CN111187843A - SNP molecular marker related to hybrid pelteobagrus fulvidraco 'Huangyou No. 1' hypoxia tolerance character and application thereof - Google Patents
SNP molecular marker related to hybrid pelteobagrus fulvidraco 'Huangyou No. 1' hypoxia tolerance character and application thereof Download PDFInfo
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Abstract
The invention discloses an SNP molecular marker related to the hypoxia-resistant character of hybrid pelteobagrus fulvidraco 'Huangyou No. 1' and application thereof, wherein the SNP molecular marker takes a cDNA sequence of von Hippel-Lindau disease promoter (Vhl gene) as a template to design a primer, and a genome sequence obtained by amplification is shown as SEQ ID NO. 1. The SNP locus is obviously related to the survival rate of the hybrid pelteobagrus fulvidraco of 'Huangyou No. 1' under the stress of hypoxia, and the hypoxia tolerance of the individual with the locus genotype TT is obviously stronger than that of the individual with the genotype AA. The SNP locus disclosed by the invention can not be limited by the age, sex and the like of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1', is suitable for screening and classifying hypoxia-resistant seedlings of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1', and can effectively improve the survival rate of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' under the hypoxia condition in culture and transportation. The method is accurate and reliable, and is simple and convenient to operate.
Description
Technical Field
The invention relates to a SNP molecular marker related to the hypoxia tolerance character of hybrid pelteobagrus fulvidraco 'Huangyou No. 1' and application thereof, belonging to the field of pelteobagrus fulvidraco genetic breeding.
Background
The hybrid pelteobagrus fulvidraco 'yellow superior No. 1' is an F1 generation obtained by taking 3 generation bred pelteobagrus fulvidraco as a female parent and 2 generation bred pelteobagrus vachelli as a male parent through artificial hybridization. Compared with parents, the feed has the characteristics of high growth speed, strong stress resistance, high feed utilization rate and the like. In the aspect of hypoxia resistance, experiments prove that the hypoxia resistance of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' is between that of the parent and the female. Therefore, the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' still faces the problem of water body hypoxia in production, culture and long-distance transportation. The low oxygen environment of the water body can obviously influence the ingestion and growth activities of the fish body and even cause the endocrine disturbance of the fish body.
The protein product of the Vhl gene is VHL protein (pVHL), the most important target protein of the β functional region is hypoxia-inducible factor (HIF), the VHL protein can regulate HIF-1 α protein from the protein level and promote HIF-1 α mRNA expression, when pVHL expression is abnormal, the HIF protein expression in cells can be obviously increased, physiological processes such as angiogenesis, apoptosis, cell cycle, cell energy metabolism and the like are influenced, and then the fish body is influenced, so the Vhl gene plays an important role in hypoxia regulation.
Single Nucleotide Polymorphism (SNP), which refers to a sequence polymorphism caused by a Single nucleotide (A, G, C, T) variation (transition, transversion, insertion and deletion) at a specific site in a genomic DNA sequence, was proposed by Lander E in 1996. SNPs are third generation genetic markers of DNA molecules developed following microsatellites. The most predominant mutations of SNPs are transitions and transversions, and occur in a ratio of 2: 1, so the SNPs are generally said to be allelic polymorphic. The SNP has the characteristics of high density, high genetic stability and the like, can be used for drawing a genetic map, finely positions a target region by starting whole genome tracking, quickly associates markers and characters, and accelerates QTL positioning. The SNP marker has wide application in plant molecular genetics due to wide whole genome distribution range, high density and adaptability of an ultrahigh-flux detection platform, and is then applied to genetic diversity research, genetic linkage map construction, molecular marker assisted breeding and excellent germplasm protection research of aquatic animals.
However, no research related to screening the hypoxia-resistant property of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' by using SNP molecular markers is disclosed in the prior art at present. Therefore, the invention utilizes SNP molecular markers to screen hypoxia-resistant related genes of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1', selects groups which are more hypoxia-resistant on the existing hypoxia-resistant level, realizes the high-efficiency breeding industry of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1', and creates benefits for farmers.
Disclosure of Invention
The purpose of the invention is as follows: in view of the above technical problems, a first object of the present invention is to provide an SNP molecular marker associated with the hypoxia tolerance trait of the hybrid pelteobagrus fulvidraco "huangyou No. 1", namely, a molecular marker selection is performed by using SNP sites significantly associated with hypoxia tolerance on the gene sequence of the hybrid pelteobagrus fulvidraco "huangyou No. 1" Vhl.
The second objective of the invention is to provide a pair of primers for detecting the SNP molecular marker.
The third purpose of the invention is to provide the application of the SNP molecular marker.
The technical scheme is as follows: in order to achieve the purpose, the invention adopts the following technical scheme:
the invention discloses an SNP molecular marker related to the hypoxia-resistant character of hybrid pelteobagrus fulvidraco ' Huangyou No. 1 ', wherein the SNP molecular marker takes a cDNA sequence of von Hippel-Lindau disease turmor supressor (Vhl gene) as a template to design a primer, the amplified genome sequence is shown as SEQ ID NO. 1, from the 5 ' end, the 495 th site of SNP g.495T > A exists in the sequence, namely the SNP site is located at the 495 th site on the SEQ ID NO. 1, and the base is T or A.
The hypoxia tolerance of the TT genotype individual with the SNPg.495T > A locus is obviously higher than that of the AA genotype individual.
The primer pair for detecting the SNP molecular marker comprises the following components:
the sequence of the upstream primer is 5'-TGAAGAGTCCGAGCGATT-3' (SEQ ID NO: 2);
the sequence of the downstream primer is 5'-ACTGCGAGGAGGATGAAC-3' (SEQ ID NO: 3).
A kit for detecting the SNP molecular marker, which comprises the primer pair.
The SNP molecular marker, the primer pair or the kit is applied to the screening of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' auxiliary hypoxia-resistant offspring seeds. A method for detecting the hypoxia tolerance character of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' comprises the steps of carrying out detection on the SNP molecular marker of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' to be detected, and determining the hypoxia tolerance capability of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' to be detected, wherein the hypoxia tolerance capability mainly comprises two parts of data of the hypoxia sensitive tolerant group of the front 100 and the hypoxia tolerant group of the back 100.
Further, the method comprises the steps of extracting the genomic DNA of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' to be detected, carrying out PCR amplification by using the primer pair or the kit, detecting whether the genotype of the SNP site is TT, TA or AA, and finally carrying out correlation analysis on different genotypes and the hypoxia resistance of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1', wherein the hypoxia resistance is the survival time under the hypoxia condition.
The method for detecting the hypoxia resistance of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' specifically comprises the following steps:
a) obtaining a hybrid pelteobagrus fulvidraco 'Huangyou No. 1' hypoxia-sensitive population and a hypoxia-tolerant population;
b) DNA extraction of tail fins of the two groups of the hybrid pelteobagrus fulvidraco of 'Huangyou No. 1';
c) based on the primer pair, carrying out PCR amplification on the genomic DNA of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' to obtain a PCR amplification product;
d) sequencing the PCR amplification product, and determining the genotype of the SNP molecular marker based on the sequencing result;
e) and (3) carrying out correlation analysis on SNPg.495T > A locus genotype and hypoxia resistance of hybrid pelteobagrus fulvidraco 'Huangyou No. 1'.
Through the extraction of the genome DNA of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1', PCR amplification, sequencing of amplification products and analysis of sequencing results, the hypoxia tolerance of TT genotype individuals with SNPg.495T > A sites is obviously higher than that of AA genotype individuals.
The invention takes the mononucleotide polymorphic site of the Vhl gene of 'Huangyou No. 1' of the hybrid pelteobagrus fulvidraco as a research target, and finds that the SNP site (SNPg.495T > A) of the Vhl gene sequence is obviously related to the hypoxia tolerance of 'Huangyou No. 1' of the hybrid pelteobagrus fulvidraco, wherein the hypoxia tolerance of TT genotype individuals of the SNPg.495T > A site is obviously higher than that of AA genotype (P <0.05) individuals. In the individual breeding process of breeding edible hybrid pelteobagrus fulvidraco 'Huangyou No. 1' which is easier to survive in an anoxic environment by taking hypoxia resistance as a breeding index, individuals of which the SNPg.495T > A locus genotype is TT can be preferentially selected as breeding objects, which has important significance for improving the breeding benefit of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1'.
The technical effects are as follows: compared with the prior art, the invention has the following advantages:
1) the SNP locus disclosed by the invention can not be limited by the age, sex and the like of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1', and can be used for selecting and classifying hypoxia-resistant seedlings of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1', so that the survival rate of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' under the hypoxia condition is improved, and the breeding economic benefit of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' is obviously increased.
2) The method for detecting the 495 th SNP locus from the 5' end of the nucleotide sequence shown in SEQ ID NO. 1 by a pair of primers shown in SEQ ID NO. 2 and SEQ ID NO. 3 is accurate, reliable and convenient to operate.
Drawings
FIG. 1 is a partial peak diagram of sequencing of the 495 th site from the 5 ' end of the AA genotype hybrid pelteobagrus fulvidraco ' Huangyou No. 1 ' Vhl gene;
FIG. 2 is a partial peak diagram of sequencing of the 495 th site from the 5 ' end of a TT genotype hybrid pelteobagrus fulvidraco ' Huangyou No. 1 ' Vhl gene;
FIG. 3 is a partial peak diagram of sequencing of the TA genotype hybrid pelteobagrus fulvidraco "Huangyou No. 1" Vhl gene from the 495 th site of the 5' end.
Detailed Description
The technical solution of the present invention is further described in detail by the following specific examples.
The invention is based on a pair of primers shown in SEQ ID NO. 2 and SEQ ID NO. 3, and performs extraction, PCR amplification, amplification product sequencing and sequencing result analysis on the genome DNA of the hybridized pelteobagrus fulvidraco, namely 'Huangyou No. 1'. The obtained SNP locus (SNPg.495T > A) of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' surviving in the hypoxia environment can be applied to the selection and cultivation of the early hypoxia-resistant juvenile fish of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1'.
Examples
a) Obtaining a hybrid pelteobagrus fulvidraco 'Huangyou No. 1' hypoxia-sensitive population and a hypoxia-tolerant population;
b) DNA extraction of tail fins of the two groups of the hybrid pelteobagrus fulvidraco of 'Huangyou No. 1';
c) carrying out PCR amplification on the genomic DNA of the hybridized pelteobagrus fulvidraco of 'Huangyou No. 1' based on the SNP primer;
d) sequencing the PCR amplification product, and determining the genotype of the SNP based on the sequencing result;
e) carrying out correlation analysis on SNP locus genotype and hypoxia resistance of hybrid pelteobagrus fulvidraco 'Huangyou No. 1';
f) the SNP locus is applied to the selection of different hypoxia tolerance groups of the hybrid pelteobagrus fulvidraco of 'Huangyou No. 1'.
The specific operation is as follows:
a) obtaining a hybrid pelteobagrus fulvidraco 'Huangyou No. 1' hypoxia-sensitive population and a hypoxia-tolerant population:
the experimental fishes are all pond-cultured fishes which are taken from the Lukou base of the research institute of aquatic science in Nanjing of Jiangsu in 2018 months, and the 5-month-old hybrid pelteobagrus fulvidraco 'Huangyou No. 1' 505 tails (the body length is 9 +/-2.1 cm, and the body mass is 12 +/-2.3 g) are counted. Before the experiment, the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' is randomly distributed into 5 aquaculture glass ponds with biofilters, wherein the water flow is 5L/min (0.8 m x0.55m x 0.4m (L x W x H); 24 +/-1 ℃; and (5) temporarily culturing the duck with the pH value of 7.5 +/-0.2. In the experiment, 445 healthy and disease-free hybrid pelteobagrus fulvidraco of 'Huangyou No. 1' is selected and placed in a 400L aquarium. At the beginning of the experiment, oxygen aeration and water inlet were closed, nitrogen gas was introduced to discharge oxygen in the water, dissolved oxygen in the water was measured by an oxygen dissolution instrument (measured by the LDO101 probe portable by the american LDO HQd), and when the dissolved oxygen in the water could not drop (about 0.9mg/L), the aquarium was sealed with a transparent film to allow natural oxygen reduction. In the experimental process, the activities of the fish are observed, timing is started when the first hybrid pelteobagrus fulvidraco 'Huangyou No. 1' loses balance and begins to sink, and 100 tails of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' which firstly sinks into the bottom of the pool and finally sinks into the bottom of the pool are taken as hypoxia-sensitive and hypoxia-tolerant groups of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1'.
b) Clipping the tail fin of the fish body, storing at-80 ℃ and extracting the genome DNA.
c) Extracting DNA of 'Huangyou No. 1' of the hybrid pelteobagrus fulvidraco:
(1) taking 15mg tail fins, adding 400 mu L ACL Solution, shearing, adding 10 mu L Proteinase K, shaking and mixing uniformly for 1 minute, and standing at 55 ℃ for about 2 hours until the lysate is clear.
(2) Then 300. mu. LExt solution and 300. mu. LAB solution were added in this order, shaken vigorously, and centrifuged at 12,000rpm for 5 minutes.
(3) The tip was passed through the upper solution deep into the lower solution and the solution was carefully aspirated into the GenClean Column, trying to avoid aspiration into the upper solution and precipitation in the middle layer.
(4) Centrifuging at 8000rpm for 1 min, taking down GenClean Column, and pouring off waste liquid in the collecting tube.
(5) GenClean Column was returned to the collection tube, 500. mu.L of land Solution was added, and centrifuged at 8,000rpm for 1 minute at room temperature.
(6) Repeating the step (5) once.
(7) The GenClean column was removed and the waste stream from the collection tube was discarded. The column was placed back into the collection tube and centrifuged at 12,000rpm for 1 minute at room temperature to remove residual Wash Solution.
(8) The column was placed in a fresh, clean 1.5mL centrifuge tube, 60. mu.l of Elution Buffer was added to the center of the column, and the column was left at room temperature for 2 minutes. Then centrifuged at 12,000rpm for 1 minute at room temperature. The liquid in the centrifuge tube is the extracted DNA, and the DNA is preserved at the temperature of minus 20 ℃.
d) Based on the SNP primer, the PCR amplification is carried out on the genomic DNA of the hybrid pelteobagrus fulvidraco' Huangyou No. 1
The amplification length of the PCR product is 213bp, and the PCR primers are as follows:
an upstream primer: 5'-TGAAGAGTCCGAGCGATT-3'
A downstream primer: 5'-ACTGCGAGGAGGATGAAC-3'
The PCR reaction system was 20. mu.L of 2 XTaq Master Max 10. mu.L, forward and reverse primers 0.8. mu.L each, DNA template 1. mu.l, and sterilized water 7.4. mu.l.
The PCR reaction is carried out for 35 cycles in total, wherein the pre-denaturation is carried out at 94 ℃ for 5min before the cycles, and each cycle comprises denaturation at 94 ℃ for 30s, annealing at 60 ℃ for 30s and extension at 72 ℃ for 30 s; after the circulation was completed, the extension was carried out at 72 ℃ for 5 min.
And carrying out electrophoresis detection on the amplification product by using 2% agarose gel, and storing the qualified PCR product at-20 ℃ for subsequent sequencing reaction.
e) Sequencing the PCR amplification product, and determining the genotype of SNP based on the sequencing result
Based on a Hiseq2000 high-throughput sequencing platform, the PCR amplification products of 200 individuals of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' are subjected to forward sequencing on an ABI3730 sequencer. Based on the sequencing result, the SNP locus of the hybrid pelteobagrus fulvidraco of 'Huangyou No. 1' is subjected to genotyping.
f) Correlation analysis of SNP locus genotype and low oxygen resistance of hybrid pelteobagrus fulvidraco' Huangyou No. 1
The SNP locus genotype and hypoxia tolerance time of 200 individuals of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' are shown in table 1: (see last Table 1)
From the data in Table 1, a linear analysis model was constructed using the SPSS (19.0) GLM program based on the traits and the characteristics of the test population to perform association analysis between gene polymorphisms and traits: where yij is u + Gi + eij, yij is a trait phenotype value, u is a population mean, Gi is a marker genotype effect (i 1, 2, 1, 4), and eij is a random residual effect. Statistical data are expressed as mean ± sem. The association of SNP site genotype with hypoxia tolerance is shown in Table 2.
TABLE 2 relativity of SNP site of "Huangyou No. 1" Vhl gene of hybrid pelteobagrus fulvidraco with hypoxia tolerance
The results show that: TT genotype and T allele of low-oxygen tolerant group in three genotypes of SNPg.495T > A are dominant, and AA genotype and A allele of low-oxygen sensitive group are dominant. The result shows that in the process of cultivating the hypoxia-resistant hybrid pelteobagrus fulvidraco 'Huangyou No. 1', the SNPg.495T > A is an SNP site which is obviously related to the hypoxia resistance.
g) The SNP locus is applied to the group screening of the hypoxia-resistant hybrid pelteobagrus fulvidraco 'Huangyou No. 1'.
On the hybrid pelteobagrus fulvidraco 'huangyou No. 1' Vhl gene as described above, snpg.495t > a sites are obviously dominant in the TT genotype individuals in the hypoxia tolerant population, and the dominant population in the hypoxia sensitive population is AA genotype individuals, and TA genotype individuals have no significant difference in both populations. Therefore, in the breeding process, individuals with the SNPg.495T > A locus genotype TT can be preferentially selected as breeding objects to culture the hypoxia-resistant hybrid pelteobagrus fulvidraco 'Huangyou No. 1'.
According to the invention, the 100 tails of the 'Huangyou No. 1' hypoxia-sensitive population of the hybrid pelteobagrus fulvidraco and the 100 tails of the hypoxia-tolerant population are selected as samples, and by analyzing the single nucleotide polymorphism sites of each individual Vhl gene, the remarkable relevance of SNPg.495T > A and the hypoxia-tolerant capability of the 'Huangyou No. 1' of the hybrid pelteobagrus fulvidraco is found, and the test result can be effectively supported. Therefore, in the selective breeding process of the hypoxia-resistant hybrid pelteobagrus fulvidraco 'Huangyou No. 1', individuals with SNPg.495T > A site genotype as TT can be preferentially selected as breeding objects, and the selection of the hypoxia-resistant hybrid pelteobagrus fulvidraco 'Huangyou No. 1' excellent variety in a short time, at low cost and with high accuracy can be effectively assisted.
Table 1:
A. hybrid pelteobagrus fulvidraco 'Huangyou No. 1' 100-tailed hypoxia-sensitive population hypoxia time and individual SNP site genotype
B. Hybrid pelteobagrus fulvidraco 'Huangyou No. 1' 100-tailed hypoxia tolerance group hypoxia time and individual SNP locus genotype
A
B
Note: the anoxic time is measured by taking the first fish losing balance and sinking to be 0min, and the time measurement unit is (min).
Sequence listing
<110> university of Nanjing university
<120> SNP molecular marker related to hybrid pelteobagrus fulvidraco 'Huangyou No. 1' hypoxia tolerance character and application thereof
<160>3
<170>SIPOSequenceListing 1.0
<210>1
<211>1249
<212>DNA
<213>Vhl(Vhl)
<400>1
acgtgatctc gatgcacact agcttcctca ccacctgcag gcaacgttcc ctcagggaat 60
acactaacac acacacacac acacgtcata tacaaaaccg tgagattaat aagagaagaa 120
gaattattac acagcctgtt tagccaaacc accctgaccc catgcactct gtgcgtgatg 180
atagttaccg ggtagcgtga tgaccactgg ggcttggttg ttgtctcgag cactgggcaa 240
atacatctcc ttattattca ccaacatcgg gtcatccgtc tgagcatcac gaaacatcca 300
cggatgtcct gtcagaaata aacagtcgtc gtcatctttc accatcatac attactcttt 360
cctcttacgt tcttaatttt taatactctt tatgctcatt ttagtagctt caacattaga 420
cttccaaagc cttattcgga tcggtttagt ttattagtct gtaaagactt gctgttgtga 480
tgatgatgat gatgatatac tctgagaggt tcaacacatc actgagaata tgattacgga 540
aaaaaaagca taatattgca cgatattgtt tcttacacta ccatgcacct tatgtacata 600
actgatcagt cctatattct gtattcatat ttaatactca ttctgtctat attgtctcac 660
atagtctgta ttgtctggtc ttgtacagta tagtgttatt tatgtctgta cttttcagag 720
tcaaaaacag ctggaaccaa gttccttgtg tgtggcagga gtgtggtagc ctagtggtta 780
atgcgtcgga ctactgcccg gaaggtcatg agttcaaatc ccagctccaa caagctgccc 840
gtgctgggcc cctgagcaag gcccttaatg ctcagttgta taaaatgaaa taaaatgtaa 900
gtcactctgt ataagtgtgt ctgcttaatg ccagaaatgt aaatgtcaat aaacctgatt 960
cagattcata tatgacaaat atatcccttt ctaatttgca tatataatat atgcacatat 1020
gaatatgcat acatatgcat gattcatgta gcagtcagtt ttccaagcta gttttctctg 1080
ccactatcta gagacggtta atcattggtc tggatgatgt tctagaaaac actggtgcac 1140
ttttacttag atgtaagtgt tggatagtct gaaaagatga atgatgatga tgatgatgat 1200
gatgagaaga agaggatcag acgtaacata gcagatttct cctcctgac 1249
<210>2
<211>18
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>2
tgaagagtcc gagcgatt 18
<210>3
<211>18
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>3
actgcgagga ggatgaac 18
Claims (8)
1. An SNP molecular marker related to the hypoxia tolerance character of hybrid pelteobagrus fulvidraco 'Huangyou No. 1', which is characterized in that the sequence of the SNP molecular marker is shown as SEQ ID NO. 1, and the SNP molecular marker is positioned in the sequence shown as SEQ ID NO:1, and the base is T or A at position 495 of the sequence shown in the specification.
2. The SNP molecular marker related to the hypoxia tolerance trait of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' as claimed in claim 1, wherein the hypoxia tolerance capability of TT genotype individuals at SNP sites is significantly higher than that of AA genotype individuals.
3. A primer pair for detecting the SNP molecular marker according to claim 1 or 2, comprising a forward primer and a reverse primer, wherein the nucleotide sequences of the primer pair are respectively as follows:
a forward primer: 5'-TGAAGAGTCCGAGCGATT-3', as shown in SEQ ID NO: 2.
Reverse primer: 5'-ACTGCGAGGAGGATGAAC-3', as shown in SEQ ID NO. 3.
4. A kit for detecting the SNP molecular marker according to claim 1 or 2, comprising the primer set according to claim 3.
5. The use of the SNP molecular marker of claim 1 or 2, the primer pair of claim 3 or the kit of claim 4 in the screening of the auxiliary hypoxia-resistant offspring seeds of hybrid pelteobagrus fulvidraco 'Huangyou No. 1'.
6. A method for detecting the hypoxia-resistant property of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1', which is characterized by comprising the step of detecting the SNP molecular marker in claim 1 or 2 on the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' to be detected, and determining the hypoxia-resistant capability of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' to be detected.
7. The method for detecting the hypoxia-resistant property of the hybrid pelteobagrus fulvidraco 'huangyou 1' according to claim 6, which is characterized by comprising the steps of extracting genomic DNA of the hybrid pelteobagrus fulvidraco 'huangyou 1' to be detected, carrying out PCR amplification by using the primer pair of claim 3 or the kit of claim 4, detecting whether the genotype of the SNP site is TT, TA or AA, and finally carrying out correlation analysis of different genotypes and the hypoxia-resistant property of the hybrid pelteobagrus fulvidraco 'huangyou 1', wherein the hypoxia-resistant capability is survival time under a hypoxia condition.
8. The method for detecting the hypoxia tolerance of the hybrid pelteobagrus fulvidraco 'Huangyou No. 1' according to claim 7, wherein the hypoxia tolerance of TT genotype individuals at the SNP sites is significantly higher than that of AA genotype individuals.
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| CN114752679B (en) * | 2022-04-06 | 2023-09-22 | 南京师范大学 | Kit for rapidly identifying hypoxia tolerance of pelteobagrus vachelli population and identification method |
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