Virus Elimination from'Shine Muscat'Grape Plantlets in vitro via Heat Treatment Combined with Shoot Tip and Axillary Bud Culture
In order to optimize the virus elimination system of grape plantlets in vitro,research on virus elimination was carried out using the'Shine Muscat'grape plantlets in vitro,which have been detected carrying five viruses including grapevine fabavirus(GFabV),grapevine rupestris stem pitting associated virus(GRSPaV),grapevine Pinot gris virus(GPGV),grapevine leafroll-associated virus-3(GLRaV-3),and grapevine virus E(GVE).The effects on virus elimination regarding heat treatment ways and duration,as well as plant materials(shoot tip or axillary bud)used for inoculation after heat treatment were studied.The results showed that the heat treatment of 38 ℃/light 8 h and 32 ℃/dark 8 h in turn generated the highest survival rate and virus elimination efficiency,followed by the treatment of 38 ℃/light 16 h and 32 ℃/dark 8 h in turn and constant 38 ℃(light 16 h+dark 8 h).Then,about 1.5 mm long shoot tips or the first or the second axillary buds under the shoot tips were cut from the plantlets cultured under the heat treatment of 38 ℃/light 8 h and 32 ℃/dark 8 h in turn for 10,15,20,25,30,and 40 days,respectively,for inoculation.The survival rate of the shoot tips was higher than that of the axillary buds.The new plantlets were detected for the viruses,and virus-free plantlets were obtained from all kinds of the materials.The virus elimination rate was 52.38％when the heat treatment duration was 10 d,and reached 100％when that was 25 d or 30 d.With the further increased duration,the survival rate of the inoculated shoot tips and the axillary buds decreased.The duration of 25 d was recommended.
grapeplantlet in vitroheat treatmentshoot tipaxillary budvirus elimination
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