Study On The Method Of Pumpkin Unfertilized Ovary Vitro Induced Plant

Pumpkin unfertilized ovary in vitro inducing regeneration plant is an important part of the pumpkin haploid breeding technology, is also an important means of pumpkin breeding.It has obvious advantages. This test is to pumpkin varieties "chestnut grey" as test materials,to study the method of pumpkin unfertilized ovary in vitro regeneration plant induced. This paper mainly discusses the influence of different types medium, different sampling time, different ovary slice thickness, different pretreatment, squash pollen stimulus to pumpkin in vitro female nucleus embryoid induction; The study of pumpkin embryonic callus bud induction; The research of pumpkin fertilized ovule in vitro culture and rooting culture. This experiment can lay a certain foundation to pumpkin nuclear culture technology in vitro female system. The main results are as follows:1.Different kinds and concentration of culture medium has different effect on pumpkin unfertilized ovary biopsy embryoid induction: the rate of ovule stigmas and turnsgreen is higher in the medium of 2,4-D+NAA+6-BA combination, attaining 89.29%--95.24%.The surface of ovule after stigmas and turnsgreen turns smooth and full. Then, ovule induce into globular embryo gradually.2.Using pumpkin fertilized ovule to test medium. The rate of embryo is the highest after three weeks pollination, attaining 61.14%.3. The effect of root induction is best in the rooting medium of 0.5mg/L GA3.The potential root is thick and long. It is conducive to improve the survival rate of smelting plants. The effect of root induction is worse in the rooting medium of NAA.4.Different sampling time has different effect on pumpkin unfertilized ovary biopsy embryoid induction: The rate of one day before flowering pumpkin ovule stigmas and turnsgreen is 77.14%.The rate of on the flowering day pumpkin ovule stigmas and turnsgreen is 71.07%.Two sampling times of the ovule stigmas and turnsgreen rate are both higher, and there was no significant difference.5.Different slice Thickness has different effect on pumpkin unfertilized ovary biopsy embryoid induction: The rate of ovule stigmas and turnsgreen is the highest in 1mm,and it isthe best slice thickness.The medium of NAA+6-BA combination failed to budding of embryonic callus induction. Callus only split without differentiation, still need to test further research.6.Different pretreatment has different effect on pumpkin unfertilized ovary biopsy embryoid induction: 35℃ dark heat shock treatment can accelerate the ovule swelled to turn green, and the best results is in 5 days.4℃low temperature treatment inhibit the ovule swelled to turn green.7.Squash pollen can improve the rate of ovule swelled to turn green after 1 day. But the rate of ovule stigmas and turnsgreen is decreased obviously after 2 days.