CN108913782A - For detecting the primer pair of trypetid Idgf4 gene expression amount - Google Patents
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Abstract
The invention discloses the primer pairs for detecting trypetid Idgf4 gene expression amount.Primer pair disclosed by the invention two single stranded DNAs shown in sequence 3 in sequence table and sequence 4 form.It is demonstrated experimentally that special, quickly and accurately quantitative detection can be carried out to the Idgf4 gene of citrus fruit fly and Bactrocera correcta provided by the present invention for the primer pair for detecting trypetid Idgf4 gene expression amount, it can be used for the evaluation of trypetid Idgf4 gene jamming effectiveness.In addition, quantitative detection solves the comparison problem of same gene expression quantity between different trypetids, and easy to operate, quick, at low cost.The present invention can also provide tool and related target for the research of Idgf4 gene and the prevention and treatment of trypetid; have great importance to the prevention and treatment of trypetid, and effectively to control the diffusion of risk trypetid, protection agricultural production and ecological safety, economic trade development in China's being promoted to provide potential target gene reference.
Description
Technical field
The present invention relates in field of biotechnology, for detecting the primer pair of trypetid Idgf4 gene expression amount.
Background technique
Citrus fruit fly Bactrocera dorsalis (Hendel) is subordinate to Diptera Diptera, Tephritidae
Tephritidae, Bactrocera Bactrocera, the worm are polyphagy pest, can 46 sections such as feeding citrus, guava, mango
250 various fruits vegetables.With larva, feeding and adult lay eggs to form hole in fruit surface inside the fruit of water fruits and vegetables
Hole causes shedding or entire fruit rot to cause harm, this not only brings serious harm to fruits and vegetables production, but also gives foreign trade band
To seriously affect.Since host range is wide for the worm, reproductive capacity is high, adaptable, and many countries are all classified as emphasis quarantine
Object (refined etc., 2008).
Citrus fruit fly originates in the torrid areas in Asia, now has a very wide distribution, and is related to the heat of the Asian-Pacific area and African Territories
Band, subtropical zone and temperate zone region.The population generation quantity of this worm is big, often causes heavy losses to the fruits and vegetables production on new invasion ground
(Chen Naizhong, 2009).Since 2012, the trypetid worker from countries and regions such as Australia from morphometry,
Science of heredity, behaviouristics, chemical ecology etc. have studied citrus fruit fly in Bactrocera dorsalis complex, pawpaw fruit fly
B.papayae Drew&Hancock, Philippine fruit fly B.philippinensis Drew&Hancock, invasion fruit fly
B.invadens Drew and Carambola fruit trypetid B.carambolae Drew&Hancock, the results show that pawpaw fruit fly, Fei Lv
Bingo trypetid and invasion fruit fly are citrus fruit fly, are its synonyms, and Carambola fruit trypetid is another species (Schutze
et al,2012；2014a；2014b).This research is reported in October, 2014 by FAO official website." four close for this
One " new discovery makes the distribution of citrus fruit fly have bigger expansion, forms global diffusion, this is to global real
The scientific research of fly type pests produces great influence, also proposed further requirement to invasion prevention and control.
Bactrocera correcta B.correcta (Bezzi) originates in Tropical Asian and subtropical zone (Wang Xingjian and Zhao Ming
Pearl, 1989), now it is distributed mainly on the countries such as Thailand, Vietnam, Burma, Nepal, India, Pakistan.It occurs only at home
Yunnan Province (beam is extensively diligent etc., 1996).Polyphagy pest, based on Perenniporia martius fruits and vegetables (beam is extensively diligent etc., 1996), including kind
More than 30 section 60 such as pomegranate, mango, cherry gourd, fruit and vegetable (White IM et al, 1992).Its endanger be Adult worms producting eggs in
On the inside of pericarp, young wormy fruit causes fungal infection and fruit rot is caused to fall off.The worm was just put into early in 1992《China
People's republic enters the territory plant quarantine danger venereal disease, worm, weed species》, and it is higher in port quarantine intercepting and capturing frequency, to guava
There are important economic significances for the important gene correlative study of fruit fly and effectively detection.
Insect cuticle is made of chitin and epidermal protein, and major function is to maintain body structure, inhibits moisture evaporation
With the influence for resisting extraneous poor environment, in terms of drug resistance, insect cuticle enters insect intracorporal as chemical insecticide
One of defence line plays very important important function.CG5210 gene (Idgf6 gene) and Idgf4 gene are under the jurisdiction of to table
Skin develops 18 family of glycoside hydrolase to play an important role, and two genes are as imaginal discs growth factor (Imaginal-disc-
Growth-factor it) plays an important role for the normal development of trypetid and emergence.In allied species drosophila Drosophila
The silencing of two genes is proved to will cause epidermis injury and the anamorphosis in drosophila larvae period, or even will cause development
Death in the process.One is presented always very in a kind of Idgf gene of important stage of silkworm Bombyx mori wing bud development
High level, and Idgf gene to it is finned at cell differentiation it is related with development.In the research of control of insect, in view of two kinds
Conservative of the gene in insect assembly and its important function in growth and development, two kinds of genes can grinding for new pesticides
The specific target gene loci of offer is provided.Producing corresponding double-strand product or preparation can be such that insect leads because epithelial barrier is impaired
Premature death is caused, and increases insect for the infection rate of pathogenic bacteria, can be used as the target of the new pesticide of exploitation control of insect
Gene.
Citrus fruit fly and Bactrocera correcta are huge to fruits and vegetables production trade harm as Important Economic quarantine pest insect.
And two kinds of trypetids all have stronger diffusivity and drug resistance, so the expression water of research CG5210 gene and Idgf4 gene
It is flat to provide important references for the invasion prevention and control of two kinds of Important Economic trypetids.
Summary of the invention
The technical problem to be solved in the present invention is to provide the real-time fluorescences of citrus fruit fly and Bactrocera correcta Idgf4 gene
PCR quantitative detecting method.
In order to solve the above technical problems, the primer pair is by sequence 3 in sequence table present invention firstly provides a kind of primer pair
With two single stranded DNA compositions shown in sequence 4.
The primer pair can be used for detecting trypetid Idgf4 gene expression amount.
The molar ratio of two single stranded DNAs in the primer pair can be 1:1.Two single stranded DNAs can independent packaging.
In order to solve the above technical problems, the present invention also provides the system for detecting trypetid Idgf4 gene expression amount, institute
State primer pair described in system.
In above system, the system also includes carry out quantitative PCR detection gene expression amount needed for other reagents and/or
Instrument.
Other reagents needed for the progress quantitative PCR detection gene expression amount can have for hundred million biotechnology of Beijing China Boulder
The Takara of limit company Premix Ex TaqTMII (Tli RNaseH Plus) seminal plasma fructose detection kit.
The instrument can be Applied Biosystems7500 Real-Time PCR System.
It is described can as the primer pair and carry out quantitative PCR detection gene expression amount needed for other reagents and/or instrument
Composition.
The system can be for only including the kit of reagent.
In order to solve the above technical problems, the present invention also provides the method for detection trypetid Idgf4 gene expression amount, the side
Method includes：The Idgf4 gene expression amount that trypetid to be measured is detected using the primer pair obtains the trypetid Idgf4 gene to be measured
Expression quantity.
In the above method, the trypetid to be measured can be citrus fruit fly and/or Bactrocera correcta.
In order to solve the above technical problems, the present invention also provides following any applications of the primer pair or the system：
X1) the application in detection trypetid Idgf4 gene expression amount；
X2) the application in preparation detection trypetid Idgf4 gene expression amount product；
X3) the application in detection trypetid Idgf4 gene；
X4) the application in preparation detection trypetid Idgf4 gene prod；
X5) the application in prevention and treatment trypetid；
X6) the application in identification trypetid；
X7) the application in preparation identification trypetid product；
X8) the application in detection trypetid Idgf4 gene jamming effectiveness；
X9) in detection trypetid prevention product to the application in trypetid Idgf4 gene jamming effectiveness；
X10) the application in detection trypetid control efficiency；
X11) in detection trypetid prevention product to the application in trypetid evaluation.
In the present invention, the trypetid can be citrus fruit fly and/or Bactrocera correcta.The trypetid concretely reality
Larva, pupa and/or the adult of fly.
It is demonstrated experimentally that can be special, fast provided by the present invention for the primer pair for detecting trypetid Idgf4 gene expression amount
Speed accurately carries out quantitative detection to the Idgf4 gene of citrus fruit fly and Bactrocera correcta, can be used for trypetid Idgf4 gene
The evaluation of jamming effectiveness.In addition, the cost during quantitative detection is low, easy to operate, solves same gene in different trypetids
Between expression quantity comparison problem, and in actual production two kinds of trypetid same genes use it is more square with a pair of of primer detection
Just, fast, it is at low cost.The present invention can also provide tool and related target for the research of Idgf4 gene and the prevention and treatment of trypetid, right
The prevention and treatment of trypetid has great importance, and for effectively control risk trypetid diffusion, protection agricultural production and ecological safety,
Economic trade development in China's is promoted to provide potential target gene reference.
Detailed description of the invention
Fig. 1 is the testing result of reference gene (18s rRNA) in citrus fruit fly (Bd) and Bactrocera correcta (Bc).
Fig. 2 is specific detection result of the CG5210-rt in citrus fruit fly (Bd) and Bactrocera correcta (Bc).
Fig. 3 is specific detection result of the Idgf4-rt in citrus fruit fly (Bd) and Bactrocera correcta (Bc).
Fig. 4 is CG5210-rt in 1 instar larvae (L1) of citrus fruit fly (Bd) and Bactrocera correcta (Bc), 3 age early larvaes
(L3-1), 3 latter stage in age larvas (L3-3), early stage pupa (P-E), mid-term pupa (P-M), latter stage pupa (P-L), just sprouted wings non-sexal maturity at
Expression quantity in worm (A1) and ten age in days sexal maturity adults (A10) tests and analyzes result.No significant difference is indicated between same letter,
It is indicated between different letters by there were significant differences (P<0.05).
Fig. 5 is Idgf4-rt in 1 instar larvae (L1) of citrus fruit fly (Bd) and Bactrocera correcta (Bc), 3 age early larvaes
(L3-1), 3 latter stage in age larvas (L3-3), early stage pupa (P-E), mid-term pupa (P-M), latter stage pupa (P-L), just sprouted wings non-sexal maturity at
Expression quantity in worm (A1) and ten age in days sexal maturity adults (A10) tests and analyzes result.No significant difference is indicated between same letter,
It is indicated between different letters by there were significant differences (P<0.05).
Specific embodiment
The present invention is further described in detail With reference to embodiment, and the embodiment provided is only for explaining
The bright present invention, the range being not intended to be limiting of the invention.Experimental method in following embodiments is unless otherwise specified
Conventional method.Material as used in the following examples, reagent, instrument etc., are commercially available unless otherwise specified.
Quantitative test in following embodiment, the technology three times that is respectively provided with repeats experiment and six groups of biology repeat to test, and is as a result averaged
Value.In following embodiments, unless otherwise specified, the 1st of each nucleotide sequence is the 5 ' end cores of corresponding DNA in sequence table
Thuja acid, last bit are the 3 ' terminal nucleotides of corresponding DNA.
The preparation of embodiment 1, primer pair for detecting trypetid CG5210 and Idgf4 gene expression amount
The primer pair for detecting trypetid CG5210 and Idgf4 gene expression amount is present embodiments provided, is denoted as respectively
CG5210-rt and Idgf4-rt.CG5210-rt two single stranded DNAs shown in sequence 1 in sequence table and sequence 2 form, this draws
The molar ratio of two single stranded DNAs of object centering is 1:1, and two equal independent packagings of single stranded DNA.Idgf4-rt is by sequence in sequence table
3 and sequence 4 shown in two single stranded DNAs composition, the molar ratio of two single stranded DNAs is 1 in the primer pair:1, and two are single-stranded
The equal independent packaging of DNA.
Embodiment 2 detects trypetid CG5210 and Idgf4 gene expression amount using the primer pair of embodiment 1
Trypetid to be measured：It picks up from the Bactrocera correcta in Yunnan and picks up from the citrus fruit fly in Guangzhou, every kind of trypetid first-instar young
50, second instar larvae 30, three age early larvaes (5 day old larva from after egg hatching) 30, three latter stage in age larvas are (from egg hatching
7 day old larva afterwards) 20, early stage pupa (1 age in days pupa after pupating certainly) 6, mid-term pupa (5 age in days pupa after pupating certainly) 6, latter stage pupa
Non- sexal maturity adult 10, ten age in days sexal maturity adults 10 after (9 age in days pupa after pupating certainly) 6, emergence.
Each trypetid is extracted using the RNAsimple total RNA extraction reagent box of TIANGEN Biotech (Beijing) Co., Ltd.
Total serum IgE, then reverse transcription obtains cDNA.The cDNA obtained using citrus fruit fly and Bactrocera correcta reverse transcription is used as template
The Takara of hundred million Bioisystech Co., Ltd of Beijing China Boulder Premix Ex TaqTMII(Tli RNaseH Plus)
Kit, be utilized respectively embodiment 1 CG5210-rt and Idgf4-rt carry out real-time fluorescence PCR quantitative detection CG5210 and
Idgf4 gene expression amount, instrument are Applied Biosystems7500Real-Time PCR System instrument, with
ddH2O replaces template as negative control.
The reaction system of real-time fluorescence PCR is as follows：
Wherein, single stranded DNA shown in sequence 1 and sequence 3 is forward primer, single stranded DNA shown in sequence 2 and sequence 4
It is reverse primer.
The response procedures of real-time fluorescence PCR are：95 DEG C of 30sec of initial denaturation；Then 95 DEG C of 5sec, 52 DEG C of 34sec circulations 40
It is secondary；95℃15sec；52℃1min；95℃30sec.
Reference gene is 18s rRNA, and primer sequence (Hu et al., 2014) is：
Forward primer 18s rRNA-rt-F：5'-GCGAGAGGTGAAATTCTTGG-3'；
Reverse primer 18s rRNA-rt-R：5'-CGGGTAAGCGACTGAGAGAG-3'.
As a result (Fig. 1-5) is shown, two primer pairs can carry out related gene in citrus fruit fly and Bactrocera correcta
Amplification it is quantitative, and can be effectively for including that the different development stages such as larva, pupa, adult be detected, and result is stable, repeats
Good, the specific height of property.The result shows that two genes are expressed in eight developmental stages of two kinds of trypetids, CG5210-rt is small in tangerine
It expresses without significant difference in trypetid except there is conspicuousness to promote remaining outer period in three latter stage in age larvas and sexal maturity adult,
And in Bactrocera correcta expression quantity conspicuousness promotion concentrate in latter stage pupa and sexal maturity adult.Two kinds of trypetid genes
The expression quantity main distinction of CG5210 is in the middle and later periods and sexal maturity adult of pupa.Idgf4-rt in citrus fruit fly early stage pupa with
There is conspicuousness promotion in sexal maturity adult, remaining period expresses without significant difference, and the expression quantity in Bactrocera correcta
Conspicuousness promotion concentrate on pupa later period and adult stage.The expression quantity main distinction of two kinds of trypetid gene Idgf4 is before pupa
In later period and early stage adult.The above result shows that the two primer pairs can detecte and compare the different worm states of two kinds of trypetids three and relate to
And the expression of eight different times related genes.
The amplified production difference sample different times successfully expanded are subjected to bidirectional sequencing, the results show that CG5210-
The sequence for the product that rt expands citrus fruit fly is sequence 5 in sequence table, expand to Bactrocera correcta
To product sequence be sequence table in sequence 7；The sequence for the product that Idgf4-rt expands citrus fruit fly is sequence
Sequence 6 in list, the sequence to the product that Bactrocera correcta is expanded are sequence 8 in sequence table.Two kinds of primer pairs
It can accurately expand the DNA fragmentation to corresponding gene.
The optimization of template quantity needed for embodiment 3, the primer pair of embodiment 1
The cDNA that embodiment 2 is obtained carries out 10 times of gradient dilutions respectively, obtains the dilution of the specific developmental stage of each trypetid
(content of cDNA is respectively 100 μ g/ μ l, 10 μ g/ μ l, 1 μ g/ μ to 10 times, 100 times, 1000 times and 10000 times of cDNA dilution
L and 0.1 μ g/ μ l), then optimal Template amount, and benefit are determined using the reaction system and response procedures of embodiment 2 as template
With water, negative control is set.
The results show that the cDNA after being 10 times of dilution using 100 μ g/ μ l is template, two kinds of primer pairs can be in the small reality of tangerine
Obtain positive amplification curve in fly and Bactrocera correcta, and the amplification curve of reference gene and target gene be it is unimodal simultaneously
Meet reference gene CT and is maintained at cDNA requisite quality and reproducible, detection target in 10 or so each sample treatments of explanation
The CT value of gene is in 20-30 zone of reasonableness；And with the cDNA of 10 μ g/ μ l, 1 μ g/ μ l and 0.1 μ g/ μ l for template, two kinds
Primer pair cannot obtain the CT value in target gene zone of reasonableness, i.e. target gene in citrus fruit fly and Bactrocera correcta
CT > 30, also no positive amplification curve in negative control.Show that cDNA concentration is that 100 μ g/ μ l are (i.e. dilute by the cDNA of embodiment 2
Release 10 times) when expanding effect it is best；The sensitivity of primer pair CG5210-rt and Idgf4-rt are 100 μ g/ μ l.
The detection of comparative example 1, other primer pairs to CG5210 and Idgf4 gene expression amount
1, it is expanded for CG5210 using other primer pairs are carried out, the primer is：
CG5210-rt-F-1：GGCTGAGGAACATAAGGAA；
CG5210-rt-R-1：GCGGGCACATCATAGAA.
Amplification system and amplification condition are the same as embodiment 2.
The primer pair is bad to expanding effect in the cDNA of citrus fruit fly and Bactrocera correcta, in sample segment and portion
Timesharing phase CT value is greater than 30, and solubility curve is not unimodal, and is unable to get successfully sequencing result.
2, it is expanded for Idgf4 using other primer pairs, the primer is to for Idgf4-rt-1 and Idgf4-rt-2.
The sequence of two primers of Idgf4-rt-1 is as follows：
Idgf4-rt-F-1：CCAAATCCCGCCAATCA；
Idgf4-rt-R-1：ATGCCGCCCAAACCTCT.
The sequence of two primers of Idgf4-rt-2 is as follows：
Idgf4-rt-F-2：TACCAAATCCCGCCAATC；
Idgf4-rt-R-2：ATGCCGCCCAAACCTCT.
The two primer pairs are bad to expanding effect in the cDNA of citrus fruit fly and Bactrocera correcta, in sample segment
It is not unimodal with some period dissolution amplification curve, and CT value is greater than 30, and is unable to get successfully sequencing result.
<110>China Agricultural University
<120>For detecting the primer pair of trypetid Idgf4 gene expression amount
<160> 8
<170> PatentIn version 3.5
<210> 1
<211> 17
<212> DNA
<213>Artificial sequence
<220>
<223>
<400> 1
cggacgagaa gagcagc 17
<210> 2
<211> 17
<212> DNA
<213>Artificial sequence
<220>
<223>
<400> 2
ggcacgcagt atgggat 17
<210> 3
<211> 17
<212> DNA
<213>Artificial sequence
<220>
<223>
<400> 3
cgcataccgt tcataaatag 17
<210> 4
<211> 20
<212> DNA
<213>Artificial sequence
<220>
<223>
<400> 4
20
atgttcctcc gccttct
<210> 5
<211> 177
<212> DNA
<213>Citrus fruit fly
<400> 5
cggacgagaa gagcagcggt catggcttgt gggtgggcta tgaagatccc gacaccgcgg 60
ctgagaaggc cggctactgt gcgtcaattc aatttgggtg gcgtggccct gtttgatttg 120
tctttcgatg acttccgtgg ttcctgcact ggcgataaat atcccatact gcgtgcc 177
<210> 6
<211> 186
<212> DNA
<213>Citrus fruit fly
<400> 6
cgcataccgt tcataaatag tgcgcactct ttggtgaaga cttacggttt cgatggtctc 60
gatttggctt ggcaattccc caagaataag gccaaaaagg tgcatagcgg catcggtaaa 120
ttgtggaagg gtttcaagaa aattttctct ggcgactttg tggtcgatga gaaggcggag 180
gaacat 186
<210> 7
<211> 176
<212> DNA
<213>Bactrocera correcta
<400> 7
cggacgagaa gagcagcggc cacggcttgt gggtgggcta tgaagatccc gacaccgcgg 60
ctgagaaggc cggctacgtg cgtcaattca atttgggtgg cgtggctctg tttgatttgt 120
ctttcgatga cttccgtggt tcctgcactg gcgataaata tcccatactg cgtgcc 176
<210> 8
<211> 186
<212> DNA
<213>Bactrocera correcta
<400> 8
cgcataccgt tcataaatag tgcgcattct ttggtgaaga cttacggttt cgatggtctc 60
gatttggctt ggcaattccc caagaacaag gccaaaaagg tgcatagcgg catcggtaaa 120
ttgtggaagg gtttcaagaa aattttctct ggcgactttg tggtcgatga gaaggcggag 180
gaacat 186
Claims (10)
1. primer pair, two single stranded DNAs shown in sequence 3 in sequence table and sequence 4 are formed.
2. primer pair according to claim 1, it is characterised in that：The primer pair is for detecting trypetid Idgf4 gene table
Up to amount.
3. the system for detecting trypetid Idgf4 gene expression amount, including primer pair of any of claims 1 or 2.
4. system according to claim 3, it is characterised in that：The system also includes carry out quantitative PCR detection gene table
Other reagents and/or instrument needed for up to amount.
5. system according to claim 4, it is characterised in that：Carry out quantitative PCR detection gene expression amount needed for other
Reagent is the Takara of hundred million Bioisystech Co., Ltd of Beijing China Boulder Premix Ex TaqTMII(Tli RNaseH
Plus) seminal plasma fructose detection kit.
6. the method for detecting trypetid Idgf4 gene expression amount, including：Reality to be measured is detected using primer pair described in claim 1
The Idgf4 gene expression amount of fly obtains the trypetid Idgf4 gene expression amount to be measured.
7. according to the method described in claim 6, it is characterized in that：The trypetid to be measured is citrus fruit fly and/or Fructus psidii guajavae immaturus
Trypetid.
8. any system or claim 6 in primer pair according to claim 1 or 2 or claim 3-5
Or method described in 7, it is characterised in that：The trypetid is citrus fruit fly and/or Bactrocera correcta.
9. primer pair according to claim 8, system or method, it is characterised in that：The trypetid be larva, pupa and/or
Adult.
10. following any applications of any system in primer pair of any of claims 1 or 2 or claim 3-5：
X1) the application in detection trypetid Idgf4 gene expression amount；
X2) the application in preparation detection trypetid Idgf4 gene expression amount product；
X3) the application in detection trypetid Idgf4 gene；
X4) the application in preparation detection trypetid Idgf4 gene prod；
X5) the application in prevention and treatment trypetid；
X6) the application in identification trypetid；
X7) the application in preparation identification trypetid product；
X8) the application in detection trypetid Idgf4 gene jamming effectiveness；
X9) in detection trypetid prevention product to the application in trypetid Idgf4 gene jamming effectiveness；
X10) the application in detection trypetid control efficiency；
X11) in detection trypetid prevention product to the application in trypetid control efficiency.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114752682A (en) * | 2022-04-14 | 2022-07-15 | 中南大学 | Pupal-stage day age inference kit for autophagic flies and application of kit |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6060590A (en) * | 1998-03-31 | 2000-05-09 | The Regents Of The University Of California | Chitinase related proteins and methods of use |
| US20020120008A1 (en) * | 2000-06-29 | 2002-08-29 | Seymour Benzer | Life extension of drosophila by a drug treatment |
-
2018
- 2018-07-04 CN CN201810721991.0A patent/CN108913782B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6060590A (en) * | 1998-03-31 | 2000-05-09 | The Regents Of The University Of California | Chitinase related proteins and methods of use |
| US20020120008A1 (en) * | 2000-06-29 | 2002-08-29 | Seymour Benzer | Life extension of drosophila by a drug treatment |
Non-Patent Citations (3)
| Title |
|---|
| LIU ET AL.: "Genome-wide identification of chitinase and chitin deacetylase gene families in the oriental fruit fly, Bactrocera dorsalis (Hendel)", 《COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY - PART D》 * |
| PESCH ET AL.: "Chitinases and Imaginal disc growth factors organize the extracellular matrix formation at barrier tissues in insects", 《SCIENTIFIC REPORTS》 * |
| 董小龙等: "橘小实蝇RNAi效应的持效期研究", 《湖北植保》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114752682A (en) * | 2022-04-14 | 2022-07-15 | 中南大学 | Pupal-stage day age inference kit for autophagic flies and application of kit |
| CN114752682B (en) * | 2022-04-14 | 2023-09-08 | 中南大学 | Cadaver fly pupa day-to-day age deducing kit and application |
Also Published As
| Publication number | Publication date |
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| CN108913782B (en) | 2021-10-15 |
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