Somatic Embryogenesis and Plant Regeneration from Immature Persimmon (Diospyros kaki Thunb.) Embryos

Abstract

Additional index words. adventitious bud formation, globular embryo, torpedo-type embryo, embryogenic callus, mass propagation Abstract. In persimmon, plant regeneration from cultured cells usually takes place through adventitious bud formation. If somatic embryogenesis were possible, the efficiency of mass propagation and genetic engineering would be greatly improved. We attempted to induce somatic embryogenesis from immature embryos and plant regeneration from the induced embryos. Hypocotyls and cotyledons from immature 'Fuyu' and 'Jiro' seeds were cultured in the dark in Murashige and Skoog medium solidified with gellan gum and supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA) at various concentrations. Callus formation started at ' '2 weeks of culture, and the callus formation rate was highest at 3 or 10 mM combinations of 2,4-D and BA. The initially formed calli gradually became brown or black from which white embryogenic calli (EC) appeared secondarily. After ' '8 weeks of culture, globular embryos were formed from these EC, and the formation proceeded until 20 weeks of culture. Formation of globular embryos was higher with 'Fuyu' than 'Jiro', especially with hypocotyls. When EC with globular embryos were transferred to fresh medium with no plant growth regulators, ' '70% developed to the torpedo-type embryo stage in 6 weeks. The torpedo-type embryos thus formed were germinated and rooted in agar medium with or without zeatin in several weeks without entering dormancy. After germination and rooting, the plantlets were transferred to the same medium and acclimatized for another 4 weeks. As the embryos germinated and rooted simultaneously, the plantlets were easy to grow in pots without transplanting shock. This is the first report on plant regeneration through somatic embryogenesis of persimmon.

Callus induction and Cytology of Vulnerable Tree Species Diospyros candolleana Wight

“…Tao and Sugiura, (1992) reported that, hard globular calli proliferated from leaf primordial of D. kaki. In the earlier stage callus showing yellowish white colour later it became brown to black, similarly leaf explants also turns to brown or black colour gradually after inoculation because of the reason of exudation of polyphenol like substances was reported by Sugiura, 2008;Choi et al, 2001;Sarathchandra and Burch, 1991.…”

“…The economic crops of genus Diospyros are interesting for researchers to get clonal rootstocks through micropropagation using tissue culture techniques to overcome ancient poor characteristics having rootstocks obtained from seed propagation (Giordani et al, 2002). Sugiura (2008) reported that85%-100% of callus was induced 4 o after 2 weeks, from the 2, 4-D+BAP (3 µM +10 µM) combination in D. kaki. Tao and Sugiura, (1992) reported that, hard globular calli proliferated from leaf primordial of D. kaki.…”

“…Diospyros is a large genus belongs to the family Ebenaceae and the species of this genushas more economic values because fruits of few species are used as economic crops. To improve, to produce specific traits, to preserve the cultivar etc., these species are propagated by tissue culture techniques (Sugiura, 2008;Kochanova et al 2011).Various explants such as leaf, shoot tips, dormant buds, adventitious bud, fruits etc., were used for callus culture and regeneration The purpose of this study is to conserve D. candolleana species using tissue culture techniques.…”

Genetic Transformation of Recalcitrant Cassava by Embryo Selection and Increased Hormone Levels

“…The present results indicate that torpedo-stage SEs can be obtained in higher abundance by shortening multiplication time on the SE inducing medium. Another torpedo-generating approach could focus on the use of plant hormone regulators or adjusted media [32,33,34,35] as suggested by recent studies with model and non-model farmer-preferred cassava cultivars [22,36]. The recent improvement of de novo shoot organogenesis from cassava also holds some promise for the development of genetic transformation methods that are not requiring the production of FEC [37].…”