Elimination of Lactobacillus plantarum, Corynebacterium spp., Staphylococcus saprophyticus and Pseudomonas paucimobillis from micropropagated Hemerocallis, Choisya and Delphinium cultures using antibiotics

Leifert, Carlo; Camotta, Helen; Wright, Sandra; Waites, Barbara; Cheyne, Valerie A.; Waites, W. M.

doi:10.1111/j.1365-2672.1991.tb03795.x

Cited by 47 publications

(38 citation statements)

References 33 publications

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“…Two studies transferred material onto non -antibiotic medium, and when no Agrobacterium grew from the explants, they were presumed to be axenic (Barghchi et al, 1994;Mylnarova et al, 1994). This technique, however, is unreliable since bacteria can remain latent within plant tissue throughout the culture period (Cassells, 1991;Leifert et al, 1991). Antibiotics have been successful in the elimination of bacteria in tissue culture in some cases (Horsch and King, 1983;Leifert et al, 1991;Barrett and Cassells, 1994), but a vital aspect to their success is to ensure the completion of drug sensitivity tests, MBC determination, stability of the antibiotic and phytotoxicity to the plant.…”

Section: Introductionmentioning

confidence: 98%

A risk assessment study of plant genetic transformation usingAgrobacterium and implications for analysis of transgenic plants

Barrett

Cobb

McNicol

et al. 1997

Plant Cell Tiss Organ Cult

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Agrobacterium transformation systems for Brassica, Solanum and Rubus, using carbenicillin, cefotaxime and ticaracillin respectively to eliminate contamination, were examined for the presence of residual Agrobacterium. The results indicated that none of the antibiotics in question, succeeded in eliminating Agrobacterium and the contamination levels increased in explants from 12 to 16 weeks to such an extent that Solanum cultures senesced and died. This may be due to the fact that four times the Minimum bactericidal concentration values (concentration to be used for elimination of contaminants in culture), for the three antibiotics, were higher than the concentrations employed in the culture medium. Contamination in shoot material decreased over 16 to 24 weeks possibly due to bacteriostatis and the use only of the apical node for further culture. The presence of the binary vector was also noted under non-selective conditions, even up to 6 months after transformation, where approx. 50% of contaminated material still harboured bacterial cells with the binary vector at levels of approx. 107 Colony forming units per gram.Abbreviations: CFU -colony forming units, GUS -fl-glucoronidase, PCR -polymerase chain reaction, MBCminimum bactericidal concentration, NPTII -neomycin phosphotransferase, RAPD's -random amplified polymorphic DNA

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Section: Introductionmentioning

confidence: 98%

A risk assessment study of plant genetic transformation usingAgrobacterium and implications for analysis of transgenic plants

Barrett

Cobb

McNicol

et al. 1997

Plant Cell Tiss Organ Cult

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“…2; determined that ampicillin, chloramphenicol, streptomycin, and tetracycline (between 32 mg L -1 and 256 mg L -1 ) efficiently inhibited the growth of endophytic bacteria or contaminants in potato tissue cultures (Solanum tuberosum). Similarly, Leifert et al (1991) cultivated the Delphinium sp. and also obtained cultures that were free of pathogens and showed in vitro growth increases when adding different antibiotics (gentamicin, streptomycin, carbenicillin, cephalothin, rifampicin and polymyxin) to their MS culture medium.…”

Section: Differementioning

confidence: 99%

Micropropagation’s Complete Protocol of Red Araçá (Psidium cattleianum, Myrtaceae) from Germinated Seeds in vitro

Freire¹,

Gardin²,

Baratto³

et al. 2018

JAS

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Red Araçá's (Psidium cattleianum) micropropagation processes have shown enormous potential both in terms of research and as a sustainable native resource to be used in the areas of food production, ecology, and pharmacology. Currently, however, despite that potential, research efforts involving this myrtaceae, native to the Brazilian Atlantic Forest, have been scarce. With that in mind, this study set out to establish micropropagation techniques that would allow the development of a feasible protocol to be used with Red Araçá, achieving its mircropropagation from in vitro germinated seeds. Different types of explants were tested for in vitro establishment. For the multiplication of nodal segments, different concentrations of BAP and IAA combinations were tested in an MS medium. Using the same medium, different concentrations of ampicillin were applied in order to determine its influence on the decontamination of the apical segments. The BAP and IAA combinations were also used to test their effects on the in vitro explants' development and rooting. During pre-acclimatization, survival of in vitro rooted plants was tested in a nebulizer chamber, using a commercial substrate and that same substrate mixed with washed sand (1:1). In essence, it was indeed possible to develop a complete protocol for the micropropagation of the Red Araçá from seedlings obtained by in vitro germination. The in vitro introduction of the Red Araçá was rather efficient, independently of the type of explants used. As the BAP and IAA concentrations increased, so did the in vitro seedlings' development (7 leaves explant -1 ) and rooting (67%). Additionally, the in vitro rooted plants exhibited a high rate of survival (80%) in the pre-acclimatization phase, independently of the substrate used.

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“…Bactérias pertencentes ao gênero Corynebacterium também têm sido relatadas como contaminantes in vitro (Leggatt et al, 1988;Leifert et al, 1991). As células bacterianas são gram-positivas, em forma de bastonetes e não esporogênicas.…”

Section: Resultsunclassified

“…Quando as condições do meio de cultura (nutrição, pH) tornam-se favoráveis ao seu desenvolvimento, as bactérias passam a competir por nutrientes minerais e carboidratos do meio de cultura, comprometendo a multiplicação e o desenvolvimento dos cultivos, podendo levá-los rapidamente à morte. Esta deterioração dos explantes está relacionada com a produção de metabólitos fitotóxicos pelas bactérias, tais como os ácidos láctico e acético, cianeto, além de certos reguladores de crescimento e antibióticos (Bakker & Schippers, 1987;Stanier et al, 1987;Kloepper et al, 1989;Leifert et al, 1991). Do ponto de vista prático, a melhor medida a ser tomada é o descarte e autoclavagem do material contaminado.…”

Section: Introductionunclassified

“…Ressalta-se, entretanto, que o sucesso do trabalho com antibióticos somente pode ser obtido após o isolamento, identificação e realização de testes de sensibilidade das bactérias. Por causa da fitotoxicidade e alto custo do tratamento, os antibióticos devem ser utilizados apenas para contaminantes específicos das culturas, pois somente as bactérias que estiverem dentro do espectro de ação de cada antibiótico serão controladas (Leifert et al, 1991;Teng & Nicholson, 1997).…”

Section: Introductionunclassified

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Identificação e controle com antibióticos de bactérias endofíticas contaminantes em explantes de batata micropropagados

Pereira

Mattos

Fortes

2003

Pesq. agropec. bras.

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Resumo -Este trabalho teve por objetivos isolar, caracterizar e identificar bactérias endofíticas contaminantes encontradas em tecidos de batata durante a micropropagação e selecionar antibióticos para o controle in vitro desses microrganismos por meio da determinação da concentração bactericida mínima inibitória. Brotações de batata apresentando contaminação bacteriana durante a etapa de multiplicação in vitro, foram superficialmente esterilizadas e os internódios transferidos para placas de Petri com ágar nutriente, onde permaneceram incubadas a 28°C por até cinco dias. Após purificação, as bactérias foram caracterizadas e identificadas por testes taxonômicos. Um total de oito estirpes bacterianas foram isoladas e identificadas como pertencentes às famílias Acetobacteriaceae (1) e Enterobacteriaceae (2) e aos gêneros Corynebacterium (3), Pseudomonas (1) e Xanthomonas (1). Os melhores resultados para a inibição do crescimento bacteriano foram obtidos com os antibióticos ampicilina, cloranfenicol, estreptomicina e tetraciclina em concentrações que variaram de 32 a 256 mg L -1 .Termos para indexação: Solanum tuberosum, contaminação, cultura de tecido, microrganismos. Identification and antibiotic control of endophytic bacteria contaminants in micropropagated potato explantsAbstract -This work aimed to isolate, characterize and identify contaminant endophytic bacteria found in potato tissues during the micropropagation and to select antibiotics for in vitro control of these microorganisms by determining the inhibitory minimal bactericidal concentration. Potato shoots presenting bacterial contamination during the in vitro multiplication were superficially sterilized and the internodes transferred to Petri dishes with nutrient agar medium for up to five days at 28°C. After subcultures the grown bacteria were purified and identified through taxonomic tests. A total of eight bacterial endophytic strains were isolated and identified as belonging to Acetobacteriaceae (1) and Enterobacteriaceae (2) families and Corynebacterium (3), Pseudomonas (1) and Xanthomonas (1) genera. The best results for bacterial growth inhibition were obtained with ampicilin, chloramphenicol, streptomycin and tetracycline antibiotics in concentrations ranging from 32 to 256 mg L -1 .

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Elimination of Lactobacillus plantarum, Corynebacterium spp., Staphylococcus saprophyticus and Pseudomonas paucimobillis from micropropagated Hemerocallis, Choisya and Delphinium cultures using antibiotics

Abstract: Elimination of Lactobacillus plantarum, Corynebacterium spp., Staphylococcus saprophyticus and Pseudomonas paucimobilis from micropropagated Hemerocallis, Choisya and Delphinium cultures using antibiotics

Cited by 47 publications

References 33 publications

A risk assessment study of plant genetic transformation usingAgrobacterium and implications for analysis of transgenic plants

A risk assessment study of plant genetic transformation usingAgrobacterium and implications for analysis of transgenic plants

Micropropagation’s Complete Protocol of Red Araçá (Psidium cattleianum, Myrtaceae) from Germinated Seeds in vitro

Identificação e controle com antibióticos de bactérias endofíticas contaminantes em explantes de batata micropropagados

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