CN106755413B - Rice nitrogen absorption and utilization site qNUE6 and molecular marking method thereof - Google Patents
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Abstract
The invention discloses waterSite of uptake and utilization of rice nitrogenqNUE6And molecular marking method thereof, the rice nitrogen absorption and utilization siteqNUE6A site located between the molecular markers NUE2 and NUE 23; the molecular marker NUE2 primer: SEQ ID NO.1, presenceqNUE6The amplified band was 135 bp. The invention can utilize the site through the absorption of the nitrogen in the riceqNUE6The molecular marker is used for detecting whether a nitrogen efficient absorption utilization variety Guanghui 998 (GH 998 for short) and a derivative variety (system) thereof contain a nitrogen absorption utilization gene locus, so that the nitrogen absorption utilization level can be predicted, and the selection efficiency of the rice with nitrogen efficient absorption utilization is greatly improved.
Description
Technical Field
The invention belongs to the field of molecular genetics, and relates to a site for absorbing and utilizing rice nitrogenqNUE6And a molecular labeling method thereof.
Background
Rice is one of the most important food crops in the world, and more than half of the world population uses rice as the main food. Nitrogen is one of the most important nutrients in rice production (Yoshida, 1981). Since the 60's of the 20 th century, the breeding of rice varieties has been progressing towards high nitrogen input with maximum grain yield (Tong, 2006). However, excessive application of nitrogen fertilizer results in decreased nitrogen utilization and environmental pollution. Therefore, the demand of the current rice nitrogen-efficient varieties becomes very urgent, the breeding of the rice varieties capable of efficiently absorbing and utilizing nitrogen is an important target in the rice breeding all over the world, and the improvement of the nitrogen utilization efficiency of the rice is an important target. Although the evaluation of the germplasm resources for the efficient absorption and utilization of the nitrogen of the rice and the research of the gene for the efficient absorption and utilization of the nitrogen have made some progress, a batch of QTLs related to the absorption and utilization of the nitrogen are positioned. However, due to the reasons of narrow original or genetic basis of materials used for research, interference of genetic background among individuals of traditional mapping groups, uncertain indexes and the like, the QTL positioning and utilization have some defects, so that no nitrogen absorption and utilization gene which can be directly applied to breeding exists so far, and no report that the nitrogen efficiency improvement is successfully obtained by means of molecular marker-assisted selection exists.
Disclosure of Invention
Aiming at the technical problems, the invention discloses a site for absorbing and utilizing rice nitrogenqNUE6And the molecular marking method can detect whether the high-efficiency nitrogen absorption and utilization variety GH998 and the derivative variety (system) thereof contain nitrogen absorption and utilization gene sites by using the molecular marking NUE2, can predict the nitrogen absorption and utilization level thereof, and greatly improve the selection efficiency of the rice with high-efficiency nitrogen absorption and utilization.
In contrast, the technical scheme of the invention is as follows:
site for absorbing and utilizing nitrogen of riceqNUE6Located at a site between the molecular markers NUE2 and NUE23, which molecular marker NUE2 primer: SEQ ID NO. 1/SEQ ID NO.2, presenceqNUE6The amplified band was 135 bp.
The primer sequence of the molecular marker NUE2 is as follows:
an upstream primer: GTTCCACATGTTGATGGG (as shown in SEQ ID NO.1)
A downstream primer: GATGAGGACGAGGAGGAG (as shown in SEQ ID NO.2)
The invention discloses a site for absorbing and utilizing rice nitrogenqNUE6The molecular marker method of (1), wherein the DNA of the rice material is molecularly marked with NUE2 primer, and if 135bp amplified fragment can be amplified, it marks the absorption of nitrogenUsing gene lociqNUE6Is present. Wherein the rice material is preferably a nitrogen-absorbing variety or a breeding material.
Screening the rice nitrogen absorption and utilization siteqNUE6The method for labeling a molecule of (1), comprising the steps of:
step S1: a set of rice chromosome fragment introgression lines BC containing 212 lines is constructed by hybridization, backcross and selfing by taking a common wild rice material Y11 as a donor and a variety GH998 as an acceptor for efficiently absorbing nitrogen4F7；
Step S2: identifying the nitrogen absorption utilization rate under the conditions of low nitrogen and high nitrogen in the field; wherein the low nitrogen is 0kg/hm of field nitrogen content2The high nitrogen is the field nitrogen content of 150 kg/hm2。
And step 3: taking 30 rice plants in an introduction system for efficient absorption of nitrogen and inefficient absorption of nitrogen, extracting DNA of the rice plants by using a CTAB method, constructing a DNA nitrogen efficient utilization pool and an inefficient utilization pool, and performing main effect gene positioning on a rice nitrogen absorption utilization gene by using a QTL-seq based on a whole genome sequencing technology; wherein, the CTAB method is adopted, which can well remove polysaccharide and is more beneficial to extracting and purifying DNA of rice plants.
And 4, step 4: uniformly selecting 40 pairs of molecular markers (Mccouch, 2002) on a 6 th chromosome of the rice; by comparing Y11 with GH998, finding out insertion or deletion of more than or equal to 6 bp between the Y11 and the GH998, and further designing an InDel marker; selecting a polymorphic good marker between two parents for analyzing 212 parts of BC4F7A population; performing linkage analysis on the polymorphic marker genotype and the nitrogen absorption utilization level of the corresponding family, and converting the Kosambi function into a genetic distance; the method comprises the steps of utilizing a Windows QTLCart V2.5 software composite interval mapping method, scanning in a genome by taking 2cM as a step length, detecting QTL by adopting a 5% total significance level, estimating a significance threshold value of corresponding LOD statistic by using a permutation test method, and repeatedly analyzing for 1000 times.
And 5: obtaining a nitrogen absorption utilization variety GH998 nitrogen absorption utilization siteqNUE6One, located near the marker NUE2, attracts nitrogen throughThe molecular marker of the major gene is collected to detect whether the site of the major gene is contained in the nitrogen uptake utilization variety GH998 and derivatives thereof, and the nitrogen uptake utilization level is predicted.
Further, in step 3, the major gene is located on chromosome 6.
Further, in step 4, uniformly selecting 40 pairs of molecular markers (Mccouch, 2002) on the 6 th chromosome of the rice; comparing Y11 and 998 through BWA software to find out insertion or deletion (more than or equal to 6 bp) between the two, and then designing an InDel marker by using Primer 5. Selecting a polymorphic good marker between two parents for analyzing 212 parts of BC4F7And (4) a group. Carrying out linkage analysis on polymorphic marker genotypes and nitrogen absorption utilization levels of corresponding families by using MAPMARKER/EXP3.0 software, and converting a Kosambi function into a genetic distance; the method comprises the steps of utilizing a Windows QTL Cart V2.5 software composite interval mapping method, scanning in a genome by taking 2cM as a step length, detecting the QTL by adopting a 5% total significance level, estimating the significance threshold value of corresponding LOD statistic by using a permutation test method, and repeatedly analyzing for 1000 times.
The rice nitrogen absorption and utilization siteqNUE6Application in rice breeding.
Further, the rice nitrogen absorption and utilization siteqNUE6The method is applied to screening of nitrogen high-efficiency absorption and utilization varieties or strains.
The nitrogen absorption and utilization site of the inventionqNUE6The molecular marker primer of (2) is applied to rice molecular breeding.
Further, the nitrogen absorption and utilization siteqNUE6The molecular marker primer is applied to the rapid screening of the nitrogen high-efficiency absorption and utilization variety or strain.
Compared with the prior art, the invention has the beneficial effects that:
by adopting the technical scheme of the invention, the BC obtained by hybridizing, backcrossing and selfing the variety GH998 which is a common wild rice material Y11 and a nitrogen element high-efficiency absorption utilization variety is utilized4F7Performing correlation analysis and genetic linkage analysis to obtain a nitrogen absorption utilizationSite of the bodyqNUE6Through reaction withqNUE6The linked molecular markers are used for detecting whether the variety GH998 and the derivative varieties (lines) thereof for high-efficiency absorption and utilization of nitrogen contain nitrogen absorption and utilization sites, so that the level of the nitrogen absorption and utilization rate can be predicted, and the selection efficiency of the rice for high-efficiency absorption and utilization of nitrogen is greatly improved. The molecular marking method of the major gene locus for absorbing and utilizing nitrogen provided by the invention has the following advantages:
(1) the invention adopts a rice QTL-seq method based on whole genome sequencing and QTL genetic linkage analysis to locate the nitrogen absorption utilization gene locus for the first time internationally.
(2) The nitrogen absorption and utilization gene locus positioned by the molecular marker has accurate position and convenient and quick identification. By detecting the molecular markers linked with the nitrogen absorption utilization gene loci, the level of the nitrogen absorption utilization rate of the rice plant can be predicted, and the method is used for detecting the genotype of rice varieties or strains to judge whether the varieties or strains have the high-efficiency nitrogen absorption utilization performance, so that the varieties or strains for the high-efficiency nitrogen absorption utilization are rapidly screened for rice breeding, the detection of the nitrogen absorption utilization gene loci is convenient and rapid, and the detection is not easily influenced by the environment.
(3) The molecular marker assisted breeding selection has definite target and high efficiency. In the conventional breeding technology, in a breeding method for improving the nitrogen absorption and utilization rate, a donor parent and a receptor parent of a gene for efficiently absorbing and utilizing nitrogen are usually hybridized, backcrossed for multiple times or polymerized and backcrossed, and the efficiency of absorbing and utilizing nitrogen is selected individually; however, the identification process of the nitrogen absorption and utilization rate of the rice is complex, the number of influencing factors is large, the identification difficulty is high, and the reliability of the phenotype identification result is low; therefore, the breeding by efficiently absorbing and utilizing the nitrogen is time-consuming, labor-consuming, difficult and high in cost. By using the molecular marker in the invention to detect the gene locus for nitrogen absorption and utilization, the single plant for nitrogen high-efficiency absorption and utilization can be identified in the seedling stage, other plants are eliminated, the cost is saved, and the selection efficiency of the rice for nitrogen high-efficiency absorption and utilization is greatly improved.
Drawings
FIG. 1 is an electropherogram of a closely linked molecular marker of the nitrogen uptake utilization gene of the present invention,
FIG. 2 shows the chromosomal location of a GH998 nitrogen uptake and utilization gene of the present invention.
The labels in fig. 1 include: 1: y11 amplified banding pattern; 2-4: the strain amplification band type of the nitrogen low-efficiency absorption utilization rate; 5-10: amplification banding pattern of heterozygous strain; 11-13: the high-efficiency absorption and utilization rate of nitrogen is increased in the form of band; 14: GH998 amplified banding pattern.
Detailed Description
Preferred embodiments of the present invention are described in further detail below.
The material and the method are as follows:
step 1: constructing a genetic analysis segregation population;
a set of rice chromosome fragment introgression lines BC containing 212 lines is constructed by hybridization, backcross and selfing by taking a common wild rice material Y11 as a donor and a variety GH998 as an acceptor for efficiently absorbing nitrogen4F7
Using ordinary wild rice material Y11 as male parent and using variety GH998 as female parent to prepare F1Backcrossing the hybrid to BC with GH998 as female parent and 998/Y11 as male parent4F1Then selfing and generation-adding seed reproduction are carried out to obtain BC4F7And (4) a group.
Step 2: carrying out nitrogen absorption rate identification by adopting field identification;
the experiment was carried out in 2013 in the rice institute field (22 o51 'N, 108o 14') of Guangxi academy of agricultural sciences. The test field is paddy soil, and the nutrient components of the paddy field are as follows: 17.05 g/kg of organic matter, 0.12 percent of total nitrogen, 0.11 percent of total phosphorus, 1.78 percent of total potassium, 90.50 mg/kg of quick-acting nitrogen, 34.00 mg/kg of quick-acting phosphorus, 198.50 mg/kg of quick-acting potassium and 6.48 of pH value.
Nitrogen (0 kg/hm) was not applied in the experimental facility2) And applying nitrogen (150 kg/hm)2) 2 treatments, nitrogen application treatment is respectively carried out at 105 kg/hm on the 5 th day after the rice transplanting2And applying 45 kg/hm on day 252. The potassium fertilizer and the phosphate fertilizer are treated the same, and the application amount of the potassium fertilizer is 100kg/hm2Applying 70 kg/hm of potash fertilizer on the 5 th day of rice transplanting respectively2Applying 30 kg/hm of potash fertilizer on the 25 th day2100kg/hm of phosphorus2All used as base fertilizer. The cell area is 20 m2The row spacing was 23.1 cm, the plant spacing was 13.2 cm, and the repetition was 3 times.
After the rice is mature, taking 10 holes of a representative plant according to the average value of the cell general survey results, cutting along the ground, separating stems and leaves from seeds, deactivating enzyme in a 105 ℃ oven for 30 min, drying for 4 days at 75 ℃ until the weight is constant, weighing, and crushing. And measuring the total nitrogen of the plants by using a semi-micro Kjeldahl method. The nitrogen absorption utilization was calculated as follows:
nitrogen uptake and utilization rate RE = (TN)F-TN0)/N×100
Wherein TN0The total nitrogen content of the plants without nitrogen application; TN (twisted nematic)FThe total nitrogen content of the normal nitrogen-applying plants.
And step 3: the nitrogen uptake of the Y11/GH998 isolated population was localized using the major gene;
(1) all the leaves of the single plant are taken in the tillering stage and stored in a refrigerator at minus 80 ℃. According to BC of 20134F7And (3) selecting 30 single plants with extremely low absorption utilization rate and 30 single plants with extremely high absorption utilization rate from the identification results of all the plants in the population, selecting leaves of the 60 single plants and 2 parents, and extracting plant genome DNA by using a CTAB method. The purity and integrity of DNA of a single sample are analyzed by agarose gel electrophoresis, and the DNA of 30 extremely low single strains with the nitrogen absorption and utilization rate is equivalently mixed to form an extremely low pool (L-pool), and the DNA of 30 extremely high single strains is equivalently mixed to form an extremely high pool (H-pool).
(2) The DNA sample is randomly broken into fragments with the length of 350bp by a Covari crusher, a TruSeqLibrary Construction Kit is adopted for library Construction, the DNA fragments are subjected to end repair, ployA tail addition, sequencing joint addition, purification, PCR amplification and other steps to complete preparation of the whole library, the constructed library is sequenced by an illumina HiSeq 2500, the sequencing depth parent is 20 ×, and the offspring is 30 ×.
(3) QTL-seq analysis
The original sequencing sequences (Sequenced Reads) or raw Reads, which contained therein the tagged, low quality Reads, were Sequenced. In order to ensure the quality of information analysis, raw reads must be filtered to obtain clean reads, and subsequent analysis is based on the clean reads. The sequencing generates Raw data 53.68 Gb and filtered Clean data53.00 Gb. The data processing steps are as follows:
① remove reads pair of tape connector (adapter);
② when the content of N contained in the single-ended sequencing read exceeds 10% of the length proportion of the read, the pair of paired reads needs to be removed;
③ when the number of low-mass (Q.ltoreq.5) bases contained in a single-ended sequencing read exceeds 50% of the length proportion of the read, it is necessary to remove this pair of paired reads.
④ calculation of SNP-index
The calculation of the SNP-index is a statistical method of the SNP in the filial generation pool, the principle is that the base of each base site is counted by sequencing reads, the number of the reads which are the same or different with the parent or the reference genome in the filial generation pool at a certain base site is counted by taking a certain parent or the reference genome as a reference, the proportion of the number of the reads which are different to the total number is calculated, namely the SNP-index of the base site, for the project with data of two filial generation pools, the points with the SNP-index of less than 0.3 in the two pools are filtered, for the filtered SNP-index, the average value of the SNP-indexes of all the SNPs in a certain window is counted by using a sliding window mode to serve as the SNP-index of the window, the common default parameter is a window with 1Mb, the sliding with 10kb, the SNP-indexes of the two filial generation pools are respectively calculated according to the method, and then the difference value of the SNP-indexes of the two filial generation pools is calculated, namely the SNP-index △ (SNP-index).
⑤ QTL location for nitrogen utilization
Calculating △ (SNP-index), namely, making a difference between two filial generation SNP-index, namely △ (SNP-index) = SNP-index (extreme character B) -SNP-index (extreme character A), carrying out 1000 times of replacement tests, selecting 95% confidence level as a screening threshold, analyzing a window which is larger than the threshold as a candidate interval under the 95% confidence level, and selecting SNP sites with significant difference between the SNP-index of two filial generations in the area, namely selecting sites with the SNP-index of 2 (extreme character B) of the filial generation being larger than or equal to 0.7 and the SNP-index of 1 (extreme character A) of the filial generation being smaller than or equal to 0.3, and selecting QTL sites which are possibly related to the nitrogen absorption utilization rate in the area meeting the characteristics.
And 4, step 4: linkage genetic mapping analysis
Uniformly selecting 40 pairs of molecular markers (Mccouch, 2002) on a 6 th chromosome of the rice; y11 and 998 are compared through BWA software to find out insertion or deletion between the two, and then Primer5 is used for designing an InDel marker. Selecting a polymorphic good marker between two parents for analyzing 212 parts of BC4F7And (4) a group.
The SSR analysis program is as follows:
① the 12. mu.l PCR reaction system includes DNA (10 ng/. mu.l), 2. mu.l, Primer (4 pmol/. mu.l), 1.5. mu.l, 2 × Taq PCR Master Mix (Zhongkoitai), 6. mu.l, ddH2O, 2.5. mu.l;
② the amplification reaction is carried out on an ETC811 amplification instrument, the PCR reaction conditions are pre-denaturation at 95 ℃ for 5min, denaturation at 95 ℃ for 35s, renaturation at 55 ℃ for 35s, extension at 72 ℃ for 1min, 35 cycles, and finally extension at 72 ℃ for 7min, the amplification product is separated by 8% non-denaturing PAGE gel, silver staining is used for color development, and a polymorphic primer is arranged between parents at BC4F7Analyzing the population to obtain genotype data of the sample;
③, the MARMAKER/EXP3.0 analysis software is used for separating and analyzing the nitrogen absorption utilization efficiency and SSR markers, and converting a Kosambi function into a genetic distance (cM). The Windows QTL Cartogrer V2.5 software Composite Interval Mapping (CIM) is used for scanning in a genome by taking 2cM as a step length, the detection of the QTL adopts a 5% overall significance level, the significance threshold of the corresponding LOD statistic is estimated by a permutation test method, and the analysis is repeated for 1000 times.
And 5: results and analysis
The nitrogen absorption utilization rates of Y11 and GH998 are respectively 7.09% and 33.18% through field identification, which shows that GH998 has higher nitrogen absorption utilization capacity, and Y11 has higher nitrogen absorption utilization capacityThe force is very low. By comparing SNP-index of low pool and high pool of nitrogen utilization rate, window larger than threshold value under 95% confidence level is analyzed as candidate interval, we find that SNP imbalance condition appears between 6,099,043 and 8,940,631 bp on 6 th chromosome, which is QTL locus for controlling nitrogen utilization rate of rice, we name it as QTL locusqNUE6。
Selecting 63 BC of 16 pairs of SSR marker pairs which show polymorphism between parents from 40 pairs of SSR markers4F7The individual plants were subjected to background detection, and as a result, they were found to have a difference in region between the rice chromosome 6 markers RM539 and RM 136. We designed 30 pairs of insertion and deletion markers by using Y11 and GH998 resequencing results, wherein 24 pairs have polymorphism, and the 212 high-generation population single strains are subjected to genotype analysis by combining the 16 pairs of SSR markers, and the range of a gene NUE6 is narrowed to 8, 051, 882-8, 990 and 567 bp by using MAPMAKER3.0 and WinQTL Cart 3.0 software. Therefore, the 6 th chromosome interval 8, 051, 882-8, 990, 567 bp of rice may contain the gene for controlling the rice nitrogen utilization character. As shown in FIG. 2, it was found that a site of nitrogen uptake and utilization in rice was detected between the molecular markers NUE2 and NUE23qNUE6LOD value of 4.96, contribution rate of 16.8%, 16.8% can account for phenotypic variation.
Molecular marker NUE2 primer sequence:
an upstream primer: GTTCCACATGTTGATGGG
A downstream primer: GATGAGGACGAGGAGGAG
The molecular marker NUE2 primer is used for amplifying DNA of varieties or breeding materials with high nitrogen absorption and utilization efficiency, as shown in figure 1, the molecular marker NUE2 can be used for amplifying amplified fragments of 135bp, and then marks sites of the varieties of rice with nitrogen absorption and utilizationqNUE6Is present.
The molecular marker linked with the gene locus is used for detecting whether the variety GH998 and the derivative varieties (systems) thereof contain the gene locus for absorbing and utilizing the nitrogen efficiently, so that the nitrogen absorbing and utilizing level can be predicted, and the selection efficiency of the rice for absorbing and utilizing the nitrogen efficiently is greatly improved.
The foregoing is a more detailed description of the invention in connection with specific preferred embodiments and it is not intended that the invention be limited to these specific details. For those skilled in the art to which the invention pertains, several simple deductions or substitutions can be made without departing from the spirit of the invention, and all shall be considered as belonging to the protection scope of the invention.
SEQUENCE LISTING (SEQUENCE LISTING)
<110> Rice research institute of Guangxi Zhuang autonomous region academy of agricultural sciences
<120> site qNUE6 for absorbing and utilizing rice nitrogen and molecular marking method thereof
<160>2
<170>PatentIn version 3.3
<210>1
<211>18
<212>DNA
<213> Artificial sequence
<400>1
gttccacatg ttgatggg 18
<210>2
<211>18
<212>DNA
<213> Artificial sequence
<400>2
gatgaggacg aggaggag 18
Claims (8)
1. Site for absorbing and utilizing nitrogen of riceqNUE6The molecular marker primer of (1), which is characterized in that: the rice nitrogen absorption and utilization siteqNUE6Is a site between molecular markers NUE2 and NUE23, the molecular marker primer is a molecular marker NUE2 primer, and the sequence is as follows: SEQ ID NO. 1/SEQ ID NO. 2.
2. Site for absorbing and utilizing nitrogen of riceqNUE6The molecular labeling method of (1), characterized in that: the method of using the site for the absorption and utilization of nitrogen in rice according to claim 1qNUE6The molecular marker primer of (a) amplifies DNA of a rice material, and if a 135bp amplified fragment can be amplified, the molecular marker primer marks a nitrogen absorption utilization gene siteqNUE6Is present.
3. Screening of rice nitrogen absorption and utilization sitesqNUE6The method for labeling a molecule of (1), which comprises: the method comprises the following steps:
step S1: a set of rice chromosome fragment introgression lines BC containing 212 lines is constructed by hybridization, backcross and selfing by taking a common wild rice material Y11 as a donor and a variety GH998 as an acceptor for efficiently absorbing nitrogen4F7；
Step S2: identifying the nitrogen absorption utilization rate under the conditions of low nitrogen and high nitrogen in the field;
and step 3: taking 30 plants of nitrogen in the introduction system for high-efficiency absorption and low-efficiency absorption, extracting DNA of rice plants by using a CTAB method, constructing a DNA nitrogen high-efficiency utilization pool and a low-efficiency utilization pool, and performing main-effect gene positioning on the rice nitrogen absorption utilization genes by using a QTL-seq based on a whole genome sequencing technology;
and 4, step 4: uniformly selecting 40 pairs of molecular markers on a 6 th chromosome of the rice; comparing Y11 and 998 to find out insertion or deletion of more than or equal to 6 bp between the two, and further designing an InDel marker; selecting a polymorphic good marker between two parents for analyzing 212 parts of BC4F7A body; performing linkage analysis on the polymorphic marker genotype and the nitrogen absorption utilization level of the corresponding family, and converting the Kosambi function into a genetic distance; the method comprises the steps of scanning in a genome by using a Windows QTL Cart V2.5 software composite interval mapping method and taking 2cM as a step length, detecting the QTL by adopting a 5% overall significance level, estimating a significance threshold value of corresponding LOD statistic by using a permutation test method, and repeatedly analyzing for 1000 times;
and 5: obtaining a high-efficiency absorption and utilization variety GH998 nitrogen absorption and utilization site of rice nitrogenqNUE6One is located near the marker NUE2, and the nitrogen absorption and utilization level of the variety GH998 and the derived varieties and/or lines thereof is predicted by detecting whether the main gene site is contained in the variety GH998 and the derived varieties and/or lines thereof through the molecular marker of the main gene.
4. The method of claim 3Screening of rice nitrogen absorption and utilization sitesqNUE6The method for labeling a molecule of (1), which comprises: in step 3, the major gene is located on chromosome 6.
5. The site of claim 2 for the absorption and utilization of rice nitrogenqNUE6The molecular marking method of (2) is applied to rice breeding.
6. The site for absorbing and utilizing rice nitrogen according to claim 5qNUE6The application of the molecular marking method in rice breeding is characterized in that: the rice nitrogen absorption and utilization siteqNUE6The method is applied to screening of nitrogen high-efficiency absorption and utilization varieties or strains.
7. The site of claim 1 for absorption and utilization of nitrogenqNUE6The molecular marker primer of (2) is applied to rice molecular breeding.
8. The nitrogen uptake utilization site of claim 7qNUE6The application of the molecular marker primer in rice molecular breeding is characterized in that: the nitrogen absorption and utilization siteqNUE6The molecular marker primer is applied to the rapid screening of the nitrogen high-efficiency absorption and utilization variety or strain.
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| CN114561403B (en) * | 2022-04-11 | 2023-07-18 | 广西壮族自治区农业科学院 | Rice nitrogen fertilizer utilization efficiency gene OsNPF3.1 and application thereof |
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| CN101805731A (en) * | 2010-04-23 | 2010-08-18 | 中国水稻研究所 | Molecular marker linked with paddy nitrogen nutrition regulation and control gene and application thereof |
| CN105039531A (en) * | 2015-07-06 | 2015-11-11 | 云南农业大学 | Expressive molecular marker for detecting rice blast resistance and nitrogenous fertilizer response, and application thereof |
| CN105112504B (en) * | 2015-07-23 | 2018-07-24 | 毕节市农业科学研究所 | A kind of molecular marker identification method of the high nitrogen use efficiency gene of rice |
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