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Production of bivalve tetraploid molluscs from diploid parents

A mollusk, tetraploid technology, applied in animal cells, climate change adaptation, animal husbandry, etc., can solve problems such as heavy weight, low competition of tetraploid larvae, and difficulty in genetic improvement.

Active Publication Date: 2010-03-24
法国海洋开发研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] 2) The need to go through triploids makes it particularly difficult and onerous to introduce at the tetraploid level any genetic improvement normally obtained at the diploid level
[0016] This will lead to low competition of tetraploid larvae, and therefore when tetraploid larvae are mixed with diploid and triploid larvae, they will disappear quickly

Method used

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  • Production of bivalve tetraploid molluscs from diploid parents
  • Production of bivalve tetraploid molluscs from diploid parents
  • Production of bivalve tetraploid molluscs from diploid parents

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Induction of polyploidy and reproduction of larvae

[0045] The diploid parents used for the experiment came from the natural wild-type diploid oyster spawning area in the French oyster Oléron Bay (Marennes Oléron Bay). Six female parents and four male parents were used for clustering and pairing through gonadal ablation. 15 million oocytes and 3 billion swimming sperm obtained from 10 parents were used for hybridization in 1 liter of filtered seawater at 25°C. The retention of the first polar body was affected by using mitostatin B (CB) dissolved in DMSO until the final concentration was 0.5 mg / L. Starting from the 10th minute after fertilization, CB was used for 15 minutes.

[0046] The protocol for inducing polyploidy is similar to the protocols for producing triploids that have been published in the literature (GUO et al., Biol. Bull., 183, 381-93, 1992; et al., Aquaculture, 174, 229-421999). However, the final concentration of CB was reduced (0.5 mg / L ins...

Embodiment 2

[0089] Example 2: Setting and growth of tetraploid oyster eggs

[0090] 1 month after placement

[0091] One month after placement, flow cytometry was used to check the presence of tetraploid oyster eggs in the successfully denatured and placed larvae population. Due to the small size of the oyster eggs at this stage, the operation was performed destructively on a sample containing some oyster eggs at some point. Put the oyster eggs into a 1.5 mL test tube. Then they were incubated in 1 mL of lysis buffer, and they were ground by using plastic stoppers. Finally, the released cell nuclei were filtered through a 30 μm mesh nylon filter (PartecCelltrics), and collected in a cell counting analysis test tube, labeled with DAPI and analyzed according to the method described in Example 1 above.

[0092] The cell count results allow us to confirm that in addition to the triploid and diploid populations, the population of tetraploid oyster eggs (29%) is also significantly present ( Figur...

Embodiment 3

[0115] Example 3: Production of tetraploid in mussels

[0116] By using the methods detailed in Examples 1 and 2, tetraploidies were effectively obtained in mussels (M. edulis and Mytilus galloprovincialis).

[0117] Among the mussels, the cell count results confirmed that in addition to the triploid and diploid populations, there is a predominant population of tetraploid oyster eggs. therefore,

[0118] -Control hybridization can provide an exclusive population of diploid nuclei ( Figure 37 , Peak "RN1");

[0119] -The induced samples maintained by GPI showed three types of nuclear populations ( Figure 38 ), corresponding to the following ploidy levels: diploid (about 10% on average, peak "RN1"), triploid (about 20% on average, peak "RN2") and tetraploid (about 70% on average, Peak "RN3"). The resulting tetraploid individual is in the process of reproduction.

[0120] All in all, by using the method detailed in the above examples, in the same manner as that used for Pacific oyste...

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Abstract

The invention relates to a method for producing viable tetraploid bivalve molluscs, by fertilisation of diploid female ovocytes with diploid male sperm, followed by the induction of the retention of the first polar body of the fertilised eggs, isolation from the obtained larvae of a larval sub-population enriched in tetraploids and selective raising of said sub-population.

Description

Technical field [0001] The invention relates to the production of tetraploid bivalve molluscs capable of feeding. Background technique [0002] The production of polyploid bivalve molluscs is currently of considerable interest, especially in the case of oysters. [0003] In fact, triploid oysters, also called "sea oysters", have several advantages. Because they have an odd number of chromosomes, they are usually non-fertile, which means that on the one hand they can grow faster (their energy is used for growth rather than for reproduction), on the other hand they do not produce semen during the summer , Which means they have constant quality throughout the year. [0004] By hybridization of the diploid parent, followed by chemical treatment (usually with cytostatin B (CB), or optionally 6-dimethyl-aminopurine (6-DMAP) treatment), or by physical treatment (in particular It is heat shock or pressure treatment) to maintain the first or second polar body to obtain triploid oysters. H...

Claims

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Application Information

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IPC IPC(8): A01K67/033
CPCA01K67/033A01K61/002A01K61/54Y02A40/81C12N5/06
Inventor 阿卜杜拉·贝纳德尔姆纳克里斯托夫·勒迪
Owner 法国海洋开发研究院
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