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Pronounced genetic differentiation in the populations of cultivable red – tailed barb Gonoproktopterus curmuca (Hamilton): Family Cyprinidae, as revealed by allozymes, microsatellites and RAPD. Musammilu, K. K., Gopalakrishnan, A*., V.S. Basheer, K.K. Lal1, V. Mohindra1, P. Punia1, Joselet Mathew2, A. G. Ponniah3 and W.S. Lakra1. National Bureau of Fish Genetic Resources (NBFGR) Cochin Unit, CMFRI Campus, Kochi – 682 018, Kerala, India; nbfgrcochin@eth.net; 1NBFGR, Canal Ring Road, Lucknow - 226 002, Uttar Pradesh, India; 2 Department of Zoology, Nirmalagiri College, Koothuparamba, Kannur 670 701, Kerala, India; 3 Central Institute of Brackishwater Aquaculture, Chennai 600 028, Tamil Nadu, India.

* Corresponding author <agopalkochi@gmail.com> (Oral Presentation No. OP014) Gonoproktopterus curmuca, a cultivable food and ornamental fish is found in the rivers originating from the biodiversity hotspot - the Western Ghats, India. It is one of the prioritized species for the region-specific aquaculture in India. G. curmuca has drastically declined in the rivers due to over-exploitation and habitat alteration. Captive breeding and milt cryopreservation of the species have been developed by the National Bureau of Fish Genetic Resources. Documentation of genetic variation at intra-specific level through genetically controlled markers is of vital importance for evolving fisheries management and aquaculture strategies; and in planning propagation assisted rehabilitation programmes. To explore the genetic variation in natural populations of this species in Periyar, Chalakkudy and Chaliyar Rivers (Kerala, India) allozymes, microsatellites and RAPD markers were employed. In allozyme analysis, 12 polymorphic enzymes yielded 29 loci with a total of 31 alleles. Only one private allele was recorded - in Chaliyar with LDH locus. For microsatellite analysis, eight single locus primers from 6 cyprinids were cross-primed in G. curmuca producing 8 presumptive microsatellite loci and a total of 53 alleles. All the 8 loci contained repeat regions (NCBI #s DQ780015, DQ780014, EF582608, EF582609, EF582610, EF582611, EF582612 and EF582613) and were polymorphic (100%). Analysis of microsatellite data using MICROCHECKER ruled out occurrence of null alleles. There were 4 private alleles in Periyar, 2 in Chalakkudy and 3 in Chaliyar. The positive FIS and lesser observed heterozygosity values in many allozymes and microsatellites loci indicated deficiency of heterozygotes and deviation from Hardy-Weinberg equilibrium. The pair wise FST and RST; and AMOVA between riverine locations indicated significant genetic heterogeneity (P<0.001) between populations. The allozyme and microsatellite loci indicated significant genetic bottleneck in all the populations of G. curmuca under the Infinite Allele Model and Two Phased Model respectively (Wilcoxon Test P<0.05). In RAPD analysis, 9 OPERON oligonucleotide decamers produced 117 amplicons in three populations. Forty one RAPD fragments were identified as stock specific markers. The coefficient of genetic differentiation (GST) (P<0.05) and Nei’s (1978) unbiased genetic distance among populations indicated significant genetic differentiation between populations. In conclusion, the genetic markers demonstrated striking genetic differentiation between G. curmuca populations. Geographic isolation is likely to be the factor for the restricted gene flow between the river systems. The inbreeding as a result of over-exploitation might be the reason for the deficiency of heterozygosity and genetic bottleneck. For propagation assisted-rehabilitation, large effective breeding population of the species may be developed with out mixing individuals from the rivers. (Abstract submitted online on 30 April 2009).

(pp. 35-36 of ISGA X Book of Abstracts 270p.)

Ref. No. OP014

18 May 2009

Biology and Biotechnology Research in India

India

Dear Dr.Gopalakrishnan A,

I would like to inform you that your abstract submission entitled:

“Pronounced genetic differentiation in the populations of cultivable red – tailed barb

Gonoproktopterus curmuca (Hamilton) : family Cyprinidae, as revealed by allozymes, microsatellites

and RAPD”

has been accepted for Oral Presentation at the 10th International Symposium on Genetics in Aquaculture

(ISGA2009) to be held on 22-26 June 2009, at the Bangkok Convention Centre and Sofitel Centara Grand

Hotel, Bangkok, THAILAND.

Your presentation details are as following:

Date: Wednesday, 24 June 2009

Time: 11:15 - 11:30

Location: Auditorium A, 5th

floor, Bangkok Convention Centre

Please note that all presenters are required to submit and test their presentation file at the Conference-

Technical Room at least 3 hours before the time of presentation. Details regarding instructions for

presentation will be announced on www.isga2009.org.

We are looking forward to meeting you at the ISGA 2009.

Presentation 2 - Poster PP140; Lakra Wazir S & Gopalakrishnan, A. Marine Molecular Biology and Biotechnology Research in India. (pp. 243 - 244 in the Abstract Book).

Yours sincerely,

Professor Dr.Uthairat Na-Nakorn

Chairperson, Organizing Committee

Secretariat Office Department of Aquaculture,

Faculty of Fisheries, Kasetsart University

Bangkok 10900, Thailand

Phone/Fax: +66-2-561-0990

Email: info@isga2009.org