Monitoring Lactobacillus plantarum in grass silages with the aid of 16S rDNA-based quantitative real-time PCR assays

Klocke, Michael; Mundt, K.; Idler, Christine; McEniry, J.; O’Kiely, P.; Barth, Susanne

doi:10.1016/j.syapm.2005.06.001

Cited by 42 publications

(37 citation statements)

References 25 publications

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“…L. plantarum is used in a variety of industrial and agricultural applications and prospers in environments containing decomposed lignocellulosic plant biomass (2). In agriculture, the acidifying properties of these organisms are employed for conservation of plant biomass for use in animal feed (3). The ability to produce lactic acid in large amounts could also be used for the production of bio-based plastics (polylactic acid) from plant biomass.…”

mentioning

confidence: 99%

Establishment of a Simple Lactobacillus plantarum Cell Consortium for Cellulase-Xylanase Synergistic Interactions

Moraïs

Shterzer

Grinberg

et al. 2013

Appl Environ Microbiol

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Lactobacillus plantarum is an attractive candidate for bioprocessing of lignocellulosic biomass due to its high metabolic variability, including its ability to ferment both pentoses and hexoses, as well as its high acid tolerance, a quality often utilized in industrial processes. This bacterium grows naturally on biomass; however, it lacks the inherent ability to deconstruct lignocellulosic substrates. As a first step toward engineering lignocellulose-converting lactobacilli, we have introduced genes coding for a GH6 cellulase and a GH11 xylanase from a highly active cellulolytic bacterium into L. plantarum. For this purpose, we employed the recently developed pSIP vectors for efficient secretion of heterologous proteins. Both enzymes were secreted by L. plantarum at levels estimated at 0.33 nM and 3.3 nM, for the cellulase and xylanase, respectively, in culture at an optical density at 600 nm (OD 600 ) of 1. Transformed cells demonstrated the ability to degrade individually either cellulose or xylan and wheat straw. When mixed together to form a two-strain cell-based consortium secreting both cellulase and xylanase, they exhibited synergistic activity in the overall release of soluble sugar from wheat straw. This result paves the way toward metabolic harnessing of L. plantarum for novel biorefining applications, such as production of ethanol and polylactic acid directly from plant biomass.

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mentioning

confidence: 99%

Establishment of a Simple Lactobacillus plantarum Cell Consortium for Cellulase-Xylanase Synergistic Interactions

Moraïs

Shterzer

Grinberg

et al. 2013

Appl Environ Microbiol

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“…The data presented here represent the first report of the use of qPCR to systematically quantify different specific lactic acid bacteria population in conserved forages in the tropics, although Klocke et al (2006) have recently used qPCR based on 16S rDNA directed primers to quantify L. plantarum in grass silage.…”

Section: Resultsmentioning

confidence: 99%

“…Other parameters were also tested, including pH using Cyberscan pH310 Eutech, lactic acid bacteria population was determined by total plate count (TPC) using MRS agar (Cappucino and Sherman, 2001). Microbial quantification analysis using real-time PCR Qiagen (Klocke et al, 2006). DNA extraction from silage samples.…”

Section: Methodsmentioning

confidence: 99%

“…The analysis of qPCR was performed as described by Klocke et al, (2006) by using the Rotorgene Q Qiagen in accordance with the manufacturer's instructions and the dsDNAbinding dye SYBR GreenI with pair specific primers of L. plantarum. The total qPCR reaction was determined in a 20 μL final volume consisted of 10 μL of Quantifast ® SYBR ® Green PCR Kit, 1 μL of each specific primer (F & R), 6 μL of pure distilled water, and 2 μL of extracted DNA sample from each treatment.…”

Section: Methodsmentioning

confidence: 99%

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The Quality of Corn Silage Product From Technopark of Banyumulek Lombok, West Nusa Tenggara

2017

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West Nusa Tenggara province is one potential area for farming of cattle and has been chosen as location for developing a Technopark in Banyumulek. Forage preservation as silage is a program to support the sustainability of forage for beef cattle. Silage was made using whole corn crop and additives of rice bran and Lactobacillus plantarum 1A-2 as inoculum under a block randomized design. Three treatments were given and 10 replications of each month. Evaluation of silage quality, based on chemical and microbiological analysis, showed that silage making during 3 months in Technopark Banyumulek is good and stable.

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“…All quantification of Real-time PCR amplification and detection were performed using Bio-Rad CFX 96 Touch TM Real time PCR Detection System.Species-specific PCR primers for L. plantarum used to amplify partial 16S rDNA regions (target DNA) were F:3'-TTACATTTGAGTGAGTGGCGAACT-5' for forward primer and R:3'AGGTGTTATCC CCCGCTTCT-5'for reversed primer (Klocke et al, 2006). The reaction was conducted inafinal volume of 20 µL, carried out in duplicate, containing the following: 10 µL SsoFast SYBR Green Real-Time PCR master mix (product of BioRad), 0.4 µL forward primer, 0.4 µL reverse primer, 7.2 µL Nucleus-Free Water (NFW), and 2 µL DNA template.…”

Section: Real-time Pcrmentioning

confidence: 99%

Survival of Lactobacillus plantarumU40 on the in vitro rumen fermentation quantified with real-time PCR

Astuti

Widyastuti

Wina³

et al. 2018

J. Indonesian Trop. Anim. Agric.

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ABSTRAKTujuan dari penelitian ini adalah untuk menganalisa daya tahan L. plantarumU40 menggunakan real-time PCR dalam fermentasi rumen in vitro. Desain penelitian yang digunakan adalah Rancangan Acak Kelompok dengan 3 perlakuan dan 4 ulangan. Perlakuan yang diberikan adalah kontrol, inokulasi dengan L. plantarumU40 danL. plantarumU40 + glukosa. Perlakuan dengan inokulasi menghasilkan populasi L. plantarumU40 yang lebih tinggi. Setelah 8 jam inkubasi, pemberian glukosa cenderung menurunkan populasi L. plantarum U40. Perlakuan kontrol menghasilkan populasi L. plantarum U40 yang paling rendah selama fermentasi in vitro.Inokulasi L. plantarum U40meningkatkan populasi Bakteri Asam Laktat secara signifikan (P<0.05) sampai dengan 12 jam inkubasi dibandingkan dengan kontrol. Perlakuan kontrol mempunyai pH yang paling tinggi selama masa inkubasi. Penambahan glukosa secara signifikan (P<0.05) menurunkan pH rumen pada akhir inkubasi (24 jam) (6.30), dibandinkgkan dengan kontrol (6.85).Inokulasi dengan L. plantarumU40 + glukosa secara signifikan (P<0.05) meningkatkan asam propionat, menurunkan asam asetat serta nisbah A/P dibanding perlakuan lainnya.Lactobacillus plantarum U40 tanpa penambahan glukosa tidak memberikan efek terhadap produksi propionat secara signifikan. Dapat disimpulkan bahwa Lactobacillus plantarumU40 dapat bertahan hidup di dalam rumen serta mengubah fermentasi rumen ketika glukosa ditambahkan sebagai sumber karbon. Kata kunci: Lactobacillus plantarumU40, daya tahan, glukosa,fermentasi rumen, quantitative real-time PCR ABSTRACTThe objective of this study was to evaluate the survival of L. plantarumU40 quantified with realtime PCR during in vitro rumen fermentation. The experiment was arranged in a randomized block design with 3 treatments and 4 replications. Treatments were control, rumen fermentation inoculated with L. plantarumU40and L. plantarumU40 + glucose solution. Population of L. plantarum U40 was higher at inoculation treatment. After 8 hours incubation, glucose addition tended to decrease L. plantarum U40 population. Control treatment showed lowest population of L. plantarum U40 along in 184 J.Indonesian Trop.Anim.Agric. 43(2):184-192, June 2018 vitro fermentation compared with other treatment. Inoculation of L. plantarumU40 significantly (p<0.05) increased population of LAB until 12 hours incubation compared with control. Control treatment had highest pH at all incubation time. Glucose addition significantly (P<0.05) decreased final rumen pH (24 hours) (6.30), compared with control treatment (6.85). Inoculation of L. plantarum U40 with glucose addition significantly (P<0.05)increased propionic acid, decreased acetic acid and A/P ratio compared with other treatments. Lactobacillus plantarum U40 without glucose addition did not affect propionic acid production significantly. As conclusion, Lactobacillus plantarum U40 can survive in rumen fluid and changes rumen fermentation when glucose is added as carbon source.

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Monitoring Lactobacillus plantarum in grass silages with the aid of 16S rDNA-based quantitative real-time PCR assays

Cited by 42 publications

References 25 publications

Establishment of a Simple Lactobacillus plantarum Cell Consortium for Cellulase-Xylanase Synergistic Interactions

Establishment of a Simple Lactobacillus plantarum Cell Consortium for Cellulase-Xylanase Synergistic Interactions

The Quality of Corn Silage Product From Technopark of Banyumulek Lombok, West Nusa Tenggara

Survival of Lactobacillus plantarumU40 on the in vitro rumen fermentation quantified with real-time PCR

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