CN110663593A - Construction method of hybrid loach with fast growth and good meat quality and suitable for paddy field cultivation - Google Patents
Construction method of hybrid loach with fast growth and good meat quality and suitable for paddy field cultivation Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a construction method of hybrid loaches which are fast in growth, good in meat quality and suitable for comprehensive rice field breeding. Taking F3-generation Paramisgurnus Dabryanus (PD) and loach (MA) groups obtained by continuous 3-generation group breeding respectively as parents, obtaining four combinations (PD, MA, PM and MP) through loach selfing and positive and negative hybridization, and screening a novel loach hybridization good combination (MP) of loach (female) multiplied by paramisgurnus dabryanus (male). The results of the control populations PD, MA, PM showed: (1) MP shows significant hypermaternal growth advantage (P<0.05), total body weight also significantly higher than PM: (0.05)P<0.05); (2) the content indexes of the MP muscle nutrient components and the flavor substances are between that of loaches and paramisgurnus dabryanus, and the deep processing and eating quality of fish meat are improved; (3) MP grows fast and has the bottom layer feeding habit similar to MA, is suitable for the comprehensive planting and breeding mode of the paddy field.
Description
Technical Field
The invention relates to a construction method of hybrid loaches, and belongs to the field of agricultural cultivation.
Background
Loaches prefer to forage on the bottom layer of a water body, but the loaches grow slowly, and have long sexual maturity periods. Paramisgurnus dabryanus grows fast, sexual maturity cycle is short, but likes to forage in the upper layer of water, is easily influenced by high temperature environment and natural enemy, and is not suitable for comprehensive planting and breeding in rice fields. Therefore, in order to adapt to the development of a novel green ecological breeding mode and widely popularize a 'rice and loach' comprehensive breeding mode, a new loach which grows fast, has good meat quality and is fond of bottom foraging needs to be bred urgently. In order to develop a new loach variety which is fast in growth, good in meat quality and suitable for comprehensive breeding in a rice field, the invention provides a construction method of the new fish variety, loaches and paramisgurnus dabryanus which are bred by continuous 3-generation population are subjected to intergeneric hybridization to obtain an excellent hybrid population with dominant characters.
Disclosure of Invention
Based on the problems mentioned in the background technology, the invention aims to provide a construction method of hybrid loaches which are fast in growth, good in meat quality and suitable for rice field cultivation, and the construction method is realized by the following technical scheme: the method comprises the following steps:
(1) respectively taking Poyang lake wild loach and Paramisgurnus dabryanus as original parents, and obtaining F3 generation loach (MA) and Paramisgurnus Dabryanus (PD) populations through continuous 3 generation population breeding;
(2) selecting excellent individuals from F3-generation loaches (MA) and large-scale paramisgurnus anguillicaudatus (PD) groups as parents (the research is recorded as P0 generation), and respectively obtaining four combinations of an inbred F1 offspring PD, an inbred F1 offspring MA, an orthogonal F1 offspring MP, a backcross F1 offspring PM and the like through loaches (female) multiplied by loaches (male), large-scale paramisgurnus anguillicaudatus (female) multiplied by male), loaches (female) multiplied by large-scale paramisgurnus anguillicaudatus (male);
(3) and screening out novel excellent hybrid combination loaches (male parent) and large-scale loaches (male parent) (MP). The process of respectively carrying out continuous 3-generation population breeding comprises the following steps:
(1) collecting 3-year-old loaches and Paramisgurnus dabryanus from Poyang lake, wherein the loaches are 100 loaches and 100 loaches with normal morphology, bright body color, good sexual maturity and strong physique, and the male-female ratio is 7: 1, constructing basic groups of loaches and paramisgurnus dabryanus, and carrying out intensive cultivation;
(2) and carrying out group breeding on loaches and paramisgurnus dabryanus through artificial induced spawning and artificial insemination to obtain two group breeding generations F1. After artificial incubation, transferring to a pond for culture, screening 5000 persons (half each female and half each male) with fast growth and good constitution according to 5 per mill of selection pressure when the persons are cultured to 1 year, and continuously carrying out intensive culture until sexual maturity;
(3) and randomly selecting 100 loaches with normal shapes, bright body colors, good sexual maturity and strong physique and 100 loaches of paramisgurnus dabryanus from the enhanced breeding F1 generation, wherein the male-female ratio is 7: 1, carrying out group breeding on loaches and paramisgurnus dabryanus through artificial induced spawning and artificial insemination to obtain two group breeding generations F2. After artificial incubation, transferring to a pond for culture, screening individuals with fast growth and good constitution according to the selection pressure of 5 per mill when the individuals grow to 1 year, forming 5000 tails of backup parent groups, wherein the males and females are half, and continuously carrying out intensive culture until sexual maturity;
(4) and randomly selecting 100 loaches with normal shapes, bright body colors, good sexual maturity and strong physique and 100 loaches of paramisgurnus dabryanus from the enhanced breeding F2 generation, wherein the male-female ratio is 7: 1, carrying out group breeding on loaches and paramisgurnus dabryanus through artificial induced spawning and artificial insemination to obtain two group breeding generations F3. After artificial incubation, transferring to a pond for culture, screening individuals with fast growth and good constitution according to the selection pressure of 5 per mill when the individuals grow to 1 year, forming 5000 tails of backup parent groups, wherein the males and the females are half respectively, and continuously carrying out intensive culture until sexual maturity.
The procedure of the four F1 generation combination is as follows:
(1) selecting excellent individuals from F3-generation loaches (MA) and Paramisgurnus Dabryanus (PD) groups, wherein the male-female ratio is 7: 1, two P0 generation populations were constructed;
(2) obtaining four combinations of loaches (male parent) and loaches (female parent) and loaches (male parent) and loaches (female parent) and loaches (male parent) to obtain four combinations of PD (male F1) offspring, MA (self F1) offspring, MP (orthogonal F1 offspring and PM (reverse cross F1) offspring, and transferring the four combinations to a pond for breeding after artificial incubation.
The step process of screening out the novel loach hybrid excellent combination loach (male parent) and large-scale loach (male parent) (MP) is as follows: (1) randomly selecting 3000 plants from the four F1-generation combinations, setting 3 parallel groups, performing a pond culture growth comparison test of two years in the period of 2 years, and comparing the growth traits of the four combinations;
(2) after the growth contrast test is finished, 10 females and 10 males are randomly selected from four F1 generation combinations respectively for muscle nutrient composition and taste and smell characteristic analysis and quality evaluation, wherein basic nutrient composition analysis, free amino acid and free nucleotide are determined according to a national standard method, and the smell component is determined by a GC-MS method.
(3) At the age of 2 months, 1.5-2.0 ten thousand of plants are randomly selected from 4F 1 combinations, and are subjected to a rice field comprehensive planting and breeding comparison test (planting area is 9 mu, breeding area is 1 mu) with medium-season rice, and the adaptability of the four F1 combinations in the rice field comprehensive planting and breeding is compared according to the yield and the output value of rice and loaches.
The invention has the advantages of
(1) The growth speed of the hybrid loach MP obtained by the invention is obviously higher than that of a female parent (MA), is close to that of a male Parent (PD), and shows growth advantages.
(2) The hybrid loach MP obtained by the invention has better muscle nutrient components and quality.
(3) The hybrid loach MP obtained by the invention is suitable for the comprehensive breeding of the rice field, can obviously improve the profit output value of the comprehensive breeding of the rice field, and simultaneously produces organic rice and high-quality loach products.
Drawings
Fig. 1 is a 4 loach population morphological parameter cluster analysis diagram.
Detailed Description
The following examples are merely examples of the technical solutions of the present invention, and the present invention is not limited to the technical solutions of the following examples.
Example 1
(1) Respectively taking Poyang lake wild loach and Paramisgurnus dabryanus as original parents, and obtaining F3 generation loach (MA) and Paramisgurnus Dabryanus (PD) populations through continuous 3 generation population breeding;
(2) selecting excellent individuals from F3-generation loaches (MA) and large-scale paramisgurnus anguillicaudatus (PD) groups as parents (the research is recorded as P0 generation), and respectively obtaining four combinations of an inbred F1 offspring PD, an inbred F1 offspring MA, an orthogonal F1 offspring MP, a backcross F1 offspring PM and the like through loaches (female) multiplied by loaches (male), large-scale paramisgurnus anguillicaudatus (female) multiplied by male), loaches (female) multiplied by large-scale paramisgurnus anguillicaudatus (male);
(3) and screening out novel excellent hybrid combination loaches (male parent) and large-scale loaches (male parent) (MP).
The process of respectively carrying out continuous 3-generation population breeding comprises the following steps:
(1) collecting 100 loaches with normal morphology, bright body color, good sexual maturity and strong physique and Paramisgurnus dabryanus from Poyang lake, wherein the male-female ratio is 7: 1, constructing basic groups of loaches and paramisgurnus dabryanus, and carrying out intensive cultivation;
(2) and carrying out group breeding on loaches and paramisgurnus dabryanus through artificial induced spawning and artificial insemination to obtain two group breeding generations F1. After artificial incubation, transferring to a pond for culture, screening individuals with fast growth and good constitution according to the selection pressure of 5 per mill when the individuals grow to 1 year, forming 5000 tails of backup parent groups, wherein the males and females are half, and continuously carrying out intensive culture until sexual maturity;
(3) and randomly selecting 100 loaches with normal shapes, bright body colors, good sexual maturity and strong physique and 100 loaches of paramisgurnus dabryanus from the enhanced breeding F1 generation, wherein the male-female ratio is 7: 1, carrying out group breeding on loaches and paramisgurnus dabryanus through artificial induced spawning and artificial insemination to obtain two group breeding generations F2. After artificial incubation, transferring to a pond for culture, screening individuals with fast growth and good constitution according to the selection pressure of 5 per mill when the individuals grow to 1 year, forming 5000 tails of backup parent groups, wherein the males and females are half, and continuously carrying out intensive culture until sexual maturity;
(4) and randomly selecting 100 loaches with normal shapes, bright body colors, good sexual maturity and strong physique and 100 loaches of paramisgurnus dabryanus from the enhanced breeding F2 generation, wherein the male-female ratio is 7: 1, carrying out group breeding on loaches and paramisgurnus dabryanus through artificial induced spawning and artificial insemination to obtain two group breeding generations F3. After artificial incubation, transferring to a pond for culture, screening individuals with fast growth and good constitution according to the selection pressure of 5 per mill when the individuals grow to 1 year, forming 5000 tails of backup parent groups, wherein the males and the females are half respectively, and continuously carrying out intensive culture until sexual maturity.
The procedure of the four F1 generation combination is as follows:
(1) selecting excellent individuals from F3-generation loaches (MA) and Paramisgurnus Dabryanus (PD) groups, wherein the male-female ratio is 7: 1, two P0 generation populations were constructed;
(2) obtaining four combinations of loaches (male parent) and loaches (female parent) and loaches (male parent) and loaches (female parent) and loaches (male parent) to obtain four combinations of PD (male F1) offspring, MA (self F1) offspring, MP (orthogonal F1 offspring and PM (reverse cross F1) offspring, and transferring the four combinations to a pond for breeding after artificial incubation.
The step process of screening out the novel loach hybrid excellent combination loach (male parent) and large-scale loach (male parent) (MP) is as follows:
(1) randomly selecting 3000 plants from the four F1-generation combinations, setting 3 parallel groups, performing a pond culture growth comparison test of two years in the period of 2 years, and comparing the growth traits of the four combinations;
(2) after the growth contrast test is finished, 10 females and 10 males are randomly selected from four F1 generation combinations respectively for muscle nutrient composition and taste and smell characteristic analysis and quality evaluation, wherein basic nutrient composition analysis, free amino acid and free nucleotide are determined according to a national standard method, and the smell component is determined by a GC-MS method.
(3) At the age of 2 months, 1.5-2.0 ten thousand of plants are randomly selected from 4F 1 combinations, and are subjected to a rice field comprehensive planting and breeding comparison test (planting area is 9 mu, breeding area is 1 mu) with medium-season rice, and the adaptability of the four F1 combinations in the rice field comprehensive planting and breeding is compared according to the yield and the output value of rice and loaches.
Experiment and analysis:
the invention relates to a method for breeding loaches, which are parents used in all experiments and are obtained from Ganxiang source ecological loach farm (27 degrees 47 '12.96' in North latitude and 115 degrees 27 '35.67' in east longitude) in New Ganxiang city, Ji city, Jiangxi, and all breeding experiments are carried out in the farm. Four F1 generation populations of selfing and positive and negative hybridization are obtained through artificial induced spawning and insemination, and analysis such as growth traits, morphological characteristics, muscle nutrient content and flavor substance content, comprehensive rice field planting and breeding applicability test and the like are respectively carried out. The specific method and results are as follows:
determination of growth traits and morphological characteristic analysis of first and fourth loaches F1 populations
1. Determination of growth traits
During the breeding test, 60 loaches were randomly selected from four loach populations of 2, 3, 4, 5, and 12 months of age, and body length and weight were measured. And (4) before measurement, wiping the water on the body surface of the loach, measuring the weight and the body length by using an electronic balance and a ruler respectively, and recording data. The average growth amounts of the four loach groups in different growth stages are shown in table 1, and the average monthly weight gain and growth gain are shown in table 2.
TABLE 1 average growth of four loach groups at different growth stages
TABLE 2 average monthly weight gain (g/M), average monthly growth (cm/M) for four loach populations
The results, combined with table 1 and table 2, show that the orthogonal (MP) F1 generation is significantly higher in growth performance than the backcross (PM) F1 generation, loach selfing (M) F1 generation (P <0.05), and significantly lower than the parapisgurnus dabryanus selfing (P) F1 generation, and the orthogonal F1 generation has uniform individual size distribution.
2. Morphological feature analysis
The traditional morphological quantifiable character (7 items) and the frame data (20 items) are measured by an electronic scale, a compass and a ruler, and the traditional morphological quantifiable character (4 items) is observed by naked eyes. Traditional morphologically quantifiable shapes include: body weight, body length, overall length, body height, body width, eye diameter, head length. The frame data includes: d1-2, D1-3, D3-5, D5-7, D7-9, D2-4, D4-6, D6-8, D8-10, D4-1, D4-3, D4-5, D6-3, D6-5, D6-7, D8-5, D8-7, D8-9, D10-7 and D10-9, and the specific measurement method is reported in literature references (2010, Hayajuan; 2017, Arisakuan). Traditional morphologically quantifiable traits include: the number of dorsal fins, the number of pectoral fins, the number of ventral fins and the number of hip fins. Calculating 25 morphological proportion parameters (full length/body length, body length/body height, body length/head length, body length/tail stalk length, tail stalk length/tail stalk height, D1-2/full length, D1-3/full length, D3-5/full length, D5-7/full length, D7-9/full length, D2-4/full length, D4-6/full length, D6-8/full length, D8-10/full length, D4-1/full length, D4-3/full length, D4-5/full length, D6-3/full length, D6-5/full length, D6-7/full length, D8-5/full length, D8-7/full length, D8-9/full length, D6-3/full length, D6-5/full length, D6-7/full length, D8-5/full length, D8, D10-7/full length, D10-9/full length) are shown in Table 3.
TABLE 3 morphological structures of four loach populations
As can be seen from Table 3, the body length/body height and the full length/body height of the positive and negative hybrids are within the corresponding proportion values of the parent and the female parents, and the hybrid characteristics are shown; the total length/body length, the body length/head length and the caudal peduncle length/caudal peduncle height are out of the corresponding proportion values of the parent and the female parent, but are deviated from the loach parent except the body length/head length. The fin type has no great difference with the parent and the mother, and the pectoral fins are biased to loaches, which shows that the influence of loach parents is great.
4 loach population morphological parameter cluster analysis
And performing cluster analysis on the four loach groups according to morphological parameters, wherein the MP and the PM are gathered into one branch, then gathered with the M, and finally gathered with the P, which shows that the morphological structure of the filial generation is more biased to the M group.
Table 44 factor load matrix in loach population principal component analysis and contribution rate of principal component to total variation
The principal component analysis is shown in table 4, and the results show that the total variance contributions are: the main component 1 is 43.000%, the main component 2 is 16.356%, the main component 3 is 7.579%, and the cumulative contribution rate is 66.935%, namely, the main components contain a large part of the total variation, which shows that the morphological differences of 4 loach populations can be summarized by using the mutually independent factors. Further research and analysis show that the contribution rate of the main component 1 to the total variation is up to 43.000 percent and mainly depends on 19 indexes such as D1-2/full length, D1-3/full length, D3-5/full length, D5-7/full length, D7-9/full length, D2-4/full length, D4-6/full length, D6-8/full length, D8-10/full length, D4-1/full length, D4-3/full length, D4-5/full length, D6-3/full length, D6-5/full length, D6-7/full length, D8-5/full length, D8-7/full length, D8-9/full length and D10-7/full length, and the 19 indexes mainly reflect the size of the loach type, the difference in morphology between the 4 loach groups is believed to be significant in the length of each segment at the front and back ends.
Discrimination analysis is carried out by SPSS21.0 software, and discrimination function formulas of 4 loach groups are obtained as follows:
MP
Y＝41319.124 A1+6866.990 A2+85.196 A3-5962.788 A4+236.492 A5+40.048 A6+72.934 A7-2510.264 A8-341.976 A9+573.425 A10-1179.177 A11-204.566 A12-356.149A13-2768.239 A14-4164.249 A15+163.825 A16+2147.579 A17+824.755A18+51.408 A19+132.292 A20-402.547 A21-3379.714 A22-1231.989 A23-528.276 A24-1937.148 A25-1879.491 A26+260.810 A27-22845.430
PM
Y＝41242.568 A1+6855.231 A2+83.021 A3-5952.299 A4+230.983 A5+40.914 A6+72.299 A7-2497.251 A8-337.371 A9+617.835 A10-1266.988 A11-178.990 A12-421.183A13-2755.862 A14-4177.782 A15+177.691 A16+2222.487 A17+800.033 A18+51.358 A19+103.266 A20-422.658 A21-3301.668 A22-1281.460 A23-565.836 A24-1941.029 A25-1862.366 A26+261.461 A27-22739.406
P
Y＝41107.434 A1+6837.236 A2+85.572 A3-5938.130 A4+229.037 A5+44.028 A6+72.676 A7-2572.865 A8-335.179 A9+511.681 A10-1220.642 A11-232.809 A12-370.395A13-2788.333 A14-4133.131 A15+175.925 A16+2296.434 A17+798.910 A18+60.460 A19+195.125 A20-434.949 A21-3309.319 A22-1225.841 A23-486.062 A24-1937.816 A25-1812.345 A26+250.248 A27-22621.181
M
Y＝41312.721 A1+6867.166 A2+85.596 A3-5963.445 A4+232.535 A5+44.049 A6+72.615 A7-2490.246 A8-322.962 A9+588.097 A10-1233.939 A11-202.314 A12-357.556A13-2738.985 A14-4168.299 A15+147.563 A16+2108.515 A17+801.962 A18+67.930 A19+119.406 A20--430.780 A21-3310.503 A22-1262.934 A23-567.895 A24-1935.786 A25-1881.208 A26+260.536 A27-22849.493
when the loach is judged to belong to, the morphological parameters of the loach are measured and respectively substituted into a judgment function, and then the loach class is judged according to the maximum Y value obtained by a certain function. To verify the utility of the above formula, the measured samples were classified according to the discriminant formula, and the results of the discriminant are listed in table 5.
Table 54 loach group discrimination analysis classification results
As can be seen from table 5, the discrimination accuracy MP was 83.33%, PM was 86.67%, P was 93.33%, and M was 83.33%; the comprehensive discrimination rate was 86.67%.
Analyzing muscle nutrient components and evaluating quality of two or four loach groups
1. Materials and methods
Sample processing
Muscle mix samples were taken from each of the above 8 combinations, and the samples were divided into 4 parts, 1 part for routine nutritional ingredient determination, 1 part for amino acid determination, 1 part for nucleotide determination, and 1 part for volatile substance determination.
Conventional nutrient content determination
The water content measuring method comprises the following steps: the method is a constant temperature drying method under normal pressure (GB5009.3-2016 (determination of moisture in food safety national standard food)).
The method for measuring the content of the crude protein comprises the following steps: the Kjeldahl method (GB5009.5-2016, measurement of protein in food safety national standard food) is adopted.
The crude fat content determination method comprises the following steps: the Soxhlet extraction method (GB5009.6-2016, determination of fat in food safety national standard food) is adopted.
The method for measuring the content of the coarse ash comprises the following steps: the method of high-temperature burning at 550 ℃ is adopted (GB5009.4-2016 determination of ash content in national food safety standards).
Total amino acid determination
The total amino acid content is detected by an automatic analyzer according to the national standard method (GB/T5009.124-2016 determination of amino acids in food safety national standard food).
Determination of free amino acids
The extraction and detection of free amino acids was performed according to the method reported by Chen et al.
Nucleotide determination
The extraction and detection of nucleotide and its related products are referred to the method reported by Zhang et al.
Volatile substance assay
The reference literature uses GC/MS measurements.
Amino acid nutritional assessment
To evaluate the quality of amino acids in muscle samples, the content of various essential amino acids in the samples was converted to milligrams of amino acids per gram of protein (mg/g), and nutritional assessments were performed according to the standard model of amino acid score per gram of nitrogen as recommended by the food and agriculture organization/world health organization (FAO/WHO) of the United nations and the amino acid model of whole egg protein. Amino Acid Score (AAS), Chemical Score (CS)[16]And the Essential Amino Acid Index (EAAI) formula is as follows:
(1) amino Acid Score (AAS) ═ sample protein amino acid content (mg/g) ]/[ corresponding essential amino acid content (mg/g) in FAO/WHO score standard model) ];
(2) chemical Score (CS) ═ sample protein amino acid content (mg/g) ]/[ corresponding essential amino acid content in whole egg protein (mg/g) ];
in the formula: amino acid content refers to the amount of amino acids in milligrams per gram of nitrogen.
(3) Essential Amino Acid Index (EAAI) [ (100a/a) × (100B/B) × (100C/C) × … × (100H/H)]1/n；
In the formula: n is the number of essential amino acids to be compared; a, b, c, …, h is the content (mg/g) of each essential amino acid in the sample; a, B, C, …, H are the essential amino acid content (mg/g) corresponding to whole egg protein.
Data processing
SPSS21.0 and Excel 2016 are used for processing, the result is represented by 'mean value plus or minus standard deviation', the data are subjected to difference significance comparison analysis according to a One-Way ANOVA method, and p <0.05 indicates that the data have significance difference.
2. Results and analysis
(1) Content comparison analysis of conventional nutrient components
The conventional nutrient components comprise four parts of water, crude protein, crude fat and crude ash, the nutritional value of the muscle is mainly determined by the protein content and the fat content of the muscle, and the conventional nutrient component content of 4F 1 loaches is shown in Table 6.
TABLE 6 conventional Nutrition composition (%) -of four loach groups
Note: the superscript letters on the same line are different, which indicates that the data have significant difference (p < 0.05); the letters are identical indicating that the difference is not significant (p > 0.05).
Note:different superscript letters on the same line indicatesignificant differences between data(p<0.05)；same letter means no significantdifference(p>0.05).
The results show that the moisture content of MP is not significantly different from that of PM, P and M (P is more than 0.05); the content of MP crude protein is respectively higher than that of PM, P and M groups, but the difference is not obvious (P is more than 0.05); the crude fat content of MP is below P, M (p >0.05), significantly below PM (p < 0.05); the MP ash content is significantly lower than M (p <0.05), and the difference from P, PM is not significant (p > 0.05).
(2) Total amino acid analysis in this experiment, 17 amino acids were detected in all 8 combinations of muscles except for the destruction of tryptophan (Trp) under acid hydrolysis conditions, including 7 essential amino acids, 4 umami amino acids, and 2 semi-essential amino acids, and the results of the total amino acid composition and content are shown in Table 7.
TABLE 7 Total amino acid composition and content (mg/g) of different combinations of loach muscle
Note: in addition, the fresh amino acid is in the opposite side; # is an essential amino acid; is a semi-essential amino acid; sigma DAA is the total amount of umami amino acid; sigma EAA is the total amount of essential amino acids; Σ NEAA is a non-essential amino acid; sigma TAA is the total amount of amino acids.
The results show that: the total amino acid content in muscle of 4 loach populations was different between female and male individuals. The total amino acid content of the female combination is MF (41.15mg/g), MPF (54.95mg/g), PMF (55.60mg/g) and PF (59.89mg/g) in sequence, the total amino acid content of the male combination is MM (48.44mg/g), PMM (53.23mg/g), MPM (53.95mg/g) and PM (58.35mg/g) in sequence, and the total amino acid content of the female MP combination muscle is higher than that of M and lower than that of P, and the total amino acid content of the male MP combination muscle is higher than that of M, PM and lower than that of P.
Of the essential amino acid contents in muscle proteins of 4 loach populations, female sequentially comprise MF (16.23mg/g), PMF (21.72mg/g), MPF (21.75mg/g) and PF (23.54 mg/g), and male sequentially comprise MM (19.22mg/g), PMM (20.92mg/g), MPM (21.10mg/g) and PM (22.75mg/g), which indicates that the essential amino acid contents in the MP combined muscle proteins of female and male are all higher than M, PM and lower than P.
The umami amino acids comprise 4 types of aspartic acid (Asp), glutamic acid (Glu), glycine (Gly) and alanine (Ala), and in the proportion of the umami amino acids in the muscle of 4 loach groups, female bodies are MPF (38%), PF (38%), MF (39%), PMF (39%) and 4 combined male bodies are 39%. The Glu content of four umami amino acids combined by 8 loaches is the highest.
The nutritional value of the food proteins was evaluated using the essential Amino Acid Score (AAS), Chemical Score (CS), and Essential Amino Acid Index (EAAI), and the results are shown in Table 8.
TABLE 8 evaluation of essential amino acid composition of different combinations of loach muscle
Note: the border label AAS is greater than 1, and the shadow labels AAS and CS minimum values.
AAS is calculated by the formula, threonine (Thr) and lysine (Lys) of MPM, threonine (Thr) and lysine (Lys) of MPF, threonine (Thr), leucine (Leu) and lysine (Lys) of PM, threonine (Thr), leucine (Leu) and lysine (Lys) of PF, lysine (Lys) of MM, lysine (Lys) of MF, threonine (Thr), leucine (Leu) and lysine (Lys) of PMF, threonine (Thr), leucine (Leu) and lysine (Lys) of PMM, respectively, each of which has a combined value of greater than 1. As can be seen in combination with AAS and CS, the methionine (Met) value for each combination was minimal, indicating that methionine is the common first limiting amino acid for 4 loach populations.
The Essential Amino Acid Index (EAAI) is one of the common indicators for evaluating the nutritional value of food protein, and takes the essential amino acids of egg protein as reference standard, and the larger the EAAI is, the higher the nutritional value is. Table 8 shows that the female EAAI of 4 loach populations are MF (51.8), MPF (68.96), PMF (69.28) and PF (75.29) from small to large, and the male EAAI is MM (61.34), PMM (66.9), MPM (67.16) and PM (72.94) from small to large, which indicates that the nutritional value of the female MP combination and the male MP combination is superior to that of M, inferior to that of P and similar to that of PM. (3) Analysis of content of muscle flavor development nucleotide of 4 loach groups
Inosinic acid (IMP) is a common tasting nucleotide that has an umami taste in itself, and IMP can be converted to adenine ribonucleotide (AMP). ATP is an energy unit of living organisms, and phosphate groups are released in the metabolic process of ATP, Adenosine Diphosphate (ADP) and AMP are sequentially formed, and AMP can also be reversibly combined with phosphate groups to form ADP and ATP. Hypoxanthine (Hx) is a catabolite of AMP, a low molecular weight intermediate of ribonucleic acid (RNA) hydrolysis. Inosine (HxR), also called inosine, is one type of ribonucleoside having Hx as a base moiety. The nucleotide content of the 8 loach combinations is shown in table 9.
TABLE 9 nucleotide content (mg/100g) of loach muscle in different combinations
In the contents of umami nucleotide IMP in muscles of 4 loach groups, female sequentially comprise MF (3.54 +/-0.02 mg/100g), PMF (3.83 +/-0.06 mg/100g), MPF (3.92 +/-0.45 mg/100g) and PF (4.26 +/-0.12 mg/100g), male sequentially comprise MPM (3.88 +/-0.08 mg/100g), MM (3.95 +/-0.09 mg/100g), PMM (3.97 +/-0.04 mg/100g) and PM (4.34 +/-0.24 mg/100g), and the contents of female combined MP are higher than M, PM and lower than P; the content of IMP in the male MP combination is lower than that of M, PM and P.
In terms of the content of picric nucleotide Hx, female sequentially comprise MPF (34.55 +/-4.18 mg/100g), PMF (44.29 +/-2.48 mg/100g), MF (44.39 +/-1.62 mg/100g) and PF (54.52 +/-3.22 mg/100g), and male sequentially comprise MPM (40.02 +/-2.54 mg/100g), MM (41.34 +/-0.96 mg/100g), PMM (43.30 +/-2.52 mg/100g) and PM (43.97 +/-5.24 mg/100g), which indicates that the Hx content of the combination of female and male MP is lower than that of M, PM and P.
In the aspect of inhibiting the content of the bitter ribonucleotide AMP, female sequentially comprise MF (7.02 +/-0.54 mg/100g), PMF (7.74 +/-0.12 mg/100g), MPF (8.73 +/-3.16 mg/100g) and PF (8.94 +/-0.77 mg/100g), and male sequentially comprise MM (6.72 +/-0.15 mg/100g), MPM (7.55 +/-0.26 mg/100g), PM (7.82 +/-0.36 mg/100g) and PMM (7.86 +/-0.19 mg/100g), thereby indicating that the female MP combination AMP content is higher than M, PM and lower than P; the male MP combination AMP content was higher than M and lower than P, PM.
(4) Analysis of free amino acid content in muscle of 4 loach groups
The free amino acid content of the combined muscle of 8 loaches is shown in table 10.
TABLE 10 free amino acid content (mg/100g) of loach muscle in different combinations
Note: "-" indicates no detection; the border marks the maximum free amino acid content.
As can be seen from Table 10, PF and PM contain threonine (Thr) in the largest amount of free amino acids, which is 24.41 + -0.26 mg/100g and 28.18 + -0.49 mg/100g, respectively; the MPF and MPM contain lysine (Lys) with the maximum content of free amino acid, which is 70.95 +/-1.77 mg/100g and 64.37 +/-5.40 mg/100g respectively, and are consistent with the distribution characteristics of the free amino acid content in MF, MM, PMF and PMM.
The 4 loach population muscle nucleotide, free amino acid taste intensity value and taste intensity value (TAV) is the ratio of the content of each taste substance in the sample to its corresponding taste threshold. When the TAV is more than 1, the substance has an important influence on the taste of the sample. Muscle TAV of 8 combination loach populations is shown in table 11.
TABLE 11 free amino acid and nucleotide content and taste Strength values (TAV) for each combination
Note: the above threshold values are all threshold values in an aqueous solution; "+": the taste is good; "-": the taste is poor; "\\ is as follows: it was not detected.
The results show that the TAV of lysine (Lys) of MPM and MPF alone is greater than 1, and the TAV of all free amino acids of MPM and the remaining amino acids and other combinations of MPF is less than 1; the nucleotide TAV of 8 loach combinations is less than 1.
(6) Determination and analysis of muscle volatile substances of 4 loach groups
Composition of volatile Compounds in Table 128 loach combination muscles (%)
As can be seen from Table 12, the total number of the PMM, PM, and MM compound species was 17 at most. Of the muscle volatile compound compositions of the 4 loach populations, there were differences between female and male individuals. The female MPF combined hydrocarbon accounts for 53.85 percent and is higher than PMF, PF and MF; the proportion of the aromatic is 23.08 percent, is lower than PMF and higher than PF and MF; the alcohol accounts for 7.69 percent, is lower than PMF and higher than PF and MF; the proportion of acid MPM is 7.69 percent and is lower than that of PMF, PF and MF; the ketone accounts for 7.69%, is lower than PMF and higher than PF and MF. The male MPM combined hydrocarbon accounts for 33.33 percent and is lower than PMM, PM and MM; the highest proportion of aromatic is 33.33 percent; the highest alcohol proportion is 11.11 percent; the acid accounts for 11.11 percent, is lower than PM and higher than PMM and MM; the proportion of ketones is the highest, 11.11%.
The MPM combination has the highest proportion of aromatics, alcohols and ketones.
Aroma substances and peak area percentage (%)
Table 13 The content of aroma substance and peak area in muscles ofeight loach combinations(％)
As can be seen from Table 13, the highest combined content of MPF, PMF, PF and PM is 2, 6-di-tert-butyl-p-cresol, which is an antioxidant, protects vitamins in muscles, prevents oxidative loss of fats and proteins, and also has a certain antibacterial effect; the MPM with the highest combination content is pentamethylbenzene; the highest content of PMM combination is 1, 6-dimethylnaphthalene; heptadecane with the highest MF combination content; the highest combined MM content was myristyl aldehyde.
Comparative analysis of three or 4 loaches on comprehensive breeding applicability of rice and loaches
And 4 loach groups are used for carrying out a comprehensive rice and loach breeding test, and the application of the test is analyzed. The total area of the rice field used in each group of tests is 10 mu, and the area of the circular ditches accounts for 10 percent. The input and output conditions of each group are summarized in tables 14 and 15.
TABLE 14 comprehensive breeding and input condition summary table for rice and loach
TABLE 15 summary table of comprehensive breeding and output conditions of rice and loaches
As can be seen from the comprehensive comparison of tables 14 and 15, the MP combination yields 5340kg of rice and 547kg of loach together, and the total profit is 12963.9 yuan, which is the highest value of the four combinations. In the four combinations, the total yield of the P combination is the highest, but the survival rate and the market unit price are lower than those of the other three combinations, and the total profit of the P combination is lower than that of the other three combinations due to the mutual restriction of multiple factors.
Compared with the method of simply planting rice by surrounding farmers in the current year, the yield per mu of rice planted is 650 kg, the rice sale is 2.6 yuan/kg according to the average price and the yield per mu value is 1690 yuan, after various production costs are removed, the profit per mu is 300-350 yuan, the comprehensive breeding and production mode of the rice and the loaches is developed, the profit per mu of MP combination reaches 1296.39 yuan, and the economic benefit is more remarkable.
In conclusion, the MP combination is suitable for the comprehensive planting and breeding mode of the rice field and can create higher economic value.
Claims (4)
1. A construction method of hybrid loaches which grow fast and have good meat quality and are suitable for paddy field cultivation is characterized by comprising the following steps:
(1) respectively taking Poyang lake wild loach and Paramisgurnus dabryanus as original parents, and obtaining F3 generation loach (MA) and Paramisgurnus Dabryanus (PD) populations through continuous 3 generation population breeding;
(2) selecting excellent individuals as parents from an F3-generation loach (MA) group and a large-scale paramisgurnus anguillicaudatus (PD) group, and marking as a P0 generation, and respectively obtaining four combinations of an inbred F1 offspring PD, an inbred F1 offspring MA, an orthogonal F1 offspring MP and a backcross F1 offspring PM through loaches (female) multiplied by loaches (male), large-scale paramisgurnus anguillicaudatus (female), large-scale paramisgurnus anguillicaudatus (male) and large-scale paramisgurnus anguillicaudatus (female) multiplied by loaches (male);
(3) and screening out novel excellent hybrid combination loach (male parent) and loach (male parent) MP.
2. The method for constructing hybrid loaches which grow fast and have good meat quality and are suitable for comprehensive rice field breeding according to claim 1, wherein the process of obtaining F3-generation loaches and paramisgurnus dabryanus respectively through continuous 3-generation population breeding comprises the following steps:
(1) collecting 3-year-old loaches and Paramisgurnus dabryanus from Poyang lake, wherein the loaches are 100 loaches and 100 loaches with normal morphology, bright body color, good sexual maturity and strong physique, and the male-female ratio is 7: 1, constructing basic groups of loaches and paramisgurnus dabryanus, and carrying out intensive cultivation;
(2) carrying out loach and paramisgurnus dabryanus group breeding through artificial induced spawning and artificial insemination to obtain two group breeding F1 generations, transferring to a pond for breeding after artificial incubation, screening individuals with fast growth and good constitution according to 5 per mill of selection pressure when the loach and paramisgurnus dabryanus are bred to 1 year to form 5000 pieces of backup parent groups, wherein each half of male and female are continuously and intensively cultivated until sexual maturity;
(3) and randomly selecting 100 loaches with normal shapes, bright body colors, good sexual maturity and strong physique and 100 loaches of paramisgurnus dabryanus from the enhanced breeding F1 generation, wherein the male-female ratio is 7: 1, carrying out loach and paramisgurnus dabryanus group breeding through artificial induced spawning and artificial insemination to obtain two group breeding F2 generations, transferring to a pond for breeding after artificial incubation, screening individuals with fast growth and good physique according to the selection pressure of 5 per mill when the loach and paramisgurnus dabryanus are bred to 1 year to form backup parent groups, wherein each 5000 individuals are female and male and half, and continuously carrying out intensive breeding until sexual maturity;
(4) and randomly selecting 100 loaches with normal shapes, bright body colors, good sexual maturity and strong physique and 100 loaches of paramisgurnus dabryanus from the enhanced breeding F2 generation, wherein the male-female ratio is 7: 1, carrying out loach and paramisgurnus dabryanus group breeding through artificial induced spawning and artificial insemination to obtain two group breeding F3 generations, transferring to a pond for breeding after artificial incubation, screening individuals with fast growth and good physique according to the selection pressure of 5 per mill when the loach and paramisgurnus dabryanus are bred to 1 year to form 5000 pieces of backup parent groups, wherein the male and female are half respectively, and continuously carrying out intensive breeding until sexual maturity.
3. The method for constructing hybrid loaches which grow fast and have good meat quality and are suitable for comprehensive rice field cultivation according to claim 1, wherein the method comprises the following steps: the step process of selecting excellent individuals from the population breeding F3 generation to form P0 generation to obtain four F1 generation combinations comprises the following steps:
(1) selecting excellent individuals from F3-generation loaches (MA) and Paramisgurnus Dabryanus (PD) groups, wherein the male-female ratio of each excellent individual is 200: 1, two P0 generation populations were constructed;
(2) obtaining four combinations of loaches (male parent) and loaches (female parent) and loaches (male parent) and loaches (female parent) and loaches (male parent) to obtain four combinations of PD (male F1) offspring, MA (self F1) offspring, MP (orthogonal F1 offspring and PM (reverse cross F1) offspring, and transferring the four combinations to a pond for breeding after artificial incubation.
4. The method for constructing hybrid loaches, which grow fast and have good meat quality and are suitable for comprehensive rice field cultivation, according to any one of claims 1 to 3, is characterized in that: the step process of screening out the novel loach hybrid excellent combination loach (male parent) and large-scale loach (male parent) (MP) is as follows:
(1) randomly selecting 3000 plants from the four F1-generation combinations, setting 3 parallel groups, performing a two-year pond culture growth comparison test, and comparing the growth traits of the four combinations;
(2) after the growth contrast test is finished, randomly selecting 10 female and 10 male individuals from four F1 generation combinations for muscle nutrient composition analysis, taste and smell characteristic analysis and quality evaluation, wherein basic nutrient composition analysis, free amino acid and free nucleotide are determined according to a national standard method, and the smell component is determined by a GC-MS method;
(3) at the age of 2 months, 1.5-2.0 ten thousand of plants are randomly selected from 4F 1 combinations, and are subjected to a rice field comprehensive planting and breeding contrast test with medium-season rice, the planting area of the rice field is 9 mu, the breeding area is 1 mu, and the adaptability of the four F1 combinations in the rice field comprehensive planting and breeding is compared according to the yield and the output value of rice and loaches.
Priority Applications (1)
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