The Studies On Main Factors To Affect Isolated Microspore Culture In Cabbage

Pollen from 19 genotypes of cabbage was cultured. The effects of cabbage genotype and affecting factors were analyzed on isolated microspore culture, meanwhile , using EW10,EF27,EF14,Qingan70,Qingan80 for materials , factors were studied that had effects on induction culture of cabbage ,such as bud length, low temperature pretreatment , high temperature treatment , AgNO₃ , active carbon , hormon and pH. The results showed that:1. Genotype was main factor that had an effect on success in cabbage pollen culture. It was significant difference in induction rate of pollen culture. Of the 19 experiment materials, 16 genotypes produced embryoids. Among them, 5 lines (EW10, EW09, EF27, Qingan70, and Qingan80) produced higher embryo yield. In addition there had 3 materials could not gain embryoids no matter what took any methods.2. Bud size had an important effect on microspore culture. According to observing bud length under microscope, found above 70% microspore of 3.1～4.5mm bud length was uninucleate period and binuclear period, this showed that under this range 3.1～4.5mm bud length was the best induction embryoids period.3. Low temperature pretreatment and proper high temperature was important fators for increasing induction rate of microspore cuture. Low temperature pretreatment had a significant effect for embryoid yield, different genotype had different pretreatment time .All the materials microspore embryoid yield had significant improvement under low temperature pretreatment. The treatment that buds were pretreated with 4℃low temperature during 48 hours before microspore cultured and microspore were cultured darkling with 33℃high temperature during 24 hours at initial stage , could obtain better inducing result.4. AgNO₃ could not promote microspore culture induction rate, and induction rate of adding AgNO₃ treatment was lower than no adding AgNO₃ treatment. Accession active carbon in medium had significant effect for embryoids induction and its formation. In the active carbon medium, embryo yield proportion had greatly advanced. It was obvious that proper active carbon played important roles on embryosis induction.5. Hormone had important affect roles on microspore induction culture, but different genotypes had different effect to hormone. According to contrast of addition hormone 6-BA and NAA five concentration proportion in the NLN-13 fluid medium to process microspore induction embryoid culture. Qingan70's embryosis rate was 3.00 embryo per bud under no hormone medium, higher than other hormone concentration treatment medium. Qingan80's embryosis rate was 3.75 embryo per bud under single addition hormone 6-BA0.05mg/l and higher than all other concentration treatment medium. EF14's embryosis rate was higher with the hormone 6-BA0.05mg/l and NAA0.5mg/l, which were 2.25 embryos per bud. While wanfeng's embryosis rate was higher with the hormone 6-BA0.5mg/l adds to NAA0.5mg/l, that was 4.25 embryos per bud. These results showed that different genotypes had different effects to hormone. It was good for embryoids induction on pH5.8, and it was not benefit for embryoids formation under very high and very low pH.