CN102586405A - Molecular marker authenticating method for related gene of cucumber fruit length - Google Patents
Molecular marker authenticating method for related gene of cucumber fruit length Download PDFInfo
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- CN102586405A CN102586405A CN2011100041027A CN201110004102A CN102586405A CN 102586405 A CN102586405 A CN 102586405A CN 2011100041027 A CN2011100041027 A CN 2011100041027A CN 201110004102 A CN201110004102 A CN 201110004102A CN 102586405 A CN102586405 A CN 102586405A
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Abstract
The invention relates to a molecular marker authenticating method for the related gene of cucumber fruit length, which comprises the following steps of: taking the deoxyribose nucleic acid (DNA) of cucumber variety or strain with unknown fruit length as a template, taking CSWCT28 as a primer to carry out polymerase chain reaction (PCR) amplification, and carrying out 3% agarose gel electrophoresis detection on an amplified product or carrying out 7.5% denaturing polyacrylamide gel electrophoresis detection on the amplified product, wherein if the amplified banding pattern is consistent with the banding pattern of cucumber Dongnong 803 with known short fruit length, the cucumber variety or strain with the unknown fruit length is the variety or strain with short fruit length , and the PCR amplification has the procedures: previously denaturating for 5min under 94 DEG C, denaturating for 30s under 94 DEG C, annealing for 1min under 47 DEG C, carrying out 35 cycles, extending for 1min under 72 DEG C, extending for 10min under 72 DEG C, and finally storing under 4 DEG C. The method is used for authenticating the related gene of the cucumber fruit length.
Description
Technical field:
The present invention relates to a kind of molecular marker identification method of cucumber fruits length genes involved, belong to plant biotechnology field.
Background technology:
The improvement of cucumber quality proterties is the important goal of breeding research, and fruit length is one of important quality proterties of cucumber, directly influences economic benefit.Along with the fast development of commodity economy and the improvement of people's living condition, the consumption pattern of cucumber becomes more diverse, as eat raw, cold and dressed with sauce, stir-fry and eat, soup food, pickles, salt marsh, sugaring, system are done, system jar etc.People require difference to the cucumber length of different purposes, need short fruit type kind like acid pickling processing, and the salt marsh slice processing then need be grown fruit type kind; Cucumber fruits with neat length can satisfy the requirement of mechanize harvesting and the processing of industrial flow-line product.Therefore, the research of cucumber fruit long correlation has great importance aspect people's consumers demand satisfying.
Cucumber fruits belongs to Peponidium, is grown in the lump by ovary and holder to form, and pericarp is the appearance of holder in fact, and edible portion is pericarp and placenta.Cucumber has the parthenocarpy ability, and the ability power is different because of kind, and pollination can improve fruit-setting rate and promote fruit development.The cucumber fruits growth curve is S-type, thanks and spends the back poor growth, quickens gradually later on, slows down gradually again after growing into to a certain degree, and the back of generally blooming reached the commodity maturation in 10～15 days.The day growth amount is bigger night, and daytime is less.The growth at night is very fast with at dusk, and dawn is slower.The growth curve of cucumber fruits meets Y=a+bX+cX
2+ dX
3Mathematical model; The first flex point place is very fast period of fruit growth speed; It is the critical period that cucumber yield and quality form; But relative growth rate the fastest period because of kind different (document that sees reference, Pan Dandan. the cytology research that cucumber fruits is grown. Northeast Agricultural University's master thesis, 2009).To the research of cucumber fruits length, focused mostly at relation, environmental factors (temperature, illumination, moisture, the CO of cytology, endogenous hormones and fruit length in the past
2, mineral nutrition) aspects such as influence that cucumber fruits is grown, and it is less that cucumber fruits length is carried out the report of genetic analysis and molecule marker correlative study.Therefore seek and the closely linked molecule marker of cucumber fruits length genes involved, have important application value for accelerating the breed cucumber process.
Summary of the invention:
The objective of the invention is to provide a kind of molecular marker identification method of cucumber fruits length genes involved to the problem of above-mentioned existence; Can utilize above-mentioned cucumber fruits length genes involved molecule marker better, and cucumber fruits length genes involved molecule marker is applied to the assisted Selection of breed cucumber.
Above-mentioned purpose realizes through following technical scheme:
The molecular marker identification method of cucumber fruits length genes involved, this method comprises the steps:
With the cucumber variety of fruit length the unknown or the DNA of strain is template, is that primer carries out pcr amplification with CSWCT28, amplified production is carried out the detection of 3% agarose gel electrophoresis perhaps amplified production is carried out 7.5% denaturing polyacrylamide gel electrophoresis detection; If the banding pattern that amplification obtains is consistent with agricultural 803 banding patterns in the cucumber of known short fruit length east, cucumber variety that then this fruit length is unknown or strain are the kind or the strain of short fruit length, and the program of said pcr amplification is: 94 ℃ of preparatory sex change 5min; 94 ℃ of sex change 30s, 47 ℃ of annealing 1min, 35 circulations; 72 ℃ are extended 1min; 72 ℃ are extended 10min, last 4 ℃ of preservations, and described CSWCT28 is:
Primer 1:5 '-GAATTCAAAAGCATTTCAAAACTA-3 ',
Primer 2: 5 '-GAATTCAATTGGGTTTTTGAACCC-3 '.
The molecular marker identification method of described cucumber fruits length genes involved; In the said pcr amplification; The PCR reaction system of per 20 μ l is following: 10 * buffer:2 μ l, 2mM dNTP:2 μ l, the Primer 1:0.8 μ l of 10pmol/ μ l; The Primer 2:0.8 μ l of 10pmol/ μ l, 25mM MgCl
2: 2.5 μ l, the TaqDNA polysaccharase of 5U/ μ l: 0.2 μ l, the template DNA of 60ng/ μ l: 1 μ l, ddH20:10.7 μ l.
The application of the molecular marker identification method of cucumber fruits length genes involved in assisted Selection breed cucumber material.
Beneficial effect:
1. the present invention directly shows with the form of DNA; Each tissue, each etap organism all can detect, and do not receive season, environmental restraint, do not exist expression whether to wait problem; Therefore; The present invention just can identify the fruit length of cucumber variety and strain and screens in seedling stage through molecule marker, reduced man power and material's waste, improved breeding efficiency.Simultaneously, the dna molecular marker technological operation is simple, and is with low cost, and the time spent is short, in one day, can accomplish whole testing processes.
2. required sample size is few, operates easylier, and efficient is high, and is with low cost.
3. the present invention is easy to grasp, and does not receive environmental influence, and repeatability is high.
Description of drawings:
Fig. 1 cucumber fruits length genes involved Markers for Detection result (east farming 804).M:DL2000DNAMarker; 1: contrast east farming 803; 2～8: agricultural 804 individual plants in east.
Embodiment:
Experimental technique among the following embodiment like no specified otherwise, is ordinary method.Used test materials among the following embodiment like no specified otherwise, is to buy from routine biochemistry reagent shop and obtains.All primers are synthetic to be accomplished by Sangon Biotech (Shanghai) Co., Ltd..The cucumber material that all are used in following examples is public.
Embodiment 1:
The molecular marker identification method of cucumber fruits length genes involved, this method comprises the steps:
With the cucumber variety of fruit length the unknown or the DNA of strain is template, is that primer carries out pcr amplification with CSWCT28, amplified production is carried out the detection of 3% agarose gel electrophoresis perhaps amplified production is carried out 7.5% denaturing polyacrylamide gel electrophoresis detection; If the banding pattern that amplification obtains is consistent with agricultural 803 banding patterns in the cucumber of known short fruit length east, cucumber variety that then this fruit length is unknown or strain are the kind or the strain of short fruit length, and the program of said pcr amplification is: 94 ℃ of preparatory sex change 5min; 94 ℃ of sex change 30s, 47 ℃ of annealing 1min, 35 circulations; 72 ℃ are extended 1min; 72 ℃ are extended 10min, last 4 ℃ of preservations, and described CSWCT28 is:
Primer 1:5 '-GAATTCAAAAGCATTTCAAAACTA-3 ',
Primer 2: 5 '-GAATTCAATTGGGTTTTTGAACCC-3 '.
Embodiment 2:
The molecular marker identification method of described cucumber fruits length genes involved; In the said pcr amplification; The PCR reaction system of per 20 μ l is following: 10 * buffer:2 μ l, 2mM dNTP:2 μ l, the Primer 1:0.8 μ l of 10pmol/ μ l; The Primer 2:0.8 μ l of 10pmol/ μ l, 25mM MgCl
2: 2.5 μ l, the TaqDNA polysaccharase of 5U/ μ l: 0.2 μ l, the template DNA of 60ng/ μ l: 1 μ l, ddH20:10.7 μ l.
Embodiment 3:
The application of the molecular marker identification method of cucumber fruits length genes involved in assisted Selection breed cucumber material.
The cucumber hybrid new breed east farming of cultivating with cucumber seminar of gardening institute of Northeast Agricultural University 804 is a material; Extracting its single plant DNA is template; With 1 cucumber fruits length molecule mark CSWCT28 provided by the invention is that primer carries out pcr amplification; The result shows; The pcr amplification banding pattern of agricultural 804 individual plants in east is in full accord with short fruit length contrast east farming 803 amplification banding patterns, and the identical rate that identify in PCR detected result and field is 100%, has proved that CSWCT28 is the molecule marker of the cucumber fruits length detection of a practicality.
Claims (3)
1. the molecular marker identification method of a cucumber fruits length genes involved is characterized in that: this method comprises the steps: that with the unknown cucumber variety of fruit length or the DNA of strain be template, is that primer carries out pcr amplification with CSWCT28; Amplified production is carried out the detection of 3% agarose gel electrophoresis or amplified production is carried out 7.5% denaturing polyacrylamide gel electrophoresis detection, if the banding pattern that amplification obtains is consistent with agricultural 803 banding patterns in cucumber east of known short fruit length, then cucumber variety of this fruit length the unknown or strain are the kind or the strain of weak point fruit length; The program of said pcr amplification is: 94 ℃ of preparatory sex change 5min; 94 ℃ of sex change 30s, 47 ℃ of annealing 1min, 35 circulations; 72 ℃ are extended 1min; 72 ℃ are extended 10min, last 4 ℃ of preservations, and described CSWCT28 is:
Primer 1:5 '-GAATTCAAAAGCATTTCAAAACTA-3 ',
Primer 2: 5 '-GAATTCAATTGGGTTTTTGAACCC-3 '.
2. the molecular marker identification method of cucumber fruits length genes involved according to claim 1; It is characterized in that: in the said pcr amplification; The PCR reaction system of per 20 μ l is following: 10 * buffer:2 μ l, 2mM dNTP:2 μ l, the Primer 1:0.8 μ l of 10pmol/ μ l; The Primer 2:0.8 μ l of 10pmol/ μ l, 25mM MgCl
2: 2.5 μ l, the TaqDNA polysaccharase of 5U/ μ l: 0.2 μ l, the template DNA of 60ng/ μ l: 1 μ l, ddH2O:10.7 μ l.
3. the application of the molecular marker identification method of an above-mentioned cucumber fruits length genes involved in assisted Selection breed cucumber material.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107072166A (en) * | 2016-03-28 | 2017-08-18 | 蔡洙湖 | maple leaf type cucumber plant |
| CN108165657A (en) * | 2018-03-05 | 2018-06-15 | 南京农业大学 | A kind of and the relevant SSR marker of cucumber fruit length character and its application |
| CN108624710A (en) * | 2018-06-19 | 2018-10-09 | 南京农业大学 | A kind of and the relevant SSR marker of cucumber fruit length character and its application |
-
2011
- 2011-01-11 CN CN2011100041027A patent/CN102586405A/en active Pending
Non-Patent Citations (4)
| Title |
|---|
| G FAZIO ET AL: "Development and characterization of pcr markers in cucumber", 《JASHS》 * |
| G. FAZIO ET AL: "Genetic mapping and QTL analysis of horticultural traits in cucumber (Cucumis sativus L.) using recombinant inbred lines", 《THEOR APPL GENET》 * |
| MATTHEW D. ROBBINS: "Molecular marker development, QTL pyramiding, and comparative analysis of phenotypic and marker-assisted selection in cucumber", 《UNIVERSITY OF WISCONSIN-MADISON 博士论文》 * |
| X. J. YUAN ET AL: "Genetic linkage map construction and location of QTLs for fruit-related traits in cucumber", 《PLANT BREEDING》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107072166A (en) * | 2016-03-28 | 2017-08-18 | 蔡洙湖 | maple leaf type cucumber plant |
| CN108165657A (en) * | 2018-03-05 | 2018-06-15 | 南京农业大学 | A kind of and the relevant SSR marker of cucumber fruit length character and its application |
| CN108165657B (en) * | 2018-03-05 | 2021-05-11 | 南京农业大学 | SSR (simple sequence repeat) marker related to cucumber fruit length traits and application thereof |
| CN108624710A (en) * | 2018-06-19 | 2018-10-09 | 南京农业大学 | A kind of and the relevant SSR marker of cucumber fruit length character and its application |
| CN108624710B (en) * | 2018-06-19 | 2021-04-30 | 南京农业大学 | SSR (simple sequence repeat) marker related to cucumber fruit length traits and application thereof |
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Application publication date: 20120718 |