Molecular cloning and characterization of auxin-regulated genes from mungbean hypocotyls during adventitious root formation

Adventitious root formation (rooting) in 'Berken' mungbean [vigna radiata (L.) Rwiclz.] cuttings is stimulated by indole-3-acetic acid (IAA). To understand the molecular events that occur during IAA-induced adventitious root initiation, a λgt11 cDNA library was made from mungbean hypocotyls treated with 500 μM IAA for 3 hours and differentially screened. Two cDNAs MII-3 and MII-4 were isolated. Southern analysis revealed that both cDNAs are encoded by different genes. Expression studies showed different patterns for Both genes. Both MII-3 and MII-4 were highly expressed in IAA treated hypocotyls. whereas MII-4 was also induced in IAA treated epicotyls. There was no expression of either MII-3 or MII-4 in control or IAA treated leaves. With increasing concentrations of IAA from 100 to 1000 μM there was an increase in the average root number per cutting as well as a stimulation in MII-3 and MII-4. Both MII-3 and MII-4 showed a stimulation in expression 4 hours following treatment with 500 μM IAA reaching a maximum from 4 to 8 hours followed by a decline thereafter. Basal expression of MII-3 was evident between 2 and 8 hours, whereas, a high degree of basal expression was found with MII-4 from 1 to 8 hours followed by a sharp decline. Cyclobeximide (50μm) dramatically reduced rooting and MII-3 expression, whereas MII-4 was only slightly affected.