PARTICLE BOMBARDMENT DRASTICALLY INCREASES THE INFECTIVITY OF CLONED DNA OF ZUCCHINI YELLOW MOSAIC POTYVIRUS

An infectious full-length cDNA clone of the RNA genome of the potyviru s zucchini yellow mosaic virus (ZYMV) was constructed under the contro l of the cauliflower mosaic virus 35S promoter. All squash, cucumber, melon and watermelon plants inoculated with the cloned cDNA of ZYMV by particle bombardment become infected. Bombardment technology is 10(6) -fold more effective than mechanical inoculation. Due to the great inc rease in efficiency, ineffective constructs now became infective (i.e. cDNA under the control of the 35S promoter without the NOS terminator ; with an addition of 127 nucleotides at the 5' end of the viral cDNA; uncapped transcripts), and the infectivity of capped-transcripts was maximized. Inoculation by particle bombardment produced visual symptom s rapidly (3-4 days), allowing the detection of viral coat protein and virions after 2 and 3 days in systemically infected leaves and inocul ated cotyledons respectively.