通过“供体-受体”原生质体融合将裸燕麦部分核基因组转移给普通小麦(英文)

TRANSFER OF PARTIAL GENOME OF AVENA NUDA L.INTO TRITICUM AESTIVUM L.BY “DONOR-RECIPIENT” PROTOPLAST FUSION

本文进行了小麦和裸燕麦悬浮细胞原生质体的电融合,并基于双亲失活(用IOA处理受体小麦原生质体,用γ-射线照射供体裸燕麦细胞系),获得可能的杂种愈伤组织。对7块愈伤组织进行了乙醇脱氢酶(Adh)同工酶筛选,发现5块表现出双亲特征酶带。对3个杂种细胞系进行5种同工酶分析,证实它们均为稳定的不对称体细胞核杂种细胞系;它们表现出小麦的完整谱带和裸燕麦的部分谱带。对2个杂种细胞系及亲本的核糖体DNA Southern分析结果表明只有一个杂种细胞系(HB 95)含有双亲的全部谱带。细胞学观察表明,2个杂种细胞系的染色体数目均显著高于双亲。从杂种细胞系HB 94中分化出叶原基等分化结构。Adh同工酶分析表明,这些分化结构具有和母体细胞系完全相同的杂种谱带。

Wheat and oat suspension protoplasts were electrofused, and putative hybrid ca- llus was recovered on the basis of double inactivation: by IOA treatment on wheat protoplasts and gamma-ray irradiation on oat cell suspension. Adh isozyme screening showed that 5 of the 7 calli were with characteristic enzymatic bands of both parents. Five isozyme analysis of 3 putative hybrid cell lines confirmed that they were all stable asymmetric nuclear hybrid cell lines: they expressed intact enzymatic bands of wheat and asymmetric bands of oat. Ribosomal DNA Southern analysis of 2 hybrid cell lines (HB94 and HB95) indicated that only one cell line (HB 95) covered bands of both parents; HB 94 only exhibited bands of wheat. Cytological observation showed that hybrid cell lines had significantly more chromosomes than either of the parents. Distinct differentiated structures, such as leaf primodia were obtained from HB 94 cell line and they expressed the same hybrid Adh isozyme activity as the cell line they derived from.