Development of high frequency multiple shoot formation in Persian clover (Trifolium resupinatum L.)

Abstract

An efficient and reliable micropropagation system for Persian clover (Trifolium resupinatum L.) was developed using different explants and media. Node, hypocotyl and cotyledonary node explants were cultured on Murashige and Skoog (MS) medium supplemented with combinations of either 6-benzyladenine (BA) and indole-3-butyric acid (IBA) or BA, Kinetin (KIN) and IBA. Direct multiple shoots developed within 6 weeks in all explants in most media tested. The best shoot multiplication capacity was obtained from cotyledonary node explants on MS medium containing 7.1 μM BA and 1 μM IBA or 14.1 μM BA and 1 μM IBA. Elongated shoots were rooted on either MS medium alone or combination with different concentrations of indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) and α-naphthaleneacetic acid (NAA). High rooting was achieved in half strength MS medium containing 8 μM IBA.