Production of bivalve tetraploid molluscs from diploid parents

A mollusk and tetraploid technology, applied in animal cells, climate change adaptation, animal husbandry, etc., can solve the problems of difficult genetic improvement, low competitiveness of tetraploid larvae, and heavy load

Active Publication Date: 2012-11-21
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 experiments were from the natural wild-type diploid oyster egg collection area in the French oyster Oléron Bay (Marennes Oléron Bay). Six female parents and four male parents were used for aggregate pairing by gonad scarification method. Fifteen million oocytes and 3 billion motile sperm from 10 parents were used for hybridization in 1 liter of filtered seawater at 25°C. The maintenance of the first polar body was affected by using cytokinostatin B (CB) dissolved in DMSO to a final concentration of 0.5 mg / L. From the 10th minute after fertilization, CB was used for 15 minutes.

[0046] This protocol for inducing polyploidy is similar to those already disclosed in the literature for the production of triploids (GUO et al., Biol. Bull., 183, 381-93, 1992; et al., Aquaculture, 174, 229-421999). However, the final concentration of CB was lowered (0.5 mg / L ...

Embodiment 2

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

[0090] 1 month after settling

[0091] One month after establishment, checks for the presence of tetraploid oyster eggs were performed by flow cytometry within successfully denatured and established larval populations. Due to the small size of oyster roe at this stage, this operation was sometimes performed destructively on samples containing some oyster roe. Put the oyster roe into a 1.5mL test tube. They were then incubated in 1 mL of lysis buffer, and they were triturated by using a plastic stopper. Finally, the released nuclei were filtered through a 30 μm mesh nylon filter (Partec Celltrics) and collected in cytometry assay tubes, labeled with DAPI and analyzed as described in Example 1 above.

[0092] Cell count results allowed us to confirm that, in addition to triploid and diploid populations, tetraploid oyster egg (29%) populations were also significantly present ( Figure 30 Mi...

Embodiment 3

[0115] Example 3: Production of tetraploids in mussels

[0116] By using the methods detailed in Examples 1 and 2, tetraploidy was efficiently obtained in mussels (M. edulis and Mytilus galloprovincialis).

[0117] In mussels, cell count results confirmed the presence of a predominant tetraploid oyster egg population in addition to the triploid and diploid populations. therefore,

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

[0119] - Induced samples maintained by GPI showed three types of nuclei populations ( Figure 38 ), corresponding to the following ploidy levels: diploid (average around 10%, peak "RN1"), triploid (average around 20%, peak "RN2") and tetraploid (average around 70%, peak "RN2") peak "RN3"). The resulting tetraploid individuals are in the process of breeding.

[0120] In summary, by using the methods detailed in the examples above, viable tetraploids can be produ...

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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 present invention relates to the production of viable tetraploid bivalve molluscs. Background technique [0002] The production of polyploid bivalve molluscs is currently of considerable interest, especially with respect to oysters. [0003] In fact, triploid oysters, also known as "season oysters," have several advantages. Because they have an odd number of chromosomes, they are usually sterile, which means that on the one hand they can grow faster (their energy is used to grow rather than reproduce) and on the other hand they do not produce semen during the summer , which means they are of constant quality throughout the year. [0004] By hybridization of diploid parents, followed by chemical treatment, usually with cytokinin B (CB), or optionally with 6-dimethyl-aminopurine (6-DMAP), or by physical treatment (especially (heat shock or pressure treatment) to maintain the first or second polar body, triploid oysters can be obtained. However, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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