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A prompt approach to generate effective mild strains of papaya ringspot virus for solving the problem of strain-specific protection in Taiwan
|關鍵字:||木瓜輪點病毒;交互保護;株系專一性;輕症病毒;papaya ringspot virus;cross-protection;strain-specific;mild strain||引用:||Abel, P. P., Nelson, R. S., De, B., Hoffmann, N., Rogers, S. G., Fraley, R. T., and Beachy, R. N. 1986. Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene. Science 232: 738-743. Anandalakshmi, R., Pruss, G. J., Ge, X., Marathe, R., Mallory, A. C., Smith, T. H., and Vance, V. B. 1998. A viral suppressor of gene silencing in plants. Proc. Natl. Acad. Sci. U S A. 95: 13079-13084. Baba, V. Y., Giampani, J. S., Tazima, Z. H., Yada, I. F., Paccola-Meirelles, L. D., and Leite Júnior, R. P., 2014. Agronomic performance of Pera and related sweet orange accessions naturally infected with Citrus tristeza virus in northern Paraná state. Braz. Trop. Plant Pathol. 39: 442–448. Bau, H. J., Cheng, Y. H., Yu, T. A., Yang, J. S., Liou, P. C., Hsiao, C. H., and Yeh, S. D. 2004. 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木瓜輪點病毒(papaya ringspot virus, PRSV)對全世界木瓜產業影響甚鉅。PRSV 夏威夷分離株HA經亞硝酸誘變產生之輕症病毒HA 5-1已被用於保護PRSV感染數十年。然而因交互保護具高度專一性，HA 5-1對台灣的病毒株無法提供完整的保護效果。我們之前的研究發現以PRSV HA 5-1作為骨架攜帶台灣YK病毒株的鞘蛋白(coat protein, CP)基因所構築的重組輕症病毒HA 5-1/YK CP與HA 5-1相比在溫室條件下可顯著提升對YK病毒株的保護效果由20%到65%。在2016-2017的田間試驗中，本研究分出兩個來自台中和彰化的PRSV病毒株TC和CH分別可擊垮HA 5-1/YK CP的保護造成感染，經序列比對此三病毒株的CP基因具94.1-95.7%的核苷酸序列相同度。為保護木瓜植株不受TC和CH病毒株感染，進一步構築以HA 5-1作為骨架攜帶TC和CH病毒株CP基因及3'端非轉譯區序列之嵌合輕症病毒，分別命名為HA 5-1/TC CP和HA 5-1/CH CP。在溫室條件下兩輕症病毒感染木瓜植株後皆未出現嚴重病徵，且接種達3個月或經3次繼代皆可維持穩定之弱病徵。在三次獨立試驗中，HA 5-1/TC CP可顯著提升對TC病毒株之保護效果達70%，高於HA 5-1/YK CP所提供之16.7%，但僅能對CH病毒株提供37.8%的保護效果；而HA 5-1/CH CP亦可對CH病毒株提供至70%的保護。本研究之結果顯示，我們發展了一個以HA 5-1作骨架來快速構築弱系病毒株的系統，藉此解決針對台灣病毒株交互保護株系專一性的問題。
Papaya ringspot virus (PRSV) restricts papaya production worldwide. PRSV HA 5-1, a mild strain derived from nitrous acid treatment of Hawaii isolate (HA), has been used to control PRSV infection in Taiwan for decades. However, HA 5-1 does not provide complete protection against different Taiwan severe strains due to strain-specific problem. In our previous study, a recombinant mild strain HA 5-1/YK CP, using PRSV HA 5-1 as a backbone to carry the heterologous coat protein (CP) gene of Taiwan YK strain, significantly increased the protection rates, from 20% to 65% against YK isolates were in papaya plants, under greenhouse condition. In this study, two PRSV strains collected from Taichung (TC) and Changhua (CH) were found able to break down the protection provided by HA 5-1/YK CP. Further analysis indicated that their CP genes share 94.1-95.7% nucleotide homology with that of YK strain. In this study, to solve the breakdown caused by TC and CH strains, chimeric PRSV HA5-1 mild strain carrying the heterologous CP gene from TC or CH, HA 5-1/TC CP and HA 5-1/CH CP, respectively, were constructed. Under greenhouse conditions, both chimeric mild strains showed infection on papaya plants without conspicuous symptoms and remained stable for 3 months or after 3 times transfer. From three independent tests, HA5-1/TC CP significantly enhanced the protection rate against PRSV TC (average of 70%), much higher than that provided by HA5-1/YK CP (average of 16.7%), but provided a relatively lower degree of protection against CH strain (average of 37.8%). Similarly, HA 5-1/CH CP provided high degree of protection against CH strain (average of 70%). Therefore, using HA 5-1 as a backbone, we have developed a prompt way to generate effective mild strains which can solve the problem of strain-specific protection in Taiwan.
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