Optimisation of Tissue Culture Conditions for Transformation Studies Using Immature Embryos of Australian Barley Cultivars

1995 ◽  
Vol 43 (5) ◽  
pp. 499 ◽  
Author(s):  
AM Walmsley ◽  
RJ Henry ◽  
RG Birch
Keyword(s):  
Tissue Culture ◽  
Plant Regeneration ◽  
Culture Medium ◽  
Immature Embryo ◽  
Culture Conditions ◽  
Medium Composition ◽  
Immature Embryos ◽  
Systematic Testing ◽  
Barley Cultivars ◽  
Key Variables

Eight Australian barley cultivars were tested for efficiency of embryonic callus initiation and plant regeneration, from immature embryo explants in tissue culture. Optimisation of tissue culture conditions was performed for cultivars Bandulla, Clipper, Schooner and Tallon in an attempt to increase regeneration frequencies to levels suitable for genetic engineering of barley. Variables tested were 2,4-D concentration, salt composition, carbon source and immature embryo explant. Optimal culture medium composition varied between cultivars. Shoot regeneration rates from culture of isolated scutellar tissues were low for all four cultivars. Halved, immature embryos produced most shoots for cultivars Clipper, Schooner and Tallon, whereas Bandulla performed best with entire immature embryo explants. Clipper (a malting barley) and Bandulla (a feed barley) are suggested as model Australian cultivars for transformation studies. Immature embryos of Bandulla produced an average of 5.3 shoots and Clipper 10.1 shoots per embryo under optimal conditions. Our results show that rates of somatic embryo and plant regeneration sufficient for use in transformation studies can be achieved for diverse Australian Barley cultivars, through systematic testing of a range of key variables including explant type and medium composition.

Paucity of somaclonal variation from immature embryo culture of barley

1989 ◽  
Vol 40 (6) ◽  
pp. 1155 ◽  
Author(s):  
DJ Luckett ◽  
D Rose ◽  
E Knights
Keyword(s):  
Tissue Culture ◽  
Somaclonal Variation ◽  
Embryo Culture ◽  
Field Trial ◽  
Immature Embryo ◽  
Immature Embryos ◽  
Golden Promise ◽  
Immature Embryo Culture ◽  
Tissue Origin ◽  
Regenerant Plants

Intact immature embryos of barley (cv. Golden Promise) and component tissues (the scutellum and embryonic axis) were cultured to produce callus. Regenerant plants were obtained from this callus and SC2 families raised. These families were examined in a field trial to search for somaclonal variation. No obvious variants were found confirming our previous unpublished results. The lack of somaclonal variation generated by barley tissue culture (which is in contrast to other species) was not a result of the tissue origin of the regeneration event.

Study on Tissue Culture of Sweet Broad Pea

2014 ◽  
Vol 644-650 ◽  
pp. 5407-5410
Author(s):  
Hui Fang Chi
Keyword(s):  
Tissue Culture ◽  
Plant Regeneration ◽  
Culture Medium ◽  
Callus Formation ◽  
Formation Rate ◽  
Adventitious Bud ◽  
Experimental Basis ◽  
Research Results

s. The cotyledons, Internodes, leaves and stems of sweet broad pea were studied on tissue culture. Research results show that: The ability of different explants for callus formation and adventitious bud differentiation in different culture medium is different. The callus formation rate and sprouting rate of Internodes is significantly higher than other explants, which is a ideal material for tissue culture. The callus formation rate of Internodes was 100% in MS +BA1.0 mg/L+NAA 1.0 mg/L and MS+ 2, 4-D 0.5 mg/L; The bud differentiation is best at the medium of MS+ 6-BA 2 mg/L, which reached 86.7%; the rooting rate was 83.3% at the medium of MS+ NAA 3mg/L. The study provides a experimental basis for further study on the plant regeneration in the sweet broad pea.

scholarly journals Factors influencing somatic embryogenesis and regeneration ability in somatic tissue culture of spring and winter rye

2008 ◽  
Vol 13 (4) ◽  
pp. 363 ◽  
Author(s):  
R. MA ◽  
S. PULLI
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Developmental Stage ◽  
Embryogenic Callus ◽  
Immature Embryo ◽  
Green Plant ◽  
Somatic Tissue ◽  
Winter Rye ◽  
Effi Ciency

Rye is an important crop in Northern and Eastern Europe. However, the application of various biotechnologies in rye breeding has been limited duo to its recalcitrant in tissue culture. In order to improve somatic tissue effi ciency, key factors affecting somatic embryogenesis and reproducible green plant regeneration of rye (Secale cereale L.) were evaluated and optimised. In this study, a total 27 rye genotypes including 10 spring and 17 winter genotypes were involved in the investigation. Genotype, culture medium, sugar, gel agent and auxin infl uenced somatic embryogenesis of immature embryo signifi cantly. One-two weeks cold pretreatment of young embryo enhanced somatic embryogenesis and green plant regeneration. In culture of immature embryos, infl orescences and leaf segments of the seedlings, explants signifi cantly infl uenced the culture effi ciency. Highest embryogenic callus yield resulted from rye immature embryo as explant compared to young infl orescence and leaf segment of seedling. Developmental stage of embryo played an important role in somatic embryogenesis. Late spherical coleoptile stage (embryo size 0.5–1mm in length) was optimal developmental stage of immature embryo for culture. Morphogenetic potential of embryogenic callus decreased with an increasing number of subcultures, and this ability could be maintained in vitro for a maximum of 8 months of culturing.;

scholarly journals Callus Induction and Plant Regeneration Efficiency According to Tissue Culture Conditions in Teff grass (Eragrostis)

2013 ◽  
Vol 19 (1) ◽  
pp. 55-62
Author(s):  
Ki-Won Lee ◽  
Jin Young Moon ◽  
Hyung Soo Park ◽  
Gi Jun Choi ◽  
Ki-Yong Kim ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Plant Regeneration ◽  
Callus Induction ◽  
Culture Conditions ◽  
Regeneration Efficiency

scholarly journals Study of Plant Regeneration by Tissue Culture from Immature Embryos of the Lacquer Tree, Toxicodendron vernicifluum

2019 ◽  
Vol 65 (3) ◽  
pp. 125-130
Author(s):  
Kentaro Tsukada ◽  
Yusuke Yamagishi ◽  
Eri Nabeshima ◽  
Michito Hosaka ◽  
Kenta Okada ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Plant Regeneration ◽  
Immature Embryos ◽  
Toxicodendron Vernicifluum

scholarly journals Design of a culture medium for optimal growth of the bacterium Pseudoxanthomonas indica H32 allowing its production as biopesticide and biofertilizer

AMB Express ◽  
2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Dayana Morales-Borrell ◽  
Nemecio González-Fernández ◽  
Néstor Mora-González ◽  
Carlos Pérez-Heredia ◽  
Ana Campal-Espinosa ◽  
...  
Keyword(s):  
Hydrogen Sulfide ◽  
Yeast Extract ◽  
Culture Medium ◽  
Nitrogen Sources ◽  
Optimal Growth ◽  
Culture Conditions ◽  
Medium Composition ◽  
Mineral Salts ◽  
Molar Ratios ◽  
Biotechnological Processes

Abstract Culture medium composition is one of the most important parameters to analyze in biotechnological processes with industrial purposes. The aim of this study was to design of a culture medium for optimal growth of the bacterium Pseudoxanthomonas indica H32 allowing its production as biopesticide and biofertilizer. The influence of several carbon and nitrogen sources and their molar ratios on P. indica H32 growth was investigated. The effect of different micronutrients such as mineral salts and vitamin on P. indica H32 growth was determined as well. A mixture design based on Design-Expert 10.0 Software was performed to optimize the culture medium concentration. Finally, in the designed medium, an attribute of the biological mechanism of action of the P. indica H32 against nematodes, was evaluated: the hydrogen sulfide production. It was found that tested carbon/nitrogen ratios were not a significant influence on P. indica H32 growth. Growth of P. indica H32 was favored with use of sucrose, yeast extract and phosphate buffer without the addition of any tested micronutrients. An optimal concentration of 10 g/L sucrose and 5 g/L yeast extract were obtained at a cost of 0.10 $/L. In this concentration, the specific growth rate (µ) and maximal optical density (Xmax) were equal to 0.439 h− 1 and 8.00 respectively. It was evidenced that under the culture conditions used, P. indica H32 produced hydrogen sulfide. The designed medium led to a 1.08 $/L reduction of costs in comparison to LB medium. These results were critical to carry on with biotechnological development of P. indica H32 as a bioproduct.

scholarly journals Optimized somatic embryogenesis and plant regeneration in elite Argentinian sugarcane (Saccharum spp.) cultivars

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Valentina Di Pauli ◽  
Paola Daniela Fontana ◽  
Dalia Marcela Lewi ◽  
Arturo Felipe ◽  
Luis Ernesto Erazzú
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Culture Conditions ◽  
Light Conditions ◽  
Saccharum Spp ◽  
Regenerated Plants ◽  
Embryogenic Calluses ◽  
Indirect Somatic Embryogenesis ◽  
Good Number

Abstract Background Biotechnological breeding of elite sugarcane cultivars is currently limited because of the difficulty of regenerating plants by tissue culture. Here, we report that commercially elite sugarcane genotypes, which are adapted to Argentinian agro-ecological conditions, are capable of being regenerated via indirect somatic embryogenesis. Leaf rolls of five elite genotypes were cultured following two callus induction protocols using different concentrations of 2,4-D as the growth regulator. Embryogenic calluses were regenerated under light conditions. Regenerated plants were subsequently acclimatized in the greenhouse under two acclimatization procedures before being transplanted to the field. Results Four of the five genotypes were able to form somatic embryos following the two induction protocols. The variables related to embryogenic callus production were influenced by the interaction between genotype and culture conditions. For plant regeneration, the embryogenic calluses were further cultured on an IBA-supplemented medium, where we observed a high genotype dependence. Calluses from the four cultivars regenerated a good number of plants. With the procedures described here, we obtained more than 90% of well-acclimatized plants both in the greenhouse and in the field. Conclusions This protocol provides a simple way to regenerate sugarcane plants through indirect somatic embryogenesis. Also, the results confirm that tissue culture ability is highly genotype-dependent in sugarcane. Our findings suggest that these elite cultivars could be good candidates for biotechnological breeding.

scholarly journals Regulation of ExoS Production and Secretion byPseudomonas aeruginosa in Response to Tissue Culture Conditions

1999 ◽  
Vol 67 (2) ◽  
pp. 914-920 ◽  
Author(s):  
Amy J. Vallis ◽  
Timothy L. Yahr ◽  
Joseph T. Barbieri ◽  
Dara W. Frank
Keyword(s):  
Tissue Culture ◽  
Cho Cells ◽  
Type Iii Secretion ◽  
Time Course ◽  
Culture Medium ◽  
Chinese Hamster ◽  
Growth Conditions ◽  
Culture Conditions ◽  
Tissue Culture Medium ◽  
Type Iii

ABSTRACT This study was initiated to characterize the regulation and secretion of ExoS by Pseudomonas aeruginosa during contact with eukaryotic cells. The production of ExoS was monitored by a sensitive ADP-ribosyltransferase activity assay, and specific activities were calculated for supernatant and cell-associated fractions. Time course analysis indicated that ExoS was produced after a lag period, suggesting that induction of the regulon is necessary for the expression of detectable amounts of enzyme activity. Under tissue culture growth conditions, ExoS was induced when P. aeruginosa was in contact with Chinese hamster ovary (CHO) cells or after growth in tissue culture medium with serum. The serum induction of ExoS appeared to result in generalized type III secretion, while induction by contact with CHO cells appeared to result in polarized type III secretion. Mutants in the type III secretory system that express a null phenotype for ExoS production in bacteriological medium produced but did not secrete the enzyme when P. aeruginosa was grown under inducing conditions in tissue culture medium. These results suggest that both induction and secretion of ExoS may differ when the bacteria are exposed to different growth environments. The putative type III translocation proteins and secretion apparatus of P. aeruginosa were required for translocation of bacterial factors that mediate changes in CHO cell morphology during infection.

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