Analysis of chromosomal and organellar DNA of somatic hybrids between Triticum aestiuvm and Haynaldia villosa Schur
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O R I GI N A L P A P E R
A. Zhou á G. Xia á X. Zhang á H. Chen á H. Hu
Analysis of chromosomal and organellar DNA of somatic hybrids between Triticum aestiuvm and Haynaldia villosa Schur
Received: 25 April 2000 / Accepted: 23 November 2000 / Published online: 21 February 2001 Ó Springer-Verlag 2001
Abstract Intergeneric somatic hybridization between wheat (cv. Jinan 177) protoplasts that have 24±28 chromosomes and Haynaldia villosa protoplasts containing 11±14 chromosomes was carried out by the polyethylene glycol (PEG) method. A high frequency of hybrid calli and plants were obtained from the fusion products, as revealed by cytological and biochemical techniques and by PCR analysis of 5S rDNA spacer sequences. GISH (genomic in situ hybridization) analysis con®rmed the presence of chromosomes from both parents in the hybrid clones and the common occurrence of translocations between them. The RFLP analysis of the organellar DNA using mitochondrionand chloroplast-speci®c probes revealed that mitochondria from both parents existed in the cells of hybrid calli and their recombination, whereas chloroplasts segregated and recombined randomly. The gross morphology of hybrid plants resembled that of wheat, but the gross morphology of their ovaries and anthers were intermediate between those of the two parents. The relationship between hybrid plant regeneration and the balance of genetic materials in hybrid clones is discussed. Key words Triticum aestivum á Haynaldia villosa á Somatic hybrids á Genomic in situ hybridization (GISH) á Organellar genes
Communicated by R. G. Herrmann A. Zhou á G. Xia (&) á H. Chen School of Life Sciences, Shandong University, Jinan 250100, People's Republic of China E-mail: [email protected] Fax: +86-531-8565610 X. Zhang á H. Hu Institute of Genetics, Chinese Academy of Sciences, Beijing 100101, People's Republic of China
Introduction The arti®cial transfer of genetic information between evolutionarily remote species can serve to create novel plants that cannot be obtained by conventional breeding. In comparison to other techniques of chromosomal and gene engineering, somatic hybridization has the unique potential to transfer both nuclear and cytoplasmic gene(s) simultaneously. Somatic hybridization of dicotyledonous plants has been developed steadily during the past 30 years, but only limited success has been achieved in Gramineae (with the exception of rice), especially in intergeneric hybridization or cell fusion experiments involving more remotely related species (Li et al. 1999). Haynaldia villosa has been chosen as a donor for intergeneric hybridization to wheat because it has several agronomically useful traits for the improvement of wheat, such as disease resistance (especially against powdery mildew), high protein and lysine content etc. Sexual and somatic hybrids between common wheat and H. villosa have been obtained previously (Liu et al. 1988; Zhou et al. 1996), but the earlier work was tedious and time consuming, and the hybrids obtained from somatic hybridization were ste
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