[Biology of two lysogenic phages from Bacillus thuringiensis MZ1]. 2007

Wei Liao, and Fan Sun, and Shao-yun Song, and Wei Shi, and Yi Pang
State Key Laboratory of Biocontrol, Sun Yat- Sen Zhongshan University, Guangzhou 510275, China. laoweimail@163.com

A Bacillus thuringiensis (Bt) fermentative strain MZ1 (subsp. kurstaki) , from a company in Meixian County of Guangdong Province, produce toxins during sporulation and are extensively used in the field to control pest insects (Lepidoptera) in China. But some unknown or random factors that inhibited or stopped B. t growth in the fermentation can be regarded as reflecting the exist of lysogenic phage. Therefore, strain MZI and its lysogenic phages were studied in this paper. With indicator strain ZK1, two kind of phage plaques, one with about 3mm diameter and the other with about 1mm diameter, can be observed after strain MZ1 cultured in plates or flasks was induced by mitomycin C. Then, two lysogenic phages, namely MZTP01 and MZTP02, were isolated and characterized in biology. They belonged to family Siphoviridae, which had icosahedral heads (MZTP01 :82nm x 85nm; MZTP02: 75nm x 55nm) and long tails (MZTP01: 220nm x 18nm; MZTP02: 183nm x 12nm) without flexibility. Host range examination showed that six and seven (including indicator strain ZKl) out of 113 B. t strains saved in our laboratory were sensitive to MZTP01 and MZTP02, respectively. MZTP01 was more stable than MZTP02 against pH value, ultraviolet and heat treatment, but contrary against organic solvents. For MZTP01 and MZTP02, K values in the neutralization reactions were 45 and 326, respectively. Both phages had no relationship in their antigenicity. Burst size of phage MZTP02 was 175, two times more than that of MZTP01. Latent time of MZTP02 was 40min, one times shorter than that of MZTP01. It was suggested that both phages DNA be linear dsDNA through the typical absorption curves, reaction with diphenylamine, DNase sensitivity and acridine orange staining. And this was in good accord with the previous findings that all tailed phages being dsDNA moleculars. The genomic DNA and their restriction maps showed that both molecular weights should be between 9.4 - 23kb. Both phage genomic DNAs were digested by Hind III, producing eight and nine bands, respectively. The results above show that strain MZl is proved a double lysogen. It is considered to be the main reason of great losses in the production. The biological information of the two lysogenic phages belonging to strain MZ1 is provided to help solve the problem and the two lysogenic phages is prepared for complete DNA sequencing in the next step.

UI MeSH Term Description Entries
D008242 Lysogeny The phenomenon by which a temperate phage incorporates itself into the DNA of a bacterial host, establishing a kind of symbiotic relation between PROPHAGE and bacterium which results in the perpetuation of the prophage in all the descendants of the bacterium. Upon induction (VIRUS ACTIVATION) by various agents, such as ultraviolet radiation, the phage is released, which then becomes virulent and lyses the bacterium. Integration, Prophage,Prophage Integration,Integrations, Prophage,Prophage Integrations
D001413 Bacillus thuringiensis A species of gram-positive bacteria which may be pathogenic for certain insects. It is used for the biological control of the Gypsy moth. Bacilan,Dipel,Thuricide
D001435 Bacteriophages Viruses whose hosts are bacterial cells. Phages,Bacteriophage,Phage

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