| Isolated microspores culture technique applied to plant genetic and breeding widely,because researchers can attain valuable double haploid for breeding by the technique with shorter time and higher efficiency. On the other hand ,the embryoid and regeneration plantlet of isolated microspores culture are used to The construction of genetic map, gene direction, trans-gene breeding , chromosome engineering, and so on.Improving microspores embryogenic ability and plantlet regeneration ability from microspore-derived embryos is the key of isolated microspores culture technique. We screened out 16 embryogenic material from sixty kinds of head cabbages, six kinds of cauliflowers, three kinds of spore cabbages and two kinds of collards, Then the genotypes of higher embryogenic ability in the 16 embryogenic materials were used to probe into the factors that effect on the embryogeny of Brassica oleracea , statistic plantlet regeneration frequency, and check the naturally diploidization frequency of regeneration plantlet by FACSCalibur and SRAP (Sequence-related amplified polymorphism) identify of the regeneration plantlet. The results were as follows:1. Microspores from late uninucleate-staged to binucleuses-staged are optimal to be bred in Brassica oleracea,It is propitious to produce embryos in head cabbage when the proportion of binucleuses-staged microspores is 50% or so.2. There were more kinds of genotypes produce embryos when the sucrose concentration of the medium up to 13% except individual genotype. Higher yield of microspore and embryos could be obtained with a method that decrease the sucrose concentration gradually in the medium during the culture, and the medium with the NAA (0.05 mg·mL-1) +BA (0.1 mg·mL-1) was suitable for the produce .If adding activated charcoal in the medium where embryoid grows, the effect is more prominent than not. |
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