Shoot regeneration seedlings from transverse thin cell layer explants excised from cotyledon, petiole, hypocotyl of [{Brassica juncea} L. Czern.] in the presence of CdCl$_2$

Abstract

The present work describes a novel neoformation process of \textit{Brassica juncea} (L.) Czern. seedlings from transverse thin cell layers in the presence of CdCl$_2$. In order to investigate the regeneration ability of this crop, the effect of CdCl$_2$ on shoot regeneration (frequency of regeneration and bud number per tTCL) was examined. The tTCL explants were excised from cotyledon, petiole and hypocotyl of 7 day-old \textit{B. juncea} seedlings and cultivated on a solid basal MS medium supplemented with $\alpha $-naphtalenacetic acid (NAA : 3.22 $\mu$M), 6-benzylamino-purine (BAP : 26.6 $\mu$M), sucrose (2 %, w/v), silver nitrate (AgNO3 : 10 $\mu$M) and various concentrations of CdCl$_2$ (0 - 250 $\mu$M). A concentration of CdCl$_2$ of 5 $\mu$M is enough to reduce significantly the percentage of regenerated tTCL from 95-100 % to 77-86 % for all organs tested. In addition, 5 $\mu$M of CdCl$_2$ reduces the bud number from 4.5 to 2.98 and 3.2 to 2.02 of hypocotyl and petiole tTCLs, but not for cotyledon tTCLs. Besides, 250 $\mu$M of CdCl$_2$ is lethal for all tTCLs whatever the organ, and 200 $\mu$M is lethal only for cotyledon and petiole tTCLs but not for hypocotyl explants which had 3.6 % of frequency of shoot regeneration. Plantlets regenerated from all shoots, whatever the treatment, developed and flowered normally 6 weeks after the transfer to pots. The regenerated plants were fertile and identical to source plants.