Revista de Biología Tropical ISSN Impreso: 0034-7744 ISSN electrónico: 2215-2075

Morphological, molecular characterization and taxonomic status of Triplophysa marmorata and Triplophysa kashmirensis (Cypriniformes: Nemacheilidae) from Kashmir valley, India
PT 64-2 JUN 2016


principal component analysis
molecular characterization
cytochrome oxidase 1
Kashmir valley.
análisis de componentes principales
caracterización molecular
citocromo-oxidasa 1
valle de Kashmir.

How to Cite

Bashir, A., Bisht, B. S., Kumar, R., Mir, J. I., & Patiyal, R. S. (2016). Morphological, molecular characterization and taxonomic status of Triplophysa marmorata and Triplophysa kashmirensis (Cypriniformes: Nemacheilidae) from Kashmir valley, India. Revista De Biología Tropical, 64(2), 473–482.


In India the distribution of genus Triplophysa has been reported only in the upper drainage of the Indus River in Jammu and Kashmir and Lahul and Spiti area of Himachal Pradesh. There is no study on the taxonomic characterization of this genus from Kashmir Himalaya. Therefore the present study was aimed to characterize two important fish species Triplophysa marmorata and T. kashmirensis from Kashmir valley, by using morphometric and molecular tools. It is difficult to discriminate these two species due to the poor quality of original descriptions, and the lack of good reviews. Keeping this in view, a morphometric and molecular study was conducted. Morphometric data were analyzed by using univariate analysis of variance (ANOVA) and multivariate analyses (Principal component analysis) and mtDNA marker Cytochrome oxidase 1 was used for molecular support. Altogether, 22 morphometric characters were used and 15 characters were found significantly variable (P < 0.05). First two components of principal component analysis (PCA) i.e. PC1 and PC2 grouped these two species into separate clusters. The Cytochrome oxidase 1 analysis showed that the mean intraspecific nucleotide divergence (K2P) was 0.001 and interspecific nucleotide divergence was 0.007. Despite having low K2P divergence, these two species got separated into two distinct clades in both Neighbour joining (NJ) and Unweighted Pair Group Method with Arithmetic Mean (UPGMA) tree building methods. But the pattern of clade formation showed that these species were recently radiated from each other and may have the same ancestor. Furthermore, these two species were found closer to Nemacheilidae than to Balitoridae family in the phylogenetic analysis. The molecular divergence between these species was also supported by variance in morphometric data. This work may build the base for the revision of taxonomic identity of these two important fishes of genus Triplophysa. The present investigation formulated that, based on morphological and mtDNA COI sequences analysis, these two taxonomic Triplophysa species should be considered as valid. The results may further assist to enhance the knowledge of the ichthyologists in understanding the ichthyofauna of Kashmir Valley and will help them in planning strategies for conservation and management of these less studied small indigenous species along their natural range of distribution.


Avise, J. C. (1994). Molecular markers, natural history and evolution. New York: Chapman and Hall, Inc.

Elliott, N. G., Haskard, K., & Koslow, J. A. (1995). Morphometric analysis of orange roughy (Hoplostethus atlanticus) off the continental slope of southern. Australian Journal of Fish Biology, 46, 202-220.

Gao, S. B., Tang, H. Y., Qiao, Y., Yang, Z., & Chen, J. S. (2011). Status of fishery resources in the main stream of the lower reaches of Jinsha River. Journal of Hydroecology, 34, 44-49 (in Chinese).

Hebert, P. D. N., Cywinska, A., Ball, S. L., & deWaard, J. R. (2003). Biological identifications through DNA barcodes. Proceedings of Royal Society of London B Biological Sciences, 270, 313-321.

Heckel, J. J. (1838). Fische aus Cashmir gesammelt und herausgegeben von Carl Freiherrn v. Hügel. Mechitaristen, Wien, 110 pp, 13 pls.

Hubert, N., Duponchelle, F., Nunez, J., Garcia-Davila, C., Paugy, D., & Renno, J. F. (2007). Phylogeography of the piranha genera Serrasalmus and Pygocentrus: Implications for the diversification of the Neotropical ichthyofauna. Molecular Ecology, 16, 2115-2136.

Hubert, N., Hanner, R., Holm, E., Mandrak, N. E., Taylor, E., Burridge, M., Watkinson, D., Dumont, P., Curry, A., Bentzen, P., Zhang, J., April, J., & Bernatchez, L. (2008). Identifying Canadian freshwater fishes through DNA barcodes. PLosONE, DOI, 10.1371/journal.pone.0002490.

Jiuxuan, Li., Kun, Y., Shengjuan, S., Xiuyue, Z., & Zhaobin, S. (2015). Complete mitochondrial genome of Triplophysa bleekeri (Cypriniformes: Balitoridae: Nemacheilinae), and analysis of mitochondrial genetic variability among Triplophysa species. Mitochondrial DNA, DOI, 10.3109/19401736.2014.1003878.

Kimura, M. (1980). A simple method of estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution, 16, 111-120.

Kullander, S. O., Fang, F., Delling, B., & Ahlander, E. (1999). The fishes of the Kashmir Valley. In L. Nyman (Ed.), River Jhelum, Kashmir Valley: Impact on the Aquatic Environment (pp. 100-167). Goteberg, Sewden: Swedmar.

Lara, A., Ponce de Leon, J. L., Rodriguez, R., Casane, D., Cote, G., Bernatchez, L., & Garcia-Machado, E. (2010). DNA barcoding of Cuban freshwater fishes: Evidence for cryptic species and taxonomic conflicts. Molecular Ecology Resources, 10, 421-430.

Li, W. J., Chen, X. C., & Hu, Y. P. (2015): A new species of the genus Triplophysa (Nemacheilinae), Triplophysa qilianensis sp. nov, from Qinghai, China. Zootaxa, 3905, 418-424.

Li, W. X., Yang, H. F., Chen, H., Tao, C. P., Qi, S. Q., & Han, F. (2008) A new blind underground species of the genus Triplophysa (Balitoridae) from Yunnan, China. Zoological Research, 29, 674-678.

Menon, A. G. K. (1987). The fauna of India and the adjacent countries. Pisces Vol. IV Teleostei Cobitoidea Part I Homalopteridae. Director, Zoological Survey of India, Calcutta.

Mir, J. I., Patiyal, R. S., & Sahoo, P. K. (2015). Length-weight relationships of 10 fish species from a Ganga River Basin tributary, Uttarakhand, India. Journal of Applied Ichthyology, 31, 431-432.

Ornelas-Garcia, C. P., Dominguez-Dominguez, O., & Doadrio, I. (2008). Evolutionary history of the fish genus Astyanax Baird & Girard (1854) (Actynopterigii, Characidae) in Mesoamerica reveals multiple morphological homoplasies. BMC Evolutionary Biology, 8, 340.

Perdices, A., Doadrio, I., & Bermingham, E. (2005). Evolutionary history of the synbranchid eels (Teleostei: Synbranchidae) in Central America and the Caribbean islands inferred from their molecular phylogeny. Molecular Phylogenetic Evolution, 37, 460-473.

Posada, D., & Crandall, K. A. (1998). MODELTEST: Testing the model of DNA substitution. Bioinformatics, 14, 817-818.

Ratnasingham, S., & Hebert, P. D. N. (2007). BOLD: The Barcode of Life Data System ( Molecular Ecology Notes, 7, 355-364.

Rendahl, E. (1933). Weitere Untersuchungen iiber den Schultergiirtel und die Brustflossenmuskulatur der Cobitiden. Arkiv för Zoologi, 25, 1-37.

Saitou, N., & Nei, M. (1987). The neighbour-joining method: a new method for reconstructing evolutionary trees. Molecular Biology Evolution, 4, 406-425.

Sambrook, J., Fritsch, E. F., & Maniatis, T. (2001). Molecular Cloning, A Laboratory Manual, 2nd Ed. New York: Cold Spring Harbor Laboratory Press.

Talwar, P. K., & Jhingran, A. G. (1991). Inland Fishes of India and Adjacent Countries. N. Delhi: Oxford and IBH publishing Co.

Tamura, K., Stecher, G., Peterson, D., Filipski, A., & Kumar, S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology Evolution, 30, 2725-2729.

Teletchea, F. (2009). Molecular identification methods of fish species: Reassessment and possible applications. Reviews in Fish Biology and Fisheries, 19, 55-85.

Thangaraj, M., & Lipton, A. P. (2010). Genetic identity of three Indian populations of three spotted seahorse, Hippocampus trimaculatus. Advanced Biological Resources, 4, 37-41.

Valdez-Moreno, M., Ivanova, N. V., Elías-Guitiérrez, M., Contreras- Balderas, S., & Hebert, P. D. N. (2009). Probing diversity in freshwater fishes from Mexico and Guatemala with DNA barcodes. Journal of Fish Biology, 74, 377-402.

Ward, R. D., Hanner, R., & Hebert, P. D. N. (2009). The campaign to DNA barcode all fishes. Journal of Fish Biology, 74, 329-356.

Ward, R. D., Zemlak, T. S., Innes, B. H., Last, P. R., & Hebert, P. D. N. (2005). DNA barcoding Australia’s fish species. Philosophical Transactions Royal Society of Biological Sciences, 360, 1847-1857.

Wilson, P. J., Woods, C. M., Maina, J. N., & White, B. N. (2000). Genetic structure of Lake Magadi tilapia populations. Journal of Fish Biology, 56, 590-603.

Zhou, W., & Cui, G. H. (1997). Fishes of the genus Triplophysa in the Yuanjiang River (upper Red River) basin of Yunnan, China, with description of a new species. Ichthyological Exploration of Freshwaters, 8, 177-183.

Zhu, S. Q. (1989). The Loaches of the Subfamily Nemacheilinae in China (Cypriniformes: Cobitidae). Nanjing: Jiangsu Science and Technology Publishing House.


Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Copyright (c) 2016 Revista de Biología Tropical


Download data is not yet available.