甘蓝与甘蓝型油菜杂交胚挽救技术研究

Study on hybrid embryo rescue technology of cabbage and Brassica napus

为恢复甘蓝(Brassica oleracea L.)雄性不育材料的育性,以甘蓝雄性不育系GL16047为母本,甘蓝型油菜N18008为父本进行杂交,对杂交后子房和胚珠进行离体培养,筛选适合甘蓝与甘蓝型油菜杂交胚挽救培养的取材时间和培养基。结果表明,授粉后的离体时间对杂种胚的发育影响很大,过早或过晚取样对子房出胚率都有影响。授粉后第5天取样进行离体子房培养,子房生长量小,出胚数少;授粉后第10天取样进行离体子房培养,随着授粉后时间的延长,杂种核和卵细胞的代谢不协调对杂种胚发育的影响增大,出胚数减少;胚龄的大小决定出胚率,种胚过小脱离母体后离体培养难度较大不容易存活,出胚率低,胚成活最短时间在授粉后第16天,超过24 d取样,受精后期异源遗传物质的导入致使胚珠内遗传不协调,胚容易出现败育,出胚率降低。NAA、6-BA两种激素对离体子房和胚珠的发育和形成起到很好的促进作用,NAA浓度相同,子房培养以1.0 mg/L 6-BA效果最好,胚珠培养以5.0 mg/L 6-BA效果最好。以授粉后第8天为子房培养适宜取样时间,授粉后第20天为胚珠培养适宜取样时间;以MS+1.0 mg/L6-BA+1.0 mg/L NAA+1.0 mg/L KT+活性炭1 000 mg/L培养基适宜子房培养,MS+5.0 mg/L 6-BA+1.0 mg/L NAA+1.0 mg/L KT+活性炭1 000 mg/L培养基适于胚珠培养。

In order to restore the fertility of male sterile materials of kale(Brassica oleracea var.capitata L.),the male sterile line of kale GL16047 was used as the female parent and the Brassica napus N18008 was used as the male parent for crossing,and the ovaries and ovules were isolated and cultured after crossing to screen the suitable sampling time and culture medium for hybrid embryo salvage culture of kale and Brassica napus.The results showed that the time of isolation after pollination had a great influence on the develop⁃ment of hybrid embryos,and sampling too early or too late had an effect on the ovary embryo rate.On the 5th day after pollination,samples were taken for isolated cohort culture,the ovary growth was small,and the number of embryos was low.On the 10th day after pollination,samples were taken for isolated cohort culture.With the prolongation of post-pollination time,the influence of the uncoor⁃dinated metabolism of the hybrid nucleus and egg cell on the development of hybrid embryo increases,and the number of embryos emerging decreases;the embryo age determines the embryo rate.If the seed embryo is too small,it is difficult to culture in vitro after being separated from the mother.The shortest time for embryo survival is the 16th day after pollination.If sampling exceeds 24 days,the introduction of heterologous genetic material at the later stage of fertilization will cause genetic incongruity in the ovule,embryos are prone to abortion,and the embryo rate will decrease.The two hormones,NAA and 6-BA,play a very good role in promoting the development and formation of the isolated ovary and ovule.At the same concentration of NAA,1.0 mg/L 6-BA was the best for ovary culture,and 5.0 mg/L 6-BA was the best for ovule culture.The 8th day after pollination is the suitable sampling time for ovary cul⁃ture,and the 20th day after pollination is the suitable sampling time for ovule culture.MS+1.0 mg/L 6-BA+1.0 mg/L NAA+1.0 mg/L KT+activated carbon 1000 mg/L medium is suitable for ovary culture;MS+5.0 mg/L 6-BA+1.0 mg/L NAA+1.0 mg/L KT+activated carbon 1000 mg/L medium is suitable for ovule culture.