CN106995846A - The detection and application of a kind of Henan south aromatic rice Badh2 gene functions mark - Google Patents
The detection and application of a kind of Henan south aromatic rice Badh2 gene functions mark Download PDFInfo
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Abstract
The invention discloses the detection and application of a kind of Henan south aromatic rice Badh2 gene functions mark, wherein the method for the southern aromatic rice in detection Henan comprises the following steps：The DNA of full-length genome is extracted using CTAB methods to the blade of the southern aromatic rice in Henan；Extracted DNA is marked to enter performing PCR amplification using Badh2 gene functions；Pcr amplification product is subjected to electrophoresis, observation result is carried out using gel imaging system.The application in the breeding of aromatic rice is marked at the invention also discloses a kind of Badh2 gene functions.The present invention identifies the corresponding mutation type of the southern aromatic rice in Henan, is laid the foundation to cultivate high-quality scented rice new varieties using molecular marker assisted selection.
Description
Technical field
The invention belongs to gene engineering technology field, specifically, it is related to a kind of southern aromatic rice Badh2 gene functions in Henan
The detection and application of mark.
Background technology
Paddy rice (Oryza sativa L.) is one of most important cereal crops in the world, and the whole world has more than half
There are 2/3rds population in population and China using rice as staple food.With the improvement of living standards, need of the people to fine quality rice
Ask and be continuously increased.But Grain Quality Traits in Rice is complicated, mainly including nutritional quality, exterior quality, attrition process quality, steaming
Boil with several aspects such as Cooking Quality, and among these qualities, consumer often more focuses on boiling and the Cooking Quality of rice.
And as the fragrant rice of cultivated rice specific type, due to fragrant aroma, rice fragrance, nutritive value is higher the features such as, be considered as rice
Treasure in rice.Traditional aromatic rice region is strong, resistance is poor and yield is relatively low, it is difficult to be widely applied, therefore scented rice valency
Lattice are higher, but people but dramatically increase in recent years to the demand of scented rice.Therefore, the genetic breeding of fragrant rice is by domestic and international paddy rice
Geneticist and the highest attention of breeding scholar.
At present, 200 kinds of volatile materials have been had more than in paddy rice to be separated and identified, wherein 2- acetyl -1-
Pyrrolin (2-Acetyl-1-pyrroline, 2-AP) is considered as one of main volatile materials in fragrant rice, and 2-AP exists
It can be just detected under relatively low concentration conditions.There are a variety of methods to be used for detecting the fragrance in rice material, wherein nozzle
It is the most commonly used method during traditional breeding method to chew method and KOH methods, but both are judged by the sense organ of people
The method accuracy of fragrance is poor.Early-stage Study shows：Recessive gene fgr on the chromosome of paddy rice the 8th, is one and perfume
The closely related gene of taste, the gene is then separated and cloned.Further research is disclosed：Gene fgr encodes betaine aldehyde chloride dehydrogenation
Enzyme (Betaine aldehyde dehydrogenase homologue 2, Badh2), suppresses the expression of fgr genes, knocks out
Fgr genes or fgr genes morph, and cause the missing of Badh2 enzyme functions, can all make the increase of 2AP precursor substances, Jin Erji
Tired 2AP makes paddy rice produce fragrance.Therefore, fgr genes and Badh2 genes are same scent gene.
For Badh2 genes, the sequencing analysis in 280 parts of common wild-rices and 242 parts of common cultivated rices are found：Except
Exist in exon 2 outside the insertion mutation of a 7-bp missing and the 8th extron in the presence of a 7-bp, in Badh2 bases
There are variant sites in the extron of the 1st, 10,13 and 14 of cause.Further study show that：The 4th and 5 in Badh2 genes are outer aobvious
Also there are insertion, missing or single in splice site, promoter region and its 5 ' the UTR areas of son, the 1st extron and the 1st introne
Nucleotide diversity site.Have now been developed multiple molecular labelings relevant with rice scent, and for new rice variety or
The seed selection of person's restorer.However, these molecular labelings are SNP molecular labeling, insertion or deletion type mostly
Molecular labeling, and the base difference of PCR primer is smaller, it is necessary to utilize polyacrylamide gel electrophoresis to carry out separation analysis.
Therefore, the process of whole experimental implementation is comparatively laborious, is unfavorable for utilizing molecular labeling to carry out detection exactly and analysis on a large scale
Work.Meanwhile, the molecular labeling based on DNA be usually using its genome sequence pleomorphism site design, often mark with
Target gene is farther out, it may occur that mark and corresponding phenotype not exclusively identical phenomenon.The functional label of gene, is for special
Gene and design, phenotype corresponding with gene fits like a glove, the molecular labeling related relative to common fragrance,
The functional label of Badh2 genes is more accurate during the genetic improvement of crop.Because Badh2 genes are in different fragrant rice product
There are different mutation types in kind, and a Badh2 gene functions mark is merely able to detect a type of mutation, is utilizing
During the molecular labeling or functional label developed carry out molecular marker assisted selection breeding, by multiple Badh2 etc.
Detection of the functional label of position gene simultaneously for the southern aromatic rice in Henan has not been reported with analysis.
The content of the invention
In view of this, the present invention may during fragrant rice genetic breeding for the related molecular labeling of rice scent gene
Occur that screening is inaccurate, screening process is comparatively laborious, be unfavorable for extensive accurate fast for fragrant rice breeding intermediate materials progress
There is provided the detection and application of a kind of Henan south aromatic rice Badh2 gene functions mark, this hair for the problem of detecting fastly with analysis
7 gene functions mark of bright utilization Badh2 Data minings, is detected and has been analyzed for aromatic rice, it is intended to be clearly fragrant
The variation type of rice resource scent gene, and then enter the molecular breeding of rice of holding or participate in a prayer service at a temple using the fragrant rice resource of molecular marker assisted selection,
Cultivation for high-quality scented rice new varieties lays the foundation.
In order to solve the above-mentioned technical problem, the fragrant rice in Henan south is detected with Badh2 gene functions mark the invention discloses one kind
The method of kind, comprises the following steps：
Step 1, PCR amplifications：The DNA of full-length genome is extracted using CTAB methods to the blade of the southern aromatic rice in Henan；
Step 2, using Badh2 gene functions extracted DNA is marked to enter performing PCR amplification；
Step 3, pcr amplification product is subjected to electrophoresis, observation result is carried out using gel imaging system.
Further, the Badh2 gene functions mark includes FMU1-2, FME2, FME4-5, FME7, FME12, FME13
With FME14 primer pairs；The FMU1-2 primer pairs include nucleotide sequence FMU1-2 F primers and core as shown in SEQ ID No.1
FMU1-2 R primer of the nucleotide sequence as shown in SEQ ID No.2；The FME2 primer pairs include nucleotide sequence such as SEQ ID
The FME2 R primers of the primers of FME2 F shown in No.3 and nucleotide sequence as shown in SEQ ID No.4；The FME4-5 primer pairs
Including nucleotide sequence as shown in SEQ ID No.5 FME4-5 F primers and nucleotide sequence as shown in SEQ ID No.6
FME4-5R primers；The FME7 primer pairs include the FME7 F1 primers and nucleosides nucleotide sequence as shown in SEQ ID No.7
FME7 R1 primer of the acid sequence as shown in SEQ ID No.8；And FME7 F2 primers and nucleosides as shown in SEQ ID No.9
FME7 R2 primer of the acid sequence as shown in SEQ ID No.10；Described FME12 primer pairs include nucleotide sequence such as SEQ ID
The FME12 R primers of the primers of FME12 F shown in No.11 and nucleotide sequence as shown in SEQ ID No.12；Described FME13 draws
Thing to including nucleotide sequence as shown in SEQ ID No.13 FME13 F primers and nucleotide sequence as shown in SEQ ID No.14
FME13 R primers；Described FME14 primer pairs include nucleotide sequence as shown in SEQ ID No.15 FME14 F primers with
FME14 R primer of the nucleotide sequence as shown in SEQ ID No.16.
Further, PCR reaction systems include DNA profiling 2 μ L, 10 × Buffer 2.5 μ L, 25mmolL-1's
MgCl21.5 μ L, 2.5mmolL-1The μ L of dNTP 2.0, forward and reverse primer (12.5 μm of olL-1) each 1.0 μ L, 5U μ
L-1The μ L of Taq enzyme 0.4, supplement sterilized water is to 20 μ L.
Further, PCR reaction conditions are 94 DEG C of pre-degenerations 5min, 94 DEG C of denaturation 30s, 55 DEG C~61 DEG C anneal 30s, 72
DEG C extension 30s~90s, reacts after 32 circulations 72 DEG C and re-extends 10min, 25 DEG C of preservations.
Further, functional label FME4-5 and FME7 PCR primer carries out electrophoresis on 2% Ago-Gel, adopts
Observation result is carried out with gel imaging system；FME2 gene functions mark PCR primer 6% non-denaturing polyacrylamide
Electrophoresis is carried out on gel, is then observed after silver nitrate silver staining.
The application in the breeding of aromatic rice is marked at the invention also discloses a kind of above-mentioned Badh2 gene functions.
Compared with prior art, the present invention can be obtained including following technique effect：
1) present invention can be directed to multiple offunctional sites of Badh2 genes in paddy rice while quickly and accurately being detected
With analysis.
2) present invention can be marked using 7 gene functions of Badh2 Data minings, identify the southern aromatic rice in different Henan
In Badh2 genes variation type and its corresponding genotype.
3) present invention can utilize the functional label of Badh2 genes, directly cultivate high-quality using molecular marker assisted selection
The fragrant rice new varieties in Henan south.
Certainly, any product for implementing the present invention it is not absolutely required to while reaching all the above technique effect.
Brief description of the drawings
Accompanying drawing described herein is used for providing a further understanding of the present invention, constitutes the part of the present invention, this hair
Bright schematic description and description is used to explain the present invention, does not constitute inappropriate limitation of the present invention.In the accompanying drawings：
Fig. 1 is 7 Badh2 gene functions marks of the present invention to 16 fragrant rice and 4 non-fragrant rice material electrophoresis detection figures；
Wherein, A is functional label FMU1-2 PCR test strips；B is functional label FME2 PCR test strips；C is functional label
FME4-5 PCR test strips；D is functional label FME7 PCR test strips；E detects for functional label FME12-13 PCR
Band；F is functional label FME14 PCR test strips；WT is 4 non-perfume rice varieties (letter round-grained rice 46, Henan agriculture 31, the and of Henan agriculture 32
Henan agriculture 9) biased sample, 1~16 all aromatic rices, it, which is numbered, corresponds to table 1.
Embodiment
Describe embodiments of the present invention in detail below in conjunction with embodiment, thereby to the present invention how application technology hand
Section can fully understand and implement according to this to solve technical problem and reach the implementation process of technology effect.
All materials are the rice varieties of japonica rice type in the present invention, wherein from the fragrant rice money in 16 parts of Henan south area
Source has widely representativeness, there is 14 parts of conventional fragrant rice and the fragrant rice of 2 parts of glutinous types, and 4 kinds of conventional japonica rice kinds of selection are used as control
(table 1).All aromatic rices and its control material are in sowing in 2016 in the same experiment crop field of Xinyang Academy of Agricultural Sciences
In, each kind plants 2 rows, often 12 plants of row, and seeding row spacing is 16.5 × 26.4cm.Carry out during be seeded into seed maturity common
Crop field conventional cultivation management.
The test material of table 1 and its source
The gene function of embodiment 11.2 is marked
There is the form of a variety of allele in natural population in scent gene Badh2, and different allele its
Fragrance difference, for Badh2 genes the extron of 5 ' UTR areas, the 2nd, 4,5,7,12,13 and 14 exist insertion, missing or
Person's single nucleotide variations site develops 7 Badh2 gene functions marks.Wherein functional label FME14 is that digestion amplification is polymorphic
Property sequence mark (The cleaved amplified polymorphic sequence marker, CAPS), remaining is
Regular-PCR molecular labeling, each Badh2 gene function marks size, the sequence of primer, annealing temperature of amplified production etc. in detail
Information is shown in Table 2.All primers are Nanjing Genscript Biotechnology Co., Ltd.'s synthesis, and are shown by trial test, and this 7
Individual Badh2 gene functions mark is clearly reliable in reaction system performance stabilization, result.
The Badh2 gene function label informations of table 2
The detection of the Henan of embodiment 2 south aromatic rice Badh2 gene functions mark
1) PCR is expanded：Henan south aromatic rice and the blade of 4 control japonica rice extract full-length genome using CTAB methods
DNA。
PCR reaction systems include 2.5 μ L, the 25m molL of μ L, 10 × Buffer of DNA profiling 2-1MgCl21.5 μ L,
2.5m mol·L-1The μ L of dNTP 2.0, forward and reverse primer (12.5 μm of olL-1) each 1.0 μ L, 5U μ L-1Taq enzyme
0.4 μ L, supplement sterilized water to 20 μ L.Reagent used is purchased in Shanghai Sheng Gong biotechnologies Co., Ltd.
PCR reaction conditions are 94 DEG C of pre-degenerations 5min, 94 DEG C of denaturation 30s, 55 DEG C~61 DEG C annealing 30s, 72 DEG C of extension 30s
~90s, 72 DEG C re-extend 10min, 25 DEG C of preservations after 32 circulations of reaction.
2) PCR primer is detected
Functional label FME14 PCR primer first passes through the restriction enzymes of Bsl I (under CutSmart buffer conditions)
Reacted 3 hours at 55 DEG C, electrophoresis is then carried out on 1.5% Ago-Gel；Functional label FME4-5 and FME7 PCR productions
Thing carries out electrophoresis on 2% Ago-Gel, and observation result is carried out using gel imaging system.Remaining Badh2 gene work(
The PCR primer that can be marked carries out electrophoresis on 6% non-denaturing polyacrylamide gel, is then observed after silver nitrate silver staining.
3) result of the test：
3.1) functional label FME2 testing result
Respectively using 16 parts of southern aromatic rices in Henan and the genomic DNA of 4 parts of conventional japonica rice kinds as template, Badh2 bases are utilized
Because functional label FME2 enters performing PCR amplification, detected using native polyacrylamide gel electrophoresis.Its testing result (Figure 1B) table
It is bright：The fragrant round-grained rice 1 of letter, often fragrant round-grained rice 805, fragrant round-grained rice 101 and the fragrant round-grained rice of agriculture have 4 parts of aromatic rices and can amplify the piece of 100bp sizes
Section, illustrates that this 4 parts fragrant rice materials all have 7bp deletion fragment at the exon 2 of Badh2 genes, they belong to
The variation type of same scent gene.The southern aromatic rice of remaining 4 parts of conventional japonica rice kind and 12 parts of Henan all amplifies one
The fragment of 107bp sizes, illustrates that this fragrant rice material in 12 parts of Henan south is not belonging to the exon 2 mutation type of Badh2 genes.
3.2) functional label FME4-5 testing result
Using 16 parts of southern aromatic rices in Henan and the genomic DNA of 4 parts of conventional japonica rice kinds as template, Badh2 gene work(is utilized
Energy flag F ME4-5 enters performing PCR amplification respectively, is detected using 2% agarose gel electrophoresis.Its testing result (Fig. 1 C) shows：
White perfume round-grained rice and fragrant round-grained rice 805 can amplify the fragment of 321bp sizes, illustrate this 2 parts fragrant rice materials the 4th and 5 of Badh2 genes
All there is 803bp deletion fragment at extron, they belong to the variation type of same scent gene.Remaining 4 parts
The southern aromatic rice of conventional japonica rice kind and 14 parts of Henan all amplifies the fragment of a 1123bp size, illustrates this fragrant rice in 14 parts of Henan south
Material is not belonging to the 4th and 5 exons mutation types of Badh2 genes.
3.3) functional label FME7 testing result
Respectively using 16 parts of southern aromatic rices in Henan and the genomic DNA of 4 parts of conventional japonica rice kinds as template, Badh2 bases are utilized
Because functional label FME7 enters performing PCR amplification, detected using 2% agarose gel electrophoresis.Its testing result (Fig. 1 D) shows：It is short
Stalk perfume rice ball, letter fragrant glutinous 933, fragrant rich 916, the fragrant rice in Huaibin County and the fragrant rice ball in Xi County can amplify size for 583bp and
169bp 2 fragments, illustrate that this 5 parts fragrant rice materials have 8bp deletion fragment at the 7th extron of Badh2 genes
With 3bp mutation, they belong to the mutation type of same scent gene.Remaining 4 parts of conventional japonica rice kind and 11 parts of Henan south
Aromatic rice amplifies 2 bands that size is respectively 591bp and 449bp, illustrates that this fragrant rice material in 11 parts of Henan south is not belonging to
7th exons mutation type of Badh2 genes.
3.4) functional label FMU1-2, FME12-13 and FME14 testing result
Using 16 parts of southern aromatic rices in Henan and the genomic DNA of 4 parts of conventional japonica rice kinds as template, Badh2 bases are utilized respectively
Because functional label FMU1-2, FME12-13 and FME14 enter performing PCR amplification, detected respectively using gel electrophoresis.Its testing result
(Fig. 1 D) shows：Using Badh2 gene function flag F MU1-2 and FME12-13, all aromatic rices and control material are all only
A band is amplified, size is respectively 163bp (Figure 1A) and 192bp (Fig. 1 E) fragment, illustrate that this 16 parts of aromatic rices exist
Mutation is not present at the extron of 5 '-UTR areas, the 12nd and 13 of Badh2 genes, they belong to this scent gene mutation class
Type.PCR amplifications are done using Badh2 gene function flag Fs ME14, then using the digestion detections of restriction enzyme Bsl I, as a result
Show that all fragrant rice materials and non-fragrant rice material can be gone out the band of 205bp sizes by the digestions of Bsl I, illustrate this 16 parts of Henan
Southern aromatic rice is at the 14th extron of Badh2 genes and in the absence of 1bp insertion mutation (Fig. 1 F).
In this test material, fragrant precious No. 2, Zheng Xiang round-grained rice 11, black fragrant rice 193, the fragrant round-grained rice 4 of agriculture, perfume glutinous 1862 and perfume are precious No. 1
Totally 6 parts of fragrant rice materials are not sent out at known Badh2 genetic mutation sites (extron of 5 '-UTR areas, the 12nd, 13 and 14) place
Now (Figure 1A, E and F) occurs for variation.Remaining 10 parts fragrant rice materials are deposited at the extron of the 2nd, 4,5 and 7 of Badh2 genes respectively
In mutation, wherein there is 7bp missing at the Second Exon of Badh2 genes in fragrant round-grained rice 805, and it is outer aobvious the 4th and 5
There is 803bp deletion fragment at son, detailed the results are shown in Table 3.
The aromatic rice Badh2 gene functions mark testing result classification of 3 16 parts of Henan of table south
4) conclusion：
4.1) detection of fragrant rice molecular labeling：The present invention is fragrant to 16 parts of Henan south using above-mentioned 7 Badh2 gene functions mark
Rice varieties have carried out detection classification, have 4 parts of fragrant rice materials and there is variation at the exon 2 of Badh2 genes；2 parts of fragrant rice
There is 803bp missing at the 4th and 5 extrons of Badh2 genes in material；There are 4 parts of fragrant rice materials the 7th of Badh2 genes
There is 8bp missing and 3bp mutation (Fig. 1 at extron；Table 3), 5 '-UTRs of remaining fragrant rice material in Badh2 genes
Variation is not found at the extron of area, the 12nd, 13 and 14 and occurs (Fig. 1).Therefore, molecule is carried out for specific aromatic rice
The detection and analysis of mark, can be greatly enhanced the breeding efficiency of fragrant rice new varieties.
4.2) exploitation and application of the fragrant rice functional label in Henan south
High-quality scented rice is commercially deep to be favored by consumers in general, and the fragrance in its rice is very important food flavor product
Matter character.Scent gene functional label always links together with specific rice fragrance character, then for the fragrant rice product in Henan south
Resource is planted, carries out the detection and analysis of perfume rice Badh2 gene functions mark, provides important by the cultivation for high-quality scented rice new varieties
With reference to reference.This result of study finds there are 10 parts of materials outside the 2nd, 4,5 or 7 of Badh2 genes in the aromatic rice of Henan south
There is variation at aobvious son, it is possible to use existing functional label carries out Marker-assisted selection to accelerate educating for high-quality scented rice new varieties
Kind.And fragrant precious No. 2, Zheng Xiang round-grained rice 11, black fragrant rice 193, the fragrant round-grained rice 4 of agriculture, perfume glutinous 1862 and fragrant precious No. 1 totally 6 parts of fragrant rice materials and 4 parts
Non- fragrant rice material is the same in the result of this 7 functional molecular marker detections.Therefore, this 6 parts fragrant rice materials are possible to
It is other mutation types for the Badh2 genes having been found that, it is necessary to further identify the functional label of these fragrant rice materials；Also have
There is the neomorph type of still undiscovered Badh2 genes in possible this 6 parts fragrant rice materials, can according to these fragrant rice materials
With in the development and utilization for positioning, separation, clone and the functional label for carrying out new gene from now on.
Some preferred embodiments of invention have shown and described in described above, but as previously described, it should be understood that invention is not
Form disclosed herein is confined to, the exclusion to other embodiment is not to be taken as, and available for various other combinations, modification
And environment, and can be carried out in invention contemplated scope described herein by the technology or knowledge of above-mentioned teaching or association area
Change., then all should be in the appended power of invention and the change and change that those skilled in the art are carried out do not depart from the spirit and scope of invention
In the protection domain that profit is required.
SEQUENCE LISTING
<110>Xinyang college of education of China
<120>The detection and application of a kind of Henan south aromatic rice Badh2 gene functions mark
<130> 2017
<160> 16
<170> PatentIn version 3.3
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Claims (6)
1. the method for the southern aromatic rice in a kind of use Badh2 gene functions mark detection Henan, it is characterised in that comprise the following steps：
Step 1, PCR amplifications：The DNA of full-length genome is extracted using CTAB methods to the blade of the southern aromatic rice in Henan；
Step 2, using Badh2 gene functions extracted DNA is marked to enter performing PCR amplification；
Step 3, pcr amplification product is subjected to electrophoresis, observation result is carried out using gel imaging system.
2. the method for the southern aromatic rice in use Badh2 gene functions mark detection Henan according to claim 1, its feature exists
In the Badh2 gene functions mark includes FMU1-2, FME2, FME4-5, FME7, FME12, FME13 and FME14 primer pair；
The FMU1-2 primer pairs include nucleotide sequence FMU1-2F primers and nucleotide sequence such as SEQ as shown in SEQ ID No.1
FMU1-2R primers shown in ID No.2；The FME2 primer pairs are drawn including nucleotide sequence FME2F as shown in SEQ ID No.3
The FME2R primers of thing and nucleotide sequence as shown in SEQ ID No.4；The FME4-5 primer pairs include nucleotide sequence such as
The FME4-5R primers of FME4-5F primers and nucleotide sequence as shown in SEQ ID No.6 shown in SEQ ID No.5；The FME7
Primer pair includes FME7F1 primer and nucleotide sequence of the nucleotide sequence as shown in SEQ ID No.7 such as SEQ ID No.8 institutes
The FME7R1 primers shown；And such as SEQ ID No.10 institutes of the FME7F2 primers and nucleotide sequence as shown in SEQ ID No.9
The FME7R2 primers shown；Described FME12 primer pairs include nucleotide sequence as shown in SEQ ID No.11 FME12F primers with
FME12R primer of the nucleotide sequence as shown in SEQ ID No.12；Described FME13 primer pairs include nucleotide sequence such as SEQ
The FME13R primers of FME13F primers shown in ID No.13 and nucleotide sequence as shown in SEQ ID No.14；Described FME14
Primer pair includes nucleotide sequence FME14F primers and nucleotide sequence such as SEQ ID No.16 institutes as shown in SEQ ID No.15
The FME14R primers shown.
3. the method for the southern aromatic rice in use Badh2 gene functions mark detection Henan according to claim 1, its feature exists
In PCR reaction systems include DNA profiling 2 μ L, 10 × Buffer2.5 μ L, 25mmolL-1MgCl21.5 μ L, 2.5mmol
L-1The μ L of dNTP 2.0, forward and reverse primer (12.5 μm of olL-1) each 1.0 μ L, 5U μ L-1The μ L of Taq enzyme 0.4, supplement
Sterilized water is to 20 μ L.
4. the method for the southern aromatic rice in use Badh2 gene functions mark detection Henan according to claim 1, its feature exists
In, PCR reaction conditions be 94 DEG C of pre-degeneration 5min, 94 DEG C of denaturation 30s, 55 DEG C~61 DEG C annealing 30s, 72 DEG C of extension 30s~
90s, 72 DEG C re-extend 10min, 25 DEG C of preservations after 32 circulations of reaction.
5. the method for the southern aromatic rice in use Badh2 gene functions mark detection Henan according to claim 1, its feature exists
In functional label FME4-5 and FME7 PCR primer carry out electrophoresis on 2% Ago-Gel, using gel imaging system
Carry out observation result；The PCR primer of FME2 gene functions mark carries out electricity on 6% non-denaturing polyacrylamide gel
Swimming, is then observed after silver nitrate silver staining.
6. the Badh2 gene functions described in claim 1 are marked at the application in the breeding of aromatic rice.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111560457A (en) * | 2020-05-26 | 2020-08-21 | 上海市农业科学院 | PCR/LDR molecular marker and method for identifying rice fragrance allele Badh2-E2 |
| CN114480706A (en) * | 2022-01-18 | 2022-05-13 | 贵州省水稻研究所 | Primer group, method and kit for identifying rice Badh2 genotype |
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| CN103725778A (en) * | 2013-12-17 | 2014-04-16 | 上海市农业科学院 | Molecular marker for detecting specific mutation of rice aroma control gene and application of molecular marker |
| KR20170002202A (en) * | 2015-06-29 | 2017-01-06 | 공주대학교 산학협력단 | Primer set for Discrimination of Aromatic Rice and Use Thereof |
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| CN103725778A (en) * | 2013-12-17 | 2014-04-16 | 上海市农业科学院 | Molecular marker for detecting specific mutation of rice aroma control gene and application of molecular marker |
| KR20170002202A (en) * | 2015-06-29 | 2017-01-06 | 공주대학교 산학협력단 | Primer set for Discrimination of Aromatic Rice and Use Thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111560457A (en) * | 2020-05-26 | 2020-08-21 | 上海市农业科学院 | PCR/LDR molecular marker and method for identifying rice fragrance allele Badh2-E2 |
| CN114480706A (en) * | 2022-01-18 | 2022-05-13 | 贵州省水稻研究所 | Primer group, method and kit for identifying rice Badh2 genotype |
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