Isolation, identification and characterization of biotechnologically important bacteria from microflora of Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae)

Figures & data

Table 1. Some morphological and biochemical characteristics of D. kuriphilus isolates.

Tests	Dk1	Dk2	Dk3	Dk4
Colony colour	Cream	Cream	Cream	Cream
Cell shape	Cocci	Rod	Rod	Rod
Gram stain	+	+	−	+
Spore stain	−	+	−	+
Motility	−	+	−	+
Catalase test	+	+	+	+
Oxidase test	−	−	+	−

+, positive; −, negative.

Table 2. API 50CH test results for D. kuriphilus isolates.

Tests	Dk2	Dk4
Glycerol	+	+
Erythritol	−	−
D-arabinose	−	−
L-arabinose	+	+
D-ribose	+	+
D-xylose	+	+
L-xylose	−	−
D-adonitol	−	−
Methyl-β-D-xylopyranoside	+	−
D-galactose	+	−
D-glucose	+	+
D-fructose	+	+
D-mannose	+	+
L-sorbose	−	−
L-rhamnose	−	−
Dulcitol	−	−
Inositol	−	−
D-mannitol	+	+
D-sorbitol	−	−
Methyl-α-D-mannopyranoside	−	−
Methyl-α-D-glucopyranoside	wp	−
N-acetylglucosamine	+	−
Amygdalin	+	+
Arbutin	+	+
Esculin (ferric citrate)	+	+
Salicin	+	+
D-cellobiose	+	+
D-maltose	+	+
D-lactose (bovine origin)	+	+
D-melibiose	+	+
D-saccharose	+	+
D-trehalose	+	+
Inulin	+	wp
D-melezitose	−	−
D-raffinose	+	+
Starch	+	+
Glycogen	+	+
Xylitol	−	−
Gentiobiose	wp	+
D-turanose	−	−
D-lyxose	−	−
D-tagatose	−	−
D-fucose	−	−
L-fucose	−	−
D-arabitol	−	−
L-arabitol	−	−
Potassium gluconate	−	−
Potassium 2-ketogluconate	−	−
Potassium 5-ketogluconate	−	−

+, positive; −, negative; wp, weak positive.

Table 3. API 20E test results for D. kuriphilus isolates.

Tests	Dk1	Dk3
β-galactosidase	+	−
Arginine dihydrolase	+	−
Lysine decarboxylase	−	+
Ornithine decarboxylase	−	−
Trisodium citrate	+	−
H2S (sodium thiosulfate)	−	−
Urease	+	wp
L-tryptophan	−	wp
Indole	−	−
VP (sodium pyruvate) test	+	+
Gelatinase	+	+
Glucose fermentation	+	−
D-mannitol fermentation	+	−
Inositol fermentation	−	−
D-sorbitol fermentation	−	−
L-rhamnose fermentation	−	−
D-saccharose fermentation	−	−
D-melibiose fermentation	−	−
Amygdaline fermentation	−	−
L-arabinose fermentation	−	−

+, positive; −, negative; wp, weak positive.

Table 4. API STAPH test results for the Dk1 isolate.

Tests
Glucose	+
Fructose	+
Mannose	+
Maltose	+
Lactose	+
Trehalose	+
Mannitol	+
Xylitol	−
Melibiose	−
Nitrate to nitrite	+
Alkaline phosphatase production	+
Acetyl methyl carbinol production	−
Raffinose	−
Xylose	−
Saccharose	+
Acid from alpha-methylglucoside	−
Acid from N-acetylglucosamine	+
Arginine dihydrolase	+
Urease	+

+, positive; −, negative.

Table 5. GenBank accession numbers and identification of bacterial isolates according to partial 16S rRNA gene sequences.

Isolates	GenBank	Conclusion	16S rRNA sequence similarity (%)	Accession numbers
Dk1	KT377432	Staphylococcus saprophyticus	97	GU197531
Dk2	KT377433	Paenibacillus sp.	99	KF465974
Dk3	KT377434	Pseudomonas fluorescens	99	GU198115
Dk4	KT377435	Paenibacillus sp.	99	EU558285

Figure 1. Neighbour-joining tree of bacterial isolates from D. kuriphilus. The dendrogram was constructed by MEGA 6.0 software based on the partial sequences of the 16S rRNA gene. Bootstrap values shown next to nodes are based on 1000 replicates. The scale on the bottom of the dendrogram shows the degree of dissimilarity.