Different Solubilizing Bacteria Ratio Of Solubilizing Ability And Solutions Of Inorganic Phosphorus Gene Cloning

Phosphorus is an important restrictive factor for plant growth. The excessive use ofphosphate fertilizer accounting for the lack of phosphate has caused so many problemssuch as resource waste,environment pollution. Phosphate-solubilizing microorganism(PSM) in soils play an important role in phosphorus cycling, which can transforminsoluble phosphate into soluble phosphate and improve using of the phosphate fertilizer.Some researches on PSM are carried out, the results are as followes:Nine phosphate-solubilizing microorganisms provided by key laboratory ofmicrobiology in Heilongjiang University, were characteristically detected by plate andphosphomolybdate-blue spectrophotometry. HDBP2,HDBP5,HDBP6,HDFP1 andHDAP1 were chosen according to genus and phosphate-solubilizing capacity, then eachmicroorganism was mixed with each other in the proportion of one to one,one to twoand two to one, and the content of dissoluble phosphorus in cultural solution wasdetermined and compared.The study shows that all of the phosphate-solubilizing strains used in theexperiment can decompose Ca₃(PO₄)₂ and leithin because they all can form colonieswhen they cultured on plates containing the media with Ca₃(PO₄)₂ or leithin as the onlyP source. HDBP5,HDBP6,HDFP1 and HDAP1 produce obvious transparency circleson inorganic-phosphate plate, HDBP5and HDAP1 produce obvious transparency circleson organic-phosphate plate. In the case of Ca₃(PO₄)₂ as P source, when mix HDBP2with HDBP5 in the proportion of two to one, the content of dissoluble phosphorous inthe cultural solution can reach 90.25μg/mL which is the highest in all of the requiredamount.Studies on inorganic-phosphate-solubilizing genes of phosphate-solubilizingbacteria are little, articles published were limited to adjusting genes of producing sour way. The representative is pqqE, the coding productions of which can promote miner-alphosphate-solubilizing of bacteria. Primers were designed by Primer 5 software basedon sequences of pqqE gene in Bacillus thuringiensis and Bacillus cereus whichpublished on GenBank, pqqE genes of HDBP1,HDBP2,HDBP4 and HDBP5 werecloned.Getting two products about 494bp by PCR with HDBP1 and HDBP2 separatelyshow 93.52% and 93.72% similarity with the Bacillus thuringiensis published onGenBank; getting two products about 494bp by PCR with HDBP4 and HDBP5separately show 95.34% and 95.55% similarity with the Bacillus cereus published onGenBank.