Supplementary Data
Identification and taxonomy of NT0401
NT0401 was identified according to traditional morphological criteria,
including characteristics of colony on the plate, morphology of substrate
and aerial mycelia, morphology of spores, pigment producing and other
characteristics. The results showed that NT0401 was a Streptomyces
strain. All biochemical and physical characteristics of NT0401 were also
consistent with the genus Streptomyces (Table I).
Table I Taxonomic and physiological characteristics used to identify the
isolate NT0401
__________________________________________________
Characteristics
Strain NT0401
___________________________________________________
Spore chain morphology
spiral
Cell wall constituent
LL-DPA*
Hydrolysis of starch
+
Liquefaction of gelatin
-
Coagulation of skim milk
H2S
+
+
Carbon utility
L-Arabinose
+
D-Fructose
+
Galactose
+
D-Mannitose
+
Sucrose
-
D-Xylose
-
Starch
+
Cellulose
-
1
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* LL-DPA, LL-diaminopimelic acid. The experiments for taxonomy of
NT0401 were designed according to Bergey's Manual of Determinative
Bacteriology (Buchanan and Gibbons 1974), the Manual of Streptomyces
Determination (Chinese Scientific Academy 1975) and the method
described by Embley and Stachebrant (1994). The composition of amino
acids and saccharides of the whole cell were analyzed using TLC as
described by Hasegawa et al. (1983).
Phylogenetic analyses
Comparison of 16S rDNA sequence of NT0401 (DQ985808) with those of
Streptomyces strains available in GenBank showed a highest sequence
similarity of NT0401 to S. violaceus (AJ519938) (Fig. I). However the
characteristics of morphology and physiology of S. violaceus were very
different from those of NT0401. NT0401 had a 16S sequence,
morphological and physiological characteristics that are more similar to S.
grisovariabilis (AY654298) (Gao et al. 2005). So NT0401 could belong to
S. grisovariabilis cluster.
Fig. 1 The phylogenic tree of 16S rDNA sequences. Genomic DNA was
prepared by a DNA extraction kit (DV810A, TaKaRa Biotechnology
Dalian Co., China), and the 16S rDNA was PCR-amplified with a forward
primer 5’-AGAGTTTGATCCTGGCTCAG-3’ and a reverse primer 5’AAGGAGGTGATCCAGCCGCA-3’. The PCR product was sequenced
by Shanghai Biotech Company (China). The resulting 16S rDNA
sequence was aligned using the Clustal-W software (Nigam et al. 2000);
the distance matrices were calculated using MEGA3.1 (Kumar et al.
2
2004), and a phylogenetic tree was constructed by the neighbor-joining
method.
Bootstrap analysis was performed with 10,000 resampling replicates and
the bootstrap values were labeled on the branches. The number followed
ever species name was accession number of its 16S rDNA in NCBI
database.
References
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Bacteriology, 8th edn. The Williams and Wilkins Co., Baltimore
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(1975)
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Kumar S, Tamura, K, Nei M (2004) MEGA3: Integrated software for
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S.hygroscopicus X79853
S.hygroscopicus AB184725
S.olivaceoviridis AB184288
S.canarius AB184396
S.spinichromogenes AB184534
S.bungoensis AB184696
S.galbus X79852
S.galbus AB184201
S.capoamus AB045877
S.capoamus AB184385
S.regensis DQ026649
S.coeruleorubidus AB184841
S.tumescens AF346483
S.tumescens AF346485
NT0401 DQ985808
S.violaceus AJ519938
S.griseovariabilis AY654298
S.triostinicus AB184519
Microbispora rosea D86936
Sup. Fig. 1
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