Arya S, Kalia R K, Arya I D
Plant Tissue Culture Lab, Division of Genetics and Tree Propagation, Forest Research Institute, Dehradun - 248006, India e-mail:
Plant Cell Rep. 2000 Jul;19(8):775-780. doi: 10.1007/s002990000197.
Embryogenic calli were initiated from embryonic explants of Pinus roxburghii using female gametophytes containing immature pre-cotyledonary embryos. Zygotic embryos were collected at different developmental stages and cultured on various media. Initiation of embryogenic calli was achieved in pre-cotyledonary zygotic embryos of a 0.1-mm to 1.2-mm embryonal head on Douglus fir cotyledon revised medium (DCR) medium supplemented with 2,4-D or NAA and BA. Embryogenic callus development was initiated from the suspensor region of immature embryos. The method of immature embryo culture was significant as rapid embryogenic callus development occurred in megagametophytes where the suspensor was stretched onto the medium from the cut micropylar end. Sixty embryogenic lines were established from 2500 explants cultured during one season. A pro-embryo with six to eight meristematic cells and suspensor of six to ten long, vacuolated cells dominated the early phase of the callus development. Cleavage polyembryony occurred in proliferating callus, constituting a method of multiplication of these somatic embryos. Somatic embryos developed to stage-I and stage-II embryos on DCR medium supplemented with 5 μM 2,4-D or 10 μM NAA.
利用含有未成熟子叶前胚的雌配子体，从喜马拉雅松的胚性外植体诱导出胚性愈伤组织。收集不同发育阶段的合子胚并在各种培养基上培养。在补充了2,4-D或NAA和BA的花旗松胚轴改良培养基（DCR）上，0.1毫米至1.2毫米胚头的子叶前合子胚实现了胚性愈伤组织的诱导。胚性愈伤组织发育始于未成熟胚的胚柄区域。未成熟胚培养方法很重要，因为在大配子体中胚性愈伤组织发育迅速，胚柄从切开的珠孔端延伸到培养基上。在一个季节培养的2500个外植体中建立了60个胚性系。一个具有六到八个分生细胞的原胚和六到十个长的、液泡化细胞的胚柄在愈伤组织发育的早期占主导。裂生多胚现象发生在增殖的愈伤组织中，构成了这些体细胞胚的一种增殖方法。体细胞胚在补充了5μM 2,4-D或10μM NAA的DCR培养基上发育到I期和II期胚。
