CN106947777B - Application of nitrogen transport gene OsNPF7.4 in rice breeding - Google Patents
Application of nitrogen transport gene OsNPF7.4 in rice breeding Download PDFInfo
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Abstract
The present invention discloses a nitrogen transport geneOsNPF7.4The application in rice breeding belongs to the field of plant gene engineering.OsNPF7.4The amino acid sequence of the gene coding protein is shown as SEQ ID NO.1, and the cDNA sequence is shown as SEQ ID NO. 2. The invention constructs riceOsNPF7.4Gene interference plant,OsNPF7.4Plants with gene overexpression, found by reductionOsNPF7.4The gene expression can increase the tillering number, the spike number and the grain filling number of normal paddy rice, therebyOsNPF7.4The gene can be used in rice breeding to improve the rice yield.OsNPF7.4The gene has important application value in the aspect of explaining that nitrogen transportation influences the growth and development process of plants.
Description
Technical Field
The invention belongs to the field of plant genetic engineering, and particularly relates to a nitrogen transport geneOsNPF7.4Application in rice breeding.
Background
Plants obtain nitrogen by absorbing ammonia, nitrate, amino acids, soluble peptides, etc. in soil; nitrogen uptake and transport is mainly accomplished by Transporters such as ammonium transport protein (AMT), nitrate transport protein (NRT), amino acid transport protein (AAT), peptide transport Protein (PTR) (Williams L, Miller A. Transporters for the uptake and purification of nitrogenes sources, Annual Review of Plant physiology and Plant Molecular Biology, 2001, 52: 659. sup. 688.). Ammonium is taken up by the Plant AMT and then glutamine and glutamate are synthesized by Glutamine Synthetase (GS) and glutamate synthase (GOGAT), which in turn further form other amino acids (Sonoda Y, Ikeda A, Saiki S, et al. feedbackslip. Ache ammonium transporter gene family AMT1 by glutamine in rice. Plant cell physiology, 2003, 44: 1396. sup. 1402.). Plants can absorb environmental nitrates via NRT2 of the High Affinity Transport System (HATS) and NRT1 of the Low Affinity Transport System (LATS), form ammonium upon reduction by Nitrate Reductase (NR) and nitrite reductase (NiR), and further form amino acids (Paungfoo-Lonhienne C, Lonhienne T G, Rentsch D, et al. plant can use protein as a nitrogen source with a sodium out of organic chemistry. Proceedings of the National Academy of Sciences, 2008, 105: 4524-4529.).
The NPF family of nitrogen transporters includes the NRT1 and PTR subfamilies, where different members transport nitrate, oligopeptides, amino acids, etc. at different sites of the plant and play different roles in plant growth and development (Rentsch D, Schmidt S, Tegeter M. transporters for uptake and allocation of organic nitrogen compounds in plants Febs Letters, 2007, 581: 2281-2289.). OsNPF2.2 mediates the unloading of xylem nitrate, affecting rice growth and seed filling and setting (Li Y, Ouyang J, Wa)ng Y Y, et al. Disruption ofthe rice nitrate transporter OsNPF2.2 hinders root-to-shoot nitrate transportand vascular development. Scientific reports, 2015, 5: 9635.）。OsNPF7.2Has low affinity transport for nitrate and can affect plant growth (Hu R, Qiu D, Chen Y, et al, Knock-down of aeroplast localized low-affinity nitrate transportOsNPF7.2affects ricegrowth under high nitrate supply. Frontiers in plant science, 2016, 7.）。
While nitrogen nutrition is known to promote plant growth and development, there is currently no systematic understanding of what types of plants nitrogen nutrition affects growth and development. In addition, more than 80 members of the rice NPF family exist, nitrogen nutrition responds through which member of the NPF gene family, nitrogen nutrition transportation is mediated at what position, and therefore what type of plant growth and development are influenced, and the existence of nitrogen nutrition is almost unknown at present. Therefore, the excavated NPF family may have nitrogen efficient transport genes, especially the nitrogen transport key genes capable of controlling the plant type of rice, and is beneficial to the cultivation of high-yield rice varieties. The invention discovers that NPF family through long-term researchOsNPF7.4The gene has important negative regulation and control effect on rice tillering, and after the gene is interfered and knocked out, the rice without transgenic markers can be cultivated and directly applied to plant type improvement, so that the yield of the rice is increased.
Disclosure of Invention
The present invention is directed to solving the problems of the prior art and to providing a nitrogen transport geneOsNPF7.4Application in rice breeding.
The purpose of the invention is realized by the following technical scheme:
the invention relates to a nitrogen transport gene of riceOsNPF7.4As a subject, a clone from rice flower 11OsNPF7.4The cDNA sequence of (1). Construction by RNAi techniqueOsNPF7.4The gene interference expression vector adopts agrobacteriumEHA105The mediated genetic transformation method is to introduce the interference expression vector into the normal japonica rice variety flower 11 to obtainOsNPF7.4Interfering plant with reduced gene expression level, tillering number, effective spike number and grain filling number of interfering plant and contrast wild type middle flower 11Compared with the prior art, the method is obviously improved. At the same time constructOsNPF7.4Gene overexpression vector, and introducing the overexpression vector into middle flower 11 to obtainOsNPF7.4Compared with the medium flower 11, the tillering number, the effective spike number and the grain filling number of the gene over-expression plant are obviously reduced. These results show that by reducingOsNPF7.4The gene expression can increase the tillering number and the effective spike number of each normal rice plant and increase the grain filling number of each plant, thereby improving the rice yield.
Based on the discovery of the present inventionOsNPF7.4The function of the gene can be used for rice breeding. The rice breeding is to improve the tillering number, the effective spike number and the grain filling number of the rice, thereby improving the rice yield. In particular can be reduced by RNAi techniquesOsNPF7.4Expression of gene or knockout by CRISPR or like gene editing technologyOsNPF7.4The gene increases the tillering number, the effective spike number and the grain filling number of the rice, and achieves the purpose of improving the rice yield. SaidOsNPF7.4The amino acid sequence of the gene-coded OsNPF7.4 protein is shown in SEQ ID NO. 1; saidOsNPF7.4The cDNA sequence of the gene is preferably shown in SEQ ID NO. 2.
It is understood that the amino acid sequence shown in SEQ ID NO.1 can be variously substituted, added and/or deleted by one or several amino acids by those skilled in the art to obtain an amino acid sequence having equivalent functions without affecting the activity of the OsNPF7.4 protein (i.e., without being in the active center of the protein). Therefore, the OsNPF7.4 protein also comprises a protein with equivalent activity obtained by substituting, replacing and/or adding one or more amino acids in the amino acid sequence shown in SEQ ID NO. 1. Furthermore, it will be appreciated that, given the degeneracy of codons and the preference of codons for different species, one skilled in the art can use codons suitable for expression in a particular species as desired.
The invention has the advantages and effects that:
(1) cloned according to the inventionOsNPF7.4The tillering capacity of rice is enhanced after gene interference expression, which shows thatOsNPF7.4The gene has obvious influence on increasing the tillering number of rice, so that the gene is reduced by the gene engineering technologyOsNPF7.4Expression of the gene can improve plant yield. This not only helps to get throughThe high-yield rice is cultivated under the normal nitrogen applying condition, and the variety improvement of the plant can be carried out by combining the gene editing technology and molecular breeding.
（2）OsNPF7.4The successful cloning of the gene further proves the important function of the NPF family in the nitrogen absorption process, has important significance for clarifying the biological function of the nitrogen transport family, and has great promotion effect on further understanding the plant nitrogen metabolic pathway and improving the nitrogen absorption efficiency.
(3) Although some genes have been cloned to improve plant yield, the molecular mechanisms for plant yield increase are still unclear. And cloned according to the inventionOsNPF7.4The gene can improve the tillering number, the effective spike number and the grain filling number of rice, and has great promotion effect on determining the key factors of plant yield increase.
Drawings
FIG. 1 shows a control flower 11,OsNPF7.42 strains of gene over-expression plants andOsNPF7.4the phenotype of the whole plant of 2 lines of the gene interference plant.
FIG. 2 shows the flower 11 in the control,OsNPF7.42 strains of gene over-expression plants andOsNPF7.4statistical histogram of tillering number of 2 lines of gene-interfered plants, data were analyzed for variables (ANOVA) using SPSS software, significance of differences was analyzed at 0.05 level using Duncan's, and asterisks (#) of different groups indicated significant differences compared to controls.
FIG. 3 shows the flower 11 in the control,OsNPF7.42 strains of gene over-expression plants andOsNPF7.4statistical histogram of effective panicles of 2 lines of gene-interfered plants, data were analyzed by variable analysis (ANOVA) using SPSS software, and differential significance was analyzed at 0.05 level using Duncan's, with different group asterisks (asterisks) indicating significant differences compared to controls.
FIG. 4 shows the flower 11 in the control,OsNPF7.42 strains of gene over-expression plants andOsNPF7.4a phenotype map of the number of grouted grains of each seed of 2 lines of gene interference plants.
FIG. 5 shows the flower 11 in the control,OsNPF7.42 strains of gene over-expression plants andOsNPF7.42 gene interference plantsStatistical plots of number of grouted grains per seed of the line, data were analyzed for variables (ANOVA) using SPSS software, and for significance of differences at the 0.05 level using Duncan's, with different group asterisks (x) indicating significant differences from the control.
FIG. 6 shows the flower 11 in the control,OsNPF7.42 strains of gene over-expression plants andOsNPF7.4statistical histograms of the expression levels of 2 lines of gene-interfered plants were obtained by performing variable analysis (ANOVA) using SPSS software and significance analysis of differences at 0.05 level using Duncan's, with different group asterisks (A) indicating significant differences compared to the control.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art; the experimental procedures used are conventional and can be carried out according to recombinant techniques already described (see molecular cloning, A laboratory Manual, 2 nd edition, Cold spring harbor laboratory Press, Cold spring harbor, N.Y.); the materials, reagents and the like used are all commercially available.
Example 1OsNPF7.4Construction of Gene-interfering plants
Extracting RNA of rice middle flower 11, reversely transcribing the RNA into cDNA, and performing primer pair:
F1：5'-GGTACCACGCCATGGAGAGAGGCCAGCA-3'（KpnI），
R1：5'-GGATCCAGGTGACCAGCACCATGCCCA-3'（BamH I）；
F2：5'-ACTAGTACGCCATGGAGAGAGGCCAGCA-3'（SpeI），
R2：5'-GAGCTCAGGTGACCAGCACCATGCCCA-3'（SacI）；
respective PCR amplificationOsNPF7.4The cDNA fragment of the gene is cut by corresponding restriction enzyme and then is connected into pTCK303 vector to constructOsNPF7.4Interfering expression vector of geneOsNPF7.4-pTCK 303. Using Agrobacterium EHA 105-mediated genetic transformation methods, the interference table was generatedThe vector is introduced into the flower 11 of the normal japonica rice variety.
Transplanting all the obtained transgenic seedlings into a basket with soil, watering and fertilizing at regular intervals, planting the seedlings in a field when the seedlings grow to be about 10cm in height, extracting genome DNA (deoxyribonucleic acid) and detecting transgenic plants through PCR (polymerase chain reaction), wherein a detection primer pair is as follows:
F3：5'-GATGTTGGCGACCTCGTATT-3'，
R3：5'-TCGTTATGTTTATCGGCACTTT-3'。
if 517bp fragments are amplified, the transgenic plants are positive plants. The positive plants are harvested and planted individually until homozygous transgenic plants are identified at T2 generation, and the transgenic plants are obtainedOsNPF7.4The gene interferes with the plant.OsNPF7.4The tillering number and the effective spike number of the gene interference plant are far more than those of the flower 11 plant in the control, and the difference is obvious, as shown in figures 1, 2 and 3; and the number of grouted grains per interfering plant was significantly increased over the control, as shown in fig. 4 and 5.
GetOsNPF7.4Gene interference of plant leaf, extracting leaf RNA and reverse transcribing to cDNA, real-time fluorescent quantitative PCR detectionOsNPF7.4The expression level of the gene in the interfering plant is shown (FIG. 6)OsNPF7.4The expression level of the gene was reduced compared to control flower 11. Primer pairs for real-time fluorescent quantitative PCR:
F4：ACGCCATGGAGAGAGGCCAGCA，
R4：ACCAGGTTCGTGGCAATGCCGT。
example 2OsNPF7.4Construction of Gene-overexpressing plants
Extracting RNA of rice middle flower 11, reversely transcribing the RNA into cDNA, and performing primer pair:
F5：5'-GGTACCATGGACGCCGGCGACGCCATGGAG-3'（kpnI），
R5：5'-TCTAGATGACACGACCGTCTTCACCTTGTA-3'（XbaI）；
amplification by PCROsNPF7.4After cDNA of the gene, bykpnI andXbai was ligated into pCAMBIA-1306 vector (pCAMBIA-1306 vector from Cambia Co.) to constructOsNPF7.4Overexpression vector of geneOsNPF7.4-p1306。By using AgrobacteriumEHA105The mediated genetic transformation method is to introduce the overexpression vector into the flower 11 of the normal rice variety.
Transplanting all the obtained transgenic seedlings into a basket with soil, watering and fertilizing at regular intervals, planting the seedlings in a field when the seedlings grow to be about 10cm in height, extracting genome DNA (deoxyribonucleic acid) and detecting transgenic plants through PCR (polymerase chain reaction), wherein a detection primer pair is as follows:
F3：5'-GATGTTGGCGACCTCGTATT-3'，
R3：5'-TCGTTATGTTTATCGGCACTTT-3'；
if 517bp fragments are amplified, the transgenic plants are positive plants. The positive plants are harvested and planted individually until homozygous transgenic plants are identified at T2 generation, and the transgenic plants are obtainedOsNPF7.4And (3) gene overexpression plants.OsNPF7.4The tillering number and the effective spike number of the gene over-expression plant are far less than those of a flower 11 plant in a control, and the difference is obvious, as shown in figures 1, 2 and 3; and the number of grouted grains per plant of the over-expressed plants was significantly reduced compared to the control, as shown in fig. 4 and 5.
GetOsNPF7.4Extracting RNA from the leaf of the plant with gene over-expression, reverse transcribing the RNA into cDNA, and detecting the cDNA by real-time fluorescent quantitative PCROsNPF7.4The expression level of the gene in the over-expressed plants was determined in the same manner as in example 1, and the results are shown (FIG. 6)OsNPF7.4The expression level of the gene was much higher than that of the control flower 11.
The above results show that it is possible to obtain,OsNPF7.4the gene can improve the tillering number, the effective spike number and the grain filling number of the rice by reducing the expression quantity, and finally improve the rice yield.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
SEQUENCE LISTING
<110> Wuhan bioengineering college
Application of <120> nitrogen transport gene OsNPF7.4 in rice breeding
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Claims (2)
1.OsNPF7.4The application of the gene in rice breeding is characterized in that: the rice breeding is to improve the tillering number, the spike number and the grain filling number of the rice; saidOsNPF7.4The amino acid sequence of the gene-coded OsNPF7.4 protein is shown as SEQIDNO.1; by reducingOsNPF7.4Expression or knock-out of genesOsNPF7.4The gene increases the tillering number, spike number and grain filling number of rice.
2. Use according to claim 1, characterized in that: saidOsNPF7.4The cDNA sequence of the gene is shown in SEQ ID NO. 2.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101027397A (en) * | 2004-04-23 | 2007-08-29 | 西尔斯公司 | Nucleotide sequences and polypeptides encoded thereby useful for modifying nitrogen use efficiency characteristics in plants |
| CN102604962A (en) * | 2011-01-20 | 2012-07-25 | 中国科学院华南植物园 | Gene OsPTR9 capable of improving nitrogen absorption efficiency and yield of rice and application thereof |
| CN104277101A (en) * | 2014-09-24 | 2015-01-14 | 中国科学院遗传与发育生物学研究所 | Application of rice nitrate transporter NRT1.1B in enhancing nitrogen utilization efficiency of plants |
| CN106222180A (en) * | 2016-07-28 | 2016-12-14 | 武汉生物工程学院 | Improve rice yield and the gene OsNPF7.3 of grain of rice protein content and purposes |
| CN106434666A (en) * | 2016-10-20 | 2017-02-22 | 武汉生物工程学院 | Application of promoter achieving specific expression in rice tiller bud base and panicle |
| CN106518993A (en) * | 2016-10-25 | 2017-03-22 | 武汉生物工程学院 | Application of amino acid transporter gene OsAAP3 in rice seed selection |
-
2017
- 2017-05-11 CN CN201710330664.8A patent/CN106947777B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101027397A (en) * | 2004-04-23 | 2007-08-29 | 西尔斯公司 | Nucleotide sequences and polypeptides encoded thereby useful for modifying nitrogen use efficiency characteristics in plants |
| CN102604962A (en) * | 2011-01-20 | 2012-07-25 | 中国科学院华南植物园 | Gene OsPTR9 capable of improving nitrogen absorption efficiency and yield of rice and application thereof |
| CN104277101A (en) * | 2014-09-24 | 2015-01-14 | 中国科学院遗传与发育生物学研究所 | Application of rice nitrate transporter NRT1.1B in enhancing nitrogen utilization efficiency of plants |
| CN106222180A (en) * | 2016-07-28 | 2016-12-14 | 武汉生物工程学院 | Improve rice yield and the gene OsNPF7.3 of grain of rice protein content and purposes |
| CN106434666A (en) * | 2016-10-20 | 2017-02-22 | 武汉生物工程学院 | Application of promoter achieving specific expression in rice tiller bud base and panicle |
| CN106518993A (en) * | 2016-10-25 | 2017-03-22 | 武汉生物工程学院 | Application of amino acid transporter gene OsAAP3 in rice seed selection |
Non-Patent Citations (5)
| Title |
|---|
| ACCESSION NO.XM_015778560，PREDICTED: Oryza sativa Japonica Group protein NRT1/ PTR FAMILY 8.3 （LOC4336852），mRNA;无;《GenBank》;20160301;FEATURES,ORIGIN * |
| Altered Xylem-Phloem Transfer of Amino Acids Affects Metabolism and Leads to Increased Seed Yield and Oil Content in Arabidopsis;L Zhang et al;《Plant Cell》;20101130;第22卷(第11期);第3603-3620页 * |
| Over-expression of OsPTR6 in rice increased plant growth at differentnitrogen supplies but decreased nitrogen use efficiency at high ammonium supply;Xiaorong Fan et al;《Plant Science》;20140608;第227卷;第1-11页 * |
| Transporters for uptake and allocation of organic nitrogen compounds in plants;Doris Rentsch et al;《FEBS Letters》;20070418;第2281-2289页 * |
| 植物寡肽运输与硝酸根运输基因家族的研究进展;蔡昭艳 等;《热带亚热带植物学报》;20110115;第19卷(第1期);第91-96页 * |
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