The Research On Hybrid Embryo Salvage Technology Of Prunus Discoidea

Prunus discoidea is a unique native cherry species in China,which is a rare flowering tree in early spring in China and has good adaptability to the climatic environment of the central-eastern region of China.In this study,P.discoidea was used as the female and P.conradinae,P.dielsiana,and P.campanulata were used as the parents in a distant cross.and used the hybrid embryo as the material,through fruit development observation,We obtained the hybrid embryo rescue technology and group culture fast propagation technology of P.discoidea by observing fruit development,culturing young embryos of different embryonic ages,screening basic culture medium and plant growth regulators,and explored the method of hybrid progeny identification,and the results of the study are as follows:(1)The fruit development observation showed that the fruit maturity time of spring cherry was 42 d,the fruit development curve was double S-shaped,the embryo abortion started 17 d after pollination,and the suitable embryo rescue time was 27 d after pollination(PF>0.5).(2)Embryo salvage tests showed that the suitable medium for P.discoidea × P.dielsiana embryo rescue is MS+1.0 mg·L-1 6-BA+1.0 mg·L-1 KT+0.1 mg·L-1 NAA;the suitable medium for P.discoidea × P.conradinae embryo rescue is MS+1.0 mg·L-1 6-BA+1.5 mg·L-1 KT+0.3 mg·L-1 NAA;the suitable medium for P.discoidea × P.campanulata embryo rescue is MS+1.5 mg·L-1 6-BA+1.5 mg·L-1 KT+0.1mg·L-1 NAA.(3)Embryo rescue seedling multiplication test showed that the suitable medium for the proliferation of P.discoidea × P.dielsiana is MS+1.0 mg·L-1 6-BA+0.2 mg·L-1 NAA+ 0.5 mg·L-1 GA3;the suitable medium for the proliferation of P.discoidea × P.conradinae is MS+1.5 mg·L-1 6-BA+0.1 mg·L-1 NAA+1.5 mg·L-1 GA3;the suitable medium for the proliferation of P.discoidea × P.campanulata is MS+1.0 mg·L-1 6-BA+0.1 mg·L-1 NAA+1.0 mg·L-1 GA3.(4)The rooting test of the proliferation seedlings showed that the suitable medium for rooting of P.discoidea × P.dielsiana is WPM+1.0 mg·L-1 NAA+0.5 mg·L-1 IBA;the suitable medium for rooting of P.discoidea × P.conradinae is WPM+1.0 mg·L-1 NAA+1.0 mg·L-1 IBA;the suitable medium for rooting of P.discoidea× P.campanulata is WPM+0.5 mg·L-1 NAA+1.0 mg·L-1 IBA.(5)The suitable time for rooting seedlings was 5 d.The suitable substrate for transplanting was peat:perlite(1:1).(6)The morphological identification of the 94 hybrid seedlings obtained from the crosses showed that the obtained hybrid seedlings could be morphologically recognized as true hybrids after comparative analysis.By ISSR molecular identification,four primers were selected to identify 18 of the 94 hybrid seedlings,and all 18 were true hybrids and genetically biased toward the parent.It showed that morphology combined with ISSR molecular identification could accurately identify the authenticity of the hybrids.