Applied Biology and Biotechnology

 ISSN:  2320 –7825  (Print); eISSN   2320 –7833  (Online)

 Volume No. 9(2)                              June, 2021

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Morphology Taxonomy DNA Barcoding and Phylogenetic Analysis of Soil Fungal isolated (JMBL-1) from Gorakhpur, Uttar Pradesh, India

Siddharth P., Prema Kumari J., Prabhuji S.K and Jonnada A.V. Prasada Rao

Abstract: Mycelium hyaline non septate sporangium globose mycelia grow slowly on PCA 7mm per day on 20-30 degree centigrade forming a rosette growth pattern without aerial mycelium. Sporangia look spherical to sub- spherical and seen terminal germinating by germ tubes. Spores are plerotic with 0.5-1.5 mm thick wall. Oogonia diclinous antheridium with acutely tipped tapering spines. Pythium species with a distinguishing feature of spherical sporangia with echinulate oogonia by slow growth rate distinctive rosette pattern of growth and production of oogonia bearing long, tapering acuminate spines infers to be P. erinaceum Robertson. The amplified product of PCR amplicons with COX II primers were subjected to BLASTn. By this, amplified products (Query sequence) shown 99% identity with the Pythium species. The BLAST analysis shows 99% identity with maximum score of 1014 with 94% query coverage for the species P. erinaceum AB362326.1. Then we did sequence homology with the wild type Pythium erinaceum COX II primer with our amplified product (Query sequence) through TCOFFEE algorithm alignment. The alignment shows very good score of 99%. This region shows 97% consensus region with P. erinaceum therefore, the fungal isolate belongs to the Pythium species. The strong homology reveals the isolate belong to the P. erinaceum. The similar studies performed with ITS universal primers for the characterization of our isolate JMBL-1 shows 98% identity with P. erinaceum. The BLAST analysis shows 98% identity with maximum score of 1559 with 95% query coverage for the species P. erinaceum HQ643534,1. Then we did sequence homology with the wild type Pythium erinaceum ITS region with our amplified product (Query sequence) through T-COFFEE algorithm alignment. The alignment shows very good score of 99%. This region shows 99% consensus region with Pythium erinaceum therefore, the fungal isolate belongs to the Pythium species. The strong homology shows Pythium erinaceum. This result also confirmed and augments the earlier result of COX II amplicons. So the morphological features augments the molecular analysis of PCR based COX II as well as ITS primers indicating that the fungal isolate undoubtedly is Pythium erinaceum Robertson. Further studied MSA using NEJ method with different other species COX ll and ITS sequences and was found out query sequence JMBL 1 showing identity with P. erinaceum followed by Pythium schmitthenneri, P. acrogynum, P. hypogynum, P. selbyi, P. rostratum, P. pulchrum and P. carolinianum. This may be the first report of Pythium erinaceum in Gorakhpur. Keywords: Pythium; morphology; taxonomy; DNA barcoding; phylogeny; India