CN101361469B - Common intertidal shellfish polymerization hybridization breeding method - Google Patents

Common intertidal shellfish polymerization hybridization breeding method Download PDF

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CN101361469B
CN101361469B CN200810013427XA CN200810013427A CN101361469B CN 101361469 B CN101361469 B CN 101361469B CN 200810013427X A CN200810013427X A CN 200810013427XA CN 200810013427 A CN200810013427 A CN 200810013427A CN 101361469 B CN101361469 B CN 101361469B
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闫喜武
张跃环
霍忠明
杨凤
张国范
王�琦
金晶宇
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Dalian Fisheries University
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Abstract

The invention discloses a breeding method of common mudflat shellfish by convergent cross. The method comprises the steps: mudflat shellfish parents from more than four different geographic populations are collected; the shellfish parents of different geographic populations are carried out eco-maturing; then dual reciprocal cross is carried between two geographic populations; a first filial generation of dual reciprocal cross combination is bred up by a three-stage method; then the convergent cross of the first filial generation of dual reciprocal cross is carried out so as to obtain a first filial generation of convergent cross; and the above steps are repeated on the basis thereof till the filial generation of convergent cross accumulates the good genes of all the geographic populations. The method overcomes the geographic isolation and the biological differences of sexual maturity and breeding among different geographic populations. Compared with the existing mudflat shellfish varieties, the shellfish integrates the good genes from more than four different geographic populations; the heterozygosity is improved greatly, thus causing the converged half-bred individual to present characteristics of fast growing and high stress resistance.

Description

The breeding method of common intertidal shellfish polymerization hybridization
Technical field
The present invention relates to a kind of shellfish genetic breeding technology, specifically a kind of breeding method of common intertidal shellfish polymerization hybridization.
Background technology
Common intertidal shellfish is the wider breed variety that distributes mostly, has culture-cycle weak point, small investment, characteristics that income is big.At present, there are bad phenomenon such as the germ plasm resource basal status is unclear, quality decline, poor, the individual miniaturization of resistance, specific yield reduction in intertidal shellfish in breed.And culture object and be wild type, almost there are not the high yield of artificial culture, degeneration-resistant improved seeds.Intertidal shellfish is carried out genetic improvement, cultivate fast growth, the new varieties of strong stress resistance are the important channels (Yan Xiwu, 2005) that addresses these problems.
Selecting breeding is the effective ways of rearing new variety, and hybridization then is the important method of cultivating new varieties, in vegeto-animal breeding, has obtained success widely.Convergent cross is meant through the excellent genes of a series of hybridization with several parental breeds and condenses together, and is divided into the combined hybrid of maximum reorganization principle and the convergent cross of ultra close reorganization principle.At present, convergent cross is mainly used in the plant breeding.
Application publication number is the breeding method that the Chinese patent of CN101103708 discloses the Patinopecten yessoensis three way cross, and the Patinopecten yessoensis of 3 geographical populations that this invention is adopted is hybridized, and it is individual to have obtained three way cross.Yet it is lower that the filial generation of adopting this method to carry out the shellfish breeding has a heterozygosity, the defective that the hybridization cycle of use is long.
Summary of the invention
The present invention is intended to the convergent cross through different geographical population intertidal shellfishs maximum reorganization principles, in the short hybridization cycle, obtains having the hybridization individuality of height heterozygosity, eliminates lethal gene, makes that excellent genes obtains expressing.It is poor finally to reach the current intertidal shellfish germplasm resistance of solution, and growth rate waits problem slowly.
Technical scheme of the present invention is: gather the intertidal shellfish parent in >=4 different geographical populations; Different geographical population shellfish parents are carried out ecological accelerating: carry out the positive and negative colony hybridization of the binary between geographical population in twos; Three-stage process is formed the first filial generation of the positive and negative hybrid combination of binary; The convergent cross of two-way cross first filial generation obtains the convergent cross first filial generation; On this basis, repeat above step, till the polymerization filial generation has added up the excellent genes of all geographical populations.
Technical process of the present invention may further comprise the steps:
Shellfish in the first step, the collection >=4 different geographical population is as the parent;
Second step, the shellfish parent of different geographical populations is carried out ecological accelerating;
The 3rd step, the positive and negative hybridization of the binary of colony in twos: after treating parent's maturation, the parent is carried out sex identification, after the discriminating; Through dry in the shade, stimulation by running water, the parent can synchronous spawning, row is smart; Hybridize according to group combination in twos then, obtain the first filial generation of several reciprocal crosss combinations;
The 4th step, three-stage process are formed the positive and negative hybridization of binary first filial generation: carry out forming of reciprocal cross first filial generation according to three-stage process, sexual gland is ripe synchronously until the hybridization first filial generation;
The convergent cross of the 5th step, two-way cross first filial generation: with third and fourth step, carry out the convergent cross of two-way cross first filial generation, obtain the convergent cross first filial generation; If the geographical population quantity of shellfish>4, need the above step of repetition, carry out secondary or convergent cross repeatedly, till the polymerization filial generation has added up the excellent genes of all geographical populations.
The described step that different proles shellfish parents are carried out ecological accelerating can adopt following method: the parent who gathers is positioned in ecological shrimp pond or the natural waters; Adopt the method for bottom sowing culture to carry out ecological accelerating, ripe synchronously until different proles shellfish parent sexual glands.
Said parent identifies and undertaken by following mode, can obtain higher resolution: the step that said parent identifies is the electric drill of 0.6mm for adopting diameter, according to the kind that will identify shellfish; Confirm the sexual gland position of shellfish parent, boring, is sterilized microsyringe to the sexual gland position until deeply on the shell of shellfish parent; Take out essence or ovum with the microsyringe of sterilizing; Adopt the water droplet method at microscopically, add a little seawater, carry out sex identification; Ovum is dispersed in seawater, is circle or pyriform at microscopically; Sperm is smoke-like in seawater, and microscopically is the tadpole shape.
During the following standard acquisition of said different geographical population shellfish, breeding effect is better: (1) 2 age, healthy harmless individuality was as the parent; (2) require its parent that enough big geographical difference and hereditary difference are arranged.
Described three-stage process is formed the positive and negative hybridization of binary first filial generation and is specially the parent that the positive and negative hybridization of binary first filial generation is gathered and is positioned in ecological shrimp pond or the natural waters, adopts the method for bottom sowing culture to carry out ecological accelerating.
Three-stage process promptly at the indoor shellfish seed breeding that carries out, is bred the juvenile mollusk (sea) in the shrimp pond that obtains and is carried out the centre and breed, and the centre is bred the young shellfish that obtains and formed at shrimp pond (sea).
The present invention adopts ecological accelerating method to the parent, has overcome the differences of Physiological of geographic isolation, sexual maturity and breeding between different geographical populations, impels sexual gland to grow smart ovum synchronously and discharges synchronously.Compare with existing intertidal shellfish kind of culturing, polymerization >=excellent genes of 4 geographical populations, its heterozygosity improves greatly, and has shortened the hybridization cycle of shellfish to the full extent.Make the convergent cross individuality show that growth is fast, the characteristics of strong stress resistance.What this method adopted is the colony hybridization mode, has avoided the single relatively drawback of monomer hybrid gene type, has enriched the individual genomic constitution of hybridization.Owing to be repeatedly positive and negative hybridization, can utilize parent's effect more fully, the most effective hybrid combination that therefrom obtains; And the growth of hybridization individuality, survival show excess of export parent effect much larger than the parent, the excellent genes of each geographical population shellfish because the convergent cross individuality has added up, and its growth rate improves 30-60%, survival rate raising 80-120%.This has established solid foundation for the exploitation of shellfish hybrid new breed.
Embodiment
Be embodiment with the Ruditapes philippinarum below, further specify the present invention, but be not construed as limiting the invention.
Embodiment 1
The first step, gather different geographical population Ruditapes philippinarum parents: in 2005, select 2 ages healthy harmless individualities as the parent, its parent comes from southern china Putian, Chinese northern Dalian, Busan, Korea, Tokyo respectively.
Second step, the young parent's of different geographical population clam ecological accelerating: the parent who gathers is positioned in the shrimp pond in Zhuanghe ocean village and carries out ecological accelerating, through 6-12 month bottom sowing culture, in autumn in 2006 sexual gland ripe synchronously.
The positive and negative hybridization of binary of the 3rd step, China's the South and the North, colony of Korean and Japanese: the employing diameter is that the electric drill of 0.6mm carries out parent's sex identification, after the discriminating; Through the stimulation of the 8h that dries in the shade, flowing water 1h, the parent can arrange smart by synchronous spawning; Carry out the positive and negative hybridization between colony according to the southern china ♀ * China north Japanese ♀ * Korea S of Chinese northern ♀ * southern china
Figure G200810013427XD00032
Korea S ♀ * Japan
Figure G200810013427XD00033
Figure G200810013427XD00034
then; Each hybrid combination obtains Chinese southern and northern reciprocal cross first filial generation and Korean and Japanese reciprocal cross first filial generation with 200 of parents.
The 4th step, three-stage process forms the positive and negative hybridization of binary first filial generation: in ecological shrimp pond, carry out forming of northern reciprocal cross first filial generation and Korean and Japanese reciprocal cross first filial generation according to three-stage process, form cycle >=1 year.Autumn in the 2007th, hybridization first filial generation sexual gland was ripe synchronously.
The convergent cross of the 5th step, two-way cross first filial generation: with the 3rd step; Carry out the hybridization between the Chinese southern and northern ♀ * Japan and Korea S of Chinese southern and northern ♀ * Korean and Japanese
Figure G200810013427XD00035
Figure G200810013427XD00036
the northern southern ♀ * Korean and Japaneses of China
Figure G200810013427XD00037
China northern southern ♀ * Japan and Korea S
Figure G200810013427XD00038
colony, obtain the Ruditapes philippinarum of the convergent cross of Chinese southern and northern Japan and Korea S, Chinese southern and northern Japan and Korea S, the northern southern Korean and Japanese of China, the northern southern Japan and Korea S of China.
Through the Ruditapes philippinarum that convergent cross obtains, its growth rate has improved 40-60%, survival rate has improved 90-120%.
Embodiment 2
The first step, gather different geographical population Ruditapes philippinarum parents: in 2005, select 2 ages healthy harmless individualities as the parent, its parent comes from southern china Putian, Chinese northern Dalian, Busan, Korea, Tokyo respectively.
Second step, the young parent's of different geographical population clam ecological accelerating: the parent who gathers is positioned in the beach in Zhuanghe ocean village and carries out ecological accelerating, through 6-12 month bottom sowing culture, in autumn in 2006 sexual gland ripe synchronously.
The positive and negative hybridization of binary of the 3rd step, China's the South and the North, colony of Korean and Japanese: the employing diameter is that the electric drill of 0.6mm carries out parent's sex identification, after the discriminating; Through the stimulation of the 8h that dries in the shade, flowing water 1h, the parent can arrange smart by synchronous spawning; Carry out the positive and negative hybridization between colony according to the southern china ♀ * China north Japanese ♀ * Korea S of Chinese northern ♀ * southern china
Figure G200810013427XD00042
Korea S ♀ * Japan
Figure G200810013427XD00043
Figure G200810013427XD00044
then; Each hybrid combination obtains Chinese southern and northern reciprocal cross first filial generation and Korean and Japanese reciprocal cross first filial generation with 240 of parents.
The 4th step, three-stage process forms the positive and negative hybridization of binary first filial generation: in ecological shrimp pond, carry out forming of northern reciprocal cross first filial generation and Korean and Japanese reciprocal cross first filial generation according to three-stage process, form cycle >=1 year.Autumn in the 2007th, hybridization first filial generation sexual gland was ripe synchronously.
The convergent cross of the 5th step, two-way cross first filial generation: with the 3rd step; Carry out the hybridization between the Chinese southern and northern ♀ * Japan and Korea S of Chinese southern and northern ♀ * Korean and Japanese
Figure G200810013427XD00045
the northern southern ♀ * Korean and Japaneses of China
Figure G200810013427XD00047
China northern southern ♀ * Japan and Korea S
Figure G200810013427XD00048
colony, obtain the Ruditapes philippinarum of the convergent cross of Chinese southern and northern Japan and Korea S, Chinese southern and northern Japan and Korea S, the northern southern Korean and Japanese of China, the northern southern Japan and Korea S of China.
Through the Ruditapes philippinarum that convergent cross obtains, its growth rate has improved 30-40%, survival rate has improved 80-90%.
Embodiment 3
The first step, gather different geographical population Ruditapes philippinarum parents: in 2005, select 2 ages healthy harmless individualities as the parent, its parent comes from southern china Putian, Chinese northern Dalian, Busan, Korea, Tokyo respectively.
Second step, the young parent's of different geographical population clam ecological accelerating: the parent who gathers is positioned in the shrimp pond in Zhuanghe ocean village and carries out ecological accelerating, through 6-12 month bottom sowing culture, in autumn in 2006 sexual gland ripe synchronously.
The positive and negative hybridization of binary of the 3rd step, China's the South and the North, colony of Korean and Japanese: through the stimulation of the 8h that dries in the shade, flowing water 1h, the parent can arrange smart by synchronous spawning; In the smart process of the row of laying eggs, can tell male and female, the male and female individuality is crawled out respectively; Carry out the positive and negative hybridization between colony according to the southern china ♀ * China north Japanese ♀ * Korea S of
Figure G200810013427XD00051
Chinese northern ♀ * southern china Korea S ♀ * Japan
Figure G200810013427XD00053
Figure G200810013427XD00054
then; Each hybrid combination obtains Chinese southern and northern reciprocal cross first filial generation and Korean and Japanese reciprocal cross first filial generation with 60 of parents.
The 4th step, three-stage process forms the positive and negative hybridization of binary first filial generation: in ecological shrimp pond, carry out forming of northern reciprocal cross first filial generation and Korean and Japanese reciprocal cross first filial generation according to three-stage process, form cycle >=1 year.Autumn in the 2007th, hybridization first filial generation sexual gland was ripe synchronously.
The convergent cross of the 5th step, two-way cross first filial generation: with the 3rd step; Carry out Chinese southern and northern ♀ * Korean and Japanese
Figure G200810013427XD0005150604QIETU
; China southern and northern ♀ * Japan and Korea S
Figure 200810013427X100002G200810013427XD0005150604QIETU
; China northern southern ♀ * Korean and Japanese
Figure 200810013427X100002G200810013427XD0005150604QIETU
; Hybridization between China northern southern ♀ * Japan and Korea S
Figure 200810013427X100002G200810013427XD0005150604QIETU
colony obtains the Ruditapes philippinarum of the convergent cross of Chinese southern and northern Japan and Korea S, Chinese southern and northern Japan and Korea S, the northern southern Korean and Japanese of China, the northern southern Japan and Korea S of China.
Through the Ruditapes philippinarum that convergent cross obtains, its growth rate has improved 30-50%, survival rate has improved 80-100%.
Embodiment 4
The first step, gather different geographical population Ruditapes philippinarum parents: in 2005, select 2 ages healthy harmless individualities as the parent, its parent comes from southern china Putian, Chinese northern Dalian, Busan, Korea, Tokyo respectively.
Second step, the young parent's of different geographical population clam ecological accelerating: the parent who gathers is positioned in the shrimp pond in Zhuanghe ocean village and carries out ecological accelerating, through 6-12 month bottom sowing culture, in autumn in 2006 sexual gland ripe synchronously.
The positive and negative hybridization of binary of the 3rd step, China's the South and the North, colony of Korean and Japanese: the employing diameter is that the electric drill of 0.6mm carries out parent's sex identification, after the discriminating; Through the stimulation of the 8h that dries in the shade, flowing water 1h, the parent can arrange smart by synchronous spawning; Carry out the positive and negative hybridization between colony according to the southern china ♀ * China north Japanese ♀ * Korea S of
Figure G200810013427XD00055
Chinese northern ♀ * southern china
Figure G200810013427XD00056
Korea S ♀ * Japan
Figure G200810013427XD00057
then; Each hybrid combination obtains Chinese southern and northern reciprocal cross first filial generation and Korean and Japanese reciprocal cross first filial generation with 300 of parents.
The 4th step, three-stage process forms the positive and negative hybridization of binary first filial generation: in the beach of shellfish culture field, Zhuanghe ocean village, carry out forming of northern reciprocal cross first filial generation and Korean and Japanese reciprocal cross first filial generation according to three-stage process, form cycle >=1 year.Autumn in the 2007th, hybridization first filial generation sexual gland was ripe synchronously.
The convergent cross of the 5th step, two-way cross first filial generation: with the 3rd step; Carry out the hybridization between the Chinese southern and northern ♀ * Japan and Korea S of Chinese southern and northern ♀ * Korean and Japanese
Figure G200810013427XD00061
the northern southern ♀ * Korean and Japaneses of China
Figure G200810013427XD00063
China northern southern ♀ * Japan and Korea S
Figure G200810013427XD00064
colony, obtain the Ruditapes philippinarum of the convergent cross of Chinese southern and northern Japan and Korea S, Chinese southern and northern Japan and Korea S, the northern southern Korean and Japanese of China, the northern southern Japan and Korea S of China.
Through the Ruditapes philippinarum that convergent cross obtains, its growth rate has improved 40-50%, survival rate has improved 80-110%.

Claims (3)

1. the breeding method of a common intertidal shellfish polymerization hybridization may further comprise the steps:
Shellfish in the first step, the collection >=4 different geographical population is as the parent;
Second step, the shellfish parent of different geographical populations is carried out ecological accelerating;
The 3rd step, the positive and negative hybridization of the binary of colony in twos: after treating parent's maturation, the parent is carried out sex identification, after the evaluation; Through dry in the shade, stimulation by running water, the parent can synchronous spawning, row is smart; Hybridize according to group combination in twos then, obtain the first filial generation of several reciprocal crosss combinations;
The 4th step, three-stage process are formed the positive and negative hybridization of binary first filial generation: carry out forming of reciprocal cross first filial generation according to three-stage process, sexual gland is ripe synchronously until the hybridization first filial generation;
The convergent cross of the 5th step, two-way cross first filial generation: with third and fourth step, carry out the convergent cross of two-way cross first filial generation, obtain the convergent cross first filial generation; If geographical population quantity＞4 of shellfish need the above step of repetition, carry out secondary or convergent cross repeatedly, till the polymerization filial generation has added up the excellent genes of all geographical populations;
The step that described shellfish parent to different geographical populations carries out ecological accelerating is specially the parent who gathers is positioned in ecological shrimp pond or the natural waters; Adopt the method for bottom sowing culture to carry out ecological accelerating, ripe synchronously until different geographical population shellfish parent sexual glands;
Described three-stage process is formed the positive and negative hybridization of binary first filial generation and is specially the parent that the positive and negative hybridization of binary first filial generation is gathered and is positioned in ecological shrimp pond or the natural waters, adopts the method for bottom sowing culture to carry out ecological accelerating;
Three-stage process, promptly at the indoor shellfish seed breeding that carries out, breed the juvenile mollusk that obtains in the shrimp pond or the sea carry out the centre and breed, the centre is bred the young shellfish that obtains and is formed in shrimp pond or sea.
2. the breeding method of common intertidal shellfish polymerization hybridization as claimed in claim 1 is characterized in that: the step of said parent's sex identification is the electric drill of 0.6mm for adopting diameter, according to the kind that will identify shellfish; Confirm the sexual gland position of shellfish parent, boring, is sterilized microsyringe to the sexual gland position until deeply on the shell of shellfish parent; Take out essence or ovum with the microsyringe of sterilizing; Adopt the water droplet method at microscopically, add a little seawater, carry out sex identification; Ovum is dispersed in seawater, is circle or pyriform at microscopically; Sperm is smoke-like in seawater, and microscopically is the tadpole shape.
3. the breeding method of common intertidal shellfish polymerization hybridization as claimed in claim 1, it is characterized in that: the different geographical population shellfishes of said collection as parent's standard are: (1), 2 ages, healthy harmless individuality was as the parent; (2), require its parent that enough big geographical difference and hereditary difference are arranged.
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CN102960272B (en) * 2012-11-09 2014-02-19 中国水产科学研究院南海水产研究所 Quantitative character gene pyramiding breeding method for pinctada fucata
CN104031974B (en) * 2014-05-14 2016-08-24 大连海洋大学 A kind of sex appraisal method of Veneridae shellfish parent
CN108931468A (en) * 2018-05-16 2018-12-04 中国海洋大学 A kind of method of easy quickly detection oyster ploidy
CN109430126B (en) * 2018-11-06 2020-09-15 广东海洋大学 Method for cultivating new hybrid scallop variety of purple cuttlefish
CN116391654B (en) * 2023-03-30 2023-11-03 中国海洋大学 Large-scale hybridization breeding method for hermaphrodite scallops

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