Soybean is the one of recalcitrant legume species for shoot induction. Shoot regeneration via direct organogenesis was investigated in five soybean cultivars, ‘Dawon’, ‘Pungsan’, ‘Daewon’, ‘Taekwang’ and ‘Chongdoo 1’ by using cotyledonary node explants. Out of 5 soybean cultivars, an efficient shoot regeneration condition was developed in the two soybean cultivars, ‘Dawon’ and ‘Pungsan’. When various kinds of plant growth regulators with different concentration were estimated, the optimum medium condition for shoot induction in both soybean cultivars was MS + B5 vitamin supplemented with BA at concentration 2 ㎎/L. In addition, shoot formation efficiency was increased with 97.09% and 93.88% by the pretreatment of BA onto the explants before in vitro culture in both cultivars. Shoot induction in ‘Dawon’ cultivar was originated from epidermal tissue and sub-epidermal layers when histological changes were investigated under shoot regeneration after culturing cotyledonary node segments on shoot induction medium for 0 to 21 days. Especially, cell dedifferentiation was observed from parenchyma cells to meristematic cell in 3-day cultured segments.
An efficient shoot regeneration condition for pea cv. ‘Sparkle’ was developed by using optimum explant, plant growth regulator concentrations, and pretreatment of BA onto explant. The average shoot number per explant showed the highest on two kinds of shoot induction media (MSB5 media containing 2 ㎎/L BA and a combination of 2 ㎎/L BA and 1 ㎎/L TDZ) when cotyledonary node explants were cultured. Moreover, the pretreatment of explant in 200 ㎎/L BA solution was found to be more effective in shoot induction than that of non-pretreatment. By histological study, cell division and proto-meristem were formed near the surface of the sub-epidermal and epidermal cell layers of cotyledonary node in earlier than 3 days after culture. The analysis of genetic stability of regenerants by using thirteen ISSR markers showed that in vitro regenerated plants showed polymorphism with 8.3% compared with their mother plants.
Plant regeneration via organogenesis and somatic embryogenesis was investigated in Korean soybean cultivars including Cheongja 3, Jinpumkong 2, Taekwangkong and Uram. Cotyledon, cotyledon+hypocotyl and hypocotyl segments of 7-day-old seedlings were cultured on MS medium containing various concentration (0, 1, 2 and 4 ㎎/L) of BA and TDZ. The results showed that MS medium supplemented with BA 2.0 ㎎/L yielded the highest shoot formation ratio of 83.3%. In 4 cultivars, Taekwangkong showed the highest ratio of shoot formation. When various sizes of immature cotyledons (S: 1∼ 2 ㎜, M: 3∼5 ㎜, L: 6∼8 ㎜) were tested on MS medium containing 2,4-D 40 ㎎/L for somatic embryogenesis, the optimum size for embryogenic callus induction was 3∼5 ㎜ in length of immature cotyledons. In 4 cultivars, Taekwangkong showed the highest percentage of embryogenic callus induction. The results indicate that Taekwangkong is the best soybean cultivar for plant regeneration via organogenesis and embryogenic callus induction among the 4 cultivars.