GENOME WIDE ANALYSIS OF DNA METHYLATION IN RESPONSE TO MACROPHOMINA PHASEOLINA (CHARCOAL ROT) IN SOYBEAN
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Date
2022-01-08
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Publisher : Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra.
Abstract
Abstract : The exploration entitled “Genome wide analysis of DNA methylation in Macrophomina phaseolina (charcoal rot) in soybean (Glycine max L.)” was carried out during 2019-20 in Biotechnology Centre, Department of Agricultural Botany, Dr. PDKV, Akola to look over the extent and pattern of genomic methylation in response to charcoal rot disease in soybean genotypes. Variation in DNA methylation enables plants to inherit traits independently of changes to DNA sequence with the help of restriction enzymes that leads to amplify small region of genome. Here, for validation purpose we screened six selected resistance and susceptible soybean genotypes in response to charcoal rot diseases.
Charcoal rot disease caused by Macrophomina phaseolina is one of the most damaging diseases of soybean resulting to 70 % losses and till date no immune genotype is known for the same. Various limitations inherent in field experiments may have hindered progress toward identifying new source(s) of resistance and breeding resistance in soybean. The Methylation Sensitive Amplified Polymorphism which is modification of Amplified Fragment Length Polymorphism (AFLP) used to investigate the epigenetic changes associated with the Charcoal rot in Soybean. The samples of genomic DNA were double digested with either HpaII or MspI and with EcoRI. To analyze epigenetic changes, 21 MSAP marker with different primer combinations screened and 11 primer combinations showed polymorphism at methylated sites in response to genotypic variation. A total of 312 and 311 clear and reproducible amplicons were amplified from control and CR treated samples, respectively. Under the control conditions, the total methylation of CCGG sequences averaged 49.03% in control and 39.22% in CR treated. The extent of DNA methylation in resistant cultivars were higher under control condition and lower in diseased condition. It means Charcoal rot decreased the percentage of total methylated amplicons in resistant cultivars, but increased it in susceptible genotype TAMS-38 (avg of 41.17%), when levels of DNA methylation were compared with those in the respective control (22.22%). These findings indicated opposite effects of disease on DNA methylation in genotypes of Soybean with different levels of resistivity to Charcoal Rot The fully methylated loci were always more abundant than the hemi-methylated ones. PCoA (Principle Co-ordination Analysis) and NJ (Neighbour Joining) radial tree diagram showed the dissimilarity between primers according to the scored data by genotypes. These results showed that there are great difference in genomic DNA of different soybean genotypes related to the response to the charcoal rot disease. The study revealed that methylation status varied substantially during the stressed or diseased condition. Polymorphic bands can be eluted and sequence further.
Description
Description : An experiment conducted during the year 2019-20 at department of biotechnology Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Dist. Akola (Maharashtra). In this experiment the investigation was carried out to study the gene expression of susceptible and resistant varieties of soybean in response to charcoal rot which reveal the extent and pattern of DNA methylation by calculating it’s methylation percentage in controlled and charcoal rot treated genotypes of Soybean using methylation sensitive amplified polymorphism.
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Citation : MADKE, MEGHA SHAMRAO. (2020). genome wide analysis of DNA methylation in response to macrophomina phaseolina (charcoal rot) in soybean. Biotechnology center, Department of Agricultural Botany. Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola. M. Sc. 2020. Print. xii, 91p. (Unpublished).