| Pumpkin double haploid breeding technology can effectively improve the speed of pumpkin breeding,the unfertilized ovule culture method is one of the main content of the technology.In order to explore the best condition of influencing factors of pumpkin unfertilized ovules culture,the ‘Beilihui’ as experimental materials to explore the different ovary slice thickness,different pre-processing,different ways of sterilization and different culture conditions,hormone concentration and the ratio of hormones,elements concentrations and types.The main results were as follows:1.Screening the best condition of influencing factors of pumpkin unfertilized ovule:(1)The rate of one day before flowering pumpkin ovule stigmas and turns green is 85%,the induction rate of callus is 32.5%.The rate of on the flowering day pumpkin ovule stigmas and turns green is 100%,the induction rate of callus is 57.5%.Therefore,the material of on the flowering day is the best.(2)The optimized thickness of the ovary slice was 1.0mm.(3)When sodium hypochlorite concentration was 7%,the rate of ovule stigmas and turns green is 100%,no pollution.(4)At the ambient temperature of 38℃,dark heat treatment 5d,the ovules was swelled and turned green.(5)When cultue temperature is 30 ℃,the light intensity is 2000 lx,illumination time of 14 h,the ovules stigmas and turns-green is 92.86%。2.Screening of ovules expanded and turn-green medium.Adding 1.0 mg/L2,4-D+1.0 mg/L6-BA+0.5 mg/LNAA on MS medium,the induction ratio was the highest attaining 89.29%--95.24%.After ovule expanded and turned green,the ovule surface turned smooth and full.Moreover ovule induced into globular embryo gradually3.Adding CaCl2 concentration of 40mg/L to the Z medium,the rate of callus inductionand turn-green are 100%.4.The medium of buds induction failed to induct budding of ovules.The experiment will continue in the future. |
