柑桔球形合子胚的离体培养

以雪柑、芦柑和柚为母本,分别用芦柑、福桔、枳壳和柚的花粉进行授粉,观察合子胚的发育.结果表明授粉后50—55d,胚囊只有1个球形合子胚,珠心胚尚未侵人胚囊,可以单独分离培养.将合子胚接种于改良White基本培养基及其附加物。蔗糖浓度为10％.待子叶胚分化明显时,移入MT培养基。即可抽芽.芽长2—3cm,再转入诱导长根培养基,便发育成为正常的植株.试验自1986年获得雪柑与福桔和芦柑授粉的合子胚苗之后,1988—1989年又先后获得雪柑与枳壳,柚与枳壳,芦柑与柚授粉的合子胚苗.小苗的GOT基因酶和过氧化物同工酶测定,证明其杂种性,通过多次杂交试验,探索柑桔球形合子胚离体培养技术,有效地排除了珠心胚的干扰.

Culture of Citrus Globose Zygotic Embryos In Vitro

In this experiment, Shekan [C. sinensis(L.) Osbeck.], Ponkan (C. reticulata Blanco) and Pummelo (C. grandis) were used as maternal parents, and Ponkan, Fuji (C.tangerina Hert.ex.Tanaka), Poncirus (Poncirus trifoliata Raf.) as paternal parents to perform pollination. It was showed from observation of embryo development that 50- 55 days after pollination, there was only one globose zygotic embryo in embryo sac and the nucellar embryos hadn't invaded the sac.The zygotic embryo can be separated alone for culture.lt was inoculated on the basal medium Improved White, supplemented with some additives and 10% sucrose.When it developed into dicotyledonous embryo, it was transferred to the basal medium MT with sucrose, concentration of which reduced to 2%, where it sprouted to 2-3 cm long. The sprout was transferred to induce root medium and then developed into small normal plant. After getting zygotic plants of Shekan x (Fuji and Ponkan) in 1986, we gained zygotic plants of Shekan x Poncirus, Pummelo x Poncirus, Ponkan x Pummelo successively in 1988-1989.The results of the determination of the gene/ enzyme system of glutamate oxaloaeetate transaminase (GOT) and zymograms of isoperoxidase suggested that these plantlets were hybrids.Through experimenting repeatly, we had explored in vitro technique of citrus globose zygotic embryo, and gotten rid of effectively the interference of the nucellar embryos in hybridization breeding.