1. General description

Bt strain: S2160-1

Isolate: 20070720S2160

Isolate source: Soil

Collected by: Wenfei Zhang and Liu Xie et al.

Collection date: July-20-2007

Collection site: Guangxi Dawangling Natural Reserves of Guangxi Province, Southwest China

Subspecies: Unknown

Serovariety: Unknown

Identified by: Xuanjun Fang, Wenfei Zhang, Liu Xie

Deposited at:The HITAR Bacillus Collections with Accession No 20070720S2160

Owned by: Hainan Institute of Tropical Agricultural Resources

2. Molecular Features

Crystal shape: Spherical

Protein profile: 140 kDa, 130 kDa, 75 kDa and 30 kDa

Plasmid profile: the plasmid profile of Bt S2160-1 was significantly different to that of Bti, and the other strains

Genotype: in contrast to Bti no cry4 or cry10 genes were identified using the S5un4/S3un4 primers (Table 1) and also that no PCR products were obtained with primers designed to amplify cry11 and cyt1 genes

Cloned gene: Cry 30Ea1 (Accession number EU503140.1); Cry 30Ga (Accession number HQ638217.1); Cry 50Ba1 (Accession number GU446675.1); Cry 54Ba1 (Accession number GU446677.1)

Target insect: Mosquitoes (Culex quinquefasciatus and Aedes albopictus)

3. Featured Photos or Table

Figure 1: Scanning electron micrograph of spores and crystals from Bt S2160-1 (Zhang et al., 2012).

Figure 2: Profiles of SDS-PAGE of Bt S2160-1 (Zhang et al., 2012).

Figure 3: PFGE banding patterns of mega plasmid from 2160-1 (Zhang et al., 2012).

Figure 4: PCR–RFLP patterns of cry-type genes from Bt S2160-1 (Zhang et al., 2012).

Figure 5: SDS-PAGE gel electrophoresis analysis of genes of cry50Ba and cry54Ba expressed in BL21 (DE3) cells using pET expression system.

Figure 1 Scanning electron micrograph of spores and crystals from Bt S2160-1 Note: SP is a spore formed during the stationary phase of growth, next to crystal protein is CP produced as regular crystal is related to spore formation

Figure 2 SDS-PAGE analyses of crystal proteins from Bt S2160-1 and Bti Note: Bt S2160-1 and Bti were cultured in G-Tris medium, The samples for SDS-PAGE analysis taken before and after sporation respectively and crystal proteins digested with 1uM trypsin were analyzed together. PM1: protein molecular weight marker, land 1, 4: strain proteins before sporation from Bti and Bt S2160-1 respectively; lane 2, 5: crystal proteins after sporation from Bti and Bt S2160-1 respectively; lane 3, 6: crystal proteins from Bti and Bt S2160-1 were treated with 1ug/ml trypsin

Figure 3 PFGE patterns of larger plasmids from Bt strains Note: lane1: B. thuringiensis subsp kurstaki HD73; lane 2: B. thuringiensis subsp kurstaki HD-1; lane 3: B. thuringiensis subsp israelensis AND508; lane 4: Bt S2160-1; B. thuringiensis subsp israelensis AND508 harboring the large plasmids of pXO16 (350 kb) and pBtoxis (128 kb), was used as size marker

Figure 4 PCR–RFLP patterns of cry-type genes from Bt S2160-1 Note: M. DNA molecular weight marker; lane1, 2, 3 are PCR amplification using primer pairs S5un4/S3un4, up30-5/up30-3 and up39.40-5/up39.40-3 respectively; lane4, 5, 6. were RFLP patterns of the corresponding PCR production digested with BanI/ HaeⅢ , BglⅡ/PstI and HindIII/HaeIII restriction enzymes

Figure 5 SDS-PAGE gel electrophoresis analysis of genes of cry50Ba and cry54Ba expressed in BL21 (DE3) cells Note: PM2: protein molecular weight marker; lane 1: E. coli BL21 (DE3) cell harboring pET-30a incubated with IPTG; lane 2, 5: E. coli BL21 (DE3) cell harboring pETcry54Ba and pETcry50Ba incubated without IPTG; lane 3, 6: E. coli BL21 (DE3) cell harboring pETcry54Ba

Table 1 Larvicidal activity of spore/crystal mixtures from Bt S2160-1 and Bti against Plutella xylostella and mosquitoes Note: a 95% FL are the 95% fiducial limits in parentheses, as determined by probit analysis

4. Related Publications

Zhang W, Crickmore N, George Z, Xie L, He YQ, Li Y, Tang JL, Tian L, Wang X, Fang X (2012) Characterization of a new highly mosquitocidal isolate of Bacillus thuringiensis--an alternative to Bti? J Invertebr Pathol 109 (2):217-222. doi:10.1016/j.jip.2011.11.003

5. Filed Patent or Related to This Strain

No.

References 

Zhang W, Crickmore N, George Z, Xie L, He YQ, Li Y, Tang JL, Tian L, Wang X, Fang X (2012) Characterization of a new highly mosquitocidal isolate of Bacillus thuringiensis--an alternative to Bti? J Invertebr Pathol 109 (2):217-222. doi:10.1016/j.jip.2011.11.003