OBJECTIVETo study the in vitro embryo culture of Epimedium wushanense and provide scientific basis for large scale production of tissue culture.

METHODCullus and buds were induced from embryo of E. wushanense on a MS medium supplemented with different 2,4-D,6-BA, NAA, IBA.

RESULTThe optimal compositions of medium that induced callus and buds from embryo were the MS medium supplemented with 2,4-D 2 mg x L(-1), IBA 2 mg x L(-1) and NAA 0.5 mg x L(-1) and the MS medium supplemented with IBA 2 mg x L(-1) and 6-BA 0.5 mg x L(-1), respectively. The optimum medium for callus differentiation was MS + 6-BA 1 mg x L(-1) + NAA 0.5 mg x L(-1) + IBA 1 mg x L(-1), and MS +6-BA 1.0 mg x L(-1) + NAA 0.5 mg x L(-1) for shoots proliferation.

CONCLUSIONUsing embryo as explants, the method of induction and culture of E. wushanense was established by the callus and buds, and the embryo of E. wushanense can be quickly propagated.

Culture Media ; Epimedium ; embryology ; Regeneration ; Tissue Culture Techniques

OBJECTIVETo study the tissue culture and rapid-proliferation techniques of Pueraria mirifica.

METHODThe tender branch were used as explants and cultivated in different media. The optimum media for inducing buds, proliferation and rooting were selected by adjusting the kinds and doses of plant hormones and special compounds.

RESULTThe medium of MS + IBA 0.05 mg L(-1) + BA 0.5 mg L(-1) was suitable for buds inducing and could be used in the first generation cultivation; MS + IBA 0. 02 mg L(-1) + BA 0.2 mg L(-1) and MS +BA 0.1 mg L(-1) were employed by turns in subculture, 25 days propagation coefficient was 3.0; and the medium of 1/2MS + IBA 0.1 mg L(-1) + IAA 0.2 mg L(-1) + C (special compound) 10 mg L(-1) was used for roots inducing, the rooting rate was 76.9%. Rooting plantlets were transplanted in spring and summer; the surviving rate was 81.0%.

CONCLUSIONThis technique system could be employed for rapid propagation of P. mirifica.

Pueraria ; growth & development ; Tissue Culture Techniques ; methods

OBJECTIVETo establish a method for the germplasm preservation of R. glutinosa.

METHODPlantlets of different cultivars obtained by tip culture were inoculated into test tubes with MS medium supplemented with 0.2 mg.L-1 BA and 0.02 mg.L-1 NAA, and preserved at 4-6 degrees C in dark. At the same time, different media (A. distilled water + 10 g.L-1 agar; B. 1/2 MS + 5 g.L-1 agar; C. MS + 10 g.L-1 agar; D. 1/2 MS + 0.5 BA + 0.02 NAA + 10 g.L-1 agar; E. MS + 0.5 BA + 0.02 NAA + 10 g.L-1 agar) were set to conserve plantlets of "85-5" on the same condition.

RESULT4 months later, the death rate of "Xinggeda" was 73%, "Tucheng" 60%, "85-5" 33%, and "Beijing 1" 9%. All of the plantlets of "85-5" in different media kept alive. 10 months later, most of the preserved plantlets browned and wilted except those on medium A.

CONCLUSIONThe low temperature endurance of R. glutinosa is cultivar-dependent. Medium A can preserve "85-5" for more than 10 months in vitro.

Agar ; Culture Media ; Culture Techniques ; Plants, Medicinal ; growth & development ; Rehmannia ; growth & development ; Temperature ; Tissue Preservation ; methods

A protocol is presented for direct and indirect regeneration of common dandelion (Taraxacum officinale Weber) from leaf and petiole explants. Multiple shoots were obtained on MS medium containing 0.2 mg/L IAA and 1 mg/L TDZ. For indirect regeneration, fragile calli were obtained from leaf and petiole explants on MS medium supplemented with 0.5 mg/L 2,4-D and 2.0 mg/L 6-BA. Regenerated plantlets were obtained when these calli were cultured on MS medium containing 1.0 mg/L 6-BA. Random amplified polymorphic DNA (RAPD) analysis of nine regenerated plantlets revealed 61 scorable bands from 10 primers, including three specific bands.
Culture Media ; Germination ; physiology ; Plant Leaves ; growth & development ; Regeneration ; Taraxacum ; growth & development ; Tissue Culture Techniques ; methods

OBJECTIVETo research the optimal conditions for the callus induction of anther culture and the plant regeneration of Angelica dahurica var. formosana.

METHODCallus was induced from the anther of A. dahurica from Sichuan province on a MS medium. The effects of callus induction and plant regeneration of different pretreatment hours under low temperature (4 degrees C), different culturing conditions under darkness and illumination, and different culture with different hormone contents and ratios were studied.

RESULTThe results showed that A. dahurica anthers without low temperature pretreatment reached the highest induction rate then under the pretreatment under low temperature (4 degrees C) for two days. The optimal culturing condition was under the darkness. The culturing efficiency reached 38.89% on the medium of MS + 2.0 mg x L(-1) 2,4-D + 1.0 mg x L(-1) 6-BA. The optimum medium for differentiate anther callus was MS + 0.5 mg x L(-1) NAA + 1.5 mg x L(-1) KT + 10 mg x L(-1) AgNO3. 1/2MS medium supplemented with 0.5 mg x L(-1) IBA could well promote seedings to take roots.

CONCLUSIONAn efficient system for callus induction of anther culture and plant regeneration of A. dahurica was preliminarily established.

Angelica ; drug effects ; growth & development ; Culture Media ; chemistry ; pharmacology ; Flowers ; Tissue Culture Techniques ; methods

OBJECTIVETo establish and optimize the rapid propagation system of Polygonum multiflorum, as well as explore method for induction and identification of autotetraploid.

METHODPropagation medium was optimized by orthogonal test. The buds were immersed in colchicine solution with different concentrations for different time to select induction conditions for autotetraploid of P. multiflorum.

RESULTThe most appropriate propagation medium was MS medium supplemented with 1.0 mg x L(-1) 6-BA, 0.3 mg x L(-1) NAA, and 0.4 mg x L(-1) PP333. That the buds were soaked in 0.2% colchicine solution for 30 h, or soaked in 0.3% colchicine solution for 18 h, was optimal condition to induce autopolyploid of P. multiflorum with induction rate as high as 16.7%.

CONCLUSIONRapid propagation of P. multiflorum could be achieved by tissue culture. Furthermore, colchicine was an effective inducer of polyploidy, and 25 tetraploid lines were obtained through chromosome identification. The experiment laid a foundation for the wild resource conservation, superior varieties breeding of P. multiflorum.

Chromosomes, Plant ; genetics ; Culture Media ; metabolism ; Polygonum ; genetics ; growth & development ; metabolism ; Tetraploidy ; Tissue Culture Techniques ; methods

The protocorm-suspension-system of Cremastra appendiculata was founded by liquid-suspension culture. The factors to effect germination rate and seedling conversion rate of C. appendiculata synthetic seeds, such as synthetic coating materials, synthetic endosperm components, storing conditions and germination materials, etc. were studied. The result showed that the germination rate and seedling conversion rate of synthetic seeds were the highest on the MS solid-medium while using 4% sodium alginate + 2% CaCl2 + 2% chitosan as synthetic coating materials, with 1/2 MS liquid-medium + 0.2 mg x L(-1) NAA + 0.1 mg x L(-1) GA3 + 0.5 mg L(-1) BA + 0.4 mg x L(-1) penicillin + 10.0 mg x L(-1) endophyte extract +0.3% carbendazim powder + 0.2% sodium benzoate + 1.0% sucrose as synthetic endosperm. And the germination rate and seedling conversion rate of synthetic seeds could attain to 68% and 65% after 20 days storing at 4 degrees C. The germination rate and seedling conversion rate of synthetic seeds decreased to a great extent with increasing the storing temperature and prolonging storing time.
Culture Media ; metabolism ; Germination ; Orchidaceae ; growth & development ; metabolism ; Seeds ; growth & development ; metabolism ; Tissue Culture Techniques ; methods

OBJECTIVETo optimize the condition of callus of Cinnamonum camphora induced.

METHODGC and plant tissue culture method were applied in the study.

RESULTThe effect of callus induced and the growth of callus were different in MS medium with different proportion of hormone. The ration of callus induced was the highest and the growth of callus was the most prosperous in the MS medium with 4 mg x L(-1) 2,4-D and 0.2 mg x L(-1) 6-BA. It is found that callus induced by young leaf contained borneol, but callus induced by young stem not.

CONCLUSIONThe optimization of callus of C. camphora induced is using the MS medium with 4 mg x L(-1) 2,4-D + 0.2 mg x L(-1) 6-BA. Callus induced by young leaf can generate borneol.

Bornanes ; metabolism ; Cinnamomum camphora ; growth & development ; metabolism ; Culture Media ; Tissue Culture Techniques

OBJECTIVEOur research studied the fast-breeding technology of Ardisia crenata sims by using tissue culture and provided the scientific foundation for industry production.

METHODThe effects of axillary buds and plant regeneration of different basic medium, hormones and additives on induction and multiplication were studied.

RESULTThe best culture medium for the induction of axillary buds, which took the stems of A. crenate were as explants, was MS + 6-BA 0.5 mg x L(-1) + NAA 0.1 mg x L(-1), and the best medium for multiplication was MS + 6-BA 2.0 mg x L(-1) + NAA 0.1 mg x L(-1) + KT 0.5 mg x L(-1), the best medium for roots generation was 1/2MS + IBA 0.2 mg x L(-1). We also found that the roots'generation, roots rate and mean number of roots can be promoted by adding 0.2% Ac, and the most suitable ground substance was river sand-perlite-vermiculite (1:1:1) or perlite-vermiculite (1:1). With axillary buds and plant regeneration methode, more than 80% A. crenata sims could be regenerated integratedly.

CONCLUSIONA. crenata sims can be regenerated integratedly and breeded fast by using axillary bud proliferation technology.

Ardisia ; growth & development ; physiology ; Culture Media ; metabolism ; Regeneration ; Tissue Culture Techniques ; methods

A line of fibroblast-type cells derived from embryos of a domestic rabbit has been cultivated continuously for over 3 years by serial passages up to the level of the moth passage. The cell line was tentatively named rabbit embryo fibroblast (REF). The establishment of primary culture, serial passages, growth rate and cytology are described in this communication. In addition some of the results of experiments on the detection of Mycoplasma contamination, on storage of the frozen cells and on its susceptibility to vaccinia virus infection are included.
Animal ; Cell Line ; Embryo/cytology* ; Fibroblasts* ; Rabbits ; Tissue Culture*