METHOD FOR MICROCLONAL REPRODUCTION OF GLADIOLUS Russian patent published in 2002 - IPC

FIELD: agriculture, flower growing, selection, biotechnology. SUBSTANCE: the method deals with moistening the corms of 5-6 mm diameter with tap water for 24 h followed by their surface sterilization with 6%-sodium hydrochloride solution for 15 min and a 5-6-fold washing with sterile distilled water for 1 h, repeated sterilization with 90%- ethanol for 10 min and a 3-fold washing. Then corms should be got rid of tunics and sterilized subsequently with 70%-ethanol for 5-7 min and 90%-ethanol for 1 min. After a 3-fold corm washing with sterile distilled water is over, apical buds with adjacent tissue portions are cut and placed upon agarized nutritive medium to determine the value of sterility quality of explants which contains 1/2 concentrations of both macro- and microsalts by Murashige and Skoog (MS), total composition of vitamins and organic supplements of MS medium, 1.0- 2.0% saccharose, 0.50-0.70 μM 1-naphthylacetic acid (NAA), after a 7-d-long cultivation upon agarized medium it is necessary to replace sterile explants into liquid nutritive medium (at medium/explants amount volume being 3:1) supplemented with 10.0-11.0 μM 6- benzylaminopurine (6-BAP). Then cultivation is conducted upon a shaker at 100 min ^-1 at continuous illumination at 25±2 C for 28-30 d in a liquid medium. Then proliferating buds are divided into single ones and clusters formed of 3-4 buds and cultivated at continuous illumination and 25±2 C upon agarized medium which contains 1/2 MS, supplements by MS: saccharose, vitamins and inositol, 0.2-0.3 μM abscisic acid (ABA), 0.50-0.70 μM NAA, 10.0-11.0 μM 6-BAP, 10.0-12.5 mg/l paminobenzoic acid (PABA) to obtain sprouts which are, 4 mo after the onset of subcultivation upon agarized medium, subjected for rooting upon 1/2 MS medium at low nitrogen content and supplemented activated carbon at photoperiod being 16 h/day at 27-28 C and 8 h/night at 19- 21 C, and after that the mentioned sprouts should be rooted. EFFECT: increased reproduction coefficient in genetically stable and infection-free plants. 3 cl