CN101884313B - Family breeding method for Penaeus chinensis 'Huanghai No.1' with high ammonia nitrogen resistance and high pH value - Google Patents
Family breeding method for Penaeus chinensis 'Huanghai No.1' with high ammonia nitrogen resistance and high pH value Download PDFInfo
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Abstract
The invention discloses a family breeding method for Penaeus chinensis 'Huanghai No.1' with high ammonia nitrogen resistance and high pH value. The method comprises the following steps of: collecting the Penaeus chinensis 'Huanghai No.1' bred in various areas to construct base groups; then, selecting parent shrimps from different groups, and establishing families according to the complete diallel cross mode to separately grow the larvae and breed; respectively carrying out a high ammonia nitrogen and high pH value stress test on each family at the larvae I stage (P1 stage) and on the day when the larvae is 150-day old, and screening the families with high tolerance by combining with the breeding survival rate to carry out breed conservation; in the following year, establishing the families by taking the conservation families as the parents and adopting a nested design, and then breeding in the families; and in the third year, carrying out small-scale demonstrated breeding on the screened families, and determining and estimating the adaptive capacities of the screened families to environment. The method of the invention has the advantages of realizing the polymerization and accumulation of the growth and stress resistance characters of the penaeus chinensis, cultivating new anti-stress strains or varieties after further breeding is performed, providing the adaptability of penaeus chinensis to different breeding environments, and having great significance for the variety replacement of the penaeus chinensis.
Description
Technical field:
The invention belongs to the breeding technique of Chinese prawn genetic improvement and improved seeds, specifically is the selection of anti-high ammonia nitrogen of a kind of Chinese prawn and high pH value family.
Background technology:
Chinese prawn is the distinctive landrace of China, also is one of principal item of current prawn culturing.Influence at present China's Chinese prawn aquaculture key constraints lasting, stable and that develop in a healthy way three aspects are arranged: seed, disease and breeding environment.The research of relevant Chinese prawn breed breeding and disease aspect, China has carried out a large amount of research works successively.The breed breeding aspect has been cultivated 2 new varieties of Chinese prawn " No. 1, the Huanghai Sea " and " No. 2, the Huanghai Sea ", has fast growth and the strong merit of anti-WSSV ability respectively.The disease control aspect is because developing rapidly of technique for gene engineering obtained breakthrough at the aspects such as epidemiology, detection technique and integrated control technique of Chinese prawn virus disease respectively.But development along with the intensification of Chinese prawn breeding production, and the accumulation of water quality deterioration, drug abuse, prawn culturing self organic pollutant has had a strong impact on the living environment of prawn in the breeding process, environment is also serious day by day to coercing of causing of Chinese prawn, but the research of the prawn culturing environment aspect of carrying out at present mainly concentrates on the influence research of the growth of environment-stress factor pair prawn, survival and the nospecific immunity factor, genetic breeding and breeding environment is coerced the factor research work of carrying out seed selection that combines at home and abroad all do not appear in the newspapers as yet.
Family selective breeding has that seed selection speed is fast, the seed selection effect is obvious and advantage such as merit is outstanding, before the present invention makes, similar research report is all arranged both at home and abroad, formulated " foundation of cultured prawn family and fine-variety breeding method " as (patent No.: ZL 03111110.6) such as the Xiang Jianhai of institute of oceanography of the Chinese Academy of Sciences, set forth from close shrimp, grown seedlings, culture and gather in the crops system of selection the cultured prawn growth traits.(Hawaii, America institute of oceanography such as Argue, 2002) adopt family selective breeding to set up growth and anti-TSV strain, and the family of adopt setting up has been estimated the heritability of 2 proterties and genetic correlation etc., but relevant report only limits to grow and the seed selection of aspect such as disease resistance research, will not culture the environment-stress factor and carry out correlative study, all not have detailed report for the family mating design in the family selective breeding process simultaneously as breeding goal.
Summary of the invention:
The purpose of this invention is to provide a kind of high yield, anti-high ammonia nitrogen and strong Chinese prawn family selective breeding method of high pH value ability of selecting.
The present invention finishes according to following operational aspect: 1, collect Chinese prawn " No. 1, the Huanghai Sea " the basis of formation colony of culturing different regions and assess; 2 select close shrimp respectively from different colonies, set up family by complete diallel pattern, grow seedlings separately and culture; 3 respectively to each family in the young shrimp I phase (P1 phase) with 150 ages in days carry out high ammonia nitrogen and high pH value is coerced test, the family of the breed survival rate screening better tolerance of each family is protected kind when going out the pond simultaneously; 4 is the parent to protect kind of a family, adopts Nested Design to set up family, adopts anti-high ammonia nitrogen of 3 methods screening and high pH value family; The 5 pairs of familys that filter out are carried out the small-scale demonstration and are cultured, and determine and assess its good anti-high ammonia nitrogen and high pH value proterties.
1, collects Chinese prawn " No. 1, the Huanghai Sea " the basis of formation colony of culturing different regions and assessing: collects the individual respectively 100 tail constructional base colonies of Chinese prawn " No. 1, the Huanghai Sea " male and female of culturing 3 above different regions, select female shrimp body length 〉=14cm, the individuality of male shrimp body length 〉=12cm is as alternative close shrimp.
2, from different colonies, select close shrimp respectively, set up family by complete diallel pattern, grow seedlings separately and culture: from alternative close shrimp, select health, anosis, energetic individuality as close shrimp, adopt technology of artificial insemination to set up family mating combination by complete diallel pattern, every kind of mating is combined as 3-5 family, female-male proportion is 1: 1, and the family of setting up is grown seedlings separately and cultured.
3, respectively to each family in the young shrimp I phase (P1 phase) with 150 ages in days carry out high ammonia nitrogen and high pH value is coerced test, the family of the breed survival rate screening better tolerance of each family is protected kind when going out the pond simultaneously: the screening of anti-high ammonia nitrogen and high pH value family is divided into three times and carries out.Growing to P1 during the phase for the first time, each family is got the 30-50 tail at random, is undertaken by different test gradients is set.The ammonia nitrogen gradient is with NH
4CL (A.R.) is dissolved in the seawater and makes, be respectively 2,4,8,16,32,64,128 and 256mg/L totally 8 the test gradients; The pH value be respectively 8.2,8.4,8.6,8.8,9.0,9.2,9.4 and 9.6 totally 8 the test gradients, adopt 0.5mol/LHCL and 1mol/LNaOH to modulate, long run test three days, calculate the deadly value of half (LD50) according to each family different tests gradient survival rate, the deadly value of average half with all familys is a screening index, the family of screening better tolerance; When each family grew to 150 ages in days, each family was got the 30-50 tail at random and is coerced test for the second time.The concentration that high ammonia nitrogen adopts is 64mg/L, and the pH value adopts 9.2 to coerce, and carries out continuously three days, adds up the death time individual in each family, is that index is carried out programmed screening with the family mean time-to-live; For the third time in conjunction with going out the pond, add up each family and culture survival rate, average breed survival rate with all familys is that index is screened, final only be chosen in the family that performance in three screenings all is higher than screening index and protect kind, parent as family foundation in 1 year, it is female shrimp 100-200 tail that the guarantor of each family plants close shrimp number, male shrimp 50-100 tail.
4, be the parent to protect kind of a family, adopt Nested Design to set up family, anti-high ammonia nitrogen of (3) method of employing screening and high pH value family: adopted Nested Design to set up family mating combination in 1 year, the 2-3 group is set up in every kind of mating combination, every group of female-male proportion is controlled at 2-3: 1, to be contrast, adopt the experimental scheme of coercing in (3) to carry out the interior seed selection continuously of family and protect kind simultaneously without the Chinese prawn of coercing test " No. 1, the Huanghai Sea "; Protect kind of close shrimp number with (3).
5, the family that filters out being carried out the small-scale demonstration cultures, determine and assess its good anti-high ammonia nitrogen and high pH value proterties: the anti-high ammonia nitrogen that filters out, high pH value family carried out the small-scale breed of demonstrating at 3 above culturing areas in the 3rd year, each demonstration pilot project cultured area is more than 50 mu, be contrast with the Chinese prawn " No. 1, the Huanghai Sea " of coercing test simultaneously, breed survival rate and per mu yield during the statistics results are estimated the family selective breeding effect.
The technology of the present invention has following characteristics:
1, the present invention carries out the family selective breeding of anti-high ammonia nitrogen, high pH value on the basis of the new varieties-Chinese prawn " No. 1, the Huanghai Sea " of growing fast, realized the polymerization of growth and degeneration-resistant proterties and added up, improved the adaptability of Chinese prawn " No. 1, the Huanghai Sea " to breeding environment, enlarged the suitable culturing area of Chinese prawn " No. 1, the Huanghai Sea ", can promote the sustainable development of China's shrimp culture industry, satisfy the growing demand of people.
2, culturing the result by demonstrating on a small scale shows, anti-high ammonia nitrogen that filters out and high pH value family have tangible effect of increasing production and stronger adaptability, having good promotion and application is worth, through small-scale demonstration test and comparative test, can obtain the high yield per mu yield more than 80kg, culture survival rate and improve 10-15% than control population.
3, can cultivate degeneration-resistant new lines or new varieties, enrich the variety source of China's Chinese prawn, improve the adaptability of Chinese prawn, China be produced the update tool significance that goes up the Chinese prawn kind the different breeding environment.
Embodiment:
Be described in detail technical method of the present invention below by embodiment:
The present invention finishes according to following operational aspect: 1, collect Chinese prawn " No. 1, the Huanghai Sea " the basis of formation colony of culturing different regions and assess; 2 select close shrimp respectively from different colonies, set up family by complete diallel pattern, grow seedlings separately and culture; 3 respectively to each family in the young shrimp I phase (P1 phase) with 150 ages in days carry out high ammonia nitrogen and high pH value is coerced test, the family of the breed survival rate screening better tolerance of each family is protected kind when going out the pond simultaneously; 4 is the parent to protect kind of a family, adopts Nested Design to set up family, adopts anti-high ammonia nitrogen of 3 methods screening and high pH value family; The 5 pairs of familys that filter out are carried out the small-scale demonstration and are cultured, and determine and assess its good anti-high ammonia nitrogen and high pH value proterties.
1, collected Chinese prawn " No. 1, the Huanghai Sea " the colony establishment underlying group that Changyi, Hebei, Jiangzhou and sunshine, cultured 4 different regions, the female shrimp of each colony and male shrimp quantity are all more than 100 tails.The body progress row of 4 colonies is measured and statistics, selected female shrimp body length 〉=14cm, the individuality of male shrimp body length 〉=12cm is as alternative close shrimp; By to of the heterosis, hybrid vigor analysis of 4 population growths, finally determine to adopt the tangible Changyi of heterosis, hybrid vigor, sunshine, 3 regional Chinese prawn " No. 1, the Huanghai Sea " colonies of culturing in Hebei as the parent who sets up family with anti-high ammonia nitrogen, high pH value proterties.
2, from the alternative close shrimp of 3 colonies, select male shrimp of 9 tails and the female shrimp of 9 tails respectively at random, 27 family full-sibses have been set up according to complete diallel pattern, the family of success of growing seedlings at last is 25, to 25 family full-sibses setting up, grow seedlings separately and form work, the same 6000 tail/mus that are controlled at of cultivation density.
3, the screening of anti-high ammonia nitrogen and high pH value family is divided into three times and carries out.
(1) concrete operations of coercing for the first time test are: treat that family grows to P1 during the phase, the test gradient is set: ammonia nitrogen concentration, and with NH
4CL (A.R.) is dissolved in the seawater and makes, gradient be respectively 2,4,8,16,32,64,128 and 256mg/L totally 8 the test gradients; The gradient of pH value is respectively 8.2,8.4,8.6,8.8,9.0,9.2,9.4 and 9.6 totally 8 test gradients, adopts 0.5mol/LHCL and 1mol/LNaOH to modulate; Adopt the test gradient configuration testing liquid that is provided with, each family is got 30 tails at random and is put into the beaker that fills the 400mL testing liquid, the trace inflation, observe every 24h, write down each family young death toll, long run test three days, adopt SPSS13.0 software to calculate and obtain family 24h, 48h and 72h are respectively 64.48 to the average LD50 of ammonia nitrogen tolerance, 31.58 and 16.37mg/L, average LD50 to pH value tolerance is respectively 10.09,9.43 and 9.14, with 24,48 and the 72h tolerance all to be higher than average LD50 be that standard is screened, filter out respectively ammonia nitrogen and pH value tolerance family 14 and 12 preferably.
(2) coercing the concrete operations of test the second time is: when treating that each family is grown to 150 ages in days, each family is got 20-30 tail individuality at random and is carried out mark, the method of mark be respectively in the left side of the 6th uromere of each individuality, the artificial fluorescent rubber (Vie mark) of right side injection different colours combination, the color combination difference of different family marks.After mark was finished, the test individuality of all familys was put into 50m
2The indoor culture pond test, the high-concentration ammonia-nitrogen that testing liquid adopts is 64mg/L, high pH value is 9.2.After on-test, pull death or frequently dead prawn every 2-3h in the pond out from culturing, write down the death time of each family individuality, the mean survival time that adopts the SPSS software statistics to obtain the anti-high ammonia nitrogen of all familys is 25.64h, and the mean survival time of anti-high pH value is 30.16h, filters out the family that is higher than all family mean time-to-live and is respectively 10 and 9.
(3) concrete operations of family screening for the third time are: arrived the breed later stage, gathered in the crops in the past in the family mating, add up the quantity of each family results, the average breed survival rate of calculating all familys is 61.25%, be combined in preceding twice test of coercing simultaneously and show all family preferably, finishing screen is selected anti-high ammonia nitrogen and high pH value family is respectively 5 and 6, and reserves seed for planting, and is used for the parent of 1 year family selective breeding.
4,11 familys to selecting adopt Nested Design to set up 55 family full-sibses, carry out seed selection continuously in the family according to the family selective breeding method in (3).In breeding process being contrast without the Chinese prawn of coercing test " No. 1, the Huanghai Sea " colony, the cultivation density unification is controlled at 6000 tail/mus, unification is raised and is managed, finishing screen is selected and is resisted respectively 24 of high ammonia nitrogen family and high pH value familys, wherein the anti-high ammonia nitrogen of 14 familys and high pH value ability are all stronger, and the family of reserving seed for planting at last is 34.
5,34 familys of reserving seed for planting are grown seedlings separately, carry out the small-scale demonstration in Haifeng county, Changyi City aquaculture Co., Ltd, the Qingdao City flourish aquatic science and technology of treasured Development Co., Ltd, three areas of the little extra large prawn seed-rearing factory in Rizhao City Donggang District Tao Luo town respectively and culture, adopt and organize in contrast without the Chinese prawn " No. 1, the Huanghai Sea " of seed selection.Wherein Haifeng county, Changyi City aquaculture Co., Ltd demonstration area is 252 mu, and average yield per mu 79.3kg realizes 160.8 ten thousand yuan of the output values, compares with control group, cultures survival rate and improves 12%; Qingdao City precious flourish aquatic science and technology Development Co., Ltd demonstration cultured area is 130 mu, and family mean per mu yield 85.4kg during results realizes 91.4 ten thousand yuan of the output values, and average survival is high by about 15% than control group; Rizhao City Donggang District Tao Luo town little extra large prawn seed-rearing factory cultured area is 200 mu, and average yield per mu is 76.8kg during results, realizes 122.6 ten thousand yuan of the output values, compares with control group, and the degeneration-resistant familial incidence of cultivation reduces by 10%.582 mu of cultured area accumulative totals, per mu yield is more than 80.5kg, and the cultured output survival rate improves 10-15% than control population.
Claims (6)
1. a Chinese prawn " No. 1, the Huanghai Sea " resists high ammonia nitrogen, high pH value family selective breeding method, it is characterized in that selection is: Chinese prawn " No. 1, the Huanghai Sea " the basis of formation colony that culture (1), collection different regions also assesses; (2), from different colonies, select close shrimp respectively, set up family by complete diallel pattern, grow seedlings separately and culture; (3), respectively to each family in the young shrimp I phase with 150 ages in days carry out high ammonia nitrogen and high pH value is coerced test, the family of the breed survival rate screening better tolerance of each family is protected kind when going out the pond simultaneously; (4), be the parent to protect kind of a family, adopt Nested Design to set up family, adopt the anti-high ammonia nitrogen of selection screening and the high pH value family of (3); (5), the family that filters out carried out small-scale demonstration culture, determine and assess its anti-high ammonia nitrogen and high pH value proterties.
2. a kind of Chinese prawn according to claim 1 " No. 1, the Huanghai Sea " resists high ammonia nitrogen, high pH value family selective breeding method, it is characterized in that Chinese prawn " No. 1, the Huanghai Sea " the basis of formation colony that culture described collection different regions and assess, be to collect individual each the 100 tail constructional base colony of Chinese prawn " No. 1, the Huanghai Sea " male and female of culturing 3 above different regions, select female shrimp body length 〉=14cm, the individuality of male shrimp body length 〉=12cm is as alternative close shrimp.
3. a kind of Chinese prawn according to claim 1 " No. 1, the Huanghai Sea " resists high ammonia nitrogen, high pH value family selective breeding method, it is characterized in that the described close shrimp that from different colonies, selects respectively, set up family by complete diallel pattern, grow seedlings separately and culture, be from alternative close shrimp, to select health, anosis, energetic individuality as close shrimp, adopt technology of artificial insemination to set up family mating combination by complete diallel pattern, every kind of mating is combined as 3-5 family, female-male proportion is 1: 1, and the family of setting up is grown seedlings separately and cultured.
4. a kind of Chinese prawn according to claim 1 " No. 1, the Huanghai Sea " resists high ammonia nitrogen, high pH value family selective breeding method, it is characterized in that described respectively to each family in the young shrimp I phase with 150 ages in days carry out high ammonia nitrogen and high pH value is coerced test, the family of the breed survival rate of each family screening better tolerance was protected kind when combination went out the pond simultaneously, the screening of anti-high ammonia nitrogen and high pH value family is divided into three times and carries out: young shrimp I coerces experiment during the phase growing extremely for the first time, long run test three days, calculate the deadly value of half according to each family different tests gradient survival rate, the deadly value of average half with all familys is a screening index, the family of screening better tolerance; When each family grew to 150 ages in days, each family was got the 30-50 tail at random and is coerced test for the second time; Carrying out continuously three days, and added up the death time individual in each family, is that index is carried out programmed screening with the family mean time-to-live; For the third time in conjunction with going out the pond, add up each family and culture survival rate, average breed survival rate with all familys is that index is screened, final only be chosen in the family that performance in three screenings all is higher than screening index and protect kind, parent as family foundation in 1 year, it is female shrimp 100-200 tail that the guarantor of each family plants close shrimp number, male shrimp 50-100 tail.
5. a kind of Chinese prawn according to claim 1 " No. 1, the Huanghai Sea " resists high ammonia nitrogen, high pH value family selective breeding method, it is characterized in that described is the parent to protect kind of a family, adopt Nested Design to set up family, adopt the anti-high ammonia nitrogen of selection screening and the high pH value family of (3): adopted Nested Design to set up family mating combination in 1 year, the 2-3 group is set up in every kind of mating combination, every group of female-male proportion is controlled at 2-3: 1, to be contrast, adopt the experimental scheme of coercing in (4) to carry out the interior seed selection continuously of family and protect kind simultaneously without the Chinese prawn of coercing test " No. 1, the Huanghai Sea "; Protecting kind of a close shrimp number is female shrimp 100-200 tail, male shrimp 50-100 tail.
6. a kind of Chinese prawn according to claim 1 " No. 1, the Huanghai Sea " resists high ammonia nitrogen, high pH value family selective breeding method, it is characterized in that describedly the family that filters out is carried out small-scale demonstration culturing, determine and assess its good anti-high ammonia nitrogen and high pH value proterties: the 3rd year anti-high ammonia nitrogen filtering out, high pH value family is carried out the small-scale demonstration at 3 above culturing areas and is cultured, each demonstration pilot project cultured area is more than 50 mu, be contrast with the Chinese prawn " No. 1, the Huanghai Sea " of coercing test simultaneously, breed survival rate and per mu yield during the statistics results are estimated the family selective breeding effect.
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| CN103250666B (en) * | 2013-05-14 | 2014-08-20 | 中国水产科学研究院黄海水产研究所 | Method for breeding blue crab families |
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| CN109526826B (en) * | 2018-12-26 | 2021-04-16 | 中国水产科学研究院黄海水产研究所 | Breeding and seed production method of salt-alkali-resistant litopenaeus vannamei |
| CN110063284B (en) * | 2019-05-23 | 2021-10-08 | 中国水产科学研究院南海水产研究所 | Method for breeding new variety (line) of penaeus monodon with crude feed resistance |
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| CN112674002A (en) * | 2020-12-26 | 2021-04-20 | 海南禄泰海洋生物科技有限公司 | Shrimp larva hatching method with high survival rate |
| CN114403068B (en) * | 2022-01-20 | 2023-08-01 | 上海市水产研究所（上海市水产技术推广站） | Breeding method of low-salt-tolerance palaemon carinicauda strain |
| CN114766401B (en) * | 2022-04-21 | 2023-06-20 | 浙江万里学院宁海海洋生物种业研究院 | Seed production method for ammonia nitrogen-resistant and high-yield sinonovacula constricta new variety |
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