| Over the last decade, Metarhizium anisopliae in the pathogenic mechanism, high-virulence strains, such as selection and application of technology has made considerable progress, and implementation of commercialization.Metarhizium anisopliae has been successfully used for prevention and control of desert locusts, grasshoppers and other oriental migratory locust and been recommended to promote the use of environmentally friendly products by UN Food and Agriculture Organization (FAO) in 1989. As microorganisms pesticide, spores are the role of infecteing insects units. cloning and identification of the Characteristics of spores and sporulation-related gene are important to clarify the characteristics of sporulation and spore formation and other important traits related to the molecular mechanism.Breeding conidiation for fast, high sporulation rate, spores of the strain characteristics of the fine provided the theoretical basis and the corresponding target gene, in order to better achieve Metarhizium anisopliae as a biological insecticide applications scale. Breeding conidiation for fast, high sporulation rate, spores of the fine strain characteristics providing the theoretical basis and the corresponding target gene to achieve applications of Metarhizium anisopliae as a biological insecticide in the scale.At present, studying conidiation in Metarhizium anisopliae, mainly from environmental factors and physical and chemical factors in terms of exploration, and its molecular mechanism of the formation of spores systematic research report also rare. Dased on the Metarhizium anisopliae CQMa102 subtracted cDNA library which constructed by suppression subtractive hybridization during the conidiation stage, we chose the EST which was highly expressed during conidiation. Using the Metarhizium anisopliae CQMa102 normalized full-length cDNA library during the conidiation stage, we have got the full-length cDNA of MaECM33 .Using the Metarhizium anisopliae CQM102 genomic DNA library,we we have got the full-length DNA of MaECM33..In addition, we use RNAi to analyse the function of MaECM33 gene.The main achievements are as follows:1. A 1396 bp DNA fragment which contained the full-length Mmc gene as well as upstream and downstream regulatory sequences was cloned and sequenced. The putative coding sequence which contained a 1167 bp ORF. And the accession number of the gene in genebank is FJ788387. 2. A protein of 388 aa with a predicted molecular mass of 40.443 kDa and a pI of 5.22 was deduced. By the http://www.cbs.dtu.dk/ forecast, including the initial methionine was found, including 19 consecutive amino acid residues for the signal peptide.3. Comparing structural gene sequences with the cDNA sequence by bl2seq software ,the results show that the structural gene containing two introns, the length of 58bp and 66bp, one intron (969-1035) has typical GT ... AG boundaries, and the other intron (73-130) of its boundary for the TG ... GG.4. southern blotting showed that, MaECM33 is the form of a single copy existing in the genome.5. MaECM33 RNA interference vector (pbar-RNAi-gpd-trp-ECM33)was constructed. pBTM plasmid as the basic skeleton containing a ammonium glufosinate selection markers, gpdA promoter and trpC promoter sequence.A multi-cloning site (MCS) is between the two promoter for the purpose of inserting target gene.6. Two transformants were verified by PCR. On 1/4SDA medium, the conidiation.speed and total yield were not significantly different between the wild-type and RNAi mutant. Meanwhile, There is no change in colony morphology.
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