Callus induction and plant regeneration from somatic tissue in spring rapeseed (<i>Brassica napus</i> L.)

Abstract

The effect of culture media, explants and genotypes on adventitious shoot regeneration in spring rapeseed (Brassica napus L.) was examined. Hypocotyls and cotyledons of the doubled haploid lines NL-611, NL-662, NL-685 were induced to form callus by culturing on the Murashi geSkoog medium supplemented with different concentrations of 6-benzylaminopurine (BAP) and α-naphtylacetic acid (NAA) or zeatin and 2,4-dichlorphenoxyacetic acid (2,4-D). Adventitious buds were regenerated from the organogenic callus on the same medium. A large variation of shoot regenerability was observed, ranging within 0-37.5% for the frequency of bud formation and within 0-3.8 for the number of buds per explant. Generally, cotyledon-derived callus exhibited a higher bud regeneration frequency than hypocotyls, however, hypocotyl-derived callus developed a higher number of buds per explant. The maximum number of adventitious buds from hypocotyl-derived callus was obtained on a medium supplemented with 4.0 mg l -1 BAP and 0.05 mg l -1 NAA. Regenerated shoots were rooted in MS medium containing 0.1 mg l -1 NAA. Well rooted plantlets were acclimatized and subsequently established in soil.

Tissue culture of Cecropia glaziovii Sneth (urticaceae): vegetative micropropagation and plant regeneration from callus

“…A combination of ZEA 0.1 mg L -1 and 2,4-D (1.0 and 3.0 mg L -1 ) also induced bud formation from calluses derived from petioles, but the bud number per callus was significantly lower (5.4 and 5.3 buds per callus, respectively). Burbulis et al (2008) obtained the maximum response of bud induction in callus of Brassica napus L. with combinations of BAP and NAA, contrasting to our results: these combinations produced few of none buds at all (Table 2). On the other hand, the same authors also obtained positive results with combinations of ZEA and 2,4-D, which is similar to the obtained here, although their calluses were derived from hypocotyls.…”

Regeneration ability analysis of closely related doubled haploids of rapeseed (Brassica napus ssp. oleifera).

“…Similar results were presented in the paper by Burbulis et al [9], where significant differences in callus induction and plant regeneration were observed among three DH lines. In these research hypocotylous explants of the lines NL-611 and NL-662 displayed greater ability to form shoots than the line NL-685 on a medium containing 4 mg/l BAP and 0.05 mg/l NAA.…”

“…Among many factors influencing the success of a regeneration process under in vitro culture, the genotype seems to play the most important role [9].…”

In vitro regeneration and overexpression of pea DNA helicase 45 (PDH45) gene through Agrobacterium-mediated genetic transformation in oilseed Brassica spp.

“…Several reports are available where IBA was used for adventitious root formation from excised regenerated shoots of Brassica (34,35) . In this study maximum root induction was achieved on half strength of MS supplemented with 13.4 µM NAA + 2 µM IBA in case of BARI Sarisha-8 (75%) and BARI Sarisha-13 (80%) whereas only 2.0 µM IBA was effective for root formation in BARI Sarisha-16 (73.33%) (Table 1).…”