The current study was focused on documentation of amphibian assemblage in North Punjab and Islamabad Capital Territory, Pakistan, by using mitochondrial gene sequences of 16S rRNA. Our study entailed 37% of the known amphibian species of the country. We provided a phylogenetic analysis based on 74 newly generated mitochondrial 16S rRNAs from nine species of genus Microlyla, Duttaphrynus, Allopaa, Nanorana, Sphaerotheca, Minervarya, Hoplobatrachus, and Euphlyctis. We employed the maximum-likelihood inference and Bayesian analysis to assess the taxonomic status of the samples obtained from Pakistan, with respect to other congeneric species from surrounding regions. Our findings confirmed the taxonomic status of South Asian anuran species Duttaphrynus stomaticus, Duttaphrynus melanostictus, Microhyla nilphamariensis, Allopaa hazarensis, Nanorana vicina, Sphaerotheca maskeyi (synonym: S. pashchima), Minervarya pierrei, Hoplobatrachus tigerinus, and Euphlyctis kalasgramensis in Pakistan. We have reported new country records of genus Minervarya (M. pierrei).Minervarya pierrei was previously misidentified as Fejervarya limnocharis, due to dearth of genetic information. We provided the first genetic records of our endemic species N. vicina. The results revealed the taxonomic placement of N. vicina with respect to its congeners and validated the taxonomic status of N. vicina from its type locality (Murree) for the first time. The findings of the present study also indicated the paraphyletic relationship of A.-hazarensis with Nanorana species. So, based on our phylogenetic inferences, morphological characters, and habitat preferences, validity of generic status of A. hazarensis is undecided. As our data were not enough to resolve this issue, we suggest sequencing of additional mitochondrial and nuclear genes in the future studies to get a better resolution. We recommend carrying out extensive surveys throughout the country for proper scientific documentation of amphibians of Pakistan. Many new species, some of them might be endemic to Pakistan, are expected to be discovered, and taxonomic status of other species would be resolved.
The genus Nanorana, also called Yunnan Slow Frogs (Frank & Ramus 1995), belongs to the family Dicroglossidae. Murree Hills Frog N. vicina, is an endemic species of Southeast Asian uplands in Pakistan and India and was first reported by Stoliczka (1872) from Murree, Punjab Province, Pakistan. Later Rais et al. (2014) recaptured it from its type locality and described morphology of one juvenile female and one adult male. The species is listed as Least Concern in IUCN Red List of Threatened Species while the EDGE (Evolutionarily Distinct and Globally Endangered) score is 2.64. In Pakistan, it is reported from brook water springs and streams of Murree, Galiat (North Punjab), Ayubia (Khyber Pakhtunkhwa), and parts of Azad Jammu and Kashmir (Rais et al. 2014). Although the adult form has been characterized (Khan 2006; Rais et al. 2014), the morphology of the tadpoles have not yet been described. The current study aimed to describe external morphology and morphometry of tadpoles of N. vicina.
The nematicidal potential of aqueous leaf extracts of ten plant species distributed in agricultural lands in Yemen were assessed against second stage juveniles of Meloidogyne incognita in laboratory. The juveniles were exposed to 6, 12, 24 and 48 hrs in three concentrations (12.5, 25 and 50%) of leaf extracts. Leaf extracts of Datura stramonium, Peganum harmala, Datura innoxia, Argemone mexicana and Nicotiana glauca were effective in causing juvenile mortality, while leaf extracts of the other five plant species, Azadirachta indica, Catha edulis, Solanum incanum, Tagetes minuta and Withania somnifera showed no or little nematode mortality (< 30 %) even at 50 % concentration and 48 hrs exposure time. The juveniles mortality was increased with increase of concentration and exposure time. Hundred percent juveniles mortality was observed at the modest concentration (25%) of leaf extracts of D. stramonium and D. innoxia after 24 and 48hrs, respectively. Whereas 100 % mortality of juveniles was found at highest concentration (50%) in leaf extract of D. stramonium after 12 hrs and in leaf extracts of D. innoxia, P. harmala and N. glauca after 24 hrs or after 48 hrs in case of leaf extracts of A. mexicana. Leaf extract of D. stramonium achieved the highest mortality percentage at different concentrations and was significantly (p≤0.05) superior over the rest of the extracts applied over time. Akhtar, M. and Farzana, B. (1996): Evaluation of nematicidal properties of some members of the family Solanaceae. Biores. Technol., 57: 95-97. Akhtar, M. and Malik, A. (2000). Role of organic soil amendments and soil organisms in the biological control of plant-parasitic nematodes: a review. Biores. Technol., 74, 35-47. Akpheokhai, I.L.; Claudius-Cole, A. O. and Fawol B. (2012). Evaluation of some plant extracts for the management of Meloidogyne incognita on soybean (Glycine max). World J. of Agric. Sci., 8 (4): 429-435. Awadh, G. M.; Saeed, M. R.; Najy, A. and Saleh, A. M. ( 2008). In-vitro nematicidal activity of some selected plants on stem nematode Ditylenchus dipsaci f. (Tylenchida: Tylenchidae). Yemeni J. of Biological Sci., 4(1):141-150. Banna, L., Darwish, R. M. and Aburjal, T. (2003). Effect of plant extracts and essential oils on root-knot nematode Phytopathol. Medit., 42: 123-128. Chaudhary, K. K.; Haile, A.; Ayresea, Z. G.; Semereab, G. and Weldegergish, T. (2013). Nematicidal activity of Eritrean weed plants against the root-knot nematode Meloidogyne incognita (Kofoid and White) Chitwood. Tahrouch, S.; Bourijate, M. and Hatimi, A. (2007). Action differents extractsts végétaux sur la mortalité des nématodes à galles du genre Meloidogyne ssp. Acta Bot. Gallica, 154 (4):503-509. Elbadria, G. A.; Leeb, D. W.; Parkc, J. C.; Yuc, H. B. and Chooc, H. Y. (2008). Evaluation of various plant extracts for their nematicidal efficacies against juveniles of Meloidogyne incognita. J. of Asia-Pacific Entomol., 11 (2): 99 -102. El-Hassan, M.; Ferji, Z. and Idrissi, H. L. (2013) Anti-nematode effect assessment of Peganum harmala based pro...
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