Polyphasic characterization of bacteria obtained from upland rice cultivated in Cerrado soil

Braga, Lívia Fabiana; Oliveira, Fênix Araújo de; Couto, Eva Aparecida Prado do; Santos, Karina Freire d'Eça Nogueira; Ferreira, Enderson Petrônio de Brito; Martin-Didonet, Claudia Cristina Garcia

doi:10.1016/j.bjm.2017.04.004

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Brazilian Journal of Microbiology

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Environmental Microbiology, Research Paper • Braz. J. Microbiol. 49 (1) • Jan-Mar 2018 • https://doi.org/10.1016/j.bjm.2017.04.004 copy

Polyphasic characterization of bacteria obtained from upland rice cultivated in Cerrado soil^☆ ☆ Part of the Master's Dissertation of the first author approved by the Molecular Science Post-Graduation Program of the Goiás State University.

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT

This work aimed to characterize 20 isolates obtained from upland rice plants, based on phenotypic (morphology, enzymatic activity, inorganic phosphate solubilization, carbon source use, antagonism), genotypic assays (16S rRNA sequencing) and plant growth promotion. Results showed a great morphological, metabolic and genetic variability among bacterial isolates. All isolates showed positive activity for catalase and protease enzymes and, 90% of the isolates showed positive activity for amylase, catalase and, nitrogenase. All isolates were able to metabolize sucrose and malic acid in contrast with mannitol, which was metabolized only by one isolate. For the other carbon sources, we observed a great variability in its use by the isolates. Most isolates showed antibiosis against Rhizoctonia solani (75%) and Sclerotinia sclerotiorum (55%) and, 50% of them showed antibiosis against both pathogens. Six isolates showed simultaneous ability of antibiosis, inorganic phosphate solubilization and protease activity. Based on phylogenetic analysis of the 16S rRNA gene all the isolates belong to Bacillus genus. Under greenhouse conditions, two isolates (S4 and S22) improved to about 24%, 25%, 30% and 31% the Total N, leaf area, shoot dry weight and root dry weight, respectively, of rice plants, indicating that they should be tested for this ability under field conditions.

Keywords:
Oryza sativa; PGPR; Associative bacteria; Bacterial diversity

Groups	Isolate	Shape	Border	Surface	Consistence	Elevation	Gram
A	S2	I	W	R	D	F	-
A	S4	I	W	R	D	F	-
A	S6A	I	W	R	D	F	-
A	S39	I	W	R	D	F	-
A	S97	I	W	R	D	F	-
B	S5	I	W	R	D	F	+
B	S6	I	W	R	D	F	+
B	S8A	I	W	R	D	F	+
B	S8B	I	W	R	D	F	+
B	S17	I	W	R	D	F	+
B	S32	I	W	R	D	F	+
B	S35	I	W	R	D	F	+
B	S37	I	W	R	D	F	+
B	S105	I	W	R	D	F	+
C	S41	C	S	G	A	H	-
C	S25	C	S	G	A	H	-
D	S22	C	S	G	A	H	+
D	S26	C	S	G	A	H	+
D	S29	C	S	G	A	H	+
D	S63	C	S	G	A	H	+

Evaluations	Isolates
	S2	S4	S5	S6	S6A	S8A	S8B	S17	S22	S25	S26	S29	S32	S35	S37	S39	S41	S63	S97	S105
Carbon sources
Sucrose	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+
Glucose	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	-	-
Fructose	-	-	-	-	-	-	+	-	+	+	-	-	-	-	-	-	-	-	-	+
Mannitol	-	-	-	-	-	-	-	-	+	-	-	-	-	-	-	-	-	-	-	-
Sorbitol	-	-	-	+	-	-	+	+	+	-	-	-	-	-	-	-	-	+	-	+
Mio-inositol	+	+	+	-	-	+	+	-	-	-	-	-	-	+	+	+	-	-	-	-
Arabinose	-	-	-	-	+	-	+	+	+	-	-	-	-	-	-	-	-	+	-	+
Maleic acid	+	+	+	+	-	-	-	-	+	-	-	-	-	-	-	-	-	+	-	-
Nicotinic acid	-	-	+	-	-	-	+	+	+	+	-	-	-	-	-	-	-	-	-	-
Malic acid	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+
Enzyme activity
Cellulase	+	-	+	-	+	+	+	-	+	-	-	-	-	-	-	-	-	-	+	+
Amylase	+	+	+	+	+	+	+	+	+	+	-	+	+	+	+	+	+	-	+	+
Protease	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+
Phosphatase	+	+	+	+	+	+	-	+	+	+	+	+	+	+	+	+	+	-	+	+
Urease	+	-	-	-	-	-	-	-	-	-	-	-	+	-	-	-	+	+	-	-
Nitrogenase	+	+	-	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	-	+
Reductase nitrate	-	+	-	+	+	+	+	-	+	+	+	-	+	+	+	-	-	-	+	+
Citrate lyase	-	+	-	-	-	+	-	+	+	-	+	+	-	+	-	-	-	+	+	-
Catalase	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+	+

Isolate	SI (%)	PEI	Antibiosis
			Ss	Rs
S2	133.86 (±16.9)	2.03 (±0.15)	-	-
S4	126.94 (±13.7)	2.52 (±0.22)	-	+
S5	133.60 (±14.5)	1.98 (±0.04)	-	-
S6	130.56 (±4.8)	3.04 (±0.06)	-	+
S6A	116.78 (±9.8)	1.88 (±0.18)	+	+
S8A	119.63 (±2.8)	1.57 (±0.10)	-	-
S8B	0	1.88 (±0.05)	-	-
S17	117.10 (±7)	2.21 (±0.22)	-	+
S22	131.02 (±7.9)	1.67 (±0.01)	+	-
S25	129.17 (±19)	1.91 (±0.17)	+	+
S26	114.95 (±4.3)	1.96 (±0.13)	+	+
S29	123.15 (±11.2)	2.36 (±0.15)	+	+
S32	119.39 (±10)	1.96 (±0.34)	+	+
S35	120.20 (±6.1)	1.61 (±0.13)	+	+
S37	117.78 (±1.9)	2.19 (±0.07)	+	+
S39	116.06 (±5.3)	1.96 (±0.13)	-	-
S41	128.52 (±5.7)	1.89 (±0.07)	+	+
S63	0	1.57 (±0.10)	-	+
S97	116.74 (±8.4)	1.73 (±0.07)	+	+
S105	126.52 (±9.1)	1.35 (±0.08)	+	+

Treatments	RDW (g plant^-1)	LA (cm² plant^-1)	SDW (g plant^-1)	N-Total (mg plant^-1)
S4	3.57 a	678.63 a	6.88 a	217.48 a
S22	2.79 a	601.01 a	5.92 a	218.02 a
S29	2.01 b	462.35 b	4.46 b	161.72 b
S105	2.05 b	561.68 a	5.44 a	219,06 a
NI	2.16 b	475.15 b	4.45 b	164.87 b
CV^a a CV = coefficient of variation. (%)	8.95	8.08	7.55	7.90

Sociedade Brasileira de Microbiologia USP - ICB III - Dep. de Microbiologia, Sociedade Brasileira de Microbiologia, Av. Prof. Lineu Prestes, 2415, Cidade Universitária, 05508-900 São Paulo, SP - Brasil, Ramal USP 7979, Tel. / Fax: (55 11) 3813-9647 ou 3037-7095 - São Paulo - SP - Brazil
E-mail: bjm@sbmicrobiologia.org.br

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[1] *Corresponding author. E-mail: enderson.ferreira@embrapa.br (E.P. Ferreira).

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Polyphasic characterization of bacteria obtained from upland rice cultivated in Cerrado soil☆ ☆ Part of the Master's Dissertation of the first author approved by the Molecular Science Post-Graduation Program of the Goiás State University.

ABSTRACT

Polyphasic characterization of bacteria obtained from upland rice cultivated in Cerrado soil^☆ ☆ Part of the Master's Dissertation of the first author approved by the Molecular Science Post-Graduation Program of the Goiás State University.