RAPDs for Identification of Somatic Hybrids,Protoplasts,Auxotrophic Mutants,Heterokaryons,Isolation,Isozymes,Proteins

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	RAPDs for Identification of Somatic Hybrids - One of the problems in production of somatic hybrids, often, is lack of appropriate methods for identification of these hybrids among the developing regenerants. Some if the methods traditionally used, include the following: (i) combining light bleached protoplasts with green leaf mesophyll protoplasts, followed by manual isolation of heterokaryons; (ii) genetic complementation of auxotrophic mutants; (iii) fusion of different antibiotic and or herbicide - resistant lines derived by plant transformation; and (iv) use of hybrid vigour, isozymes, proteins and/or RFLPs.
These methods, however, are time consuming and costly. RAPDs can be utilized for the same purpose, where a small DNA sample from developing regenerant can be used to determine hybridity. Primers may be used which give polymorphic PCR products from each of the parents involved in fusion, so that the hybrids can be identified by the presence of molecular profiles, which represented a combination of the profiles of the two parents. This has been achieved successfully in potato (Baird et al. 1992), and the approach will certainly be utilized in future for identification of somatic hybrids in a variety of plant materials.