保護能力的輕症病毒株，本研究以亞硝酸對野生型PVMV-Tn病毒株進行誘變，利用病毒會於奎藜 (Chenopodium quinoa) 葉片上產生明顯的局部壞疽病斑特性，篩選不會形成壞疽病斑之突變病毒株，並測試其交互保護效果。本研究共進行了十次亞硝酸誘變試驗，獲得1769個單斑，篩選出三個表現最為穩定的輕症突變株，代號分別為m4-8、m10-1和m10-11，可於奎黎葉片上產生不明顯的褪綠單斑，在菸草 (Nicotiana benthamiana) 和番茄上造成輕微的系統性感染。m4-8、m10-1和m10-11於菸草及番茄上之鞘蛋白 (coat protein，CP) 累積速率較野生型病毒株PVMV-Tn緩慢，m4-8則對野生型PVMV-Tn具有最佳的交互保護能力。經過基因組序列分析後得知m4-8突變株與野生型PVMV-Tn之間具有20個核苷酸的差異，導致有18個胺基酸的改變。本研究已於溫室條件下證實m4-8突變株對PVMV-Tn具有交互保護能力，未來將於田間測試其保護效果。 Tomatoes (Solanum lycopersicum L.) and peppers (Capsicum annuum L.), including bell peppers and chili peppers, are the most globally important vegetable and fruit solanaceous crops, with a total annual planting area of about 38.8 million ha in the world, including over 10,000 ha in Taiwan. The cultivation of tomatoes and peppers in Taiwan is commonly affected by aphid-borne potyviruses, including potato virus Y (PVY), pepper mottle virus (PepMoV) and pepper veinal mottle virus (PVMV), among which PVMV is the most prevalent. To obtain mild virus strains with the cross protection ability against severe PVMV strains, nitrous acid-induced mutagenesis was performed in this study. The tomato isolate of PVMV (PVMV-Tn), inducing necrotic lesions on inoculated Chenopodium quinoa leaves, was used for mutagenesis. Attenuated mutants were screened based on milder host responses. Three stable attenuated PVMV mutants, denoted m4-8, m10-1 and m10-11, were selected from 1769 local lesions in ten nitrous acid treatments. The mutants induced inconspicuous chlorotic spots on C. quinoa leaves, mild mottles on Nicotiana benthamiana leaves and asymptomatic infection with lower accumulation in tomato plants. m4-8 showed the highest protective efficacy against wild-type (WT) PVMV-Tn in N. benthamiana and tomato plants in greenhouse experiments. Comparison of whole-genome sequences of m4-8 and WT PVMV-Tn indicated that twenty nucleotide substitutions leading to eighteen amino acid changes occurred in the m4-8 genome. Our results showed that m4-8 has great potential for protecting Solanaceae crops from PVMV. The protective effect of m4-8 in the field will be further evaluated.
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