Digoxigenin(DIG)标记RNA探针检测侵染花生的Potyviruses

DIGOXIGENIN(DIG)-LABELED RNA PROBES FOR THE DETECTION OF POTYVIRUSES INFECTING PEANUT(ARACHIS HYPOGAEA)

以花生条纹病毒(PStV)和花生斑驳病毒(PMV)克隆cDNA为模板,体外转录合成Digo-xgenin(DIG)标记PStV—I_9、PStV-I_(131)、PStV—I_(57-7)和PMV—A_(15)四种RNA探针。在点杂交反应中,PStV—I_9RNA探针能检测出0.25ng提纯PStV和花生病汁液62500倍稀释液中PStV。PMV—A_(16)RNA探针能检测到0.67ng提纯PMV和菜豆病汁液5120倍稀释液中PMV。两种病毒RNA探针均有高度特异性。PStV—I_(57-7)和PStV—I_(131)两种RNA探针具有与PStV—I_9RNA探针相同敏感性,但特异性稍差。

Three Digoxigenin (DIG)labeled PStV RNA Probes and one PMV RNA Probes were synthesised by use of PStV and PMV cloned cDNA as templates in vitro transcription.PStV-I9 RNA probe detected as little as 0.25 ng of purified PStV and 1; 62 500 dilution of PStV infected peanut leaf sap.PMV A15 RNA probe detected as little as 0.67ng of purified PMV and PMV infected extracts of bean up to a dilution of 1:5120.Both RNA probes had high specificity.PStV-I57-7 and PStV-I131 RNA probes were as sensitive as PStV-I9 RNA-probe for detection of homologous viruses.However,its specificity was less than PStV I9 RNA probe.