Pressure and cold shock induction of meiotic gynogenesis and triploidy in the European sea bass, Dicentrarchus labrax L.: relative efficiency of methods and parental variability
Identifieur interne : 000C63 ( Istex/Corpus ); précédent : 000C62; suivant : 000C64Pressure and cold shock induction of meiotic gynogenesis and triploidy in the European sea bass, Dicentrarchus labrax L.: relative efficiency of methods and parental variability
Auteurs : Stefano Peruzzi ; Beatrice ChatainSource :
- Aquaculture [ 0044-8486 ] ; 2000.
Abstract
The optimal conditions for the retention of the second polar body in sea bass eggs were investigated by altering the timing, intensity and duration of application of pressure and cold shocks. Treatment optima for cold shocks were 0–1°C for 15–20 min at 5 min after fertilisation (a.f.) and 8500 psi for 2 min at 6 min a.f. for pressure shocks. Meiogenesis was obtained by fertilising eggs with UV-irradiated homologous sperm (32,000 erg mm−2) and pressure or cold shocking eggs as above. 100% triploidy was induced following definition of liable periods for the disruption of the meiotic spindle obtained in gynogenesis. Ploidy investigations were performed on experimental groups by flow-cytometry. Verification of uniparental transmission in meiogens was carried out by microsatellite marker loci analysis. This work highlights the degree of variation in individual responses of selected broodstock to these agents. Finally, some preliminary results on heterologous fertilisation in sea bass with potential applications for gynogenetic studies are also provided.
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DOI: 10.1016/S0044-8486(00)00355-0
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<front><div type="abstract" xml:lang="en">The optimal conditions for the retention of the second polar body in sea bass eggs were investigated by altering the timing, intensity and duration of application of pressure and cold shocks. Treatment optima for cold shocks were 0–1°C for 15–20 min at 5 min after fertilisation (a.f.) and 8500 psi for 2 min at 6 min a.f. for pressure shocks. Meiogenesis was obtained by fertilising eggs with UV-irradiated homologous sperm (32,000 erg mm−2) and pressure or cold shocking eggs as above. 100% triploidy was induced following definition of liable periods for the disruption of the meiotic spindle obtained in gynogenesis. Ploidy investigations were performed on experimental groups by flow-cytometry. Verification of uniparental transmission in meiogens was carried out by microsatellite marker loci analysis. This work highlights the degree of variation in individual responses of selected broodstock to these agents. Finally, some preliminary results on heterologous fertilisation in sea bass with potential applications for gynogenetic studies are also provided.</div>
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<notesStmt><note type="content">Fig. 1: Survival of D. labrax eggs fertilised with UV irradiated sperm. Means with a common superscript are not significantly different by χ2 test (P<0.05). Results observed at 4 h after fertilisation. (a) Mean and standard error of survival rates for the four tested couples. 2n: diploid status; n: haploid status. (b) Survival rate in each couple relative to control. At this stage, survival rates in control groups were of 90%, 83%, 98% and 90% for couples 1, 2, 3 and 4, respectively. Results observed at 72 h after fertilisation. (c) Mean and standard error of survival rate for the four tested couples. 2n: diploid status; n: haploid status. (d) Survival rate in each couple relative to control. At this stage, survival rates in control groups were of 56%, 70%, 69% and 53% for couples 1, 2, 3 and 4, respectively.</note>
<note type="content">Fig. 2: D. labrax haploid, diploid and triploid larvae and their respective nuclear DNA content measured by flow cytometry (M1, M2, M3). DNA values are reported in arbitrary units expressed as fluorescence channel numbers (FL2-Area). Scale bars represent 1 mm.</note>
<note type="content">Fig. 3: Survival of D. labrax eggs activated with homologous and heterologous sperm, from fertilisation to hatching. Spawning were issued from two different females (female 1: –; female 2: ---). • D. labrax sperm. ○ D. labrax irradiated sperm (total UV dose of 32.000 erg mm−2). ▴ Sparus aurata sperm.</note>
<note type="content">Fig. 4: Survival of D. labrax eggs fertilised with UV irradiated sperm (total UV dose of 32.000 erg mm−2). Eggs were submitted to a pressure shock (▩, 8000 psi for 2 min) or to a cold shock (□, 1°C for 20 min) applied at different moments after fertilisation. Results represent survival rates observed 72 h after fertilisation. Means with a common superscript are not significantly different by χ2 test (P<0.05). (a) Mean and standard error of survival rates for the four tested couples. 2n: diploid status; n:haploid status; ▨, diploid control groups. (b) Survival rate in each couple relative to control. At this stage, survival rates in control groups were of 69%, 66%, 97% and 70% for couples 1, 2, 3 and 4, respectively.</note>
<note type="content">Fig. 5: Observed Labrax 17 DNA banding pattern in control and gynogenetic progenies of D. labrax. Meiotic gynogens homozygous for one of the maternal alleles are indicated by arrows. F: female parent; M: male parent; bp: allele sizes given in base pairs.</note>
<note type="content">Fig. 6: Survival of D. labrax eggs fertilised with non-irradiated sperm. Eggs were submitted to pressure or cold shocks of different levels. Pressure shocks (▩) were applied 6 min after fertilisation for 2 min. Cold shocks of 1°C (□) were applied 5 min after fertilisation. Results represent the survival rates observed 72 h after fertilisation. Means with a common superscript are not significantly different by χ2 test (P<0.05). (a) Mean and standard error of survival rates for the three tested couples. 2n: diploid status; 3n: triploid status; ▨, diploid control groups. (b) Survival rates in each couple relative to control. At this stage, survival rates in control groups were of 76%, 95% and 88% for couples 1, 2, 3 and 4, respectively.</note>
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<ce:abstract-sec><ce:simple-para>The optimal conditions for the retention of the second polar body in sea bass eggs were investigated by altering the timing, intensity and duration of application of pressure and cold shocks. Treatment optima for cold shocks were 0–1°C for 15–20 min at 5 min after fertilisation (a.f.) and 8500 psi for 2 min at 6 min a.f. for pressure shocks. Meiogenesis was obtained by fertilising eggs with UV-irradiated homologous sperm (32,000 erg mm<ce:sup>−2</ce:sup>
) and pressure or cold shocking eggs as above. 100% triploidy was induced following definition of liable periods for the disruption of the meiotic spindle obtained in gynogenesis. Ploidy investigations were performed on experimental groups by flow-cytometry. Verification of uniparental transmission in meiogens was carried out by microsatellite marker loci analysis. This work highlights the degree of variation in individual responses of selected broodstock to these agents. Finally, some preliminary results on heterologous fertilisation in sea bass with potential applications for gynogenetic studies are also provided.</ce:simple-para>
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<ce:keyword><ce:text>Triploidy</ce:text>
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<ce:keyword><ce:text>Sea bass</ce:text>
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<ce:keyword><ce:text><ce:italic>Dicentrarchus labrax</ce:italic>
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<ce:keyword><ce:text>Pressure and cold shocks</ce:text>
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<abstract lang="en">The optimal conditions for the retention of the second polar body in sea bass eggs were investigated by altering the timing, intensity and duration of application of pressure and cold shocks. Treatment optima for cold shocks were 0–1°C for 15–20 min at 5 min after fertilisation (a.f.) and 8500 psi for 2 min at 6 min a.f. for pressure shocks. Meiogenesis was obtained by fertilising eggs with UV-irradiated homologous sperm (32,000 erg mm−2) and pressure or cold shocking eggs as above. 100% triploidy was induced following definition of liable periods for the disruption of the meiotic spindle obtained in gynogenesis. Ploidy investigations were performed on experimental groups by flow-cytometry. Verification of uniparental transmission in meiogens was carried out by microsatellite marker loci analysis. This work highlights the degree of variation in individual responses of selected broodstock to these agents. Finally, some preliminary results on heterologous fertilisation in sea bass with potential applications for gynogenetic studies are also provided.</abstract>
<note type="content">Fig. 1: Survival of D. labrax eggs fertilised with UV irradiated sperm. Means with a common superscript are not significantly different by χ2 test (P<0.05). Results observed at 4 h after fertilisation. (a) Mean and standard error of survival rates for the four tested couples. 2n: diploid status; n: haploid status. (b) Survival rate in each couple relative to control. At this stage, survival rates in control groups were of 90%, 83%, 98% and 90% for couples 1, 2, 3 and 4, respectively. Results observed at 72 h after fertilisation. (c) Mean and standard error of survival rate for the four tested couples. 2n: diploid status; n: haploid status. (d) Survival rate in each couple relative to control. At this stage, survival rates in control groups were of 56%, 70%, 69% and 53% for couples 1, 2, 3 and 4, respectively.</note>
<note type="content">Fig. 2: D. labrax haploid, diploid and triploid larvae and their respective nuclear DNA content measured by flow cytometry (M1, M2, M3). DNA values are reported in arbitrary units expressed as fluorescence channel numbers (FL2-Area). Scale bars represent 1 mm.</note>
<note type="content">Fig. 3: Survival of D. labrax eggs activated with homologous and heterologous sperm, from fertilisation to hatching. Spawning were issued from two different females (female 1: –; female 2: ---). • D. labrax sperm. ○ D. labrax irradiated sperm (total UV dose of 32.000 erg mm−2). ▴ Sparus aurata sperm.</note>
<note type="content">Fig. 4: Survival of D. labrax eggs fertilised with UV irradiated sperm (total UV dose of 32.000 erg mm−2). Eggs were submitted to a pressure shock (▩, 8000 psi for 2 min) or to a cold shock (□, 1°C for 20 min) applied at different moments after fertilisation. Results represent survival rates observed 72 h after fertilisation. Means with a common superscript are not significantly different by χ2 test (P<0.05). (a) Mean and standard error of survival rates for the four tested couples. 2n: diploid status; n:haploid status; ▨, diploid control groups. (b) Survival rate in each couple relative to control. At this stage, survival rates in control groups were of 69%, 66%, 97% and 70% for couples 1, 2, 3 and 4, respectively.</note>
<note type="content">Fig. 5: Observed Labrax 17 DNA banding pattern in control and gynogenetic progenies of D. labrax. Meiotic gynogens homozygous for one of the maternal alleles are indicated by arrows. F: female parent; M: male parent; bp: allele sizes given in base pairs.</note>
<note type="content">Fig. 6: Survival of D. labrax eggs fertilised with non-irradiated sperm. Eggs were submitted to pressure or cold shocks of different levels. Pressure shocks (▩) were applied 6 min after fertilisation for 2 min. Cold shocks of 1°C (□) were applied 5 min after fertilisation. Results represent the survival rates observed 72 h after fertilisation. Means with a common superscript are not significantly different by χ2 test (P<0.05). (a) Mean and standard error of survival rates for the three tested couples. 2n: diploid status; 3n: triploid status; ▨, diploid control groups. (b) Survival rates in each couple relative to control. At this stage, survival rates in control groups were of 76%, 95% and 88% for couples 1, 2, 3 and 4, respectively.</note>
<subject><genre>Keywords</genre>
<topic>Meiotic gynogenesis</topic>
<topic>Triploidy</topic>
<topic>Sea bass</topic>
<topic>Dicentrarchus labrax</topic>
<topic>Pressure and cold shocks</topic>
<topic>Parental variability</topic>
</subject>
<relatedItem type="host"><titleInfo><title>Aquaculture</title>
</titleInfo>
<titleInfo type="abbreviated"><title>AQUA</title>
</titleInfo>
<genre type="journal">journal</genre>
<originInfo><dateIssued encoding="w3cdtf">20000925</dateIssued>
</originInfo>
<identifier type="ISSN">0044-8486</identifier>
<identifier type="PII">S0044-8486(00)X0113-5</identifier>
<part><date>20000925</date>
<detail type="volume"><number>189</number>
<caption>vol.</caption>
</detail>
<detail type="issue"><number>1–2</number>
<caption>no.</caption>
</detail>
<extent unit="issue pages"><start>1</start>
<end>196</end>
</extent>
<extent unit="pages"><start>23</start>
<end>37</end>
</extent>
</part>
</relatedItem>
<identifier type="istex">137D45329C8625CEAF8D45F18F986669F4B64709</identifier>
<identifier type="DOI">10.1016/S0044-8486(00)00355-0</identifier>
<identifier type="PII">S0044-8486(00)00355-0</identifier>
<accessCondition type="use and reproduction" contentType="copyright">©2000 Elsevier Science B.V.</accessCondition>
<recordInfo><recordContentSource>ELSEVIER</recordContentSource>
<recordOrigin>Elsevier Science B.V., ©2000</recordOrigin>
</recordInfo>
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