摘要
用酶解-研磨法分离烟草与大叶烟草受精后的胚囊,继之以显微解剖分离合子与二胞原胚。将3—5个合子或二胞原胚置于微室的琼脂糖小滴中,以预先培养3—4天、分裂1—2次的烟草、大叶烟草或黄花烟草叶肉原生质体饲养。培养基为KM8p与其它附加成分。在25℃与黑暗下静置培养。培养3—4天,约60%合子完成第一次分裂。多数行不等分裂产生大小两个细胞。12天后形成少数原胚或多细胞团。二胞原胚培养亦分裂为多细胞原胚。研究了合子分离方法、合子发育时期、饲养细胞种类与培养天数等因素对合子离体发育的影响。
Fertilized embryo sacs of Nicotiana tabacum cultivar G - 80 and N. tabacum var. macrophylla were isolated by enzymatic maceration and grinding, then the zygotes or two-celled proembryos were isolated by microdis-section. Three to five zygotes or two-celled proembryos were embedded in agnrose in mi-crochambers (Millicell-CM, Millipore) feeded with 3- or 4- day- old mesophyll protoplasts of Nicotiana tabacum , N. tabacum var. macrophylla and N. rustica and cultured in KM8p basic medium supplemented with various addi-
tives at 25℃ in darkness. The zygotes underwent first division after 3d culture and the division frequency was approximately 60%. Microscopical observations showed that the first cell division was unequal in most of the zygotes. Few proembryos and multicellular structures were acquired after 12d culture . The two-celled proembryos also were induced to form multicellular proembryos. We also studied the effects of the isolation method, zy-gote stage and the genotype and age of feeder cells on in vitro development of zygotes.
出处
《实验生物学报》
CSCD
1998年第3期309-315,共7页
Acta Biologiae Experimentalis Sinica
基金
国家自然科学基金
高等学校博士点基金
并获得中国与欧盟合作项目(IC18CT960072)部分资助~~
关键词
烟草
合子
二胞原胚
离休培养
发育
Zygote. Two-celled proenlbryo. Microcullure. Nicotiana.
