In Vitro Tissue Culture And Virus-Free Detection Of Potato

The main reason of potato degeneration is virus. The main way of control potato virus is used no virus seedling, stem/shoot tip culture is the main way of potato virus-free. The principium of stem/shoot tip culture is utilization of heterogenicity of virus distribute in plant tissue, which no or only a few nucleocapsid in tissue of tip of plant stem or shoot. If we cut off the tip tissue which no nucleocapsid in stem, then we could get virus-free tube seedling by issue culture., if we could ensure no nucleocapsid in tube seedling by detection, then we could cut the seedling to segment with bud, put them in cuttage medium in greenhouse or netshed, get cuttage seedling, mini-tuber of potato after someday culturing, these are first original seed. We study on the effect of growth regulator to potato issue culture seedling, the effect of cuttage medium and nutrition solution, which irrigate in cuttage medium to cuttage seedling, and detection of virus-free seedling using RT-PCR in this dissertation. The main conclusion as following:1 There are different effects of different concentration of NAA and KT to potato issue culture seedling, and there are interaction of this two growth regulator.2 The model functions of different NAA and KT concentration influence towards potato issue culture seedlings fresh weight, root weight, root number, bud number were:(Z₁,Z₂,Z₃,Z₄)^T=(seedlings weight, root weight, root number, bud number)^T, x was NAA, y was KT3 The best concentration of NAA and KT were 0.0649ml/L and 0.6020ml/L, for root weight, the best concentration of NAA and KT were 0.0904ml/L and 0.4260ml/L, for root number, the best concentration of NAA and KT were 0.0728ml/L and 0.5193ml/L, for bud number, the best concentration of NAA and KT were 0.0561ml/L and 0.2420 ml/L. So the best concentration of NAA and KT were 0.05-0.08ml/L and 0.4-0.6ml/L for potato organization culture seedlings.4, when the cuttage medium prescription was vermiculite/deeply decayed sheep dung/ muck from forest land=3/l/l(V), the survival rate and raising status of variety Kexin-1 and Anshu56 were best. When the prescription was vermiculite/ muck from forestland=3/l (V), the survival rate and raising status of variety Kexin-1 and Anshu56 take second place. When the prescription was vermiculite/deeply decayed sheep dung =3/1 (V), the survival rate and raising status of variety Kexin-1 and Anshu56 take third place. But them all better than control(only vermiculite), the survival rate of Kexin-1 enhance 8.78%, 13.67% and 11.89% respectively, and the survival rate of Anshu56 enhance6.78%, 11% and 10.67 % respectively,When add deeply decayed sheep dung in cuttage medium, it's favorable to foot stretch, and when add muck from forestland in cuttage medium, it's favorable to seedling stretch. But mechanism need more study deeply.5 The effect of different compound nutrition solution to the survival rate and raising status of cuttage seedling was no distinct different. It's mean needn't nutrition solution that contain microelement to water cuttage seedling in practice. The nutrition solution only contains NH4NO3 and KH2PO4 could appease the need of cuttage seedling growing.6 The virus had been detected rate from cuttage seedling was 6.7% in total used RT-PCR, and the PVX> PVY^ PLRV had been detected rate from Kexin-1 cuttage seedling was 0%, 6.7% and 6.7% respectively, they had been detected rate from Anshu56 cuttage seedling was 13.33%, 6.7% and 6.7% respectively. There are 6 time had been detected distinctly, obviously and especially belt in total 90 time detection, these especially belt just same as the belt that no free-virus seedling,other 84 no especially belt. The especially belt length were 750bp,810bp and 510bp respectively. It's coincide with we wish 750bp, 812bp and 512bp. so we could said that 6 seedling was free-virus but no success seedling. It's mean respective primer to P\O^ PVY> PLRV are characteristic.