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International Journal of Biological Technology
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ISSN: 0976-4313 Print
Abstract
Isolate JMBL-5 of Oomycete was isolated from soil samples taken in wheat field of “Jungle Dhusan ” in Gorakhpur, Uttar Pradesh, India. The oomycete grows well on hemp seed halves as well as on solid media like PCA, CMA and PDA producing a chrysanthemum pattern and colony show rosette growth of medium density. It also grows luxuriantly on hemp seed halves in water. The daily growth rate of this oomycete on PCA at 25 °C is 35 mm. On hemp seed halves in water, the oomycete produces whitish colonies. The oomycete sporangia terminal or occasionally intercalary globose and sub spherical.. However it produces hyphal bodies of different dimensions. These hyphal bodies germinate by producing germ tubes to give rise to new mycelium. The oomycete JMBL-5 produces male gametangia (antheridia) and female (oogonia) gametangia plentifully in water on hemp seeds and on solid media. The antheridia are usually monoclinous, 1 to 3 per oogonium. At times the antheridia are sessile. The female gametangia are smooth walled, mostly terminal, at times intercalary, globose, measuring between 17 to 22 µm in diameter (average 18 µm). A fertilization tube is developed between the antheridia and oogonia and after fertilization the antheridia quickly disappear. At times the remains of the antheridial cell is visible even after fertilization and the formation of zygote (oospore which are mostlyaplerotic). These morphological characteristics of our isolate JMBL-5 comes very close to P. ultimum Trow. Which is a highly parasitic oomycete. Agarose gel electrophoretic analysis of the purified DNA of JMBL-5 showed approximate molecular weight of 40 kb. The cytochrome oxidase II (COX II) gene of JMBL-5was PCR amplified and the amplicon was approximately 600 bp. The sequences of COX II gene region of our isolate JMBL-5 was subjected to BLASTn. The analyses shows our isolate is having more closeness and similar to Pythium ultimum Trow. By this amplified product show 99% identity with Pythium species. Blast result show 99% identity and maximum score 1048 with 97% query coverage for the species. The alignment show very good score 98% .This region show99% consensus region with pythium. The amplified COX II of our isolates JMBL 5 was also subjected to MSA through CLCbio Main work bench while constructing the phylogenetic tree by using NEJ Method Further studied MSA and NEJ method with different other species COX II and it was found our query sequence is JMBL 5 identity with P. longisporangium,P. pulchrum , P. selby and P.multidporum. The result of Phylogentic analysis using COX II indicated Pythium is very close to P. longisporangium and P. selby.
Keywords: Pythium; morphology; taxonomy; DNA barcoding; phylogeny; India
Isolate JMBL-5 of Oomycete was isolated from soil samples taken in wheat field of “Jungle Dhusan ” in Gorakhpur, Uttar Pradesh, India. The oomycete grows well on hemp seed halves as well as on solid media like PCA, CMA and PDA producing a chrysanthemum pattern and colony show rosette growth of medium density. It also grows luxuriantly on hemp seed halves in water. The daily growth rate of this oomycete on PCA at 25 °C is 35 mm. On hemp seed halves in water, the oomycete produces whitish colonies. The oomycete sporangia terminal or occasionally intercalary globose and sub spherical.. However it produces hyphal bodies of different dimensions. These hyphal bodies germinate by producing germ tubes to give rise to new mycelium. The oomycete JMBL-5 produces male gametangia (antheridia) and female (oogonia) gametangia plentifully in water on hemp seeds and on solid media. The antheridia are usually monoclinous, 1 to 3 per oogonium. At times the antheridia are sessile. The female gametangia are smooth walled, mostly terminal, at times intercalary, globose, measuring between 17 to 22 µm in diameter (average 18 µm). A fertilization tube is developed between the antheridia and oogonia and after fertilization the antheridia quickly disappear. At times the remains of the antheridial cell is visible even after fertilization and the formation of zygote (oospore which are mostlyaplerotic). These morphological characteristics of our isolate JMBL-5 comes very close to P. ultimum Trow. Which is a highly parasitic oomycete. Agarose gel electrophoretic analysis of the purified DNA of JMBL-5 showed approximate molecular weight of 40 kb. The cytochrome oxidase II (COX II) gene of JMBL-5was PCR amplified and the amplicon was approximately 600 bp. The sequences of COX II gene region of our isolate JMBL-5 was subjected to BLASTn. The analyses shows our isolate is having more closeness and similar to Pythium ultimum Trow. By this amplified product show 99% identity with Pythium species. Blast result show 99% identity and maximum score 1048 with 97% query coverage for the species. The alignment show very good score 98% .This region show99% consensus region with pythium. The amplified COX II of our isolates JMBL 5 was also subjected to MSA through CLCbio Main work bench while constructing the phylogenetic tree by using NEJ Method Further studied MSA and NEJ method with different other species COX II and it was found our query sequence is JMBL 5 identity with P. longisporangium,P. pulchrum , P. selby and P.multidporum. The result of Phylogentic analysis using COX II indicated Pythium is very close to P. longisporangium and P. selby.
Keywords: Pythium; morphology; taxonomy; DNA barcoding; phylogeny; India
Volume No.12(3) December,2021
Table and Contents
Forth Coming Issue
Archives
Morphology Taxonomy DNA Barcoding and Phylogenetic Analysis of Soil Fungal Isolate JMBL - 5 From Gorakhpur Uttar Pradesh India
1J. Prema Kumari, C. Vijayakumar 2 and Jonnada A.V. Prasada Rao*
1Molecular Biology Laboratory, DDU Gorakhpur University, Gorakhpur, Uttar Pradesh, India
1St. Ann’s College for Women Visakhapatnam, Andhra Pradesh, India
2St.Andrew’s College Gorakhpur Uttar Pradesh, India
*Correspondence:drjonnada_avpr@rediffmail.com; Mob.: +919450420142
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