关键词: 聚合酶链反应（PCR）, 地高辛标记探针, 斑点杂交, ZYMV, WMV, CMV, PRSV-W, SqMV

Abstract: Dot-blot hybridization was developed to detect 5 viruses infecting cucurbitaceous crops：Zuccini yellow mosaic virus（ZYMV），Watermelon mosaic virus（WMV）， Cucumber mosaic virus（CMV），Papaya ringspot virus- watermelon strain（PRSV-W）and Squash mosaic virus（SqMV）, and to provide a good alternative for detection of infected seeds in cucurbitaceous crops and for routine detection of 5 viruses in epidemiological and transgenic research.【Method】Using the clones of coat protein gene of five viruses, digoxigenin- labelled cDNA probes were synthesized by PCR with the specific primers designed according to the nucleotide sequences of the coat gene of the five viruses. The probes were applied in dot-blot hybridization to detect ZYMV、WMV、CMV、PRSV-W and SqMV in simple extraction of infected leaves.【Result】The sensitivity for detection of the simple extraction of infected leaves by ZYMV、WMV、CMV、PRSV-W and SqMV was 160、160、320、160、320 times respectively.【Conclusion】The digoxigenin-labelled probes prepared by PCR could be used for accurate and rapid identification of 5 viruses infecting cucurbitaceous crops in the growth stage with good stablities, sensitivities and specificness.

Key words: PCR, digoxigenin-labelled cDNA probe, 斑点杂交, ZYMV, WMV, CMV, PRSV-W, SqMV