Embryo rescue and plant regeneration in vitro of selfed chickpea (Cicer arietinum L.) and its wild annual relatives
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The main constraint to the transfer of desired traits into cultivated chickpea from wild Cicer relatives is the presence of post-zygotic barriers which result in abortion of the immature embryo following interspecific hybridisation. Rescue of hybrid embryos in vitro and regeneration of hybrid plantlets could allow chickpea breeders to transfer desirable traits from wild relatives of chickpea. The development of embryo rescue techniques using selfed chickpea and selfed wild relatives is being used as a first step to protocols for wide hybrids. Optical microscopy studies of embryogenesis, in both selfs and hybrids, identified deleterious changes in the fertilised hybrid seed as early as 2–4 days after pollination in some crosses. These observations suggest that the appropriate time to rescue chickpea × C. bijugum hybrids is at the early globular stage of embryogenesis (2–7 days old), which requires the development of a complex tissue culture medium. In contrast hybrids between chickpea × C. pinnatifidum abort later (up to 15–20 days old) at the heart-shaped or torpedo stages, and are easier to rescue in vitro. Genotype also plays a significant role in the ability of immature selfed ovules to germinate in vitro. In this paper we report on the optimisation of␣protocols for rescueing immature embryos using selfed chickpea and its wild relatives in ovule, and subsequently to regenerate plantlets.
Keywordsgrain legume improvement interspecific hybridisation tissue culture wild relatives
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This work is supported by the Grains Research and Development Corporation of Australia and CLIMA. We are grateful to the curators of genebanks, at the International Center for Agricultural Research in Dry Areas in Syria, the International Crops Research Institute for the Semi Arid Tropics in India and the Australian Temperate Field Crops Collection in Australia, for providing germplasm used in the study.
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