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禾本科属间体细胞杂交:高羊茅(Festuca arundinacea Schreb.)与多花黑麦草(Lolium multiflorum Lam.)之间的体细胞杂种植物。

Intergeneric somatic hybridization in Gramineae: somatic hybrid plants between tall fescue (Festuca arundinacea Schreb.) and Italian ryegrass (Lolium multiflorum Lam.).

作者信息

Takamizo T, Spangenberg G, Suginobu K, Potrykus I

机构信息

National Grassland Research Institute, Tochigi, Japan.

出版信息

Mol Gen Genet. 1991 Dec;231(1):1-6. doi: 10.1007/BF00293814.

Abstract

Tall fescue (Festuca arundinacea Schreb.) protoplasts, inactivated by iodoacetamide, and non-morphogenic Italian ryegrass (Lolium multiflorum Lam.) protoplasts, both derived from suspension cultures, were electrofused and putative somatic hybrid plants were recovered. Two different genotypic fusion combinations were carried out and several green plants were regenerated in one of them. With respect to plant habitus, leaf and inflorescence morphology, the regenerants had phenotypes intermediate between those of the parents. Southern hybridization analysis using a rice ribosomal DNA probe revealed that the regenerants contained both tall fescue- and Italian ryegrass-specific-DNA fragments. A cloned Italian ryegrass-specific interspersed DNA probe hybridized to total genomic DNA from Italian ryegrass and from the green regenerated somatic hybrid plants but not to tall fescue. Chromosome counts and zymograms of leaf esterases suggested nuclear genome instability of the somatic hybrid plants analyzed. Four mitochondrial probes and one chloroplast DNA probe were used in Southern hybridization experiments to analyze the organellar composition of the somatic hybrids obtained. The somatic hybrid plants analyzed showed tall fescue, additive or novel mtDNA patterns when hybridized with different mitochondrial gene-specific probes, while corresponding analysis using a chloroplast gene-specific probe revealed in all cases the tall fescue hybridization profile. Independently regenerated F. arundinacea (+) L. multiflorum somatic hybrid plants were successfully transferred to soil and grown to maturity, representing the first flowering intergeneric somatic hybrids recovered in Gramineae.

摘要

用碘乙酰胺灭活的高羊茅(Festuca arundinacea Schreb.)原生质体和来自悬浮培养物的非形态发生型意大利黑麦草(Lolium multiflorum Lam.)原生质体进行了电融合,并获得了假定的体细胞杂种植物。进行了两种不同基因型的融合组合,其中一种组合再生出了几株绿色植株。就植株习性、叶片和花序形态而言,再生植株的表型介于双亲之间。使用水稻核糖体DNA探针进行的Southern杂交分析表明,再生植株同时含有高羊茅和意大利黑麦草特有的DNA片段。一个克隆的意大利黑麦草特有的散布DNA探针与意大利黑麦草和绿色再生体细胞杂种植物的总基因组DNA杂交,但不与高羊茅杂交。叶片酯酶的染色体计数和酶谱表明所分析的体细胞杂种植物的核基因组不稳定。在Southern杂交实验中使用了四个线粒体探针和一个叶绿体DNA探针来分析所获得的体细胞杂种的细胞器组成。当与不同的线粒体基因特异性探针杂交时,所分析的体细胞杂种植物显示出高羊茅、累加或新的线粒体DNA模式,而使用叶绿体基因特异性探针进行的相应分析在所有情况下都显示出高羊茅的杂交图谱。独立再生出的高羊茅(+)意大利黑麦草体细胞杂种植物成功移栽到土壤中并生长至成熟,这是禾本科中首次获得的开花属间体细胞杂种。

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