摘要
多年生黑麦草(Lolium perenne L.)悬浮培养细胞来源的原生质体和小麦(Triticumaestivum L.)含叶绿体的悬浮培养细胞来源的原生质体间,用直流方波脉冲进行电融合,获得了体细胞杂种愈伤组织。小麦的原生质体经过碘乙酰胺失活。愈伤组织的形态和颜色被用作识别预期杂种的标记。为了对杂种愈伤组织进行同工酶分析,观察了亲本的9种同工酶谱,其中3种在亲本间表现出差异(ADH、GOT 和 SDH)。酒精脱氢酶(ADH)的分析结果表明,有6个细胞系表现出杂种带。这些细胞系经过其他两种同工酶分析和 rDNA 探针杂交试验表明,一个细胞系表现出基本完全的亲本基因组间的组合,其余5个细胞系是部分杂种。
Somatic hybrid calli were recovered following electrofusion of protoplasts from a chloro- plast-containing cell suspension culture of whear(Triticum aestivum L)and cell suspen- sion culture of ryegrass(Lolium perenne L.).The protoplasts of wheat were inactivated by io- doacetamide;in addition morphology and colour were used as markers to aid selection of pu- tative hybrid calli.For isozyme analysis of putative hybrids,nine isozymes were tested for di- fferences in bands between the parental lines.Of these,three showed differences(ADH,GOT, SDH),Analysis of ADH bands of calli indicated that six lines were hybrids.These lines were analysed with the other isozymes,and at the DNA level by Southern hybridisation with a wheat ribosomal DNA probe.The overall results indicated that one line was an almost complete com- bination of the genomes of the parental lines,but the other 5 lines were probably partial hy- brids.In the latter,some loss of the wheat genome had probably occurred.
关键词
黑麦草
小麦
多年生黑麦草
电融合
Wheat
Perennial ryegrass
Protoplast
Electrofusion
Isozyme
RFLP
