Induction, maintenance and preservation of embryogenic competence of Gentiana cruciata L. cultures

• Autorzy: Mikula A , Fiuk A. , Rybczynski J.J.
• Języki publikacji: EN

Abstrakty

EN

The paper describes structural and ultrastructural changes in primary explants, induction of embryogenesis, somatic embryo development, and four protocols for cryopreservation of cell suspensions. The changes during tissue culture of hypocotyl and cotyledon explants from 10-day-old seedlings and fragments of leaf explant of Gentiana cruciata L. were studied. Seedling explants were cultured on MS medium supplemented with 1.0 mg/l dicamba + 0.1 mg/l NAA + 2.00 mg/l BAP + 80.0 mg/l adenine sulphate. The hypocotyl callus tissue was initiated by cell divisions of the vascular cylinder, but in cotyledons only parenchyma cells took part in callus formation. The leaf blade expiants usually responded only by proliferation of the wounded surface. The effect of auxins (2,4-D, NAA, DIC) and cytokinins (kinetin, zeatin, BAP) in various concentration and combinations on leaf explant response was examined. Generally, embryos were formed sporadically on media containing NAA (1.64% responding explants) or 2,4-D (0.38%), but were not produced in the presence of dicamba. Production of somatic embryos was more effective from suspension culture than from agar medium. Liquid culture made it possible to maintain the cell suspension’s embryogenic competence for 5 years. For preservation of proembryogenic masses, four protocols of cryopreservation were studied: direct cooling, sorbitol/DMSO treatment, vitrification, and encapsulation. Direct cooling and sorbitol/DMSO treatment was unsuccessful. Vitrified tissue required a minimum 3 weeks of culture on solid medium for cell proliferation to reach the proper fresh weight for manipulation. Alginate beads with PEMs were transferred directly to liquid medium for post-freezing culture. Vitrification and encapsulation maintained high viability of post-freezing PEM, but encapsulation ensured faster restoration of G. cruciata cell suspension.

Słowa kluczowe

EN

suspension culture; leaf explant; plant reproduction; Gentiana cruciata; hypocotyl; cryopreservation; differentiation

• Wydawca: -
• Czasopismo: Acta Biologica Cracoviensia. Series Botanica
• Rocznik: 2005
• Tom: 47
• Numer: 1
• Opis fizyczny: p.227-236,fig.,ref.

Twórcy

autor

Mikula A

• Polish Academy of Sciences, Prawdziwka 2, 02-976 Warsaw, Poland

autor

Fiuk A.

autor

Rybczynski J.J.

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