CN103451290A - Primer pair, kit and detection method for detecting abamectin resistance of tetranychus urticae - Google Patents
Primer pair, kit and detection method for detecting abamectin resistance of tetranychus urticae Download PDFInfo
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Abstract
The invention relates to the field of molecular detection, and in particular relates to a primer pair, a kit and a detection method for judging abamectin resistance of tetranychus urticae. The primer pair comprises (1) upstream primers for detecting resistant and susceptible populations, which are RF:5'-AAGCTGTTGACATTTGGACGA-3' and SF:5'-AAGCTGTTGACATTTGGACGG-3', and (2) a common downstream primer which is paired with the upstream primers for use. According to the primer pair, the kit and the detection method, the resistance or susceptibility of tetranychus urticae to abamectin is judged by detecting GluCl (Glutamate-gated chloride channel) gene mutation in tetranychus urticae. The primer pair, the kit and the detection method have the advantages of high speed, effectiveness, high sensitivity, capability of realizing high-flux detection of a large number of samples and the like, and have the effects of monitoring the resistance gene frequency and resistance development trend of the field population of tetranychus urticae to abamectin, timely adjusting a prevention and control strategy for tetranychus urticae, guiding rational drug use and delaying the resistance development.
Description
Technical field
The present invention relates to the Molecular Detection field, be specifically related to for detection of the drug-fast primer pair of the Avrmectin of Tetranychus urticae, test kit and detection method.
Background technology
Tetranychus urticae Tetranychus urticae belongs to spider guiding principle Acari Acariformes Tetranychidae, is commonly called as red spider, is global Important Agricultural Mites, and the host of causing harm is various, comprises important farm crop such as fruit tree, vegetables and cotton, wheat etc.Cause harm if tetranychid is stung to inhale with the back side that becomes mite and children mite to be gathered in plant leaf, cause on blade and the chlorosis spot occurs, when serious, cause blade to dry up and come off, greatly affect the yield and quality of crop.
Because the tetranychid build is small, the generation generation is many, reproduction speed is fast, development duration is short, this mite very easily develops immunity to drugs.Avrmectin is as the antibiotics insecticidal/acaricidal agent, by stimulating polypide to produce γ-aminobutyric acid, the transmission of blocking-up motorius information, make insect paralysis, food refusal, slow-action or motionless rapidly in several hours, therefore, this medicament was once becoming the first-selected medicament of control Tetranychus urticae.But experiment shows 20 generations of two classes of Avrmectin subculture seed selections tetranychid, Tetranychus urticae can produce higher resistance to Avrmectin.And there is larger difference in the medicining condition of China each department, cause Tetranychus urticae also different to the resistance of Avrmectin, in order to instruct better peasant's rational use of drug, just need the resistance of clear and definite Tetranychus urticae to Avrmectin.Yet, traditional Insecticide Resistance of Spider Mites monitoring adopts the slide pickling process, the small operation of examination worm is difficult for, and need test worm amount very large (at least 700), ability observation test result after at least 24 hours, by the contrast with relative sensitive strain, just known its Detection of insecticide resistance to medicament, therefore expend time in very long.
L-glutamic acid is important neurotransmitter in vertebrates and invertebrates neural system, L-glutamic acid is after postsynaptic receptor is combined, open chloride channel, be called the chloride channel (Glutamate-gated chloride channel, GluCl) of L-glutamic acid gate.Former studies shows, L-glutamic acid gate chloride channel is one of action target of avermectins, the second cross-film district downstream appearance point sudden change (9299S) of GluCl causes fruit bat to produce 10 times of resistance (Kane et al. to ivermectin, 2000), also found the point mutation (Njue et al., 2004) of this gene different loci on nematode.Kwon et al. (2010, Insect Molecular Biology, 19 (4): the molecular mechanism that 583-591) in report Tetranychus urticae resistant population, the point mutation (G323D) of GluCl causes it to develop immunity to drugs to Avrmectin.
On this basis, the present invention is directed to this point mutation, the specific fragment that comprises this point mutation site in design Tetranychus urticae resistance and sensitive population, adopt specific alleles pcr amplification technology (PCR amplification of specific alleles, PASA, also claim allele-specific PCR, AS-PCR) amplify specific band respectively from resistance and sensitive population, according to direct differentiation Tetranychus urticae of having or not of band individual (population), whether Avrmectin has been produced to resistance.The method is easy, quick, can realize that the Tetranychus urticae population is to the drug-fast high throughput testing of Avrmectin.
Summary of the invention
The purpose of this invention is to provide for detection of Tetranychus urticae individual (population) whether Avrmectin has been produced to drug-fast primer pair.
A further object of the present invention is to provide for detection of Tetranychus urticae individual (population) whether Avrmectin has been produced to drug-fast test kit.
A further object of the present invention is to provide detection Tetranychus urticae individual (population) whether Avrmectin has been produced to drug-fast method.
The invention provides for detection of Tetranychus urticae whether Avrmectin has been produced to drug-fast primer pair, it comprises:
(1) detection of resistance and sensitive population is respectively with upstream primer: RF:5 '-AAGCTGTTGACATTTGGAC
g
a-3 ' and SF:5 '-AAGCTGTTGACATTTGGAC
g
g-3 ';
And the shared downstream primer used with the pairing of above-mentioned upstream primer (2), preferably, during pcr amplification and public downstream primer (3'405F) match and increased respectively.RF and 3'405F(TGA ATC CTT GGC GGT GTC AAA) the constitutive resistance primer pair, SF and 3'405F(TGA ATC CTT GGC GGT GTC AAA) form responsive primer pair.
Also provide for detection of Tetranychus urticae according to the present invention whether Avrmectin has been produced to drug-fast test kit, described test kit comprises:
(1) detection of resistance and sensitive population is respectively with upstream primer: RF:5 '-AAGCTGTTGACATTTGGAC
g
a-3 ' and SF:5 '-AAGCTGTTGACATTTGGAC
g
g-3 ';
And the shared downstream primer used with the pairing of above-mentioned upstream primer (2), preferably, during pcr amplification and public downstream primer (3'405F) match and increased respectively.RF and 3'405F(TGA ATC CTT GGC GGT GTC AAA) the constitutive resistance primer pair, SF and 3'405F(TGA ATC CTT GGC GGT GTC AAA) form responsive primer pair.
Whether detection Tetranychus urticae according to the present invention has produced drug-fast method to Avrmectin, and described method comprises uses following primer pair to carry out the step of pcr amplification:
(1) resistance primer pair, comprise upstream primer RF:5 '-AAGCTGTTGACATTTGGAC
g
a-3 ' and shared downstream primer, preferred 3'405F(TGA ATC CTT GGC GGT GTC AAA); And
(2) responsive primer pair, comprise upstream primer SF:5 '-AAGCTGTTGACATTTGGAC
g
g-3 ' and shared downstream primer, preferred 3'405F(TGA ATC CTT GGC GGT GTC AAA).
GluCl sequencing result to Avrmectin resistance and sensitive population adopts DNAMAN to carry out sequential analysis relatively, upstream, site to the origination point sudden change arranges the PASA primer, and the detection of resistance and sensitive population is respectively with upstream primer: RF:5 '-AAGCTGTTGACATTTGGAC
g
a-3 ' and SF:5 '-AAGCTGTTGACATTTGGAC
g
g-3 '
The present invention is by analyzing the GluCl specific fragment of 405bp, GluCl sequencing result to Avrmectin resistance and sensitive population adopts the DNAMAN5.0 version to carry out sequential analysis relatively, discovery a point mutation (G in sensitive population sport resistant population in A) occurs at 251 in this amplified fragments, namely variation (by G, sporting A) has occurred in 968 Nucleotide in the open reading frame of Tetranychus urticae GluCl complete sequence (ORF), this point mutation causes the aminoacid sequence of 323 of its ORF to sport aspartic acid (D) by glycine (G), the 84th amino acids in increased specific fragment sports aspartic acid (D) by glycine (G), mutational site identifies with square frame in two groups of aminoacid sequences below.
The aminoacid sequence of the GluCl specific fragment before sudden change is as shown in SEQIDNo.1
GEYSCLKVELVFKREFSYYLFLIYVPCCMLVIVSWVSFWIDPNSAAARVLLGVTSLLTMSRQISGINASLPPVSYTKAVDIWT
CCLIFVFGALIEFAIVNYVSRTDTIKADKHRRRRPQGIVGARRKFDTAKDS
The aminoacid sequence of the GluCl specific fragment after sudden change is as shown in SEQIDNo.2
GEYSCLKVELVFKREFSYYLFLIYVPCCMLVIVSWVSFWIDPNSAAARVLLGVTSLLTMSRQISGINASLPPVSYTKAVDIWT
CCLIFVFGALIEFAIVNYVSRTDTIKADKHRRRRPQGIVGARRKFDTAKDS
Two pairs of Specific primer pairs provided by the invention and amplification method thereof can fast and effeciently identify the G323D sudden change whether the Tetranychus urticae population L-glutamic acid gate chloride channel (GluCl) gene has occurred, detailed process is: the Tetranychus urticae genomic dna to be measured of take is template, adopt respectively responsive Auele Specific Primer to the specific resistance primer pair, carrying out pcr amplification, if adopt responsive primer pair amplification to obtain the amplified fragments of 176bp, and adopt the resistance primer pair not obtain the amplified production of 176bp simultaneously, Tetranychus urticae to be measured is the GG genotype sensitive individual that isozygotys, if adopt the resistance primer pair to obtain the amplified production of 176bp, and adopt responsive primer not obtain this amplified production simultaneously, Tetranychus urticae to be measured is the AA genotype Resistant Individuals that isozygotys, if adopt responsive primer pair and resistance primer pair all to obtain respectively the amplified production of 176bp, Tetranychus urticae to be measured is GA gene hybridizing type (during the GA gene hybridizing type, being resistance to the phenotype of Avrmectin).
By the detection to GluCl transgenation in the Tetranychus urticae body, thereby the judgement Tetranychus urticae is for resistance or the susceptibility of Avrmectin, have quick, effective, highly sensitive, can realize the advantages such as high throughput testing to gross sample, for monitoring Tetranychus urticae field population, resistance gene frequency and the development of drug resistance of Avrmectin are adjusted to the control strategy of Tetranychus urticae dynamically, in time, instruct the effect of rational use of drug, delaying drug resistance development.
The accompanying drawing explanation
Fig. 1 shows the glutamate receptor gene fragment of primer amplified sensitivity and resistance Tetranychus urticae population, Marker:Marker I; 4 S swimming lanes represent sensitive population; 4 R swimming lanes represent resistant population; CK represents the negative control that clear water is template.
Fig. 2 shows the detected result in mutational site in resistance and sensitive population, M:Marker I; The 1-3 swimming lane is the indoor sensitive population of Tetranychus urticae, and amplimer is to being SF+5 ' 405R, and 4 swimming lanes are blank; The 5-7 swimming lane is the Tetranychus urticae resistant population, and amplimer is to being RF+5 ' 405R, and 8 is blank; 9-11 is the indoor sensitive population of Tetranychus urticae, and amplimer is to being RF+5 ' 405R, and the 12-14 swimming lane is the Tetranychus urticae resistant population, and amplimer is to being SF+5 ' 405R.
Annotate: in above-mentioned test, each population detects respectively 30 individualities, and the electrophoresis picture here only shows 3.
Embodiment
Materials and methods
1. try worm, medicament and reagent
The indoor sensitive population of Tetranychus urticae gathered from Jinan, Shandong Province in June, 2009, and indoor employing Avrmectin medicament carries out being defined as sensitive population after biological assay.From 2009 so far, this population adopts sponge leaf dish method in indoor insect growth case, by " Bi Feng " kind carry out the subculture raising without worm Kidney bean seedling leaf as the host, during do not contact any sterilant.The raising condition is 26 ± 1 ℃, and the photoperiod is L:D=16h:8h.The field resistance population gathers respectively the eggplant field from Haidian, Beijing, Huairou, after an indoor feeding generation, is tested.
Miticide is 1.8% abamectin emulsifiable concentrate, by Beijing ZhongNon Da Biology Technology Co., Ltd, is produced.
The primer is synthetic by Shanghai Ying Jun bio-engineering corporation, and the sequence order-checking is held up biotechnology company limited of section by Beijing and carried out.The polysaccharase used in pcr amplification is that 2 * Taq10MASTER Mix (0.1U TaqE/ μ l) is purchased from Beijing skill Development Co., Ltd of Olympic Competition Boke.The genome DNA extracting reagent kit of tetranychid is purchased from Beijing hundred Tyke Bioisystech Co., Ltd.
2, the biological assay of the female one-tenth mite of tetranychid
The standard method that the Toxicity Determination of tetranychid is recommended with reference to Food and Argriculture OrganizationFAO-slide pickling process (Slide-dip method) (FAO, 1980) is carried out.Double sticky tape is cut into to 2～3cm long, be attached to an end of slide glass, throw off the scraps of paper on double sticky tape with tweezers, with No. zero writing brush select in the same size, body colour is bright-coloured, active female one-tenth mite takes action, its back is bonded on double sticky tape, do not cling mite foot, mite palpus and mouthpart, every sticky 3 row of adhesive tape, approximately sticky 15～20 head lobe mites of every row.In temperature, be 26 ± 1 ℃, photoperiod 16h:8h(L:D), place 4h in the insect biochemical cultivation case of relative humidity 60% after, with Olympus binocular anatomical lens, observe, strictly reject dead or not too active and the inappropriate tetranychid individuality of position.Each medicament dilutes 5～7 concentration on the basis of pilot study, the end with the mite slide is soaked into the liquid, takes out after shaking gently 5s, blots mite body and unnecessary liquid on every side thereof with thieving paper rapidly.The moistening gauze of paving one deck is in white Porcelain Jar, and the slide that soaked liquid is positioned in large porcelain dish, covers the moistening gauze of one deck on Porcelain Jar again, then covers sheet glass to reach the effect of moisturizing, and prevents that the gauze downslide from falling on slide.With band mite slide dipping clear water in contrast, each is processed and repeats 3～4 times.After 24h, by binocular anatomical lens check result, record is survived and dead tetranychid quantity respectively.Touch the mite body with writing brush, take the motionless person of mite foot as dead, the control treatment mortality ratio is being efficiency test below 10%.Testing data adopts Probit software (Feng et al., Pest Management Science, 2010,66 (3): 313-318.) processed, calculate slope, SE value, LC50 value and 95% fiducial limit thereof etc. of medicament, calculate the resistant multiple of each resistant population.
1 pair of Avrmectin resistance of embodiment and sensitive population detect the acquisition with Auele Specific Primer
1, the extraction of tetranychid genomic dna
The genome DNA extracting method of the female one-tenth mite of single head adopts the DNA extraction test kit (solution-type) of Beijing hundred Tektronix Ltd., and method reference reagent box specification sheets carries out.The concentration of single head tetranychid genomic dna is measured as 6.60ng/ μ l by Nanodrop2000c.
2, the genomic pcr amplification of tetranychid and order-checking
According to Kwon et al(2010) report Tetranychus urticae L-glutamic acid gate chloride channel (GluCl) point mutation site and sequence, design packet contains the primer pair of the GluCl fragment of this point mutation: 5'405F:GGG GAA TAC AGC TGT CTC AAA, 3'405F:TGA ATC CTT GGC GGT GTC AAA, indoor and genomic dna field resistance Tetranychus urticae population increases.The pcr amplification system is: 2 * Taq10MASTER Mix is 10 μ l, and DNA profiling is 1 μ l, and upstream and downstream primer (being respectively 10 μ M) adds respectively 1 μ l, and last water is supplemented to 20 μ l.The pcr amplification program of setting is: 95 ℃ of denaturation 3min at first; Then carry out 35 circulations (72 ℃ are extended 1min for 95 ℃ of sex change 30s, 58 ℃ of annealing 30s), finally at 72 ℃ of condition downward-extension 10min.Get PCR product 8 μ l detects on 1.2% agarose gel electrophoresis.The specific PCR product obtained after Qiagen purification kit purifying, send Beijing Qing Ke Bioisystech Co., Ltd to adopt upstream and downstream primer (5'405F and 3'405F) to carry out two-way sequencing through product.
3, the point mutation detection of resistance and sensitive population
GluCl sequencing result to Avrmectin resistance and sensitive population adopts DNAMAN to carry out sequential analysis relatively, upstream, site to the origination point sudden change arranges the PASA primer, and the detection of resistance and sensitive population is respectively with upstream primer: RF:5 '-AAGCTGTTGACATTTGGAC
g
a-3 ' and SF:5 '-AAGCTGTTGACATTTGGAC
g
g-3 '.
During pcr amplification and public downstream primer (3'405F) match and increased respectively.RF and 3'405F constitutive resistance primer pair, SF and 3'405F form responsive primer pair.The pcr amplification system is as follows: upstream and downstream primer (10 μ M) adds respectively 0.5 μ l, DNA profiling 1 μ l, and 2 * Es Mix (0.1U TaqE/ μ l) is 10 μ l, finally adds ddH
2o complements to 25 μ l.Amplification program is: 94 ℃ of denaturation 3min, carry out 30 circulations (72 ℃ are extended 30s for 94 ℃ of sex change 20s, 62 ℃ of annealing 30s), subsequently finally at 72 ℃ of condition downward-extension 3min.Sample the result of 8 μ l at the gene DNA of 1.5% electrophoresis detection resistance primer pair and responsive primer pair amplification different population Tetranychus urticae individuality.
2 pairs of Avrmectin resistances of embodiment and sensitive population detect to be analyzed
1, the biological assay of different population Tetranychus urticae
Table 1 Tetranychus urticae sensitivity and the resistant population susceptibility measurement result to Avrmectin
| Population | b±SE | LC 50(mg/L) | 95% fiducial interval | The resistant multiple |
| Sensitive population | 1.12(±0.17) | 0.093 | 0.0486-0.176 | 1.00 |
| The Huairou population | 0.81（±0.14） | 343.54 | 144.73-815.44 | 3693.98 |
| Haidian population | 1.61(±0.19) | 201.13 | 142.60-283.68 | 2162.69 |
As can be seen from Table 1, Huairou population and Haidian population have produced high horizontal resistance to Avrmectin respectively, to the female of Huairou and Haidian tetranychid population, become the LC50 value of mite to be respectively 343.54mg/L and 201.13mg/L, with indoor relative sensitive population, compare, the resistant multiple reaches respectively 3693.98 times and 2162.69 times.Because the resistant multiple of Huairou population is higher, so the resistant population in next step Fast Detection Technique research and development test adopts the Huairou population to carry out.
2, resistance and sensitive population GluCl gene fragment amplification and sequence alignment
Adopt the genomic dna of the Auele Specific Primer of above-mentioned amplification GluCl to amplification Tetranychus urticae sensitivity and resistant population, all obtain the specific fragment (Fig. 1) of a 405bp size.
GluCl sequencing result to Avrmectin resistance and sensitive population adopts the DNAMAN5.0 version to carry out sequential analysis relatively, discovery a point mutation (G in sensitive population sport resistant population in A) occurs at 251 in this amplified fragments, namely variation (by G, sporting A) has occurred in 968 Nucleotide in the open reading frame of Tetranychus urticae GluCl complete sequence (ORF), and this point mutation causes the aminoacid sequence of 323 of its ORF to sport aspartic acid (D) by glycine (G).
3, the detection in resistance and sensitive population mutational site
The above-mentioned RF primer and the SF primer that detect resistance and responsive Tetranychus urticae population are matched with public downstream primer (3'405F) respectively, and resistance and the sensitive population of amplification Tetranychus urticae, the results are shown in Figure 2.As seen from Figure 2, primer pair SF+5 ' 405R during to the amplification of indoor sensitive population, can stablize the band that a 176bp left and right occurs, and during the amplification resistant population without band; During primer pair RF+5 ' 405R amplification resistant population, the band of a 176bp can appear, and during the amplification sensitive population without band.Further sequencing result also shows that this 176bp band is a part of sequence of tail end in above-mentioned 405bp sequence, is also the band of expection amplification.The primer pair for resistance and sensitive population and amplification condition that above-mentioned design is described are all feasible and stable, can detect for the resistance gene frequency of next step different population.
4, resistant population gene mutation site frequency detecting
To the Tetranychus urticae resistant population in Huairou, Beijing and Haidian, randomly draw 30 head lobe mite samples, adopt two pairs of mutational sites detection primer pairs of above-mentioned foundation to carry out respectively pcr amplification, detect the mutation frequency of resistant gene in each resistant population, the results are shown in Table 2.
GluCl gene mutation frequency detected result in table 2 Tetranychus urticae different population
As can be seen from Table 2, in indoor responsive Tetranychus urticae population, adopt can the increase specific band of 176bp of responsive primer pair, during the amplification of resistance primer pair, have no band, resistant gene do not detected.In Huairou, Beijing, two, Haidian resistant population, the resistance gene frequency of Tetranychus urticae sample there are differences, and the resistance gene frequency of Huairou population reaches 96.66%, and the resistance gene frequency of Haidian population is 90%.This result has also obtained checking by order-checking, illustrates that accuracy rate can reach 100%.
Claims (6)
1. for detection of the drug-fast primer pair of mite Avrmectin of two variegated leafs, it is characterized in that, described primer pair comprises:
(1) the detection upstream primer of resistance and sensitive population, be respectively RF:5 '-AAGCTGTTGACATTTGGAC
g
a-3 ' and SF:5 '-AAGCTGTTGACATTTGGAC
g
g-3 ';
And the shared downstream primer used with the pairing of above-mentioned upstream primer (2).
2. the drug-fast primer pair of the Avrmectin for detection of Tetranychus urticae according to claim 1, is characterized in that, described downstream primer is primer 3'405F:TGA ATC CTT GGC GGT GTC AAA.
3. for detection of the drug-fast test kit of the Avrmectin of Tetranychus urticae, it is characterized in that, described test kit comprises:
(1) detection of resistance and sensitive population is respectively with upstream primer: RF:5 '-AAGCTGTTGACATTTGGAC
g
a-3 ' and SF:5 '-AAGCTGTTGACATTTGGAC
g
g-3 ';
And the shared downstream primer used with the pairing of above-mentioned upstream primer (2).
4. the drug-fast test kit of the Avrmectin for detection of Tetranychus urticae according to claim 3, is characterized in that, described downstream primer is primer 3'405F:TGA ATC CTT GGC GGT GTC AAA.
5. detect the drug-fast method of Avrmectin of Tetranychus urticae, it is characterized in that, described method comprises uses following primer pair to carry out the step of pcr amplification:
(1) resistance primer pair, comprise upstream primer RF:5 '-AAGCTGTTGACATTTGGAC
g
a-3 ' and shared downstream primer; And
(2) responsive primer pair, comprise upstream primer SF:5 '-AAGCTGTTGACATTTGGAC
g
g-3 ' and shared downstream primer.
6. the drug-fast method of the Avrmectin of detection Tetranychus urticae according to claim 5, is characterized in that, described downstream primer is primer 3'405F:TGA ATC CTT GGC GGT GTC AAA.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104032028A (en) * | 2014-07-01 | 2014-09-10 | 中国农业科学院蔬菜花卉研究所 | Rapid molecular detection method for drug resistance of tetranychus urticae to pyrethroid insecticide |
| CN104789642A (en) * | 2014-01-20 | 2015-07-22 | 北京乐普医疗科技有限责任公司 | Primer for detecting polymorphism of mononucleotide, and kit and detection method thereof |
| CN107267632A (en) * | 2017-07-18 | 2017-10-20 | 西南大学 | The new small peaceful mite of Pasteur is to the molecular labeling of AVM resistance and its application and detection method |
| CN107841539A (en) * | 2017-12-13 | 2018-03-27 | 中国农业科学院蔬菜花卉研究所 | A kind of CAPS labeling methods for detecting tetranychid and being mutated to AVM resistance related gene |
| CN114480316A (en) * | 2022-04-02 | 2022-05-13 | 中国农业科学院蔬菜花卉研究所 | Cytochrome P450 and application thereof in degrading pesticide residues |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104789642A (en) * | 2014-01-20 | 2015-07-22 | 北京乐普医疗科技有限责任公司 | Primer for detecting polymorphism of mononucleotide, and kit and detection method thereof |
| CN104032028A (en) * | 2014-07-01 | 2014-09-10 | 中国农业科学院蔬菜花卉研究所 | Rapid molecular detection method for drug resistance of tetranychus urticae to pyrethroid insecticide |
| CN107267632A (en) * | 2017-07-18 | 2017-10-20 | 西南大学 | The new small peaceful mite of Pasteur is to the molecular labeling of AVM resistance and its application and detection method |
| CN107267632B (en) * | 2017-07-18 | 2019-11-01 | 西南大学 | The new small peaceful mite of Pasteur is to the molecular labeling of avermectin resistance and its application and detection method |
| CN107841539A (en) * | 2017-12-13 | 2018-03-27 | 中国农业科学院蔬菜花卉研究所 | A kind of CAPS labeling methods for detecting tetranychid and being mutated to AVM resistance related gene |
| CN114480316A (en) * | 2022-04-02 | 2022-05-13 | 中国农业科学院蔬菜花卉研究所 | Cytochrome P450 and application thereof in degrading pesticide residues |
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