The project results included in a book (in Romanian)

Click here:

http://www.editura.bioflux.com.ro/docs/ceex140electro.pdf

01.11.2008. 16:30

Details regarding CEEX 140/2006

Research Program: Research of Excelence

Financed by: National Authority for Scientific Research

Monitor: Centrul National de Management Programe - Biotech

Contractor: University of Agricultural Sciences ad Veterinary Medicine Cluj-Napoca

Research Manager: Petrescu-Mag Ioan Valentin

Title of the project: Development of RAPD markers associated to low water-temperature resistance in Poecilia reticulata Peters, 1859

Number of the project: 140/2006

Acronym: RAPDTEMP

Parteners:

UAIC Iasi

USAMV B Timisoara

USAMV Iasi

Period of the contract: 15 September 2006 – 15 September 2008

Initial team of the project:

USAMV Cluj-Napoca

Petrescu-Mag Ioan Valentin; Manager

Bud Ioan; Researcher

Pamfil Doru; Researcher

Grigoras Mircea Adrian; Asistant Manager

Vlaic Augustin; Researcher

Petrescu R. Malina; Researcher

Cosier Viorica; Researcher

Pop Rodica; Researcher

Georgescu Bogdan; Researcher

Ladoşi Daniela; Researcher

Boaru Anca; Researcher

Bâlteanu Valentin; Researcher

Raica Paul; Researcher

Dombi Istvan; Researcher

Vladau Vlad Viorel; Researcher

Pop Sorin; Researcher

Reka Stefan; Researcher

Petrescu Crina; Researcher

Botha Miklos; Researcher

Coroian Cristian; Researcher

Husovschi Adriana Maria; Economist

Florea Mariana; Economist

Molnar Florin; Student

Sabados Lavinia; Student

Hărşan Radu; Student

Maxim Elisabeta; Tehnician

UAIC Iasi

Gorgan Lucian; Group Leader

Cîmpeanu Mirela Mihaela; Researcher

Gherasim Raluca Sorina; Student

Buburuzan Laura; Student

USAMV B Timisoara

Bura Marian; Group Leader

Grozea Adrian; Researcher

Dumitrescu Gabi; Researcher

Patruica Silvia; Researcher

Vintilă Teodor; Researcher

Bănăţean-Dunea Ioan; Researcher

Muscalu-Nagy Radu; Researcher

Muscalu-Nagy Cristina; Researcher

Cozma Liliana; Tehnician

Korbuly Bogdan; Tehnician

Chiriţoiu Adrian; Tehnician

Ciobotaru Emilia; Economist

Popescu Ioan; Database Administrator

USAMV Iasi

Păsărin Benone; Group Leader

Stan Traian; Researcher

Gâlcă Ioan; Researcher

Creangă Steofil; Researcher

Usturoi Marius; Researcher

Confederat Margareta; Researcher

Marciuc Vasile; Researcher

Macovei Valerica; Researcher

Hoha Gabriel; Researcher

Diac Alexandrina; Researcher

Gruber Adrian; Researcher

Stănescu Roxana; Researcher

Radu Rusu Răzvan; Researcher

01.11.2008. 16:03

CEEX

There are, in aquaria, coldwater fish species and tropical fishes, the guppy being one of the latter (Bud 2002). However, the guppy has the ability to survive in cooler waters that would certainly kill most tropical fishes (Mag & Bud 2006; Mag et al 2006; Păsărin et al 2007). In their paper “Detection of a low temperature-resistant gene in guppy (Poecilia reticulata), with reference to sex-linked inheritance”, Fujio et al (1990) reported an X-linked gene, responsible of low water-temperature resistance in guppy. Sixteen years later, in the paper: “Effect of temperature on sex ratio in guppy Poecilia reticulata (Peters 1860)”, Karayϋcel et al (2006) emitted the hypothesis according to which the same gene could be involved in high water-temperature resistance of guppy. In that moment, the above mentioned gene became a very important one, especially for guppyculture and generally for aquaculture, because of extrapolation possibility of these studies to many cases of economical and commercial important species.

In a CEEX-Biotech project (Research of Excelence Program, no 140/2006) our team tested a number of 150 random primers (Biosearch Technologies Inc.) in order to identify RAPD markers which could be associated to low temperature resistance in guppy. However, 71 random primers have not amplified DNA at all, 35 primers have not generated DNA polymorphisms, and 44 of them generated (more or less) polymorphisms. Only one random primer (no 77) was associated with cold tolerance (5'd(CCAACGACCA)3'; see Figure 1) in the Red Blond variety.

Figure 1. http://www.bioflux.com.ro/docs/vol1/2008.1.99-110.pdf  Polymorphism, induced by the primer 77, associated with cold tolerance in the Red Blond guppy. The supplementary DNA fragment of 878 bp is associated to coldwater resistance (11-20), and the supplementary fragment of 839 bp is associated to coldwater sensitivity (1-10).

UPGMA (Unweighted Pair Group Method with Arithmetic Mean) was used to illustrate the two distinct fish clades: coldwater resistant group and coldwater sensitive group (see Figure 2). However, using the same primer (77), our repeated RAPD analyses failed to discriminate between coldwater resistant and sensitive individuals in other guppy strains. The primer was useless even for identification of coldwater resistant fishes from an F_B generation whose father and both grandfathers were Red Blond males.

After the identification of coldwater resistant individuals, the next step forward of the research in the project CEEX 140/2006 was obtaining of a homozygote guppy strain for coldwater resistance gene, and this was possible due to the X-linkage of Nigrocaudatus II (Ni) which was used as color marker gene. The breeding program of the research was based on a ♂Red Blond X ♀Half-Black Black cross, followed by a backcross of F₁ females with the initial Red Blond male.

One hundred of individuals, homozygote for the coldwater resistance gene, were further tested for resistance to heat. The results indicated a significant higher tolerance to heat of these ones compared to control lot. These data supported the hypothesis of Karayϋcel et al (2006) and indicated the common identity of coldwater resistance gene and heat tollerance gene. This X-linked gene (for resistance) seems to encode a protein with multiple role, a molecular chaperone, a protein formally included in HSPs group. We concluded at least some of the HSPs were involved in both coldwater resistance and heat tollerance, modulating animal metabolism under different environmental conditions. 

Figure 2. http://www.bioflux.com.ro/docs/vol1/2008.1.99-110.pdf  The two phenotypic distinct groups illustrated by UPGMA (when genomic DNA was amplified with the primer no 77).   

References

Bud I., 2002 [Aquariophilia]. Academicpres, Cluj-Napoca, Romania. [in Romanian]

Fujio Y., Nakajima M., Nagahama Y., 1990 Detection of a low temperature-resistant gene in the guppy (Poecilia reticulata), with reference to sex-linked inheritance. The Japanese Journal of Genetics 65(4):201-207.

Karayücel I., Ak O., Karayücel S., 2006 Effect of temperature on sex ratio in guppy Poecilia reticulata (Peters 1860) Aquaculture Research 37:139–150.

Mag I. V., Bud I., 2006 Environmental factors’ impact upon sex-ratio in guppies (Poecilia reticulata Peters, 1859). In: 41st Croatian and 1st International Symposium on Agriculture - Opatija, pp. 523-524, Osijek, Croatia. 

Mag I. V., Bud I., Petrescu R. M., 2006 Sex-dependent mortality in the guppyfish. Acta Ichtiologica Romanica 1:153-164.

Păsărin B., Gorgan L., Bura M., Botha M., Petrescu-Mag I. V., 2007 The invasive potential of the exotic guppyfish (Poecilia reticulata Peters 1859) in temperate zone. Studia Universitatis Babes-Bolyai, Ambientum 1-2:179-187.

31.10.2008. 09:25