Abstract
BackgroundCowpea (Vigna unguiculata L.) is an important grain and forage legume grown throughout sub-Saharan Africa primarily by subsistence farmers on poor, drought prone soils. Genetic improvement of the crop is being actively pursued and numerous functional genomics studies are underway aimed at characterizing gene controlling key agronomic characteristics for disease and pest resistances. Unfortunately, similar to other legumes, efficient plant transformation technology is a rate-limiting step in analysis of gene function in cowpea.ResultsHere we describe an optimized protocol for the rapid generation of transformed hairy roots on ex vitro composite plants of cowpea using Agrobacterium rhizogenes. We further demonstrate the applicability of cowpea composite plants to study gene expression involved in the resistance response of the plant roots to attack by the root parasitic weed, Striga gesnerioides. The utility of the new system and critical parameters of the method are described and discussed herein.ConclusionsCowpea composite plants offer a rapid alternative to methods requiring stable transformation and whole plant regeneration for studying gene expression in resistance or susceptibility responses to parasitic weeds. Their use can likely be readily adapted to look at the effects of both ectopic gene overexpression as well as gene knockdown of root associated defense responses and to the study of a broader range of root associated physiological and aphysiological processes including root growth and differentiation as well as interactions with other root pests, parasites, and symbionts.
Highlights
Cowpea (Vigna unguiculata L.) is an important grain and forage legume grown throughout subSaharan Africa primarily by subsistence farmers on poor, drought prone soils
An optimized protocol for the generation of ex vitro composite cowpea plants The creation of ex vitro composite plants consisting of wild-type shoots and transgenic roots induced by transformation with A. rhizogenes has been described for several species [35,36,37,38]
Plants showing any signs of disease are discarded, and only vigorous, healthy looking cowpea plants are carried forward for the production of composite plants
Summary
Cowpea (Vigna unguiculata L.) is an important grain and forage legume grown throughout subSaharan Africa primarily by subsistence farmers on poor, drought prone soils. Genetic improvement of the crop is being actively pursued and numerous functional genomics studies are underway aimed at characterizing gene controlling key agronomic characteristics for disease and pest resistances. Similar to other legumes, efficient plant transformation technology is a rate-limiting step in analysis of gene function in cowpea. Walp.) is the most important grain legume grown in sub-Saharan Africa [1,2]. 12.5 million tons of cowpea grains are produced worldwide each year with a majority (over 64%) of the production taking place on low-input, subsistence farms in West and Central Africa [3]. Improvement of cowpea as a multifunctional crop is a key breeding concern and significant efforts are currently aimed at its genetic improvement [2]
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