Plant regeneration from Crotalaria spectabilis hairy roots which showed inhibited growth of root-knot nematodes (Meloidogyne hapla and M. incognita) in vitro

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Hairy roots were induced from leaf segments of Crotalaria spectabilis, which is used as a green manure crop antagonistic to nematodes, by infection with a mikimopine type wild strain of Agrobacterium rhizogenes A13 (MAFF02-10266). For initial proliferation of hairy roots after induction, addition of gibberellin A3 to the medium was indispensable. These roots exhibited vigorous growth and abundant lateral branching on half-strength Murashige and Skoog (1/2MS) medium without plant growth regulators. The adventitious shoots were induced from 24% of root segments 2 months after transfer onto B5 medium containing 5 mg l−1 6-benzyladenine. The axillary buds excised from these shoots produced roots 1 month after transfer onto 0.4% (wv−1) gellan gum-solidified 1/2MS medium containing 30 mM fructose without plant growth regulators. Regenerated plants were successfully grown under greenhouse conditions. Infection of hairy roots of C. spectabilis with root-knot nematodes Meloidogyne hapla and M. incognita led to the growth inhibition of these nematodes.