Issn 305-9397. Ғылым және білім. 2022. №1-1 (66) issn 2305-9397
ISSN 2305-9397. Ғылым және білім. 2022. № 3-3 (68)
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| ISSN 2305-9397. Ғылым және білім. 2022. № 3-3 (68)____ _
173 Identification of causative agents of black and basal bacteriosis of wheat was carried out using markers ITS, rpoD, dnaK, fyuA, recA, gyrB for Xanthomonas and ITS, rpoD, gyrB, gapA for Pseudomonas. Tandem repeats for both pathogens were characterized using FastPCR software. Phylogenetic analysis was performed by concatenation of rpoD, dnaK, fyuA, recA, gyrB markers and rpoD, gyrB, gapA markers. ITS markers and tandem repeats were analyzed separately; the overall results of phylogenetic analysis for all markers will be combined. Amplification of marker regions. The sequence of the ITS region of DNA, which includes two spacers (ITS), as well as the 5S rRNA gene between them with adjacent regions of the 16S rRNA and 23S rRNA genes, was amplified using universal primers. The rpoD, dnaK, fyuA, recA, gyrB, gltA, and gapA sequences were amplified using known primers [19, 20]. The DNA nucleotide sequence was determined on an ABI 310 genetic analyzer (Applied Biosystems). Molecular genetic identification of bacteria was carried out with respect to available nucleotide sequences deposited in the GeneBank databases ( https://www.ncbi.nlm.nih.gov/ ) using the BLAST algorithm. Amplification of tandem repeats. To identify tandem repeats in bacterial genomes, as well as to select PCR primers, bioinformatic analysis was performed using the FastPCR program ( http://primerdigital.com/fastpcr.html ). Alternatively, PCR primers are optimized for fragment analysis to detect small insertions/deletions in target gene fragments. Methods for collecting primary information. The collection of primary information was carried out by methods of analysis of scientific, and statistical sources, as well as analysis of genetic databases. Primary information on the problem of genetic analysis and diagnosis of wheat bacterioses was obtained from scientific papers. The data obtained were processed by conventional analytical and statistical methods. All results obtained during the implementation of this work are reliable and reproducible since international standards will be used to obtain and process them. жүктеу/скачать 7.12 Mb. Достарыңызбен бөлісу:
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