Monitor lizards are Varanus species widely distributed, endangered reptile in the IUCN red data list. In India, based on the morphological and ecological characteristic, it is divided into four species viz. Bengal monitor lizard, Yellow monitor lizard, Desert monitor lizard and Water monitor lizard. These four species listed as Schedule I species in Indian Wildlife (Protection) Act 1972. This paper first attempt to present Forensically Informative Nucleotide Sequencing (FINS) for the Indian Varanus based on three mitochondrial genes. The molecular framework will be useful for the identification of Indian Varanus species and trade products derived from monitors and as such, have important applications for wildlife management and conservation. Here, we used known 14 individual skin pieces of four species of monitor lizards; the partial fragment of three mitochondrial genes (Cyt b, 12S rRNA, and 16S rRNA) were amplified for genetic study. In Cyt b, 12S rRNA and 16s rRNA, we observed, 5, 5 and 4 Haplotypes; 71, 69, and 43 Variables sites; 90, 89, and 50 Parsimony Informative sites within four species of Indian monitor lizards, respectively. Despite it, the nucleotide composition was T 26.4, C 32.8, A 29.2 and G11.6; T 18.8, C 29.7, A 34.0 and G 17.5; T 21.7, C 27.3, A 32.5 and G 18.5 in Cyt b, 12S rRNA and 16S rRNA, respectively. The neighbor joining phylogenetic tree and maximum parsimony tree of three mitochondrial genes, showed similar results and reveal that, there are two major clades are present in Indian monitor lizards.
The present study represents first genetic record of single spider species Araniella cucurbitina, genus Araniella from Uttarakhand, India. There are 12 identified species under Araniella genus and they are distributed in Palearctic region. Here, we used known N = 47 (2 from present study and 45 from GenBank) cytochrome oxidase 1 (CO1) sequences of A. cucurbitina representing seven different geographical groups, additionally 23 sequences of eight Araniella species were used for phylogenetic relationship. The CO1 (561 bp) sequences of A. cucurbitina consisted of n = 14 haplotypes, where haplotype 14 (Hap 14) represents Indian species, while all 13 haplotypes (Hap1-13) shared between six A. cucurbitina groups. The overall 'h' and 'π' diversities among seven groups of A. cucurbitina were 0.85291 and 0.00888, respectively, while overall evolutionary divergence was 0.04. The Indian Hap 14, showed minimum sequence divergence (0.02) from Italy and Czech Republic haplotypes (Hap 6), it means it is the closest group compared to others. Evolutionary divergence among eight species of Araniella ranges from 0.003 to 0.114. The maximum likelihood (ML) topology based on 14 haplotypes of A. cucurbitina was divided into two major clades and further two subclades. Furthermore, ML topology between eight species of Araniella was divided into three major clades, where A. cucurbitina and A. proxima clustered together in clade 'A', while six others were present together in clade 'B' and 'C'. This study helps to identify the Indian species from the rest of Araniella species and other cucurbitina population across the world. This study further needs to be on a large scale to know the exact status distribution and molecular phylogeography of this single species of genus Araniella from India.
Certain articles of worship are commonly sold in Uttarakhand, India by the name Hatha Jodi, a root of a rare plant found only in a few parts of central India. The present work provides genetic proof that the Hatha Jodi sold in three local markets of Uttarakhand contained material from the Varanus species, species protected under the Indian Wildlife (Protection) Act, 1972 . A total of eight samples were bought, two each from the local markets in Haridwar and Rishikesh, three from Dehradun and one from an online source (Amazon). The initial inspection confirmed that two of the samples were made of plastic material. Therefore only the other six samples were subjected to DNA analysis. DNA sequences were successfully obtained and matched with reference sequences available in NCBI Genbank database through BLAST search tool for species identification. All the six samples matched 100% with the Indian monitor lizard. The findings indicate how commercialization and the wildlife trade are playing a role in decline of the population of the Indian monitor lizard. If strong protection measures are not taken as soon as possible, the Indian monitor lizards will go Extinct very soon. Therefore, we suggest that the Government and Wildlife enforcement agencies take serious action against the illegal articles available in the local markets of Uttarakhand under the name Hatha Jodi. Further, the government needs to take legal action against offenders in other states in which the product is available for sale.
In DNA barcoding, mitochondrial gene cytochrome c oxidase I recommended as a tool for the rapid identification and discovery of species. Genus Eurema, Family Pieridae is a highly diverse and distributed along wide geographic ranges in the world as well as India including approximately 70 species throughout the world. The present study is preliminary approach, we included n=12 specimen (3 samples per species) of four different Eurema species, listed in IUCN as Least concern species, were collected from Uttarakhand (India), to give the DNA barcodes and examine patterns of gene evolution through molecular phylogenetics with publicly available sequences of other 17 Eurema species present in different countries.The generated (n=12) COI sequences compared with the sequences of the conspecifics submitted from different geographic regions, all four species were correctly identified. The obtained COI barcodes clearly showed the intraspecific and interspecific distance among four Eurema species by using a K2P technique. In spite NJ, clustering analysis also successfully discriminated all four species. In phylogenetic topology, included 21 Eurema species in the Bayesian and MaximumLikelihood analysis recovered all species in two monophyletic clades with strong support and resolved taxonomic position within species groups. Although higher-level relationships among other Eurema species groups require additional study.Keywords: Eurema; Cytochromec oxidase (COI); DNA barcoding; Phylogeny; Uttarakhand DNA barcoding (http://www.barcodinglife.org) has gained so much popularity during the last decade in documenting the global biodiversity (Hebert et al. 2003ab, Ebach and Holdrege 2005, Abdo and Golding 2007, Meusnier et al. 2008, Monaghan et al. 2009, Hajibabaei et al. 2011, Haye et al. 2012, Leray and Knowlton 2015. Cytochrome c oxidase subunit I (COI) is a widely used standard mitochondrial gene for DNA barcoding to facilitate identification of biological specimens (Hebert et al. 2003). DNA Barcoding is also using as a tool for discrimination between cryptic species and to discover new species based on nucleotide sequence divergence. Availability of large amount of sequences in public databases will indeed speed up species description and identification for taxonomist (Blaxter 2004, Ratnasingham andHebert 2007). Overall the goal of DNA barcoding is explicitly to aid species identificapeer-reviewed) is the author/funder. All rights reserved. No reuse allowed without permission.The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/242263 doi: bioRxiv preprint first posted online Jan. 3, 2018; tion, it has frequently been used for phylogenetic inference at multiple taxonomic levels (Tautz et al. 2003, Savolainen et al. 2005 prompting many scientist to contemplate the phylogenetic value of DNA barcode datasets (Tautz et al. 2003, Savolainen et al. 2005, Hajibabaei et al. 2007, Wahlberg and Wheat 2008 The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/242263...
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