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Photosystem I P700 chlorophyll a apoprotein A1 as PCR marker to identify diatoms and their associated lineage
Authors:Vandana Vinayak  Mohd Jahir Khan  Anupam Nath Jha  Harish
Affiliation:1. Diatom Nanoengineering and Metabolism Laboratory (DNM), School of Applied Sciences, Dr. Harisingh Gour Central University, Sagar, India;2. Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur, India;3. Department of Botany, Mohanlal Sukhadia University, Udaipur, India
Abstract:The morphological characteristics of diatoms are useful for studying their taxonomy. However, the distinction between closely related diatom taxa can be very difficult, especially when the morphological characters are modified by environmental constraints. In the present study, 13 fresh water diatoms were identified morphologically and cultured under axenic conditions. To check this, PCR primers specific for multilocus genes were designed to amplify and screen 13 fresh water diatom monocultures. Multilocus PCR primers (DRR3, scfcpA, Lhcf11, SIT1, SIT3, SIT4, LOC101218388, COI-5P, rbcL, rbcL-3P, LSU D2/D3, UPA, psaA, and 18S rRNA) were tested. It was found that psaA gene, a plant pigment chlorophyll-based PCR marker, amplified in all the diatoms. Out of 13 diatom amplicons, only two fresh water diatoms DNA were sequenced. This included Cyclotella meneghiniana and Sellaphora pupula. The Sanger sequencing results thus established that morphologically identified diatom, Sellaphora pupula, exhibited close phylogeny to Sellaphora whereas fresh water Cyclotella meneghiniana has close lineage to marine diatom Thallosiosira.
Keywords:diatoms  DNA  multilocus PCR markers  phylogeny  psaA
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