[Establishment of in vitro culture, plant regeneration and genetic transformation of Camelina sativa].

[Establishment of in vitro culture, plant regeneration and genetic transformation of Camelina sativa].

TSitologiia i genetika (2013-07-05)

A I Emets, Iu N Boĭchuk, E N Shisha, D B Rakhmetov, Ia B Blium

PMID23821950

ABSTRACT

The results on in vitro culture establishment, plantlet regeneration and rooting of Camelina sativa cultivar sample Peremozhets and cultivar Mirazh are presented. Effective concentrations of sterilizing agents and duration of plant material treatment were estimated. Phytohormone ratio, sucrose concentration in nutrient medium that induce effective formation of C. sativa shoots and NAA concentration for plantlet rooting have been established. The method of Agrobacterium-mediated transformation of Camelina by using binary vector pGH217 carrying reporter beta-glucoronidase (gus) gene driven under 35S CaMV promoter and nos-terminator, and selective marker hpt gene conferring hygromycin-resistance in transgenic plant was elaborated.

MATERIALS

Product Number

Brand

Product Description

H5527

Sigma-Aldrich

Hygromycin B solution from Streptomyces hygroscopicus, ≥60% (HPLC), 45-60 mg/mL in H₂O, γ-irradiated

G0876

Sigma-Aldrich

β-Glucuronidase from Helix pomatia, Type H-2, aqueous solution, ≥85,000 units/mL

G0762

Sigma-Aldrich

β-Glucuronidase from Helix pomatia, Type H-3AF, aqueous solution, ≥60,000 units/mL

G7770

Sigma-Aldrich

β-Glucuronidase from Helix pomatia, Type HP-2S, aqueous solution, ≥90,000 units/mL

G7017

Sigma-Aldrich

β-Glucuronidase from Helix pomatia, Type HP-2, aqueous solution, ≥100,000 units/mL

G1512

Sigma-Aldrich

β-Glucuronidase from Helix pomatia, Type H-5, lyophilized powder, ≥400,000 units/g solid

G8885

Sigma-Aldrich

β-Glucuronidase from Helix pomatia, Type H-3, aqueous solution, ≥90,000 units/mL

G0751

Sigma-Aldrich

β-Glucuronidase from Helix pomatia, Type H-1, partially purified powder, ≥300,000 units/g solid

H0654

Sigma-Aldrich

Hygromycin B solution from Streptomyces hygroscopicus, ≥60% (HPLC), 45-60 mg/mL in H₂O

G8132

Sigma-Aldrich

β-Glucuronidase from limpets (Patella vulgata), Type L-II, lyophilized powder, 1,000,000-3,000,000 units/g solid

G8420

Sigma-Aldrich

β-Glucuronidase from Escherichia coli, ≥20,000 units/mg protein, recombinant, expressed in E. coli overproducing strain, lyophilized powder

H9773

Sigma-Aldrich

Hygromycin B from Streptomyces hygroscopicus, suitable for plant cell culture, BioReagent, ≥60% (HPLC), lyophilized powder

H3274

Sigma-Aldrich

Hygromycin B from Streptomyces hygroscopicus, powder, BioReagent, suitable for cell culture, suitable for insect cell culture

G4259

Sigma-Aldrich

β-Glucuronidase from Helix aspersa (garden snail), Type HA-4

G0376

Sigma-Aldrich

β-Glucuronidase from bovine liver, Type B-3, ≥2,000,000 units/g solid

G0251

Sigma-Aldrich

β-Glucuronidase from bovine liver, Type B-1, ≥1,000,000 units/g solid

G8162

Sigma-Aldrich

β-Glucuronidase from Escherichia coli, aqueous glycerol solution, ≥5,000,000 units/g protein, pH 6.8 (biuret)

G7646

Sigma-Aldrich

β-Glucuronidase from Escherichia coli, Type VII-A, lyophilized powder, 5,000,000-20,000,000 units/g protein, pH 6.8 (30 min assay)

G7396

Sigma-Aldrich

β-Glucuronidase from Escherichia coli, Type IX-A, lyophilized powder, 1,000,000-5,000,000 units/g protein (30 min assay)

H7772

Sigma-Aldrich

Hygromycin B from Streptomyces hygroscopicus, lyophilized powder

G8295

Sigma-Aldrich

β-Glucuronidase from Escherichia coli, ≥10,000,000 units/g protein (30 min assay), recombinant, expressed in E. coli overproducing strain, lyophilized powder

G0799

Sigma-Aldrich

β-Glucuronidase from Escherichia coli, >20,000,000 units/g protein, recombinant, expressed in E. coli, aqueous glycerol solution