CN107974456B - Control gene and its application of corn Nitrogen response - Google Patents
Control gene and its application of corn Nitrogen response Download PDFInfo
- Publication number
- CN107974456B CN107974456B CN201610910526.2A CN201610910526A CN107974456B CN 107974456 B CN107974456 B CN 107974456B CN 201610910526 A CN201610910526 A CN 201610910526A CN 107974456 B CN107974456 B CN 107974456B
- Authority
- CN
- China
- Prior art keywords
- gene
- nitrogen
- corn
- response
- nitrogen response
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/13—Plant traits
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Botany (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Mycology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention provides a kind of genes for controlling corn Nitrogen response, control the amino acid sequence of coded sequence coding of the gene of corn Nitrogen response as shown in SEQ ID NO:2.Preferably, the nucleotide sequence of the coded sequence of the gene of control corn Nitrogen response is as shown in SEQ ID NO:1.Additionally provide application of the gene of control corn Nitrogen response in control corn Nitrogen response.The gene of control corn Nitrogen response of the invention can play regulatory role the expression of the transcript regulated and controled by nitrogenous fertilizer, it lays a good foundation for research corn Nitrogen response regulatory mechanism, promote corn nitrogen utilization efficiency (NUE) progress of research, is suitable for large-scale promotion application.
Description
Technical field
The present invention relates to molecular genetic technique fields, special more particularly, to the gene technology field of control Nitrogen response
Do not refer to a kind of gene for controlling corn Nitrogen response and its preparation method and application.
Background technique
Corn is extremely sensitive as the big crop Nitrogen response in China first, and the height high degree of yield depends on extraneous nitrogenous fertilizer
Amount of application.However excessive apply of nitrogenous fertilizer not only increases a large amount of consumption that economic cost brings resource；And it is unemployed
Nitrogen will generate great negative effect to environment.Therefore research corn Nitrogen response molecular regulation mechanism improves sustainability beautiful
Rice yield is of great significance.
Nitrogen (Nitrogen, N) has to pass the growth and development of plant as macronutrient necessary to plant
Important role, NLP (NIN-like proteins) gene family is the distinctive a kind of transcription factor of plant, to plant nitrogen
Signal transduction has important regulating and controlling effect.The formation of legume nodule depends on the presence of the gene, such as NIN in crowtoe
(Suzuki W,Konishi M,Yanagisawa S.2013.The evolutionary events necessary for
the emergence of symbiotic nitrogen fixation in legumes may involve a loss of
nitrate responsiveness of the NIN transcription factor.Plant signaling&
behavior 8(10),e25975)；The research of arabidopsis NLP gene is more detailed in non-leguminous plant, and 2009 years
Castaings etc. has found that Atnlp7 Arabidopsis Mutants, mutant n-signal transduction and assimilation approach are damaged and show
A series of phenotypic character that nitrogens lack finally speculates that the AtNLP7 being located in core may be transcription factor to nitrogen by test
Assimilation have important regulating and controlling effect (Castaings L, Camargo A, Pocholle D, Gaudon V, Texier Y,
Boutet-Mercey S,Taconnat L,Renou JP,Daniel-Vedele F,Fernandez E,Meyer C,Krapp
A.2009.The nodule inception-like protein 7modulates nitrate sensing and
metabolismin Arabidopsis.Plant Journal 57,426-435).The discovery such as Marchive in 2013, outside
Nitrogen signal stimulus several minutes of rear arabidopsis transcription factor AtNLP7 in boundary's are responded rapidly to using core retention mechanism and to nitrogen metabolism keys
Gene is regulated and controled, the mechanism be directly controlled by extraneous n-signal and independently of the regulation of transcriptional level (Marchive C,
Roudier F,Castaings L,Brehaut V,Blondet E,Colot V,Meyer C,Krapp
A.2013.Nuclear retention of the transcription factor NLP7orchestrates the
early response to nitrate in plants.Nature Communications 4,1713).The above evidence is said
Bright AtNLP7 is the major regulatory person of arabidopsis initial stage Nitrogen response.
However, research report of the NLP gene in corn is very few.Therefore, it is necessary to study the initial regulation of corn Nitrogen response
Element, the function of the Xie Ming gene, in order to control corn Nitrogen response state in source to promote corn to the efficient of nitrogen
It is absorbed and utilized and finally improves corn yield.
Summary of the invention
In order to overcome the disadvantages of the prior art mentioned above, it is an object of the present invention to provide a kind of control corn nitrogen to ring
The gene answered can play regulatory role the expression of the transcript regulated and controled by nitrogenous fertilizer, to study corn Nitrogen response regulatory mechanism
It lays a good foundation, promotes corn nitrogen utilization efficiency (NUE) progress of research, be suitable for large-scale promotion application.
Another object of the present invention is to provide a kind of preparation methods of gene for controlling corn Nitrogen response, pass through this method
The gene of control corn Nitrogen response can be obtained, and simple to operate, be suitable for large-scale promotion application.
Another object of the present invention is to provide a kind of applications of gene for controlling corn Nitrogen response, thus to by nitrogenous fertilizer tune
The expression of the transcript of control plays regulatory role, and lays a good foundation for research corn Nitrogen response regulatory mechanism, promotes corn nitrogen
Plain utilization rate (NUE) progress of research is suitable for large-scale promotion application.
To achieve the above objectives, in the first aspect of the present invention, a kind of gene for controlling corn Nitrogen response is provided, it is special
Point is that the amino acid sequence of the coded sequence coding of the gene of the control corn Nitrogen response is as shown in SEQ ID NO:2.
Encode the amino acid sequence as shown in SEQ ID NO:2 nucleotide sequence can there are many, preferably, described
The nucleotide sequence of the coded sequence of the gene of corn Nitrogen response is controlled as shown in SEQ ID NO:1.
In the second aspect of the present invention, a kind of preparation method of gene for controlling corn Nitrogen response is provided, its main feature is that,
The preparation method the following steps are included:
(1) gene containing NLP family functional domain, NLP family functional structure are screened in the corn genome
Domain includes RWP-RK structural domain and PB1 structural domain；
(2) gene to described containing NLP family functional domain carries out Nitrogen response analysis, and it is most sensitive to obtain Nitrogen response
Gene.
In the step (1), screening the gene containing NLP family functional domain in the corn genome can be used
Any suitable method, preferably, being screened in the Maize genome in the step (1) using Pfam-HMM model
The gene containing NLP family functional domain.
In the step (1), any conjunction can be used by being screened in the Maize genome using Pfam-HMM model
Suitable specific steps, more preferably, the step (1) specifically include:
(11) use Pfam database sharing NLP family Hidden Markov Model file: RWP-RK.hmm and
PB1.hmm；
(12) Maize genome is scanned for comparing according to the RWP-RK.hmm and the PB1.hmm, thus
Obtain the gene containing NLP family functional domain.
In the step (12), carries out described search and compare any suitable method that can use, further,
In the step (12), described search comparison is carried out using the hmmersearch program in HMMER3.0 software package.
In the step (12), the Maize genome can be any suitable Maize genome, further,
In the step (12), the Maize genome is corn B73V3 editions with reference to genome.
In order to guarantee the accuracy of result, preferably, being further comprised the steps of: between the step (1) and the step (2)
The gene containing NLP family functional domain is verified using public database.
The public database can be any suitable public database, and more preferably, the public database is NCBI
Conserved Domains Database database.
The step (2) can use any suitable specific steps, preferably, the step (2) specifically includes:
(21) plant grown under identical environment is divided into processing group and control group, wink is carried out to the processing group
When nitrogen handle；
(22) gene containing NLP family functional domain described after the instantaneous nitrogen is handled is monitored in different time points
Expression situation of change, expression quantity raises maximum gene after filtering out the instantaneous nitrogen processing, rings to obtain the nitrogen
Answer most sensitive gene.
In the step (22), the base containing NLP family functional domain described after the instantaneous nitrogen is handled is monitored
Because expression situation of change in different time points can use any suitable method, more preferably, in the step (1) and described
The genomic locations information for obtaining the gene containing NLP family functional domain is further comprised the steps of: between step (2),
Utilize the genomic locations information design specific primer；In the step (22), used using the specific primer
Real time fluorescence quantifying PCR method carries out the monitoring.
In the step (21), the instantaneous nitrogen processing can use any suitable condition, more preferably, in the step
Suddenly in (21), the plant is corn B73 self-mating system, and the concentration of the instantaneous nitrogen processing is 15mM KNO3, the control
Group can be handled using 15mM KCl, and the plant is in V3 phase (third leaf is fully deployed).
In the step (22), the different time points can be any suitable time point, more preferably, the difference
Time point includes 0h, 0.5h, 1h, 1.5h and 2h.
For the efficacy of the most sensitive gene of the clearly described Nitrogen response, preferably, after the step (2), institute
The preparation method of the gene for the control corn Nitrogen response stated further include: monitor different nitrogen and handle lower mutant Zea mays plant and wild
The situation of change of type plant express spectra determines the character mutation and Nitrogen response state of the mutant Zea mays plant, clear
The efficacy of the most sensitive gene of the Nitrogen response.
It monitors different nitrogen and handles the situation of change of lower mutant Zea mays plant and wild-type corn plant express spectra with can be with
Using any suitable method, more preferably, using RNA-seq sequencing technologies monitor different nitrogen handle lower mutant Zea mays plant and
The situation of change of wild-type corn plant express spectra.
Determine that the character mutation of the mutant Zea mays plant can be using any suitable method, more preferably, to mutation
The character mutation of body plant and wild-type corn plant the V3 phase plant under sufficient nitrogen and low nitrogen environment carries out Physiology and biochemistry point
Analyse the character mutation to determine mutant plants.The nitrogen concentration of sufficient nitrogen and low nitrogen environment can be respectively such as 15mM KNO3With
0.15mM KNO3, phenotype includes that such as plant total nitrogen content, amino acid content, third piece fully expanded leaves length, plant are dry
Weight, initial root length etc..
In the third aspect of the present invention, the gene or beautiful according to above-mentioned control of above-mentioned control corn Nitrogen response is provided
Application of the gene for the control corn Nitrogen response that the preparation method of the gene of rice Nitrogen response obtains in control corn Nitrogen response.
The beneficial effects of the present invention are:
A. the amino acid sequence such as SEQ ID NO:2 of the coded sequence coding of the gene of control corn Nitrogen response of the invention
It is shown, the expression of the transcript regulated and controled by nitrogenous fertilizer can be played regulatory role, be established for research corn Nitrogen response regulatory mechanism good
Good basis promotes corn nitrogen utilization efficiency (NUE) progress of research, is suitable for large-scale promotion application.
B. the gene of control corn Nitrogen response of the invention preparation method the following steps are included: (1) in Maize genome
Middle gene of the screening containing NLP family functional domain, NLP family functional domain includes RWP-RK structural domain and PB1
Structural domain；(2) gene to described containing NLP family functional domain carries out Nitrogen response analysis, and it is most sensitive to obtain Nitrogen response
Gene the gene of control corn Nitrogen response therefore can be obtained by this method, and it is simple to operate, be suitable for big rule
Mould promotes and applies.
C. the gene of control corn Nitrogen response of the invention or the preparation method for the gene for controlling corn Nitrogen response obtain
Application of the gene of corn Nitrogen response in control corn Nitrogen response is controlled, so that the expression to the transcript regulated and controled by nitrogenous fertilizer rises
Adjustment effect lays a good foundation for research corn Nitrogen response regulatory mechanism, promotes corn nitrogen utilization efficiency (NUE) research
Progress is suitable for large-scale promotion application.
These and other objects of the invention, feature and advantage, by following detailed descriptions, drawings and claims are obtained
To fully demonstrate, and can be achieved by means, device and the their combination specially pointed out in appended claims.
Detailed description of the invention
Fig. 1 is that the base containing NLP family functional domain after instantaneous nitrogen processing is monitored using real time fluorescence quantifying PCR method
Because of expression situation of change in different time points, crt gene is corn housekeeping gene UPF1.
Fig. 2 is the growth conditions figure that two kinds of nitrogen handle lower zmnlp5 mutant and wild type V3 phase.
Fig. 3 is the comparison that two kinds of nitrogen handle lower zmnlp5 mutant and wild type V3 phase leaf length, dry weight and initial root length
Figure.
Fig. 4 A is the enrichment analysis chart that Nitrogen response funtion part loses gene in zmnlp5 mutant.
Fig. 4 B is the expression analysis figure of N marker gene in zmnlp5 mutant and wild type.
Specific embodiment
In order to be more clearly understood that technology contents of the invention, spy lifts following embodiment and is described in detail.
The gene (ZmNLP5) of control corn Nitrogen response of the invention can control corn Nitrogen response state, the gene
The nucleotide sequence of coded sequence is as shown in SEQ ID NO:1, and the amino acid of coding is as shown in SEQ ID NO:2.
The gene of the control corn Nitrogen response can be prepared by the following:
1.1 is literary using the Hidden Markov Model of Pfam database (http://pfam.xfam.org/) building NLP family
Part (RWP-RK.hmm, PF02042；PB1.hmm, PF00564) (wherein PF02042 and PF00564 is respectively in Pfam database
The number of NLP family two functional domains RWP-RK and PB1), then using HMMER3.0 software package (http: //
Hmmer.janelia.org/static/binaries/hmmer3.0_windows.zip the hmmersearch program pair in)
Corn B73 scans for comparing (E value < 1E-5) with reference to genome, to obtain all comprising above 2 conserved domains
NLP gene, is named as ZmNLP1~ZmNLP9, that is, the above-mentioned base containing NLP family functional domain by totally 9
Cause, then using public database (NCBIConserved Domains Database (CDD) (http: //
Www.ncbi.nlm.nih.gov/cdd) database) said gene is verified and (ID of each gene is numbered and is input to number
According to detecting whether the gene contains the two structural domains in library), the NLP gene family after verifying passes through the Perl journey voluntarily write
Sequence parses and releases genomic locations information (being shown in Table 1) (Ge Min, Lv Yuan great, Li Tan, Zhang Tifu, Zhang Xiao of each member
Woods, full-length genome identification and the analysis Scientia Agricultura Sinica of Zhao Han .2014. corn Dof transcription factor family, 47 (23):
4563-4572)。
The genomic locations information of gene of the table 1 containing NLP family functional domain
1.2 design specific primer using the location information of each member of NLP gene family, wherein the forward primer of ZmNLP1
With reverse primer respectively as shown in SEQ ID NO:3 and SEQ ID NO:4, the forward primer and reverse primer of ZmNLP2 is respectively such as
Shown in SEQ ID NO:5 and SEQ ID NO:6, the forward primer and reverse primer of ZmNLP3 is respectively such as SEQ ID NO:7 and SEQ
Shown in ID NO:8, the forward primer and reverse primer of ZmNLP4 respectively as shown in SEQ ID NO:9 and SEQ ID NO:10,
The forward primer and reverse primer of ZmNLP5 respectively as shown in SEQ ID NO:11 and SEQ ID NO:12, draw by the forward direction of ZmNLP6
Object and reverse primer are respectively as shown in SEQ ID NO:13 and SEQ ID NO:14, the forward primer and reverse primer point of ZmNLP7
Not as shown in SEQ ID NO:15 and SEQ ID NO:16, the forward primer and reverse primer of ZmNLP8 is respectively such as SEQ ID NO:
Shown in 17 and SEQ ID NO:18, the forward primer and reverse primer of ZmNLP9 is respectively such as SEQ ID NO:19 and SEQ ID NO:
Shown in 20, the forward primer and reverse primer of UPF1 is respectively as shown in SEQ ID NO:21 and SEQ ID NO:22.Certainly by corn
Friendship is B73 (being attained at Jiangsu Province Agriculture Science Institute agricultural biotechnologies research institute crop molecular breeding research room) plantation in abundant
In the sandy soil mixed thoroughly, instantaneous nitrogen processing (the 15mM KNO of V3 phase progress is grown to3, compare as 15mM KCl), and using glimmering in real time
The variation of the expression quantity of each time point NLP gene of 0h, 0.5h, 1h, 1.5h and 2h after the processing of Fluorescent Quantitative PCR (qPCR) technical monitoring
Situation, two kinds of processing of each time point guarantee that 3 biology repeat.
1) extraction and reverse transcription of RNA: each fresh sample of time point plant root tissue is collected, utilizes TRIzol after liquid nitrogen grinding
Total serum IgE is extracted described in method, utilizes Prime ScriptTM RT Reagent after every part of quantification of uniform concentration of sample RNA
RNA reverse transcription is cDNA by kit (Takara) kit.
2) qPCR reaction system: total reaction volume is 10.0 μ l, comprising:Premix Ex
TaqTMII (TaKaRa, Dalian, China), 1.0 μ l templates (cDNA concentration is 10ng/ μ l), 1.0 10 μm of μ l ol/L
Primer,3μl ddH2O.Reaction is in RocheIt is carried out on the full-automatic fluorescence quantitative PCR instrument of 96 types.
3) qPCR reaction condition: 95 DEG C of 3min；95 DEG C of 10s, 58 DEG C of 20s, 72 DEG C of 20s, 40 circulations；72℃5min.
As a result shown in Figure 1, through qPCR analysis shows after nitrogen processing the expression of 0.5h corn NLP gene have not
With the up-regulation of degree, wherein ZmNLP4, ZmNLP5, ZmNLP6, tetra- genes of ZmNLP8 raise super when 0.5h expression quantity is compared with 0h
2 times are crossed, is nitrogen sensitive gene.Up-regulation is the most significantly ZmNLP5 gene (0.5h expression quantity is raised more than 5 times), and at nitrogen
4 times when ZmNLP5 expression quantity still raises about 0h when 1h after reason, ZmNLP5 gene is the most sensitive gene of Nitrogen response.
Application of the 2.ZmNLP5 gene on regulation corn Nitrogen response
The influence of 2.1 ZmNLP5 gene pairs seedling stage plant phenotypes
First in the website MaizeGDB UniformMu mutant library (http://www.maizegdb.org/
Uniformmu the Maize mutant material zmnlp5 (number: UFMu-01175, mutant and wild of screening ZmNLP5 gene in)
Type seed is attained at american corn genetic resources stored in association center-UniformMu transposons resources bank Maize
Genetics Cooperation Stock Center UniformMu Transposon Resource).Detect wild profile
Expect (W22) and mutant material (zmnlp5) in sufficient nitrogen (+N:15mM KNO3) and low nitrogen environment (- N:0.15mM KNO3) under grow
To the character mutation situation of V3 phase plant (see Fig. 2).As a result, it has been found that: under two kinds of environment of sufficient nitrogen and low nitrogen, mutant material
Physical signs (initial root length, leaf length, dry weight, plant total nitrogen content, amino acid content) is lower than wild type (see Fig. 2 and figure
3, table 2).Mutant is compared with the long decline 30.68% of WT lines main root, the decline of plant total nitrogen content especially under low nitrogen environment
11.70%, mutant material obviously weakens in the ability for responding extraneous nitrogen concentration variation.
The content of the total nitrogen and amino acid of 2 wild type of table and zmnlp5 mutant under two kinds of nitrogen environments
The influence of 2.2 ZmNLP5 gene pairs seedling stage plant express spectras
By zmnlp5 mutant and WT lines plantation in 15mM KNO3Environment, planting conditions are as described in 1.2.10 days
Afterwards, plant carries out nitrogen hunger processing (0mM KNO3) processing, after 10 days after re-start 15mM KNO3Or KCl is handled, after 0.5h
(WT+N, WT-N, zmnlp5+N, zmnlp5-N) is sampled to plant, using RNA-seq sequencing technologies, is monitored at different nitrogen
Manage the situation of change of lower mutant and wild-type corn plant express spectra.It refers to shown in Fig. 4 A, as a result, it has been found that: nitrogen is handled in WT
Sharing 9690 genes afterwards, there are the differential expressions of conspicuousness, are the Nitrogen response gene of wild type；However nearly 44.37% (4299/
9660) significant differential expression is not present in Nitrogen response gene in zmnlp5 mutant, i.e., the Nitrogen response gene of close half exists
Some lost Nitrogen response function in mutant, enrichment analysis (specifically utilize agriGO (http://bioinfo.cau.edu.cn/
AgriGO/analysis.php, Du et al., 2010) singular enrichment analysis (SEA) analysis is flat in
Platform carries out, and inputs gene set) find that such genetic enrichment is metabolized phase in N such as protein modified and Argine Monohydrochloride phosphorylations afterwards
The access of pass.In addition, to whole nitrogen biomarker genes listed in the articles such as Yang in 2011 (Yang XS, Wu J,
Ziegler TE,Yang X,Zayed A,Rajani MS,Zhou D,Basra AS,Schachtman DP,Peng M,
Armstrong CL,Caldo RA,Morrell JA,Lacy M,Staub JM.2011.Gene expression
biomarkers provide sensitive indicators of in planta nitrogen status in
Maize.Plant Physiology 157,1841-1852) analysis find: in wild type and mutant significant difference express
Nitrogen marker gene number be respectively 632 and 582, and in 81% (514/632) WT express significant changes N marker gene
There is weaker or opposite Nitrogen response state (referring to Fig. 4 B) in zmnlp5.Illustrate that the missing of ZmNLP5 gene seriously affects
The Nitrogen response state of corn.
The preparation method of the gene of the above-mentioned control corn Nitrogen response of the present invention through the invention obtains control corn
The gene of Nitrogen response is verified by Nitrogen response analysis and gene mutation body heredity, shows the gene of the control corn Nitrogen response
The expression of the transcript regulated and controled by nitrogenous fertilizer can be played regulatory role.
In conclusion the gene of control corn Nitrogen response of the invention can play the expression of the transcript regulated and controled by nitrogenous fertilizer
Adjustment effect lays a good foundation for research corn Nitrogen response regulatory mechanism, promotes corn nitrogen utilization efficiency (NUE) research
Progress is suitable for large-scale promotion application.
In this description, the present invention is described with reference to its specific embodiment.But it is clear that can still make
Various modifications and alterations are without departing from the spirit and scope of the invention.Therefore, the description and the appended drawings should be considered as illustrative
And not restrictive.
Claims (3)
1. a kind of gene for controlling corn Nitrogen response, which is characterized in that the code sequence of the gene of the control corn Nitrogen response
The amino acid sequence of coding is arranged as shown in SEQ ID NO:2.
2. the gene of control corn Nitrogen response according to claim 1, which is characterized in that the control corn Nitrogen response
Gene coded sequence nucleotide sequence as shown in SEQ ID NO:1.
3. application of the gene of control corn Nitrogen response according to claim 1 or 2 in control corn Nitrogen response.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610910526.2A CN107974456B (en) | 2016-10-19 | 2016-10-19 | Control gene and its application of corn Nitrogen response |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610910526.2A CN107974456B (en) | 2016-10-19 | 2016-10-19 | Control gene and its application of corn Nitrogen response |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107974456A CN107974456A (en) | 2018-05-01 |
| CN107974456B true CN107974456B (en) | 2019-03-08 |
Family
ID=62003637
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610910526.2A Active CN107974456B (en) | 2016-10-19 | 2016-10-19 | Control gene and its application of corn Nitrogen response |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107974456B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110468146A (en) * | 2018-05-11 | 2019-11-19 | 中国科学院上海生命科学研究院 | Regulate and control the method for legume nodule nitrogen fixing capacity |
| CN111217895B (en) * | 2018-11-26 | 2022-05-06 | 江苏省农业科学院 | Corn NLP transcription factor ZmNLP5 and application thereof |
| CN111454346B (en) * | 2020-06-04 | 2022-04-08 | 山东农业大学 | Transcription factor HvNLP2 from barley and participating in nitrate nitrogen regulation and application thereof |
| CN113832218A (en) * | 2021-09-28 | 2021-12-24 | 江苏省农业科学院 | Method and primer group for predicting nitrogen state of corn plant |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103343132A (en) * | 2013-07-25 | 2013-10-09 | 中国科学技术大学 | Application of plant nitrogen high-efficiency utilization gene and protein coded thereby |
-
2016
- 2016-10-19 CN CN201610910526.2A patent/CN107974456B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103343132A (en) * | 2013-07-25 | 2013-10-09 | 中国科学技术大学 | Application of plant nitrogen high-efficiency utilization gene and protein coded thereby |
Non-Patent Citations (7)
Also Published As
| Publication number | Publication date |
|---|---|
| CN107974456A (en) | 2018-05-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Beyer et al. | Loci and candidate genes controlling root traits in wheat seedlings—a wheat root GWAS | |
| Li et al. | Validation and comparison of reference genes for qPCR normalization of celery (Apium graveolens) at different development stages | |
| CN107974456B (en) | Control gene and its application of corn Nitrogen response | |
| Briones et al. | Influence of different cultivars on populations of ammonia-oxidizing bacteria in the root environment of rice | |
| Bernhard et al. | Loss of diversity of ammonia‐oxidizing bacteria correlates with increasing salinity in an estuary system | |
| Etchells et al. | Plant vascular cell division is maintained by an interaction between PXY and ethylene signalling | |
| Yang et al. | Expression Profiling of Cassava Storage Roots Reveals an Active Process of Glycolysis/Gluconeogenesis F | |
| Shiono et al. | Microarray analysis of laser-microdissected tissues indicates the biosynthesis of suberin in the outer part of roots during formation of a barrier to radial oxygen loss in rice (Oryza sativa) | |
| Hayashi et al. | Rj (rj) genes involved in nitrogen-fixing root nodule formation in soybean | |
| Anderson et al. | Ecological genetics and genomics of plant defences: evidence and approaches | |
| Francisco et al. | Genome wide association mapping in Arabidopsis thaliana identifies novel genes involved in linking allyl glucosinolate to altered biomass and defense | |
| Juenger et al. | Exploring genetic and expression differences between physiologically extreme ecotypes: comparative genomic hybridization and gene expression studies of Kas‐1 and Tsu‐1 accessions of Arabidopsis thaliana | |
| Norton et al. | Genome wide association mapping of grain and straw biomass traits in the rice Bengal and Assam Aus panel (BAAP) grown under alternate wetting and drying and permanently flooded irrigation | |
| Tian et al. | De novo transcriptome assembly of common wild rice (Oryza rufipogon Griff.) and discovery of drought-response genes in root tissue based on transcriptomic data | |
| Rampino et al. | Acquisition of thermotolerance and HSP gene expression in durum wheat (Triticum durum Desf.) cultivars | |
| Hoang et al. | Identification of conserved gene-regulatory networks that integrate environmental sensing and growth in the root cambium | |
| Ludwig-Müller et al. | Manipulation of auxin and cytokinin balance during the Plasmodiophora brassicae–Arabidopsis thaliana interaction | |
| Wang et al. | Belowground microbial communities respond to water deficit and are shaped by decades of maize hybrid breeding | |
| Alegria Terrazas et al. | Defining composition and function of the rhizosphere microbiota of barley genotypes exposed to growth-limiting nitrogen supplies | |
| Qin et al. | Wildfire drives the transition from deterministic-to stochastic-dominated community assembly of abundant bacterial in forest soils | |
| Dumack et al. | What drives the assembly of plant-associated protist microbiomes? Investigating the effects of crop species, soil type and bacterial microbiomes | |
| Ferreira | Molecular analysis of genebanks for sustainable conservation and increased use of crop genetic resources. | |
| CN101760462A (en) | Clone of plant disease resistance genes | |
| Jiang et al. | Research advances and prospects of orphan genes in plants | |
| CN105602948B (en) | The gene and method of Upland Cotton resisting verticillium are identified using fluorescent quantitative PCR technique |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |