Transgenic Tobacco Lines Expressing Yam Mosaic Virus Coat Protein-Derived dsRNA Are Resistant to Yam Mosaic Virus  

作　　者：Effiom Eyo Ita Edak Aniedi Uyoh Ikuo Nakamura Valentine Otang Ntui Effiom Eyo Ita;Edak Aniedi Uyoh;Ikuo Nakamura;Valentine Otang Ntui(Department of Genetics & Biotechnology, University of Calabar, Calabar, Nigeria;Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University, Chiba, Japan;International Institute of Tropical Agriculture, Nairobi, Kenya)

机构地区：[1]Department of Genetics & Biotechnology, University of Calabar, Calabar, Nigeria [2]Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University, Chiba, Japan [3]International Institute of Tropical Agriculture, Nairobi, Kenya

出　　处：《American Journal of Plant Sciences》2020年第9期1487-1504,共18页美国植物学期刊（英文）

摘　　要：Yam mosaic virus (YMV), a Potyvirus, is a highly destructive pathogen of yam accounting for yield losses up to 40%. Apart from causing significant reduction in tuber size and quality, it restricts international exchange of germplasms. It thus becomes crucial to get resistant or at least virus-free planting materials for farmers. This study was aimed at inducing resistance to YMV in tobacco by RNA silencing. An RNAi construct containing 161 bp fragment of <span style="font-family:Verdana;">YMV-coat protein (CP) </span><span style="font-family:Verdana;">gene was developed and used to produce transgenic tobacco lines expressing </span><span style="font-family:Verdana;">YMV-coat protein (CP)</span><span style="font-family:Verdana;"> derived </span><span style="font-family:Verdana;">double stranded RNA (dsRNA)</span><span style="font-family:""><span style="font-family:Verdana;"> via </span><i><span style="font-family:Verdana;">Agrobacterium</span></i><span style="font-family:Verdana;">-mediated transformation. Of the eight T</span>_{<span style="font-family:Verdana;">1</span>}<span style="font-family:Verdana;"> transgenic lines inoculated with YMV, six (L1, L2, L3, L5, L7 and L8) showed immunity to YMV as no symptoms were detected, whereas two (L4 and L10) exhibited high resistance with mild symptoms limited to inoculation portions. No virus could be detected in uninoculated new leaves of the transgenic lines after RT-PCR and qPCR analyses of </span></span><span style="font-family:Verdana;">YMV-coat protein (CP)</span><span style="font-family:Verdana;">. </span><span style="font-family:Verdana;">The presence of small interfering RNAs in transgenic</span><span style="font-family:MinionPro-Capt;"> </span><span style="font-family:Verdana;">lines after virus challenge indicates</span><span style="font-family:Verdana;"> that the resistance was acquired through RNA silencing.</span>Yam mosaic virus (YMV), a Potyvirus, is a highly destructive pathogen of yam accounting for yield losses up to 40%. Apart from causing significant reduction in tuber size and quality, it restricts international exchange of germplasms. It thus becomes crucial to get resistant or at least virus-free planting materials for farmers. This study was aimed at inducing resistance to YMV in tobacco by RNA silencing. An RNAi construct containing 161 bp fragment of <span style="font-family:Verdana;">YMV-coat protein (CP) </span><span style="font-family:Verdana;">gene was developed and used to produce transgenic tobacco lines expressing </span><span style="font-family:Verdana;">YMV-coat protein (CP)</span><span style="font-family:Verdana;"> derived </span><span style="font-family:Verdana;">double stranded RNA (dsRNA)</span><span style="font-family:""><span style="font-family:Verdana;"> via </span><i><span style="font-family:Verdana;">Agrobacterium</span></i><span style="font-family:Verdana;">-mediated transformation. Of the eight T</span>_{<span style="font-family:Verdana;">1</span>}<span style="font-family:Verdana;"> transgenic lines inoculated with YMV, six (L1, L2, L3, L5, L7 and L8) showed immunity to YMV as no symptoms were detected, whereas two (L4 and L10) exhibited high resistance with mild symptoms limited to inoculation portions. No virus could be detected in uninoculated new leaves of the transgenic lines after RT-PCR and qPCR analyses of </span></span><span style="font-family:Verdana;">YMV-coat protein (CP)</span><span style="font-family:Verdana;">. </span><span style="font-family:Verdana;">The presence of small interfering RNAs in transgenic</span><span style="font-family:MinionPro-Capt;"> </span><span style="font-family:Verdana;">lines after virus challenge indicates</span><span style="font-family:Verdana;"> that the resistance was acquired through RNA silencing.</span>

关 键 词：Yam Mosaic Virus DSRNA PTGS Coat Protein Gene RNAI 

分 类 号：O17[理学—数学]