CN116751885A - Identification method and application of key genes expressed under influence of sunlight in rice - Google Patents
Identification method and application of key genes expressed under influence of sunlight in rice Download PDFInfo
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Abstract
The invention discloses an identification method and application of key genes expressed under the influence of sunlight in rice, which adopts rice seedlings to treat two different conditions, namely indoor (incandescent lamp) and outdoor (natural sunlight); setting different treatment time points, and sampling seedlings; transcriptome analysis is carried out on the sampled sample, and the key genes which are differentially expressed under the influence of sunlight are identified. The invention completes the functional verification of RTN1 gene, defines the feasibility of the identification method, and lays a foundation for further excavating key genes regulated and controlled by illumination, improving the plant type of crop rice and increasing the yield. Wherein the RTN1 gene (Seq ID No: 1) is expressed by sun light induction and participates in photosynthesis of rice, and the gene encodes a fructose-1, 6-bisphosphate aldolase (Seq ID No: 2). RTN1 participates in regulating and controlling the elongation of rice tillering buds, so that the tillering number and the rice yield are influenced, and a certain theoretical basis is provided for the research on the formation of the rice yield.
Description
Technical Field
The invention relates to the field of plant genetic engineering, in particular to an identification method and application of a key gene expressed in rice under the influence of sunlight.
Background
Light is one of the most important environmental signals affecting the growth and development of plants, and the plants form a plurality of light receptor systems in the historical evolution process, mainly comprising photosensitive pigment, cryptochrome, phototropin, light-like receptor, UV-B receptor and the like, and the light receptor systems are used for receiving the light signals and transmitting the light signals in plants, and the light receptor systems are involved in regulating and controlling the normal operation of important vital activities and physiological reactions of the plants in seed germination, photomorphogenesis, flowering results, plant aging and the like.
Photosynthesis refers to the biological process by which green plants assimilate carbon dioxide and water by absorbing light energy, synthesize organic matter, and release oxygen. The product of photosynthesis is mainly sugars, which are used to store energy in plants. Light energy is the most important energy source for plant photosynthesis, however, plants have different absorption amounts for solar energy with different wavelengths, and photosynthesis products are different. Studies have shown that leaves are prone to produce abundant carbohydrates with less protein synthesis under red light irradiation: and under the irradiation of blue light, the carbohydrate is obviously reduced and the protein content is obviously increased.
Photosynthesis is a photo-biochemical reaction, in which the rate of photosynthesis increases with the increase of the intensity of light, but after the intensity of light exceeds a certain range, the rate of photosynthesis does not increase, which is called a light saturation phenomenon. The light saturation phenomenon mainly aims at C3 plants, the light saturation point of the C4 plants is far higher than that of the C3 plants, and the representative plants include corn and sorghum, and the biological yield of the representative plants is also higher than that of the C3 plants, so that the light energy is efficiently utilized to improve the photosynthetic efficiency, and the plant yield is increased, and the light saturation phenomenon is an important subject in the field of food crop research.
In addition, under the influence of illumination conditions, the gene expression and the function of the coded protein molecules in the plant body are inevitably changed significantly, and although some functional genes are reported to be influenced by the illumination conditions and regulate the growth and development of plants, no related research reports on the use of sunlight and incandescent lamps for treating plant materials, particularly monocotyledonous plants, for identifying important differentially expressed genes influenced by different illumination and how to influence plant morphogenesis and the like exist at present.
Rice is one of the most important food crops in the world, and is served to nearly half of the population in the world. Therefore, the rice yield is crucial to grain safety, and the continuous and stable increase of the rice yield is the basis of grain safety. The plant type is an important agronomic character affecting the yield, so that the breeding of the ideal plant type rice is an important way for solving the problem of the rice yield. Plant types generally refer to morphological characteristics of all tissues of the overground parts and spatial arrangement modes of the plant types, and different plant types have great influence on the viability, yield and environment adaptability of plants. The plant type of the rice mainly depends on the number and angle of tillers, plant height, leaf shape, spike shape and the like. The number of tillers is an important index for determining the plant type of the rice, and can directly influence the effective spike number of the rice, so that the final yield of the rice is influenced.
Therefore, the identification method of the key genes regulated and controlled by illumination is still to be further developed, and a brand new identification method and application of the key genes expressed by the influence of sunlight in the rice can be created, so that more key genes expressed by the influence of sunlight can be excavated, a foundation is laid for improving the plant type of the crop rice and increasing the yield, and the method and the application of the identification method are very needed to be improved in the industry at present.
Disclosure of Invention
The invention aims to solve the technical problem of providing a brand-new identification method and application of key genes expressed under the influence of sunlight in rice, so that the method can excavate more key genes expressed under the influence of sunlight, and lay a foundation for improving the plant type of crop rice and increasing the yield.
In order to solve the technical problems, the invention adopts the following technical scheme:
in one aspect, the invention provides a method for identifying key genes expressed in rice under the influence of sunlight, comprising the following steps:
(1) Adopting rice seedlings to perform indoor and outdoor two different condition treatments, wherein the indoor illumination mode is an incandescent lamp, and the outdoor illumination mode is natural sunlight;
(2) Setting different treatment time points, and sampling seedlings;
(3) Transcriptome analysis is carried out on the sampled sample, and the key genes which are differentially expressed under the influence of sunlight are identified.
As a further improvement of the invention, in the step (1), the illumination intensity and the relative wavelength of two different conditions, namely indoor and outdoor, show significant differences; wherein the coverage of the relative spectral value of the outdoor sunlight in the relative wavelength range is significantly higher than the indoor incandescent lamp condition; the outdoor illumination intensity was also significantly higher than the indoor incandescent lamp conditions at the three illumination intensity detection time points of 9:00,11:00, and 17:00.
In the step (1), the rice seedlings are rice seedlings growing indoors for 21 days, the seeds with the urged buds are sowed into a box filled with field soil, and the box is placed in an indoor incandescent lamp for 21 days to grow under the treatment condition of the indoor incandescent lamp.
In the step (1):
setting indoor conditions: incandescent light illumination time, 5 am: 00 to 19 pm: 00; dark time, afternoon 19:00 to 5 am the next day: 00;
outdoor condition setting: natural sunlight illumination time, 5 am: 00 to 19 pm: 00; dark time, afternoon 19:00 to 5 am the next day: 00.
in the step (2), the treatment time points of 0 day, 3 days, 7 days and 15 days or the treatment time points of 3 days, 7 days and 15 days are set, and the stem base tissues of the indoor and outdoor treated rice seedlings are sampled respectively.
And (3) after the transcriptome analysis and identification of the key genes with differential expression in the step (3), metabolic pathway analysis is carried out, and functional verification is carried out on the differential expression genes participating in photosynthesis.
On the other hand, the invention also provides application of the key genes expressed under the influence of sunlight in the rice, wherein the key genes are involved in photosynthesis of the rice and are used for controlling the growth and development of the rice and the plant type of the rice.
The key gene is a gene RTN1 for encoding fructose-1, 6-bisphosphate aldolase, and the sequence of the RTN1 gene is represented by the sequence of Seq ID No:1, the protein sequence of the RTN1 gene is as shown in Seq ID No:2, the gene is used for positive regulation:
(1) Elongation of rice tillering buds;
(2) Number of tillers of rice;
and/or (3) rice yield.
In yet another aspect, the present invention provides a mutant gene of a key gene expressed under the influence of sunlight in rice, which is expressed in the sequence of Seq ID No:1, or at Seq ID No:1, and a deletion of 1 base C at position 154 of the sequence shown in FIG. 1.
On the other hand, the invention also provides application of the mutant gene of the key gene expressed under the influence of sunlight in the rice, and the mutant gene is used for:
(1) Inhibiting the rice tillering bud elongation;
and/or, (2) reducing the tiller number of rice.
The invention utilizes different indoor and outdoor illumination conditions to treat upper rice seedlings, and identifies different functional genes regulated and controlled by illumination through transcriptome technology, wherein a plurality of genes participate in photosynthesis. Wherein RTN1 gene codes fructose-1, 6-bisphosphate aldolase, RTN1 participates in regulating and controlling the elongation of rice tillering buds, thereby affecting the number of rice tillers and the rice yield, and providing a certain theoretical basis for the research on the formation of the rice yield. The invention designs an identification method of key genes expressed in rice under the influence of sunlight, completes functional verification of RTN1 genes, defines the feasibility of the method, and lays a foundation for further excavating key genes regulated and controlled by illumination, improving plant types of crops and improving yield.
Drawings
The foregoing is merely an overview of the present invention, and the present invention is further described in detail below with reference to the accompanying drawings and detailed description.
FIG. 1 is a graph showing rice material growth and different light treatment modes;
FIG. 2 is spectral information for outdoor sunlight and indoor greenhouse incandescent conditions, where A is relative spectral information under sunlight; b is relative spectrum information under indoor greenhouse (incandescent lamp) condition.
FIG. 3 is a graph showing the differential expression of genes after indoor and solar treatment, wherein A is the differential expression of genes after indoor and solar treatment; b is the co-expression result of genes indoors and under the sun.
FIG. 4 is a diagram showing the induction expression of RTN1 in photosynthesis and under sunlight, wherein A is a photosynthesis differential expression gene; and B is the expression of RTN1 induced by sunlight.
FIG. 5 is a graph showing a phenotype of fewer tillers of an RTN1 mutant, wherein A is RTN1 mutant gene information; B-D is the plant phenotype of wild type and mutants rtn1-1 and rtn 1-2; e is the statistical result of the tillering number of the wild type and RTN1 mutants.
FIG. 6 is a graph showing the effect of RTN1 mutation on rice tillering bud elongation, wherein A, B and C represent wild type ZH11, mutants RTN1-1 and RTN1-2, respectively, in shoot basal part tillering bud tissue structure; d, E and F respectively represent the tissue structures of tillering buds of wild ZH11, mutants rtn1-1 and rtn 1-2; g represents wild-type ZH11, mutants rtn1-1 and rtn1-2, tillering bud elongation.
Detailed Description
The following examples further illustrate the invention but should not be construed as limiting the invention. Modifications and substitutions to methods, procedures, or conditions of the present invention without departing from the spirit and nature of the invention are intended to be within the scope of the present invention. The technical means used in the examples are conventional means well known to those skilled in the art unless otherwise indicated. Unless otherwise indicated, all biochemical reagents, carriers, consumables and the like used in the examples were commercially available products.
The invention relates to a method for identifying key genes expressed under the influence of sunlight in rice, which comprises the following specific technical steps:
1. indoor group and outdoor group growth environments are used for preparing experimental materials
(1) The rice grows without leaving the sun, and researches find that the rice growing under the sun promotes tillering and morphogenesis of the whole plant, so that stable grains are ensured to be obtained for eating by people. However, the number of rice tillers grown under the incandescent lamp of the greenhouse is significantly reduced, the spike development is incomplete, and the yield of enough rice cannot be obtained. At present, no report is yet made on how sunlight influences gene expression in rice plants so as to influence the development of rice tillering buds and the formation of yield.
The invention adopts two completely different growing environments of indoor groups and outdoor groups, and the main difference is the difference of lighting modes. The indoor lighting is mainly incandescent lamps, while the outdoor lighting is natural sunlight (fig. 1). The indoor and outdoor growth conditions showed significant differences in illumination intensity and relative wavelength (fig. 2). The relative spectral coverage in the relative wavelength range of outdoor sunlight is significantly higher than that of indoor incandescent lamp conditions. The outdoor illumination intensity was also significantly higher than the indoor incandescent lamp conditions at the three illumination intensity detection time points of 9:00,11:00, and 17:00.
(2) Sowing the seeds with the sprouted seeds into 4 blue boxes filled with field soil, placing the seeds in indoor conditions for growth, and using the rice seedlings growing for 21 days for the subsequent treatment experiment of the invention, wherein the seedlings in the period are in a stage with proper size of tillering buds, and after being treated by sunlight and an incandescent lamp, the seedlings are favorable for identifying differential expression genes related to the development of the tillering buds, and the result is shown in figure 1.
(3) 2 blue boxes with rice seedlings grown were transferred outdoors, two remained indoors, all materials continued to grow in culture, and samples were taken at four growth time nodes (0, 3, 7 and 15 days) covering all time points from formation to elongation of rice tillers for subsequent study (fig. 1).
2. Transcriptome analysis and differential expression Gene identification
And (3) sending the prepared experimental material to a Person company for extracting RNA, carrying out second-generation high-throughput RNA sequencing, and completing corresponding bioinformatics technical analysis. The identified differentially expressed genes were analyzed by transcriptome data for 3, 7 and 15 days of indoor and outdoor growth, found that there were a large number of identical differentially expressed genes under indoor incandescent and outdoor sunlight at three different time points, and found that 1062 genes were up-regulated after 3 days of treatment and 926 genes were down-regulated after sunlight treatment; it was found that 1766 genes were up-regulated after 7 days of treatment and 1158 genes were down-regulated after solar light treatment; it was found that 826 genes were up-regulated after 15 days of treatment and 1566 genes were down-regulated after solar light treatment; furthermore, by co-expression analysis, 237 genes were found to be differentially expressed at three times simultaneously, and the results are shown in FIG. 3.
The key genes which are differentially expressed under the influence of sunlight can be primarily identified by the identification method.
In order to conduct intensive studies thereon and confirm the feasibility of the identification method, the following analysis may be further conducted.
3. Metabolic pathway analysis
Through metabolic pathway analysis, the above-mentioned majority of differentially expressed genes are found to be involved in the pathways of carbon fixation, citrate cycle, tyrosine metabolism, glycolysis/gluconeogenesis, fructose and mannose metabolism, pentose phosphate pathway, photosynthesis, and the like of photosynthetic organisms.
4. Identification of genes differentially expressed involved in photosynthesis
By profiling the differentially expressed genes, it was identified that multiple genes involved in the photosynthetic pathway of rice exhibited significant differences in RNA transcription levels under both indoor and outdoor environmental conditions, including genes encoding fructose-1, 6-bisphosphate aldolase RTN1, fructose-1, 6-bisphosphate aldolase FBA2 and FBA4, fructose-1, 6-bisphosphate MOC2, ribulose 1,5 bisphosphate carboxylase/oxygenase encoding gene OsRBSC1-4, and phosphoribulokinase encoding gene OsPRK. Wherein, MOC2 expression difference is most obvious, RTN1 expression difference is less, and other genes expression difference is slightly less. In addition, we performed expression verification of the identified RTN1 gene by qRT-PCR analysis technique, and the results indicate that the gene is expressed by solar light induction (fig. 4).
5. Functional verification of genes differentially expressed in participation in photosynthesis
(1) The invention speculates that the genes regulated by light are possibly involved in the growth and development of rice, therefore, we select RTN1 gene as target gene, edit it by CRISPR/Cas9 gene editing technology, create mutant materials RTN-1 and RTN-2 under the background of japonica rice variety ZH11, and the result of gene sequencing shows that insertion and deletion of one base respectively occur in the second exon of the gene, the coded protein sequence changes, and the mutated protein shifts the frame, resulting in protein function change (figure 5A).
(2) Through plant tillering number phenotype analysis, the tillering numbers of both mutants were found to be significantly reduced compared to wild-type ZH11, indicating that the RTN1 gene regulates the tillering numbers of rice (fig. 5B, 5C and 5D). Through histological section experiments and bud dissection experiments, axillary buds can be normally formed in the development process of two mutated tillering buds, but the elongation of the tillering buds is abnormal, and the result is shown in fig. 6. Wherein FIG. 6A, FIG. 6B and FIG. 6C show the structure of the tillering bud tissue at the base of the stem of wild type ZH11, mutant rtn1-1 and rtn1-2, respectively; FIG. 6D, FIG. 6E and FIG. 6F show the tissue structure of tillering buds from longitudinal sections of the shoot tips of wild-type ZH11, mutants rtn1-1 and rtn1-2, respectively; FIG. 6G shows the elongation of the tillered shoots of wild type ZH11, mutants rtn1-1 and rtn 1-2.
The following is a detailed description of the embodiments.
Example 1. Two growth environments of indoor and outdoor group for preparation of Rice Material
(1) The present embodiment includes two growth environments, indoor and outdoor: setting indoor group growth conditions: lighting mode, incandescent lamp; illumination time: 5 am: 00 to 19 pm: 00; dark time: afternoon 19:00 to 5 am the next day: 00. setting of outdoor group growth conditions: the illumination mode is natural sunlight; illumination time, 5 am: 00 to 19 pm: 00; dark time, afternoon 19:00 to 5 am the next day: 00. temperature and humidity conditions the indoor and outdoor groups of two growth environments are kept as consistent as possible. The illumination intensity is measured by adopting a spectrometer HR-550 to measure the illumination intensity of different indoor and outdoor light sources, and the measurement time is 9:00,11:00 and 17:00.
(2) Four blue boxes (length X, width X, height: 75cmX, 50 and cmX cm) are prepared in advance, field soil is filled in the blue boxes, the blue boxes are uniformly mixed, the soil is guaranteed to be moist, and the blue boxes are placed in a room for later use. The japonica rice seeds are soaked in water for 48 hours, then sprouted for 24 hours at 35 ℃, and then the sprouted seeds are sowed in the soil of four blue boxes, wherein a certain distance is kept between each seed, and the row spacing is 5cm and 5cm, so that the seedlings can grow conveniently. Seedlings of rice grown for 21 days after sowing were used as experimental materials prepared in advance.
(3) Sampling the rice seedlings after 0 day, 3 days, 7 days and 15 days of indoor and outdoor treatment, pulling the rice seedlings out of the soil, cleaning the soil on the roots with water, and sucking the water with paper for later use. The sampling part is stem basal tissue of 0.3cm of rice seedling, the stem basal tissue is cut off by a surgical knife and wrapped in tinfoil paper, indoor and outdoor independent sampling is carried out, each sample comprises stem basal tissue of 10 seedlings, three samples are repeated, and then the samples are rapidly placed in liquid nitrogen for freezing and then placed at the temperature of minus 80 ℃ for standby.
Example 2 outdoor solar light influences expression of plant genes
(1) Rice tissue RNA extraction
Taking out the standby tissue sample, grinding the tissue sample by liquid nitrogen, then loading the powder sample into a centrifuge tube with 1.5mL frozen by liquid nitrogen, extracting total RNA of rice by adopting a TRIZOL method, measuring the concentration of the extracted RNA by using a Nanodrop2000 instrument, and sending the sample with the required RNA extraction quality (OD 260/280 is more than 1.9) and concentration (total concentration is more than 5 micrograms) to a company for completing warehouse establishment and sequencing.
(2) Transcriptome sequencing analysis
Transcript pair-wise terminal sequencing was performed on an Illumina HiSeq 2500 instrument, low quality reads were filtered, the remaining sequenced sequences aligned to the reference genome, japan, and the complete file on the alignment was processed by Cufflinks software. The level of transcript significance for differential expression was set at q <0.05.
Example 3 RTN1 mutants exhibit a reduced tillering phenotype
In the early research of the invention, differential expression gene information and metabolic pathway analysis are utilized to identify that a plurality of genes are regulated by sunlight and participate in photosynthesis, so that the genes are deduced to participate in regulating and controlling the plant types of rice, wherein a target gene RTN1 is locked, and genetic material creation and function verification are carried out.
Based on the whole genome and cDNA sequence of Nipponbare (Oryza sativa L cv. Nipponbare) provided in NCBI, a general website is designed through targetshttp://crispr.dbcls.jp/The invention combines the characteristics of expression vector and RTN1 gene sequence, and adds specific joint sequences at two ends of specific target.
The specific design primer requirements are as follows: the adaptor sequence CAG is added to the 5 'end of the forward primer (RTN 1-F), the adaptor sequence ACC is added to the 5' end of the reverse primer (RTN 1-R), and the specific primer sequence information is as follows:
RTN1-F forward primer: 5' -cagGAACGTGGAGCCCAACCGCC-3’
RTN1-R reverse primer: 5' -aacGGCGGTTGGGCTCCACGTTC-3’
Using the primers (RTN 1-F and RTN 1-R) designed and synthesized above, the primers were annealed to form primer dimers by dissolving in 50uL of double distilled water at a final concentration of 10. Mu.M, and the specific annealing procedure was as follows: slowly cooling at 95 deg.C for 3 min and at 95 deg.C to 25 deg.C, programming at-1 deg.C/10 sec, naturally cooling to room temperature, and at last 25 deg.C for 5 min.
Prepared primer IIThe polymer is connected with a target carrier, and the specific reaction system is as follows: 1 microliter of Cas9/gRNA vector; primer dimer 1 microliter; t (T) 4 Ligase 1 microliter; t (T) 4 1. Mu.l of ligation buffer. The temperature of the water bath kettle is set at 16 ℃ and incubated overnight (more than 12 hours), then competent cells of escherichia coli DH5a are transformed, positive clones are screened through kana resistance, and the positive clones are sent to Hangzhou qing department company to finish sequencing, and finally, successfully connected expression plasmid Cas9-RTN1 is obtained. And transferring the Cas9-RTN1 expression plasmid into agrobacterium tumefaciens (Agrobacterium tumefaciens) EHA105 by adopting a liquid nitrogen freeze thawing method for standby.
Callus of 11 seeds of flowers in preparation was prepared in advance, the seeds were dehulled, then washed with 75% alcohol, then sterilized in 3% hypochlorous acid solution for 45 minutes, washed with sterilized water for 3 times, blotted with sterilized filter paper and the seeds were sown in NB medium. The agrobacterium carrying Cas9-RTN1 expression plasmid is then infected with the medium flower 11 callus, then screened on NB medium with 50mg concentration of hygromycin, seedlings are differentiated on differentiation medium with 50mg concentration of hygromycin, positive plants are screened on rooting medium with 50mg concentration of hygromycin, and finally transgenic seedlings (30 positive seedlings are ensured) are obtained and transplanted to the field for identification analysis.
By using transgenic seedlings, about 0.1 gram of leaves are taken from a single plant in the seedling stage for extracting total DNA of plants, sequencing the target sequence of RTN1 genes and judging whether the genes are edited or not. Specific sequencing primers are shown below:
RTN1-seq-F：ACACTGTAGCGGCCACTG
RTN1-seq-R：CCTCCACTGCTCCACATCTT
the amplification procedure was as follows: pre-denaturation at 94 ℃ for 4 min; denaturation at 94℃for 30 seconds, annealing at 55℃for 30 seconds, elongation at 72℃for 30 seconds, 40 cycles; finally, the extension was carried out at 72℃for another 10 minutes. And separating the PCR product by 5% agarose gel electrophoresis, and sequencing by a company's engine, thus finally obtaining the RTN1 target sequence of each transgenic seedling mutation. The created RTN1 mutant is finally used for statistical analysis of the tiller number of rice.
In conclusion, the invention designs an identification method of key genes expressed in rice under the influence of sunlight, completes functional verification of RTN1 genes, defines the feasibility of the method, and lays a foundation for further excavating key genes regulated and controlled by illumination, improving plant types of crops and improving yield.
The above description is only of the preferred embodiments of the present invention, and is not intended to limit the invention in any way, and some simple modifications, equivalent variations or modifications can be made by those skilled in the art using the teachings disclosed herein, which fall within the scope of the present invention.
Claims (10)
1. The identification method of the key genes expressed in the rice under the influence of sunlight is characterized by comprising the following steps:
(1) Adopting rice seedlings to perform indoor and outdoor two different condition treatments, wherein the indoor illumination mode is an incandescent lamp, and the outdoor illumination mode is natural sunlight;
(2) Setting different treatment time points, and sampling seedlings;
(3) Transcriptome analysis is carried out on the sampled sample, and the key genes which are differentially expressed under the influence of sunlight are identified.
2. The method for identifying a key gene expressed under the influence of sunlight in rice according to claim 1, wherein in the step (1), the illumination intensities and the relative wavelengths of two different conditions, indoor and outdoor, show a significant difference; wherein the coverage of the relative spectral value of the outdoor sunlight in the relative wavelength range is significantly higher than the indoor incandescent lamp condition; the outdoor illumination intensity was also significantly higher than the indoor incandescent lamp conditions at the three illumination intensity detection time points of 9:00,11:00, and 17:00.
3. The method for identifying key genes expressed under the influence of sunlight in rice according to claim 1, wherein in the step (1), the rice seedlings are rice seedlings grown indoors for 21 days, and the bud-promoting seeds are sown in a box containing field soil and placed in indoor incandescent lamp treatment conditions for 21 days.
4. A method for identifying a key gene expressed under the influence of sunlight in rice according to any one of claims 1 to 3, wherein in the step (1):
setting indoor conditions: incandescent light illumination time, 5 am: 00 to 19 pm: 00; dark time, afternoon 19:00 to 5 am the next day: 00;
outdoor condition setting: natural sunlight illumination time, 5 am: 00 to 19 pm: 00; dark time, afternoon 19:00 to 5 am the next day: 00.
5. the method for identifying key genes expressed by solar light influence in rice according to any one of claims 1 to 3, wherein in the step (2), the basal tissues of the seedlings of rice treated indoors and outdoors are sampled at the treatment time points of 0 day, 3 days, 7 days and 15 days or at the treatment time points of 3 days, 7 days and 15 days, respectively.
6. The method for identifying a key gene expressed by solar light influence in rice according to any one of claims 1 to 5, wherein after the transcriptome analysis and identification of the key gene differentially expressed in step (3), metabolic pathway analysis is performed and functional verification is performed on the differentially expressed gene involved in photosynthesis.
7. The application of the key genes expressed under the influence of sunlight in rice is characterized in that the key genes are involved in photosynthesis of the rice and are used for controlling the growth and development of the rice and the plant type of the rice.
8. The use of a key gene expressed under the influence of sunlight in rice according to claim 7, wherein the key gene is RTN1 which codes for fructose-1, 6-bisphosphate aldolase gene, and the sequence of the RTN1 gene is as set forth in Seq ID No:1, the protein sequence of the RTN1 gene is as shown in Seq ID No:2, the gene is used for positive regulation:
(1) Elongation of rice tillering buds;
(2) Number of tillers of rice;
and/or (3) rice yield.
9. A mutant gene of a key gene expressed under the influence of sunlight in rice, characterized in that the mutant gene is expressed in Seq ID No:1, or at Seq ID No:1, and a deletion of 1 base C at position 154 of the sequence shown in FIG. 1.
10. Use of a mutant gene of a key gene expressed under the influence of sunlight in rice as claimed in claim 9, wherein said mutant gene is used for:
(1) Inhibiting the rice tillering bud elongation;
and/or, (2) reducing the tiller number of rice.
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