Co-culture of Bacillus amyloliquefaciens ACCC11060 and Trichoderma asperellum GDFS1009 enhanced pathogen-inhibition and amino acid yield

Abstract Background Retention of agricultural bio-mass residues without proper treatment could affect the subsequent plant growth. In the present investigation, the co-cultivation of genetically engineered T. asperellum and B. amyloliquefaciens has been employed for multiple benefits including the enrichment of lignocellulose biodegradation, plant growth, defense potential and disease resistance. Results The Vel1 gene predominantly regulates the secondary metabolites, sexual and asexual development as well as cellulases and polysaccharide hydrolases productions. Overexpression mutant of the Trichoderma asperellum Vel1 locus (TA OE-Vel1) enhanced the activity of FPAase, CMCase, PNPCase, PNPGase, xylanase I, and xylanase II through the regulation of transcription regulating factors and the activation of cellulase and xylanase encoding genes. Further, these genes were induced upon co-cultivation with Bacillus amyloliquefaciens (BA). The co-culture of TA OE-Vel1 + BA produced the best composition of enzymes and the highest biomass hydrolysis yield of 89.56 ± 0.61%. The co-culture of TA OE-Vel1 + BA increased the corn stover degradation by the secretion of cellulolytic enzymes and maintained the C/N ratio of the corn stover amended soil. Moreover, the TA OE-Vel1 + BA increased the maize plant growth, expression of defense gene and disease resistance against Fusarium verticillioides and Cohilohorus herostrophus. Conclusion The co-cultivation of genetically engineered T. asperellum and B. amyloliquefaciens could be utilized as a profound and meaningful technique for the retention of agro residues and subsequent plant growth.

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Sequence of the N-terminal half of Bacillus amyloliquefaciens α-amylase

Biochemical Journal ◽

10.1042/bj1850387 ◽

1980 ◽

Vol 185 (2) ◽

pp. 387-395 ◽

Cited By ~ 26

Author(s):

H Chung ◽

F Friedberg

Keyword(s):

Amino Acids ◽

Bacillus Subtilis ◽

Amino Acid ◽

Aspartic Acid ◽

Primary Structure ◽

Bacillus Amyloliquefaciens ◽

Proteolytic Enzymes ◽

Alpha Amylase ◽

Sequence Determination ◽

N Terminus

Bacillus amyloliquefaciens alpha-amylase (1,4-alpha-D-glucan glucanohydrolase. EC 3.2.1.1), which is commercially supplied as ‘Bacillus subtilis alpha-amylase’ does not cross-react immunologically with B. subtilis alpha-amylase. This enzyme (from B. amyloliquefaciens) was cleaved by treatment with CNBr into seven fragments. Peptide A was selected for sequence determination. It is the longest one, containing 185 amino acids (i.e. approx. 50% of the total molecule) and connects to the hexapeptide of the N-terminus. Its primary structure was aligned by use of various proteolytic enzymes. The sequence of amino acids 181-184 is identical with that of amino acids 14-17 of the alpha-amylase isolated from B. subtilis (except that amino acid 183 is asparagine rather than aspartic acid).

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Effectiveness of the Biological Control of Garlic (Allium sativum L.)

Acta Horticulturae et Regiotecturae ◽

10.1515/ahr-2016-0017 ◽

2016 ◽

Vol 19 (s1) ◽

pp. 15-17

Author(s):

Jacek Nawrocki ◽

Anna Pogodzińska

Keyword(s):

Biological Control ◽

Health Status ◽

Root Rot ◽

Bacillus Amyloliquefaciens ◽

Allium Sativum ◽

Field Experiments ◽

First Year ◽

Trichoderma Asperellum ◽

Pythium Oligandrum ◽

Second Year

Abstract In two-year field experiments (2014 and 2015), the effect of used preparations on health status of leaves and roots and bulbs on two cultivars of garlic: ‘Arkus’ and ‘Garpek’ was studied. During investigations: Polyversum WP (Pythium oligandrum), Trifender WP (Trichoderma asperellum) and RhizoVital 42 (Bacillus amyloliquefaciens) and standard fungicide Topsin M 500 SC (tiophanate methyl) were used. Unprotected plants presented control. The obtained results showed that in the first year of the studies, all the tested formulations effectively protected the roots and bulbs of garlic against rot, except RhizoVital 42 for ‘Arkus’ variety and Trifender WP for cultivar ‘Garpek’. In 2015, all tested preparations, without exception, limited root rot and the rot of basal part of bulbs both cultivars of garlic. The applied biological preparations had no significant effect on health of the leaves of garlic in 2014 and in 2015 for cultivar ‘Garpek’, while in the second year of studies, all the tested formulations effectively limited the dieback of leaves of the garlic cultivar ‘Arkus’.

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Amino-acid Sequence of Extracellular Ribonuclease (Barnase) of Bacillus amyloliquefaciens

Nature New Biology ◽

10.1038/newbio235015a0 ◽

1972 ◽

Vol 235 (53) ◽

pp. 15-16 ◽

Cited By ~ 44

Author(s):

ROBERT W. HARTLEY ◽

EDWARD A. BARKER

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Bacillus Amyloliquefaciens

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Amino acid sequence of alpha-amylase from Bacillus amyloliquefaciens deduced from the nucleotide sequence of the cloned gene.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)33151-x ◽

1983 ◽

Vol 258 (2) ◽

pp. 1007-1013 ◽

Cited By ~ 5

Author(s):

K Takkinen ◽

R F Pettersson ◽

N Kalkkinen ◽

I Palva ◽

H Söderlund ◽

...

Keyword(s):

Amino Acid ◽

Nucleotide Sequence ◽

Amino Acid Sequence ◽

Bacillus Amyloliquefaciens ◽

Alpha Amylase ◽

Cloned Gene

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Characterization of a Novel PepF-Like Oligopeptidase Secreted by Bacillus amyloliquefaciens 23-7A

Applied and Environmental Microbiology ◽

10.1128/aem.72.1.968-971.2006 ◽

2006 ◽

Vol 72 (1) ◽

pp. 968-971 ◽

Cited By ~ 4

Author(s):

Shiou-Huei Chao ◽

Tzu-Hao Cheng ◽

Chin-Ying Shaw ◽

Meng-Hwan Lee ◽

Yuan-Hsun Hsu ◽

...

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Structural Gene ◽

Bacillus Amyloliquefaciens ◽

Enzymatic Activities ◽

Secreted Protein

ABSTRACT An oligopeptidase from Bacillus amyloliquefaciens 23-7A was characterized along with its biochemical activities and structural gene. The protein's amino acid sequence and enzymatic activities were similar to those of other bacterial PepFs, which belong to metallopeptidase family M3. While most bacterial PepFs are cytoplasmic endopeptidases, the identified PepFBa oligopeptidase is a secreted protein and may facilitate the process of sporulation.

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Isolation and Purification of Two Novel Streptomycete RNase Inhibitors, SaI14 and SaI20, and Cloning, Sequencing, and Expression in Escherichia coli of the Gene Coding for SaI14

Journal of Bacteriology ◽

10.1128/jb.180.6.1582-1585.1998 ◽

1998 ◽

Vol 180 (6) ◽

pp. 1582-1585 ◽

Cited By ~ 3

Author(s):

Daniela Krajcikova ◽

Robert W. Hartley ◽

Jozef Sevcik

Keyword(s):

Escherichia Coli ◽

Amino Acid ◽

Complex Formation ◽

Bacillus Amyloliquefaciens ◽

Significant Similarity ◽

Isolation And Purification ◽

Rnase Inhibitor ◽

Expression In Escherichia Coli ◽

Gene Coding ◽

Sequencing And Expression

ABSTRACT Two new RNase inhibitors, SaI14 (M r, ∼14,000) and SaI20 (M r, ∼20,000), were isolated and purified from a Streptomyces aureofaciensstrain. The gene sai14, coding for SaI14 protein, was cloned and expressed in Escherichia coli. The alignment of the deduced amino acid sequence of SaI14 with that of barstar, the RNase inhibitor from Bacillus amyloliquefaciens, showed significant similarity between them, especially in the region which contains most of the residues involved in barnase-barstar complex formation.

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Glyceolin and phaseolin induction in soybean and beans as a function of biocontrol agents application

Científic - Multidisciplinary Journal ◽

10.29247/2358-260x.2019v6i1.p113-118 ◽

2019 ◽

Vol 6 (1) ◽

pp. 113-118

Author(s):

Marilia Gervásio da Costa Ferreira ◽

Neilson De Oliveira Borges ◽

Ricardo Francischini ◽

Jair Pereira de Melo Júnior ◽

Antônio Jussiê Da Silva Solino ◽

...

Keyword(s):

Bacillus Amyloliquefaciens ◽

Integrated Management ◽

Longitudinal Section ◽

Lower Surface ◽

Resistance Mechanisms ◽

Biocontrol Agents ◽

Cultivated Plants ◽

Trichoderma Asperellum ◽

Regression Test ◽

Soybean Plants

Induction of plant resistance is a tool that can be incorporated into the integrated management of diseases of cultivated plants. In this way, this work aims to evaluate the elicitor action of biocontrol agents in the induction of glyceolin. In this study were used concentrations about 0; 0.5; 1; 1.5 and 2% diluted in water; spore suspension of Trichoderma asperellum BV10, Bacillus subtilis BV02 and Bacillus amyloliquefaciens BV03 microorganisms. To determine the glyceolin, cotyledons were grown in sand, weighed and cut in longitudinal section on the lower surface. Subsequently, these were deposited 50 μL of the concentrations in the cuts and the extraction performed in H2O and concentration determined by absorbance at wavelength 285nm. The results were submitted to analysis of variance and compared by the regression test (p <0.05). When measuring the glyceolin in soybean cotyledons, there was an increase in the accumulation of this phytoalexins as the concentration of B. subtilis BV02 cells increased. The 4% concentration of B. subtilis BV02 promoted 237% more glyceolin accumulation in soybean cotyledons than the control. The accumulation of glyceolin was increased as the concentration of T. asperellum BV10 cells increased, so that 4% concentration promoted 228% more glycerol accumulation than the control. Increasing the concentration of Bacillus amyloliquefaciens BV03 cells increased the accumulation of this phytoalexins. The 4% concentration promoted 129% more accumulation of glycerol in soybean cotyledons than the control. The microorganisms T. asperellum, B. subtilis and B. amyloliquefaciens are indicated as activators of resistance mechanisms of soybean plants.

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Resposta da Fisális (Physalis peruviana L.) à inoculação e coinoculação de microrganismos promotores de crescimento de plantas

Revista em Agronegócio e Meio Ambiente ◽

10.17765/2176-9168.2020v13n3p915-932 ◽

2020 ◽

Vol 13 (3) ◽

pp. 915-932

Author(s):

Felix Cidade do Prado ◽

Rodrigo Ferraz Ramos ◽

Eloi Evandro Delazeri ◽

Cristiano Bellé ◽

Daniel Joner Daroit ◽

...

Keyword(s):

Bacillus Amyloliquefaciens ◽

Azospirillum Brasilense ◽

Trichoderma Asperellum ◽

Chl A ◽

Physalis Peruviana

Objetivou-se avaliar a resposta de Physalis peruviana L. frente à inoculação e coinoculação de diferentes microrganismos promotores de crescimento de planta (MPCP) em condição de campo. O delineamento foi em blocos casualizados, sendo quatro blocos e oito tratamentos. Os tratamentos do experimento referem-se à inoculação individual de Azospirillum brasilense (A), Bacillus amyloliquefaciens (B) e Trichoderma asperellum (T), diferentes combinações com esses MPCPs (T+B; T+A; B+A e T+A+B) e um tratamento controle (C). Aos 15 dias após o transplantio das mudas (DAT) avaliou-se os índices SPAD de clorofilas (Chl a, Chl b e Chl total) e aos 50 e 150 DAT avaliou-se a altura da parte aérea (APA). Os frutos foram colhidos aos 75 DAT até 120 DAT, sendo determinado o número de frutos por planta (NFP) e a massa média de frutos (MF). Amostras dos frutos foram submetidas a análises físico-químicas, com determinação de luminosidade (L*), cromaticidade (c*) e coloração externa (ºhue) dos frutos. Aos 270 DAT determinou-se a biomassa fresca da parte aérea (BFPA) e biomassa seca da parte aérea (BSPA). Não foram observadas diferenças significativas para as variáveis analisadas. As coinoculações de B+A e A+T apresentam as maiores médias para Chl a, enquanto que a combinação de T+A+B apresentou maiores médias para Chl total, ambos diferindo estatisticamente da inoculação de A. brasilense. Para a variável ºhue os tratamentos A, B e T+B apresentaram as menores médias, resultando melhor aspecto visual da coloração de frutos.

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A Single Mutation in the Carbohydrate-Binding Module Enhances Cellulase Activity in Bacillus Amyloliquefaciens Mutant

Walailak Journal of Science and Technology (WJST) ◽

10.48048/wjst.2021.23985 ◽

2021 ◽

Vol 18 (18) ◽

Author(s):

Nitipol POLSA ◽

Chomphunuch SONGSIRIRITTHIGUL ◽

Wasana SUYOTHA ◽

Sugunya SUEBSAN ◽

Somboon ANUNTALABHOCHAI ◽

...

Keyword(s):

Amino Acid ◽

Bacillus Amyloliquefaciens ◽

Cellulase Activity ◽

Carbohydrate Binding ◽

Carbohydrate Binding Module ◽

Single Mutation ◽

Plasma Technology ◽

Lignocellulosic Wastes ◽

Cellulose Binding ◽

Hydrolysis Activity

From our earlier work, we modified the carbohydrate-binding module (CBM) of Bacillus amyloliquefaciens to increase cellulase activity using cold plasma technology. The cellulase gene (BglC-M) from the mutant was expressed in Escherichia coli BL21(DE3) under the T7 promoter. The hydrolysis activity of the cellulase mutant (BglC-M) was approximately 2.5-fold higher than the control (BglC-W) over a wide range of pH and temperature conditions. The amino acid sequence of the mutant BglC-M contained 471 residues that were almost identical to the control BglC-W. Only a single amino acid, lysine, was replaced by glutamic acid at position 370 (K370E) within the carbohydrate-binding module (CBM). Structure prediction and substrate docking of BglC-M indicated that the single mutation (K370E) might involve cellulose binding of the β-sandwich facilitated by hydrogen bonding. The docking study of cellopentaose with the model structure of BglC-M indicated that the replacement of lysine-370 led to the formation of a hydrogen bond with 436Y, which has a shorter distance (2.6 Å) compared with the control (5.4 Å). As a result, the structure becomes more compact and stable, resulting in increased catalytic efficiency. Finally, the biomass hydrolysis ability of cellulase was investigated on lignocellulosic wastes such as pineapple peel, corncob, and durian peel. The BglC-M enzyme showed a more significant amount of reducing sugar released from all lignocellulosic wastes than the control. This was the first evidence that altering the base composition of the cellulose binding module enhanced the catalytic activity. HIGHLIGHTS Increasing cellulase activity of Bacillus amyloliquefaciens using plasma technology Mutation at cellulose-binding module enhance cellulase hydrolysis activity Greater cellulase activity in the hydrolysis on lignocellulosic wastes GRAPHICAL ABSTRACT

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