CN103255148B - Application of paddy rice transcription factor Os01g18440 gene - Google Patents
Application of paddy rice transcription factor Os01g18440 gene Download PDFInfo
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Abstract
The invention relates to an application of a paddy rice transcription factor Os01g18440 gene. A constitutive transcription factor is constructed by fusing a transcription factor activation motif VP64 and the paddy rice transcription factor Os01g18440 gene and transformed into paddy rice, the plant height is changed, and the blooming period of the paddy rice is regulated, so that the output of the paddy rice is increased. The application disclosed by the invention has important theoretical value for detailly illuminating paddy rice blooming, fruit ripening and upgrowth regulating and controlling mechanisms, can be used for increasing the output of the paddy rice through a transgenic technology and has important significance in production practice.
Description
Technical field
The present invention relates to genetically engineered field, specifically, relate to the application of rice transcription factor Os01g18440 gene.
Background technology
Paddy rice (Oryza sativa L.) is one of most important food crop of depending on for existence of the mankind, also be the pattern species of an important functional gene research, relative genetics and molecular biology research receive much attention always, wherein, the regulation and control of transcriptional level are the important way of gene expression regulation.The research of current increasing production of rice depends on limited Rice Germplasm Resources, and traditional cross-breeding advantage weakens gradually, and Transgenic Rice technology becomes the research means of increasing production of rice gradually.
In vegitabilia, the plant that can form seed accounts for the more than 2/3rds of plant total, and as important organ of multiplication, seed is simultaneously also for people provide food source, and paddy rice is exactly important representative wherein, and seed source is in the ovule of after fertilization.From molecular biological angle, the Development and germination of seed be one orderly, optionally genetic expression process.And transcription factor has played critical effect in the accuracy controlling of gene betwixt.
MADS-box gene is extensively present in (Alvarez-Buylla etc., 2000 in animal, plant and fungi; Becker and Theissen, 2003; Kaufmann etc., 2005).To MADS-box gene studies the earliest be to start from the floral shape mutant of Common Snapdragon and Arabidopis thaliana., there is a large amount of gene redundancy events in MADS-box genes encode transcription factors, thereby formed a multigene family (Becker and Theissen, 2003 in plant (particularly angiosperm) evolutionary process; Irish, 2003).Nearly 107 MADS-box genes in Arabidopis thaliana, show from current genomic data, nearly more than 70 MADS-box genes (Litt and Irish, 2003 in paddy rice; Martinez-Castilla and Alvarez-Buylla, 2003; Parenicova etc., 2003; Nam etc., 2004; Causier etc., 2005; Irish and Litt, 2005).
MADS-box gene is a sequence-specific regulatory gene family, and coded albumen is transcription factor, is mainly made up of wherein MADS box high conservative 5 parts such as MADS box, K box, I district, C-terminal, N-terminal.MADS-box transcription factor is with its conserved domain and the specific DNA sequence dna regulate gene expression that combines by Dimerized form.Large quantity research shows, MADS-box gene has important effect in the growth of plant flowers, has cloned the MADS-box gene of many ABC classes relevant to development of floral organs, and found many MADS-box genes that can regulation florescence.At present, increasing research shows, MADS-box gene also has important effect in the growth of fruit, maturation.
The ripening process of fruit is subject to the regulation and control of external environmental condition and gene.2002, Vrebalov etc. found that the maturation of MADS-box gene in tomato and fruit is closely related.If Ripening-Inhibitor(RIN in the discovery tomatoes such as Vrebalov) site undergos mutation, and tomato fruit can not be ripe, and there will be sepal to become large phenomenon.Find to exist on rin site the MADS-box gene of 2 series connection by positional cloning, respectively called after LeMADS-RIN and LeMADS-MC, the former regulating fruit ripening process, latter affects sepal and first educates with inflorescence decisive.The ripening process of fruit and output have direct relation, therefore, to blooming for understanding on the further research theory of this transcription factor, the regulation and control of fruit maturation and output provide new thread, on putting into practice, also will provide fundamental basis for high-yield breeding of crops.
VP16 finds in animal virus gene, has now been widely applied in plant, is mainly used in the research of transcriptional control of plant gene.VP64 is merged the class enhanser forming by 4 VP16 functional domain motifs.Transcription factor effect in vivo can be divided into two kinds substantially: a kind of is transcriptional enhancer, another kind of for transcribing inhibition.After transcription factor and the fusion of VP16 functional domain motif, it will strengthen the function of transcription factor, thereby in transfer-gen plant, occurs that more obvious phenotype changes.
Summary of the invention
The object of this invention is to provide the application of rice transcription factor Os01g18440 gene.
In order to realize the object of the invention, the invention provides rice transcription factor Os01g18440 gene in the application improving in rice yield and the high bar proterties of adjusting and controlling rice, the sequence of described rice transcription factor Os01g18440 gene is: the i) nucleotide sequence shown in Seq ID No.1; Ii) under stringent condition with the nucleotide sequence of sequence hybridization shown in Seq ID No.1.Described stringent condition is containing 0.1 × SSPE of 0.1%SDS or containing in 0.1 × SSC solution of 0.1%SDS, hybridization at 65 DEG C, and wash film with this solution.
The aminoacid sequence of rice transcription factor Os01g18440 coded by said gene albumen is as shown in Seq ID No.2.
According to aforesaid application, it is the downstream of the CDS sequence construct to 4 of a rice transcription factor Os01g18440 gene transcription factor being activated to motif VP16, rice transformation, screening the final transgenic paddy rice that obtains.Described VP16 is from hsv (Herpes simplex virus), and the nucleotide sequence of 4 transcription factor activation motif VP16 is as shown in Seq ID No.3.
According to aforesaid application, it is the downstream of the CDS sequence of rice transcription factor Os01g18440 gene being activated to motif VP16 by Gateway system constructing to 4 transcription factor, rice transformation, screening the final transgenic paddy rice that obtains.
Particularly, aforesaid application comprises the following steps:
1) according to rice transcription factor Os01g18440 gene order design pcr amplification primer pair, it is forward primer F:5 '-CAAAAAAGCAGGCTTCATGGCTCGCAAGAAGATCGT-3 ' and reverse primer R:5 '-CAAGAAAGCTGGGTCTTACATGGGAGGGAAAGATGG-3 ';
2) as template, utilize above-mentioned primer taking wild Japanese fine ' kitaake ' rice cDNA, obtain Os01g18440 gene complete sequence by pcr amplification;
3) by PCR product cloning to pDONER cloning vector, correct through checking order;
4) sequence comprising taking border, binary expression vector pCAMBIA1300 left and right is as frame sequence, pass through vitro recombination, ubi promoter-VP64-Gateway is expressed to unit, 35S promoter-asRED expression unit and 35S promoter-bar expression unit and construct with it, obtain the complete sequence of carrier nVP64-bar-asRED as shown in Seq ID No.4;
5) reacted goal gene Os01g18440 is building up to carrier nVP64-bar-asRED above by LR, obtain plant expression vector ubi:VP64-Os01g18440, its sequence is as shown in Seq ID NO.5;
6) adopt the conventional biotechnological meanss such as agrobacterium-mediated transformation, directly delivered DNA, microinjection or electroporation method, by carrier ubi:VP64-Os01g18440 Introduced into Rice, screening the final transgenic paddy rice that obtains.
Further, step 6) adopts agriculture bacillus mediated method, utilize pCAMBIA1301 that carrier ubi:VP64-Os01g18440 is proceeded in Rice Callus, with transforming containing the AAM conversion fluid of inductor and Agrobacterium, material after conversion through cultivating altogether, de-bacterium, screening, break up, take root, hardening and transplanting, with Basta screening transgenic rice plant.
The present invention also provides a kind of plant expression vector ubi:VP64-Os01g18440, and its nucleotide sequence is as shown in Seq ID NO.5.
The present invention also provides the engineering bacteria that contains carrier ubi:VP64-Os01g18440.
The present invention also provides carrier ubi:VP64-Os01g18440 in the application improving in rice yield and the high bar proterties of adjusting and controlling rice.Adopt the conventional biotechnological meanss such as agrobacterium-mediated transformation, directly delivered DNA, microinjection or electroporation method, by carrier ubi:VP64-Os01g18440 Introduced into Rice, screening the final transgenic paddy rice that obtains.
Further, adopt agriculture bacillus mediated method, utilize pCAMBIA1301 that carrier ubi:VP64-Os01g18440 is proceeded in Rice Callus, with transforming containing the AAM conversion fluid of inductor and Agrobacterium, material after conversion through cultivating altogether, de-bacterium, screening, break up, take root, hardening and transplanting, with Basta screening transgenic rice plant.
The present invention utilizes transcription factor to activate i.e. 4 the transcription factors activation motif VP16 of motif VP64(first) build and obtain composing type transcription factor with rice transcription factor Os01g18440 gene fusion, and be transformed in paddy rice, change plant height and regulate flowering period of paddy rice, thereby improving rice yield.There is important theory value for illustrating in detail Rice Flowering, fruit maturation and developmental regulation mechanism, and can pass through transgenic technology, improve the output of paddy rice, also significant in production practice.
Brief description of the drawings
Fig. 1 is nVP64-bar-asRED carrier collection of illustrative plates in the embodiment of the present invention 1.
Fig. 2 is ubi:VP64-Os01g18440 carrier collection of illustrative plates in the embodiment of the present invention 1.
Fig. 3 is the PCR qualification result that in the embodiment of the present invention 3, VP64-Os01g18440 composing type transcription factor is crossed the transgenic positive strain of expression; Wherein, WT is wild-type paddy rice ' kitaake ', ubi:VP64-Os01g18440 transgenic paddy rice strain.
Fig. 4 is that in the embodiment of the present invention 3, Western blot detects the positive strain result of transgenosis VP64-Os01g18440; Wherein, WT is wild-type paddy rice ' kitaake ', and MADS-43, MADS-09 are VP64-Os01g18440 transgenic paddy rice strain.
Fig. 5 is the species test analytical results that VP64 in the embodiment of the present invention 4 (4 × VP16) activates Os01g18440 rice high yield; Wherein, WT is wild-type paddy rice ' kitaake ', and MADS-09, MADS-13, MADS-43 are VP64-Os01g18440 transgenic paddy rice strain.
Fig. 6 is that VP64 in the embodiment of the present invention 4 (4 × VP16) activates Os01g18440 paddy rice phenotype analytical result.
Embodiment
Following examples are used for illustrating the present invention, but are not used for limiting the scope of the invention.If do not specialize, the conventional means that in embodiment, technique means used is well known to those skilled in the art, the raw materials used commercial goods that is.
Separation and the plant expression vector construction of embodiment 1 Os01g18440 gene
Login http://rice.plantbiology.msu.edu/analyses_search_locus.shtml website, find Os01g18440 gene, according to its sequences Design pcr amplification primer, forward primer F:5 '-CAAAAAAGCAGGCTTCATGGCTCGCAAGAAGATCGT-3 ' and reverse primer R:5 '-CAAGAAAGCTGGGTCTTACATGGGAGGGAAAGATGG-3 '; Taking the total cDNA of wild Japanese fine ' kitaake ' paddy rice as template, carry out pcr amplification, obtain Os01g18440 complete sequence, its nucleotide sequence is as shown in Seq ID NO.1, and the aminoacid sequence of its coded albumen is as shown in Seq ID No.2.Carry out PCR reaction according to PrimeSTAR polymeric enzymatic amplification system and response procedures.In this process, comprise two-wheeled PCR, the primer of first round PCR adopts the gene primer that has added part adaptor attB joint, and the PCR product of the first round for the second template of taking turns, and complete adaptor attB primer for primer.PCR product cloning, to connecting on pDONER cloning vector, is obtained and the identical sequence of goal gene through order-checking qualification.Obtain and the identical sequence of goal gene through order-checking qualification.React the plant expression vector nVP64-bar-asRED upper (plasmid map is shown in Fig. 1) that the gene constructed Os01g18440 end of the N to its goal gene has been merged to VP64 label by LR, obtain carrier ubi:VP64-Os01g18440(plasmid map and see Fig. 2, carrier complete sequence is as shown in Seq ID NO.5).
Wherein, the building process of plant expression vector nVP64-bar-asRED is: the sequence comprising taking border, binary expression vector pCAMBIA1300 left and right is as frame sequence, pass through vitro recombination, ubi promoter-VP64-Gateway is expressed to unit, 35S promoter-asRED expression unit and 35S promoter-bar expression unit and construct and obtain with it, the complete sequence of carrier nVP64-bar-asRED is as shown in Seq ID No.4.
The acquisition of embodiment 2 transgenic rice plants
Get Mature seed of rice, artificial or mechanical dejacketing, select the full bright and clean seed without bacterial plaque is inoculated on inducing culture after sterilization, selection outward appearance is good, the Rice Callus that growing ability is good is acceptor material, adopt agriculture bacillus mediated method to utilize pCambia1301 that carrier ubi:VP64-Os01g18440 is proceeded in Rice Callus, with transforming containing the AAM conversion fluid of inductor and Agrobacterium, material after conversion is through exercise and the transplanting of cultivation-Tuo bacterium altogether-screen-break up-take root-transgenic seedling, with Basta screening transgenic paddy rice, after growing 4 spires, start to spray Basta (1:1000/V:V), every spray in 1 day 1 time, spray altogether 3 times.
The qualification of embodiment 3 transgenic paddy rices
Whether be integrated in rice genome in order to detect Os01g18440 gene, extract respectively wild-type and transgenic paddy rice leaf DNA, on the plant expression vector at gene two ends, design one section of primer and carry out PCR reaction, qualification transfer-gen plant, PCR product is detected, found that, transgenic paddy rice all amplifies object band (Fig. 3), and in wild-type paddy rice without this object band, preliminary definite Os01g18440 gene is inserted in rice genome by agriculture bacillus mediated method, and PCR product is correct goal gene sequence through sequencing analysis.
Utilize VP64 antibody can identify whether goal gene expresses in transgenic paddy rice, on protein level, identify by Western blot technology, after process SDS-PAGE protein electrophoresis-immune marking-Immunofluorescence Reactions, Western blot qualification result shows that transfer-gen plant hybridizes target protein, and wild-type plant does not hybridize target protein (Fig. 4).
Embodiment 4 transgenic paddy rice phenotype analyticals and species test analysis
Transgenosis and wild-type paddy rice are compared, phenotype, can significantly find out that transfer-gen plant is higher than wild-type, and on the cycle, show flower in suitable evening spending.Can significantly find out the single plant yield of transgenic paddy rice be significantly improved (Fig. 5) by the data results of species test compared with wild-type.
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (8)
1. rice transcription factor Os01g18440 gene is in the application improving in rice yield and the high bar proterties of adjusting and controlling rice, and the nucleotide sequence of described rice transcription factor Os01g18440 gene is as shown in Seq ID No.1.
2. application according to claim 1, is characterized in that, it is the downstream of the CDS sequence construct to 4 of a rice transcription factor Os01g18440 gene transcription factor being activated to motif VP16, rice transformation, screening the final transgenic paddy rice that obtains;
Described VP16 is from hsv (Herpes simplex virus), and the nucleotide sequence of 4 transcription factor activation motif VP16 is as shown in Seq ID No.3.
3. application according to claim 2, it is characterized in that, it is the downstream of the CDS sequence of rice transcription factor Os01g18440 gene being activated to motif VP16 by Gateway system constructing to 4 transcription factor, rice transformation, screening the final transgenic paddy rice that obtains.
4. application according to claim 3, is characterized in that, comprises the following steps:
1) according to rice transcription factor Os01g18440 gene order design pcr amplification primer pair, it is forward primer F:5'-CAAAAAAGCAGGCTTCATGGCTCGCAAGAAG ATCGT-3' and reverse primer R:5'-CAAGAAAGCTGGGTCTTACATGGGAG GGAAAGATGG-3';
2) taking the fine rice cDNA of wild Japanese as template, utilize above-mentioned primer, obtain Os01g18440 gene complete sequence by pcr amplification;
3) by PCR product cloning to pDONER cloning vector, correct through checking order;
4) sequence comprising taking border, binary expression vector pCAMBIA1300 left and right is as frame sequence, pass through vitro recombination, ubi promoter-VP64-Gateway is expressed to unit, 35S promoter-asRED expression unit and 35S promoter-bar expression unit and construct with it, obtain the complete sequence of carrier nVP64-bar-asRED as shown in Seq ID No.4;
5) reacted goal gene Os01g18440 is building up to carrier nVP64-bar-asRED above by LR, obtain plant expression vector ubi:VP64-Os01g18440, its sequence is as shown in Seq ID NO.5;
6) adopt agrobacterium-mediated transformation, directly delivered DNA, microinjection or electroporation method by carrier ubi:VP64-Os01g18440 Introduced into Rice, screening the final transgenic paddy rice that obtains.
5. application according to claim 4, it is characterized in that, step 6) adopts agriculture bacillus mediated method, utilize pCAMBIA1301 that carrier ubi:VP64-Os01g18440 is proceeded in Rice Callus, with transforming containing the AAM conversion fluid of inductor and Agrobacterium, material after conversion through cultivating altogether, de-bacterium, screening, break up, take root, hardening and transplanting, with Basta screening transgenic rice plant.
6. plant expression vector ubi:VP64-Os01g18440 is in the application improving in rice yield and the high bar proterties of adjusting and controlling rice, and wherein, the nucleotide sequence of carrier ubi:VP64-Os01g18440 is as shown in Seq ID NO.5.
7. application according to claim 6, is characterized in that, adopts agrobacterium-mediated transformation, directly delivered DNA, microinjection or electroporation method by carrier ubi:VP64-Os01g18440 Introduced into Rice, screening the final transgenic paddy rice that obtains.
8. application according to claim 7, it is characterized in that, adopt agriculture bacillus mediated method, utilize pCAMBIA1301 that carrier ubi:VP64-Os01g18440 is proceeded in Rice Callus, with transforming containing the AAM conversion fluid of inductor and Agrobacterium, material after conversion through cultivating altogether, de-bacterium, screening, break up, take root, hardening and transplanting, with Basta screening transgenic rice plant.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009127443A2 (en) * | 2008-04-17 | 2009-10-22 | Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften E.V | Transcription factors involved in salt stress in plants |
| CN102775498A (en) * | 2012-08-01 | 2012-11-14 | 中国农业科学院作物科学研究所 | Application of rice transcription factor Os05g41070 genes |
| CN102786599A (en) * | 2012-08-01 | 2012-11-21 | 中国农业科学院作物科学研究所 | Application of rice transcription factor Os05g39950 gene |
-
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009127443A2 (en) * | 2008-04-17 | 2009-10-22 | Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften E.V | Transcription factors involved in salt stress in plants |
| CN102775498A (en) * | 2012-08-01 | 2012-11-14 | 中国农业科学院作物科学研究所 | Application of rice transcription factor Os05g41070 genes |
| CN102786599A (en) * | 2012-08-01 | 2012-11-21 | 中国农业科学院作物科学研究所 | Application of rice transcription factor Os05g39950 gene |
Non-Patent Citations (6)
| Title |
|---|
| Anuradha Kotla et al.Quantitative trait loci and candidate genes for yield and related traits in Madhukar x Swarna RIL population of rice.《J.Crop Sci.Biotech.》.2013,第16卷(第1期),35-44. |
| Quantitative trait loci and candidate genes for yield and related traits in Madhukar x Swarna RIL population of rice;Anuradha Kotla et al;《J.Crop Sci.Biotech.》;20130331;第16卷(第1期);35-44 * |
| Simona Masiero et al.The Emerging Importance of Type I MADS Box Transcription Factors for Plant Reproduction.《The Plant Cell》.2011,第23卷865-872. |
| The Emerging Importance of Type I MADS Box Transcription Factors for Plant Reproduction;Simona Masiero et al;《The Plant Cell》;20110331;第23卷;865-872 * |
| 两个水稻MADS盒基因cDNA的克隆与分析;袁自强 等;《自然科学进展》;20001231;第10卷(第2期);129-134 * |
| 袁自强 等.两个水稻MADS盒基因cDNA的克隆与分析.《自然科学进展》.2000,第10卷(第2期),129-134. |
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