摘要
用饥饿预处理分离烟草（Ｎｉｃｏｔｉａｎａ ｔａｂａｃｕｍ Ｌ．）品系Ｎ３６４ Ｋｍ ＋ 二核早中期的花粉原生质体，用ＰＥＧ－高钙高ｐＨ 法诱导其与黄花烟草（Ｎ．ｒｕｓｔｉｃａ Ｌ．）叶肉原生质体融合。通过抗性筛选再生的４ 株小植株，经过氧化物酶同工酶、根尖染色体计数鉴定均为配子－体细胞三倍体杂种。未经筛选处理再生的２１株小植株，经鉴定有６ 株为配子－体细胞杂种。
Isolated pollen protoplasts of Nicotiana tabacum L.N364 Km + were fused with mesophyll protoplasts of N.rustica L.using PEG high Ca/pH method.The cells resulted from fusion between immature (early middle bicellular) pollen protoplasts and mesophyll protoplasts could divide to produce microcalli and regenerated plantlets when cultured in a selection KM8p medium containing 50 g/L kanamycin.Four plantlets were regenerated.The isoenzyme patterns of leaf peroxidases of these plantlets had bands characteristic of both parents.Root tip squash showed that the gameto somatic hybrids had the expected triploid chromosome number. Aside from these kanamycin selected plantlets,six of the twenty one plantlets that had not undergone selection were also evidenced to be gameto somatic hybrids.
基金
国家自然科学基金
关键词
烟草
花粉
原生质体
原生质体融合
叶肉
Nicotiana tabacum
Nicotiana rustica
Pollen Protoplast
Protoplast fusion
Gameto somatic hybridization
