CN111518184B

CN111518184B - Novel application of SiTGAL6 protein

Info

Publication number: CN111518184B
Application number: CN202010407063.4A
Authority: CN
Inventors: 陈明; 马有志; 黎毛毛; 张玥玮; 唐文思; 周永斌; 徐兆师; 陈隽
Original assignee: Institute of Crop Sciences of Chinese Academy of Agricultural Sciences
Current assignee: Institute of Crop Sciences of Chinese Academy of Agricultural Sciences
Priority date: 2020-05-14
Filing date: 2020-05-14
Publication date: 2022-06-07
Anticipated expiration: 2040-05-14
Also published as: CN111518184A

Abstract

The invention discloses a new application of SiTGAL6 protein. The invention discloses application of SiTGAL6 protein in regulation and control of plant stress tolerance and yield-related traits. By adopting a minimum expression frame transformation method, the rice callus is infected by the DNA fragment only containing the promoter, the target gene SiTGAL6 and the terminator (the transformation fragment does not have any vector skeleton sequence, so that the safety risk possibly brought by the vector skeleton sequence is reduced to the maximum extent), and the SiTGAL6 rice is obtained. Experiments prove that: under the low nitrogen stress, the yield-related traits (spike number, spike length, grain number, thousand grain weight, straw weight, rice grain weight and the like) and nitrogen content of the SiTGAL 6-transformed rice are higher than those of wild rice. The SiTGAL6 protein has the function of regulating and controlling the stress tolerance and yield-related traits of plants, particularly improves the low-nitrogen tolerance and yield of the plants, and lays a foundation for cultivating stress-tolerant and high-yield plant varieties.

Description

Novel application of SiTGAL6 protein

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a new application of SiTGAL6 protein.

Background

With the rapid growth of the world population, food supply is becoming an increasingly serious global problem. In recent years in China, with the rapid growth of industrial land and residential land, the agricultural land area is rapidly reduced. In addition, with the improvement of life of people, the occupation ratio of non-food production agricultural land in the total agricultural land is increased day by day, which further influences the self-sufficiency of domestic food and aggravates the domestic food safety problem.

Rice is one of the most important crops in the world and is also one of the main grains in China. The rice yield of the rice is about 50 percent of the total amount of the commercial grains in China, so the key influence of the rice yield on the grain yield in China is caused. Effectively improving the rice yield has important significance for improving the total grain yield in China and ensuring the national grain safety.

Disclosure of Invention

The invention aims to provide application of SiTGAL6 protein and related biological materials thereof in regulation and control of plant stress tolerance and/or plant yield related traits.

In order to achieve the above object, the present invention firstly provides a novel use of the sital 6 protein.

The invention provides application of SiTGAL6 protein in regulation and control of plant stress tolerance and/or plant yield related traits;

the SiTGAL6 protein is derived from millet (Setaria italica) and is a protein represented by A1) or A2) or A3) or A4):

A1) a protein consisting of an amino acid sequence shown in a sequence 2 in a sequence table;

A2) a fusion protein obtained by connecting labels to the N end or/and the C end of the protein shown in the sequence 2 in the sequence table;

A3) the protein with the same function is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown in the sequence 2 in the sequence table;

A4) a protein having a homology of 99% or more, 95% or more, 90% or more, 85% or more, or 80% or more with the amino acid sequence defined in any one of A1) -A3) and having the same function.

Wherein, the sequence 2 in the sequence table is composed of 401 amino acid residues.

The labels are specifically shown in table 1.

TABLE 1 sequences of tags

The protein represented by any one of A1) -A4) above may be artificially synthesized, or may be obtained by synthesizing the encoding gene and then performing biological expression.

In order to achieve the above object, the present invention also provides a novel use of a biomaterial related to the sital 6 protein.

The invention provides application of biological materials related to SiTGAL6 protein in regulation and control of plant stress tolerance and/or plant yield related traits;

the biological material is any one of the following B1) -B10):

B1) a nucleic acid molecule encoding a sital 6 protein;

B2) an expression cassette comprising the nucleic acid molecule of B1);

B3) a recombinant vector comprising the nucleic acid molecule of B1);

B4) a recombinant vector comprising the expression cassette of B2);

B5) a recombinant microorganism comprising the nucleic acid molecule of B1);

B6) a recombinant microorganism comprising the expression cassette of B2);

B7) a recombinant microorganism containing the recombinant vector of B3);

B8) a recombinant microorganism containing the recombinant vector of B4);

B9) a transgenic cell line comprising the nucleic acid molecule of B1);

B10) a transgenic cell line comprising the expression cassette of B2).

In the application, the nucleic acid molecule B1) is any one of the following C1) -C4):

C1) DNA molecule shown in sequence 1 in the sequence table;

C2) DNA molecule shown in sequence 3 in the sequence table;

C3) a DNA molecule having at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% homology to the sequence of a DNA molecule defined by C1) or C2) and encoding a sital 6 protein;

C4) a DNA molecule which hybridizes with a DNA molecule defined by C1) or C2) or C3) under stringent conditions and encodes a SiTGAL6 protein.

Wherein, the sequence 1 in the sequence table is composed of 1206 nucleotides.

The nucleotide sequence encoding the SiTGAL6 protein of the present invention can be easily mutated by one of ordinary skill in the art using known methods, such as directed evolution and point mutation. Those nucleotides which are artificially modified to have 75% or more identity to the nucleotide sequence encoding the sital 6 protein are derived from and identical to the nucleotide sequence of the present invention as long as they encode the sital 6 protein and have the same function.

The term "identity" as used herein refers to sequence similarity to a native nucleic acid sequence. "identity" includes a nucleotide sequence that is 75% or greater, or 85% or greater, or 90% or greater, or 95% or greater identical to the nucleotide sequence of a protein consisting of the amino acid sequence set forth in coding sequence 2 of the present invention. Identity can be assessed visually or by computer software. Using computer software, the identity between two or more sequences can be expressed in percent (%), which can be used to assess the identity between related sequences.

The above-mentioned identity of 75% or more may be 80%, 85%, 90% or 95% or more.

In the above application, the stringent conditions are hybridization and membrane washing 2 times at 68 ℃ for 5min in a solution of 2 XSSC, 0.1% SDS, and hybridization and membrane washing 2 times at 68 ℃ for 15min in a solution of 0.5 XSSC, 0.1% SDS; alternatively, hybridization was carried out at 65 ℃ in a solution of 0.1 XSSPE (or 0.1 XSSC), 0.1% SDS, and the membrane was washed.

In the application, the expression cassette sequentially comprises a promoter, the encoding gene of the SiTGAL6 protein and a terminator. In a specific embodiment of the invention, the expression cassette consists of a constitutive promoter of ubiquitin, the gene encoding the above-mentioned sital 6 protein and the terminator nos 3' in that order.

In the above application, the vector may be a plasmid, a cosmid, a phage, or a viral vector.

The recombinant vector is obtained by inserting the nucleic acid molecule into an expression vector to express the protein. When the nucleic acid molecule is used for constructing a recombinant vector, any one of enhanced, constitutive, tissue-specific or inducible promoters can be added in front of the transcription initiation nucleotide, and can be used alone or combined with other plant promoters; in addition, when recombinant expression vectors are constructed using the nucleic acid molecules, enhancers, including translational or transcriptional enhancers, may be used, and these enhancer regions may be ATG initiation codons or adjacent regions initiation codons, etc., but must be in the same reading frame as the coding sequence to ensure proper translation of the entire sequence. The translational control signals and initiation codons are widely derived, either naturally or synthetically. The translation initiation region may be derived from a transcription initiation region or a structural gene. In order to facilitate the identification and screening of transgenic plant cells or plants, plant expression vectors to be used may be processed, for example, by adding a gene encoding an enzyme or a luminescent compound which can produce a color change (GUS gene, luciferase gene, etc.), an antibiotic marker having resistance (gentamicin marker, kanamycin marker, etc.), or a chemical-resistant marker gene (e.g., herbicide-resistant gene), etc., which can be expressed in plants. From the safety of transgenic plants, the transgenic plants can be directly screened and transformed in a stress environment without adding any selective marker gene. In a specific embodiment of the invention, the recombinant vector is obtained by cloning the SiTGAL6 gene fragment shown in sequence 1 into the SpeI and BamH I enzyme cutting sites of the vector LP 0471118-Bar-ubi-EDLL.

In the above application, the microorganism may be yeast, bacteria, algae or fungi, such as Agrobacterium. The recombinant microorganism is a microorganism containing the expression cassette or the recombinant vector. In a specific embodiment of the present invention, the recombinant microorganism is agrobacterium EHA105 containing the above expression cassette.

The invention also provides application of the SiTGAL6 protein or the biological material in cultivating transgenic plants with improved stress tolerance and/or yield.

The invention also provides application of the SiTGAL6 protein or the biological material in plant breeding. The breeding aims to breed plants with high stress tolerance and/or high yield.

Further, the modulation is an increase.

Further, the stress tolerance is low nitrogen tolerance.

The regulation and control of the plant stress tolerance are specifically embodied in that: when the content and/or activity of the SiTGAL6 protein in a plant is reduced, the low nitrogen tolerance of the plant is reduced; when the content and/or activity of the SiTGAL6 protein in the plant is increased, the low nitrogen tolerance of the plant is increased.

Said plant yield-related traits comprise dry grain weight and/or ear number and/or ear length and/or kernel number per ear and/or total kernel number per ear and/or thousand kernel weight and/or rice weight and/or straw weight.

The plant yield-related traits are specifically embodied as follows: when the content and/or activity of the SiTGAL6 protein in a plant is reduced, the dry grain weight and/or the number of ears and/or the ear length and/or the number of grains per ear and/or the total number of grains per ear and/or the thousand-grain weight and/or the rice weight and/or the straw weight of the plant is reduced; when the content and/or activity of the sital 6 protein in a plant is increased, the dry grain weight and/or the number of ears and/or the ear length and/or the number of grains per ear and/or the total number of grains per ear and/or the thousand-grain weight and/or the rice weight and/or the straw weight of the plant is increased.

To achieve the above object, the present invention finally provides a method for breeding transgenic plants with improved stress tolerance and/or yield.

The method for cultivating the transgenic plant with improved stress tolerance and/or yield comprises the steps of increasing the content and/or activity of SiTGAL6 protein in a receptor plant to obtain a transgenic plant; the transgenic plant has higher stress tolerance and/or higher yield than the recipient plant.

Further, the method for increasing the content and/or activity of the SiTGAL6 protein in the receptor plant is to overexpress the SiTGAL6 protein in the receptor plant. The overexpression method is to introduce a gene encoding the SiTGAL6 protein into a recipient plant.

Further, the nucleotide sequence of the encoding gene of the SiTGAL6 protein is any one of the C1) -C4). In a specific embodiment of the invention, the gene encoding the sital 6 protein is introduced into the recipient plant via the expression cassette described above.

The stress tolerance is low nitrogen tolerance.

The transgenic plant has higher stress tolerance and/or yield than the acceptor plant and is embodied in any one of the following Y1) -Y10):

y1) the transgenic plant has a higher dry grain weight than the recipient plant;

y2) the transgenic plant has more ears than the recipient plant;

y3) the spike length of the transgenic plant is longer than that of the recipient plant;

y4) the transgenic plant has more kernels per ear than the recipient plant;

y5) the transgenic plant has more total grains per ear than the recipient plant;

y6) the transgenic plant has a thousand kernel weight higher than that of the recipient plant;

y7) the transgenic plant has a higher straw weight than the recipient plant;

y8) the transgenic plant has a higher rice weight than the recipient plant;

y9) the transgenic plant has a higher content of nitrogen in rice than the recipient plant;

y10) the transgenic plant has a higher nitrogen content in the straw than in the recipient plant.

In any of the above uses or methods, the plant may be a monocot or a dicot. Further, the monocotyledon may be a gramineae. Further, the gramineous plant is specifically rice (e.g., rice variety Kitaake) or millet (e.g., millet variety yu-gu No.).

The invention provides SiTGAL6 protein related to plant stress tolerance and yield traits, and rice callus is infected by a DNA fragment only containing a promoter, a target gene SiTGAL6 and a terminator (no vector skeleton sequence exists in a transformation fragment, so that the safety risk possibly brought by the vector skeleton sequence is reduced to the maximum extent) by adopting a transformation method of a minimum expression frame, so that the transformed SiTGAL6 rice is obtained. Experiments prove that: under the low nitrogen stress, the yield-related traits (spike number, spike length, grain number, thousand grain weight, straw weight, rice grain weight and the like) and nitrogen content of the SiTGAL 6-transformed rice are higher than those of wild rice. The SiTGAL6 protein has the function of regulating and controlling the stress tolerance and yield-related traits of plants, particularly improves the low-nitrogen tolerance and yield of the plants, and lays a foundation for cultivating stress-tolerant and high-yield plant varieties.

Drawings

FIG. 1 shows molecular characterization of rice transgenic for SiTGAL 6. Note: marker: DL1000 Marker; negative control: kitaake; positive control: the SiTGAL6 plasmid.

Fig. 2 is 2018 field data arrangement. Wherein CK is wild rice Kitaake; OE2, OE7 and OE17 are trans-sital 6 rice.

FIG. 3 shows the identification result of low nitrogen stress of SiTGAL6 rice transferred from Jiangxi in 2019. Wherein, the receptor (Kitaake) is wild rice Kitaake; m23193/17 is SiTGAL6 transformed rice.

FIG. 4 shows the results of the test data of 2019. FIG. 4A shows the results of the test data of the normal treatment group (nitrogen fertilizer application group). FIG. 4B shows the results of the test data of the low nitrogen treatment group (nitrogen fertilizer non-application group). Wherein, WT is wild rice Kitaake; OE7 and OE17 are trans-SiTGAL 6 rice.

Fig. 5 shows the 2019 bio-yield measurement results. Wherein, WT is wild rice Kitaake; OE7 and OE17 are trans-SiTGAL 6 rice.

Detailed Description

The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are all conventional ones unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.

The LP 0471118-Bar-ubi-EDLL expression vector in the following examples is described in the literature: the influence of over-expressed millet SiANT1 on the salt tolerance of rice [ J ] in China agricultural science, 2018,51(10): 1830-.

The marker gene bar expression vector pSBAR in the following examples is described in the literature: threepo, drought-resistant transgenic wheat [ D ] obtained by using an improved minimum expression cassette technology, university of inner mongolia agriculture, 2012, the public can be obtained from the institute of crop science of the Chinese academy of agricultural sciences, and the biomaterial is only used for repeating the relevant experiments of the present invention and cannot be used for other purposes.

Example 1 acquisition of SiTGAL6 Gene

1. RNA extraction

Total RNA of two-week-old seedlings of millet (millet variety Yu Gu I) was extracted using RNA extraction kit (cat # ZP405K-1) of Jiang Union Internationality Biotech Co., Ltd, and the procedure was performed according to the instruction.

2. Obtaining cDNA

Mu.l of the total RNA extracted was subjected to reverse transcription using a reverse transcription kit (cat 311-02) of the all-fashioned gold Biotechnology Ltd according to the instructions to obtain cDNA.

3. PCR amplification

PCR amplification was carried out using 2. mu.l of the reverse-transcribed cDNA as a template using KOYOTO KOD FX (cat # KFX-101) which is a high fidelity enzyme to obtain a PCR amplification product (coding sequence of SiTGAL 6). The primer sequences are as follows:

F：5’-ATGATGGAGCTCTACCAT-3’；

R：5’-CTAGCTAATCGCTGCCTCGC-3’。

4. PCR amplification product detection and sequencing

Carrying out 1% agarose gel electrophoresis detection on the obtained PCR amplification product, recovering and purifying to obtain a DNA fragment with the size of 957bp, recovering the PCR product by using a Takara glue recovery kit (the product number is 9760) and according to the instruction after the reaction is finished, sending the recovered product to Okosan biotechnologies GmbH for sequencing, and cloning the coding sequence of SiTGAL6 to a pBlunt vector (the vector is carried by the kit) according to the instruction by using a zero background cloning kit (the product number is CB501-01) of the whole gold biotechnology GmbH according to the instruction after the sequencing is correct, wherein the pBlunt-SiTGAL6 is named.

The sequencing result shows that: the amplified product with the size of 1206bp is obtained by PCR amplification and is named as SiTGAL6 gene, the nucleotide sequence of the amplified product is shown as sequence 1 in the sequence table, and the amino acid sequence of SiTGAL6 protein coded by the SiTGAL6 gene is shown as sequence 2 in the sequence table.

Example 2 acquisition of transgenic SiTGAL6 Rice and analysis of stress tolerance and yield-related traits

One-time, SiTGAL6 obtaining of rice

1. Construction of recombinant vector psSiTGAL6

The SiTGAL6 gene fragment shown in sequence 1 was cloned between the Spe I and BamH I cleavage sites of vector LP 0471118-Bar-ubi-EDLL using the seamless cloning kit (cat # 639649) of Clotech as described in the following protocol to obtain recombinant vector psSiTGAL 6.

2. Obtaining a Linear minimum expression Box

(1) The recombinant vector psSiTGAL6 obtained in the step 1 is subjected to double enzyme digestion by Hind III and EcoRI to obtain a linear minimum expression frame fragment of the SiTGAL6 gene, and the linear fragment of the minimum expression frame of the SiTGAL6 gene sequentially comprises a ubiquitin constitutive promoter (1.8kb), the SiTGAL6 gene (1206bp) and a terminator nos 3' (300 bp).

(2) And (3) digesting the marker gene bar expression vector pSBAR by using Hind III to obtain a linear minimum expression frame fragment of the bar gene, wherein the linear minimum expression frame fragment of the bar gene sequentially consists of a maize ubiquitin promoter, a marker gene bar and a terminator nos 3'.

3. Obtaining of recombinant bacteria

And (3) jointly transforming the linear minimum expression frame segment of the SiTGAL6 gene obtained in the step (2) and the linear minimum expression frame segment of the bar gene into agrobacterium EHA105 (purchased from Beijing Onggaku New Biotechnology Co., Ltd.) to obtain a recombinant strain.

4. Obtaining of transgenic Rice

And transforming the recombinant strain into a rice variety Kitaake callus by an agrobacterium infection method to obtain T0 generation transgenic rice. The specific transformation steps are as follows:

soaking immature rice embryos 12-14 days after pollination in 70% ethanol for 1 minute, then disinfecting the immature rice embryos for 15 minutes by 10% sodium hypochlorite, washing the immature rice embryos for 3-5 times by using sterile water, taking out the immature rice embryos on a super clean bench, and inoculating the immature rice embryos to SD (secure digital)₂Inducing young embryo callus on a culture medium (MS basic culture medium (containing no vitamin) +2 mg/L2, 4-D +1mg/L VB1+150mg/L Asn asparagine +30g/L sucrose +2.4g/L plant gel with pH being 5.8) for 7 days, transferring the induced callus to a hypertonic culture medium (MS basic culture medium +5 mg/L2, 4-D +0.4mol/L mannitol +3g/L plant gel) to carry out hypertonic treatment for 4-6 hours, carrying out agrobacterium infection after the hypertonic treatment, and obtaining the infected callusContinuously culturing the injured tissue on a hypertonic culture medium for 16-18 hours, and then transferring the infected callus to SD₂Dark culture on medium for two weeks, followed by placing calli on selection medium (at SD) containing 2-3mg/L herbicide Bialaphos₂Adding 2-3mg/L herbicide Bialaphos on the basis of the culture medium, and carrying out callus screening, differentiation and seedling strengthening on the mixture with the pH value being 5.8).

5. Identification of transgenic Rice

And (3) carrying out PCR identification on the T0 transgenic plants obtained in the step (4), wherein the plants with 186bp obtained by PCR amplification are positive transgenic SiTGAL6 rice (figure 1), and 5 positive transgenic SiTGAL6 rice are obtained in total, and the positive rate is 2%. After greenhouse generation adding, screening and identification, the SiTGAL6 rice strains M23193/02 (OE 2 for short), M23193/07 (OE 7 for short) and M23193/17 (OE 17 for short) are selected for the following functional verification experiments.

The specific steps of PCR identification are as follows:

1) extracting DNA of rice leaves by an SDS method: 100-500mg of fresh rice leaves are put in liquid nitrogen for full grinding, transferred into a 50mL centrifuge tube, added with 15-20mL of CTAB lysate (0.1mM Tris-HCl (pH8.0), 0.02M EDTA (pH8.0), 1.5M NaCl, 2% PVP-4, 2% CTAB) and subjected to water bath at 65 ℃ for 1-2 h. Adding equal volume of phenol, chloroform and isoamyl alcohol (the volume ratio of phenol, chloroform and isoamyl alcohol is 25:24:1), mixing, standing for 5-10min, and centrifuging at 12000rpm for 10min (if the rotation speed can not be reached, the centrifugation time is prolonged). 1mL of the supernatant was taken into a 2mL centrifuge tube, and 2/3 volumes of isopropanol were added to precipitate the DNA, which was left to stand at-20 ℃ for 0.5-1h, centrifuged at 12000rpm for 5min, and the supernatant was decanted (care was taken not to decant the precipitate). Washing with 1mL of absolute ethanol, centrifuging at 12000rpm for 5min, and removing the supernatant. Adding 70% ethanol, washing once, centrifuging at 12000rpm for 5min, and pouring off the supernatant. Removing residual liquid in the tube by using the tip of a gun, drying the tube by using a fume hood, and adding a proper amount of ddH₂And dissolving the O.

2) And (3) PCR amplification: and (3) designing primers of amplification part sequences according to the sequence of the SiTGAL6 gene by using the DNA obtained in the step (1) as a template for PCR amplification, wherein the primer sequences are as follows:

F：5’-CCGTCGGAAGTGCTCAAGAT-3’；

R：5’-GGGGAGTCCGAATGGGTC-3’。

the PCR conditions were as follows: denaturation at 94 deg.C for 5 min; 50sec at 94 ℃, 50sec at 62 ℃, 1min at 72 ℃ and 35 cycles; extension at 72 ℃ for 10 min.

The PCR reaction system is shown in Table 1.

TABLE 1 PCR reaction System

Composition (I)	Dosage (ul)
		10 XPCR buffer (containing Mg)²⁺)(Takara)	2.50
25mM Mg²⁺(Takara)	0.05
		2.5mM dNTP Mixture(Takara)	2.00
Primer 1	0.80
		Primer 2	0.80
r-Taq DNA Ploymerase(5U/ul)(Takara)	0.25
		Template	1.00
ddH₂O	17.6
		TOTAL	25

3) And (3) detecting a PCR product: PCR products were detected on 0.8% agarose gel and photographed by UV.

Second, analysis of stress tolerance and yield-related traits of SiTGAL 6-transgenic rice

And (3) analyzing the application of SiTGAL6 in regulating and controlling the low nitrogen tolerance and yield-related traits of the rice by taking the transformed SiTGAL6 rice strains M23193/02 (OE 2 for short), M23193/07 (OE 7 for short) and M23193/17 (OE 17 for short) obtained in the step one as test materials and taking wild rice Kitaake as a control.

1. 2018 test analysis method and result

Test site: transgenic test base of rice institute of agricultural science institute of Jiangxi province in Gao' an city of Jiangxi province. Seedling bed management is the same as field production. Each test material was able to grow normally.

The test method comprises the following steps: two treatment groups of nitrogen fertilizer and nitrogen fertilizer are used for testing facility, and each treatment group is provided with two repetitions. Planting 6 rows of each material, 8 pockets of each row, 5 multiplied by 6 inches of row spacing, and planting a single material. The test area is 4.0 mu. Each group of specific treatment methods is as follows:

nitrogen fertilizer application treatment group (normal treatment group): from the transplanting to the harvest time, 12 kg of pure nitrogen is applied to each mu. The nitrogen fertilizer application time is respectively a green-turning period (6 days after transplanting), a tillering period (15 days after transplanting) and a heading period. The applied nitrogen fertilizer is urea.

Nitrogen fertilizer treatment group (low nitrogen treatment group): no nitrogen fertilizer was applied from the time of transplanting to the time of harvest.

And (3) test results:

the dry grain weight of each material was measured separately at the rice maturity stage. Each treatment was repeated three times with 8 plants each.

The effective number of ears per plant, the number of grains per ear and the total number of grains per ear of each material were determined at the rice maturity stage. 3 plants were taken for each treatment and tested indoors for three replicates.

The results show that: under the treatment of no nitrogen fertilizer application, the average dry grain weight values of wild rice Kitaake, a SiTGAL 6-transformed rice line OE2, a SiTGAL 6-transformed rice line OE7 and a SiTGAL 6-transformed rice line OE17 are 150.50g, 160.42g, 157.30g and 146.29g respectively; the average values of the effective spike numbers of the single plants of wild rice Kitaake, a SiTGAL 6-transformed rice strain OE2, a SiTGAL 6-transformed rice strain OE7 and a SiTGAL 6-transformed rice strain OE17 are respectively 7.00, 9.33, 7.33 and 10.33; the average grain number per ear of wild type rice Kitaake, SiTGAL-transgenic 6 rice line OE2, SiTGAL-transgenic 6 rice line OE7 and SiTGAL-transgenic 6 rice line OE17 are 41.05, 56.71, 58.59 and 45.42 respectively; the average total number of grains per ear of wild type rice Kitaake, SiTGAL-transfected 6 rice line OE2, SiTGAL-transfected 6 rice line OE7 and SiTGAL-transfected 6 rice line OE17 were 42.71, 59.00, 60.55 and 50.03, respectively.

Compared with wild rice Kitaake, the yield-related traits of the SiTGAL6 transgenic rice line, such as dry grain weight, effective spike number per plant, solid grain number per spike, total grain number per spike and the like, are all higher than those of the wild rice Kitaake (figure 2).

2. 2019 test analysis method and result

Nitrogen fertilizer treatment group (low nitrogen treatment group): from the transplanting to the harvest stage, no nitrogen fertilizer is applied.

And (3) test results:

photographs were taken during the rice maturity stage. The phenotypic results are shown in FIG. 3 (the phenotype of 3 strains in the low nitrogen treatment group). The results show that: compared with wild rice Kitaake, SiTGAL6 rice lines OE02 and OE17 grow better and flourish.

The effective ear number, ear length, seed number per ear, total seed number per ear, seed setting rate and thousand seed weight of each plant of each material are measured in the rice mature period. 3 plants were taken for each treatment and tested indoors for three replicates. The results are shown in table 2, table 3 and fig. 4. The results show that: under the treatment of applying nitrogen fertilizer and not applying nitrogen fertilizer, the biological yield traits of the SiTGAL6 rice line, such as spike length, total grain number per spike and the like, are all higher than that of wild rice kitakke.

The straw weight and the rice weight of each material were measured at the rice maturity stage. 20 plants were taken for each treatment and repeated three times. The results are shown in table 4, table 5 and fig. 5. The results show that: under the nitrogen fertilizer application treatment, the average rice straw weight values of wild rice Kitaake, a SiTGAL-transferred 6 rice line OE7 and a SiTGAL-transferred 6 rice line OE17 are 390g, 440g and 413.3g respectively, and the average rice straw weight values of wild rice Kitaake, a SiTGAL-transferred 6 rice line OE7 and a SiTGAL-transferred 6 rice line OE17 are 243.3g, 366.7g and 316.7g respectively; under the condition of no nitrogen fertilizer application, the average rice straw weight values of wild rice Kitaake, SiTGAL-transferred 6 rice line OE7 and SiTGAL-transferred 6 rice line OE17 are respectively 203g, 253g and 263.3g, and the average rice straw weight values of wild rice Kitaake, SiTGAL-transferred 6 rice line OE7 and SiTGAL-transferred 6 rice line OE17 are respectively 173.3g, 173.3g and 173.3 g. Under the treatment of applying nitrogen fertilizer, the rice strain OE7 and OE17 of the SiTGAL6 has higher straw weight and paddy weight than wild rice kitakke. Under the condition of no application of nitrogen fertilizer, the weight average of straws of rice lines OE7 and OE17 of the SiTGAL6 is higher than that of wild rice kitakke, and the weight of rice lines OE7 and OE17 of the rice lines of SiTGAL6 is equal to that of wild rice.

And respectively measuring the nitrogen content of the straw and the nitrogen content of the paddy in the rice mature period by using a Kjeldahl method. 20 plants were taken for each treatment and repeated three times. The method for detecting nitrogen content is specifically described in the references "comparison of total nitrogen in plants by Yangyi, Chuitanlin. flow analysis and Kjeldahl method [ J ]. proceedings of Yuxi institute of academy of sciences, 2016,32(08): 51-54" and "Dahonglin, Wu Xiaojun. determination of nitrogen content in dried samples of plants by Kjeldahl method [ J ]. proceedings of Jiangsu institute of academy of agriculture, 1995(03): 70"). The results are shown in Table 6. The results show that: under the treatment of applying nitrogen fertilizer and not applying nitrogen fertilizer, the nitrogen content of the straw and the nitrogen content of the paddy rice of a SiTGAL6 rice line OE17 are both higher than that of wild rice kitakke.

TABLE 2 results for the Normal treatment groups (12 kg of pure nitrogen per mu application)

TABLE 3 results for low nitrogen treatment group (no nitrogen fertilizer application)

TABLE 4 weight of rice straw and weight of rice straw in nitrogen fertilizer application group

TABLE 5 weight of rice straw and rice without nitrogenous fertilizer

TABLE 6 Total nitrogen content of nitrogenous fertilizer application group and no nitrogenous fertilizer application group

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the technical principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Sequence listing

<110> institute of crop science of Chinese academy of agricultural sciences

Novel use of <120> SiTGAL6 protein

<160> 3

<170> PatentIn version 3.5

<210> 1

<211> 1206

<212> DNA

<213> Artificial Sequence

<400> 1

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ggcagcgccg cctccccgcc ggagttcatg acctctgcgt ccgcggtgca gttcgcggcg 120

gtgcctgtca gaatgggggt gtacgagcgg ccggcgccgg cgcccgtgat gggcatgtgg 180

aacagcgatc cgttcaaggt cgatagcggc caggccacca gcggctccac cgtcatggaa 240

gcggacaaga agttcgacaa taggctagaa gatgttccac aggtagcact agagccggcg 300

agaagtacgg atcaggaaac aagcaggccg ccagagaggg tcctgagaag acttgcacag 360

aacagagaag ctgctcggaa gagtcgcctg cgaaaaaagg cttacatcca gcagctagag 420

acaagccgga tgaagctggc acaactagag caagagcttc aacgggctcg acaacagggt 480

gcatatgcaa atgggaacat gggagactcg actctcggat tcacaggacc aatggatcca 540

ggtgttgctg ggttcgagat agactacagc aactgggtcg aagagcagaa caggcacaca 600

gccgagctga ggtctgctct ccaggggcag ccatcggagc tggagctccg cacgctcgtg 660

gagaccgggc tcaacaacta cgagcacctc ttcaggatca aggcgctggc cgcgaacgcc 720

gacgttttct acgtcatgtc cggcatgtgg aagacacccg ccgagcggtt cttcctgtgg 780

atcggcgggt tccggccgtc ggaagtgctc aagatcctaa ggccgcagct ggagccactg 840

acggagccgc agctcatggc cgtgggcggc ctgcagcaca cctcgacgca ggccgaggac 900

gccctgtcgc agggcatgga gaagctgcag cagaacctcg ccgagaccct gacggccgcg 960

gccgacccat tcggactccc cgacggctac atgctgcaga tggcgaccgc cgtggagaag 1020

ctgaaagagc tcgtcggctt cgtcacccag gcggaccatc tccggcagac gacgatgcag 1080

caaatgcata agatcctgac gacgcggcag gcggcgaggg gcctcctggc gctcggcgac 1140

tacttccagc gcctccgcgc gctgagccat ctgtgggcga cgcggcgcga ggcagcgatt 1200

agctag 1206

<210> 2

<211> 401

<212> PRT

<213> Artificial Sequence

<400> 2

Met Met Glu Leu Tyr His Gly Tyr Leu Glu Asp His Phe Asn Leu His

1 5 10 15

Lys Leu Ser Ile Gly Ser Ala Ala Ser Pro Pro Glu Phe Met Thr Ser

20 25 30

Ala Ser Ala Val Gln Phe Ala Ala Val Pro Val Arg Met Gly Val Tyr

35 40 45

Glu Arg Pro Ala Pro Ala Pro Val Met Gly Met Trp Asn Ser Asp Pro

50 55 60

Phe Lys Val Asp Ser Gly Gln Ala Thr Ser Gly Ser Thr Val Met Glu

65 70 75 80

Ala Asp Lys Lys Phe Asp Asn Arg Leu Glu Asp Val Pro Gln Val Ala

85 90 95

Leu Glu Pro Ala Arg Ser Thr Asp Gln Glu Thr Ser Arg Pro Pro Glu

100 105 110

Arg Val Leu Arg Arg Leu Ala Gln Asn Arg Glu Ala Ala Arg Lys Ser

115 120 125

Arg Leu Arg Lys Lys Ala Tyr Ile Gln Gln Leu Glu Thr Ser Arg Met

130 135 140

Lys Leu Ala Gln Leu Glu Gln Glu Leu Gln Arg Ala Arg Gln Gln Gly

145 150 155 160

Ala Tyr Ala Asn Gly Asn Met Gly Asp Ser Thr Leu Gly Phe Thr Gly

165 170 175

Pro Met Asp Pro Gly Val Ala Gly Phe Glu Ile Asp Tyr Ser Asn Trp

180 185 190

Val Glu Glu Gln Asn Arg His Thr Ala Glu Leu Arg Ser Ala Leu Gln

195 200 205

Gly Gln Pro Ser Glu Leu Glu Leu Arg Thr Leu Val Glu Thr Gly Leu

210 215 220

Asn Asn Tyr Glu His Leu Phe Arg Ile Lys Ala Leu Ala Ala Asn Ala

225 230 235 240

Asp Val Phe Tyr Val Met Ser Gly Met Trp Lys Thr Pro Ala Glu Arg

245 250 255

Phe Phe Leu Trp Ile Gly Gly Phe Arg Pro Ser Glu Val Leu Lys Ile

260 265 270

Leu Arg Pro Gln Leu Glu Pro Leu Thr Glu Pro Gln Leu Met Ala Val

275 280 285

Gly Gly Leu Gln His Thr Ser Thr Gln Ala Glu Asp Ala Leu Ser Gln

290 295 300

Gly Met Glu Lys Leu Gln Gln Asn Leu Ala Glu Thr Leu Thr Ala Ala

305 310 315 320

Ala Asp Pro Phe Gly Leu Pro Asp Gly Tyr Met Leu Gln Met Ala Thr

325 330 335

Ala Val Glu Lys Leu Lys Glu Leu Val Gly Phe Val Thr Gln Ala Asp

340 345 350

His Leu Arg Gln Thr Thr Met Gln Gln Met His Lys Ile Leu Thr Thr

355 360 365

Arg Gln Ala Ala Arg Gly Leu Leu Ala Leu Gly Asp Tyr Phe Gln Arg

370 375 380

Leu Arg Ala Leu Ser His Leu Trp Ala Thr Arg Arg Glu Ala Ala Ile

385 390 395 400

Ser

<210> 3

<211> 8498

<212> DNA

<213> Artificial Sequence

<400> 3

gtaccttgcc tacagcctac ctagtggatg ctgctgcttc ggggactagc tgccgtccaa 60

ggcccaggca aggttggccc accagattag cagtacaaca aaaccagtgt tcgtttccag 120

cagagcatcg gatgcagtca acccaattct tgctccgctt ctccaatcta ccaccaccta 180

cttctgcctg ccgaaaggta gccagccgga atcctcgacg cagaagaaat ctgaagttgc 240

atctacgtag caagtaagta attccttttt ttcagatcat acagagtacg accttcactt 300

tatgatctgt tgcttcccag aaatcaaacc gctgcatttg atgtttgagc ggattatttg 360

atcaacccgc tgcccttcct gaactattat attcttggga tatatttaag gccccaatta 420

actcgagggg gctataaaat taccctaact tttttaccag aaatttccct gaaatcatac 480

tcaacatcat gaactctgga aagggggcaa aaggaggtcg ggaaaagaag ggtgcagacc 540

aatcaggcca aaaggaagac aaggcgacga aggaatcaag cagccacagc cacgactatt 600

gctgctacgc attctcactc cagaaagagg aaagggaaaa atagacgcac cgtgatgtgg 660

acatgcatga acagcttcaa taatgctgcc atcagcaagc aggggcccgt ttgcattaag 720

gttcagtagt ttgcatatat tagcaggact gcatcattgt ggcctaatta tactactaat 780

tgatccctca tagtgcttta tcttatctct tcttttttag gttgcccgtc caaaggttcc 840

tcttcttttg cgcctttatg gcaaaggaga attcttcgtc gtcgcgcacc agggttacgc 900

aattcacaaa agagtaggtg cccccactaa gcgtaatata ttcctcttcg ttttgttttg 960

ctttggatca gattattatg cagtgtaatt tcctcttgct tttggaaaga tatgggcaat 1020

gattgcacaa tagcgggacg acttagactc gtaaatgagt agacgacgta gcattacaga 1080

atcaaataaa aattcgtaac agatttgtac cttctttcca gttttcaaca atgtttagtt 1140

tcttgttacc aagataggtt tatctactgt atacttaaat gactgatcaa catcatgtgt 1200

tcttctcaac aacattgtta ggcatcactg ggttaacaaa aaaaaatgtt tatacaccac 1260

tgtggaaagc accgaggaat tggagacgag tagcatttga gcaaaaagga aagatgtaaa 1320

ggcttctttt tgggaaggtt gaggaggtct agtgatcagg agggataccc cagaaatggg 1380

aagttccacc atgcaccaac ttggtgaatc tagtgatgtc ccttacttgc atcacggaat 1440

ctaatgggac cccattatag ggtcatcaca acgatgagca ttacttaaat ccttatggcc 1500

ttcttgaggt tccaaaggca ctatagcaat tgaggtaagt catgataagc tcactttata 1560

atttcgcatg aacctagtgc tgtcctagtg tcctccgctc cattatactc cttaaccatt 1620

tatgtttaga tagtttcatc tgtatctgtg agtaacaagc tcacaaatgc aaggaccttt 1680

aatctgcagg agttcttgtg ccataagaaa ctccctttgc actaaaatct tctcatctat 1740

gagtaactgt gtatcatgcc tgcagagttt tcttgcgttc acctttctgc ctaatcagta 1800

aatgacagtg gatgctgtgt atttaaccat ctttgcttgg tggaattgca gtacaatgta 1860

cgtggaaact actcggagtc ttgcctttcc agggcactgc caatctgttg tgttatctga 1920

tactgttgag cacctaatgt gtcgatctct ctaactaacc aagtgtatgc ctgcaatgca 1980

ggcttaaaag catgcttctt ttacacctga aagttaccgt ctttcttctc tgtcagaatc 2040

atgtgcatac ttttctggct ctcacttgtt aggtgatgat ggatgcgtat atgtattcct 2100

gtgtgcaagc aacttcgcac atatccaact agtttacgta aagaataatt agtgcatcct 2160

aagcatgagg cccaggtgat aatattagat actaaaagct ctttatactt ggctcctcct 2220

tttaaaggga aggcagctgg ctcctatata accctgcctc acttactccg gccaactggt 2280

ggtgtccttt ggagctgagc tagagcacat ctgtggaggc ccttacgttc agtgattcga 2340

tgtctcccgt catccttaac tgaagaaact gcaatcaggt actaagtttc cctacctttt 2400

gcttgttcag tctgtgttga tcttagtcat gaacaagtag aattcagtta tttgtctgct 2460

cttcatgtat gctagagatt tgatagctgg agttttggta gaaattgaca acaacaaagg 2520

catcgcacta gagcttgcca agcatagctt gtttacctct ccattcctgc tggcttgaaa 2580

gaaatagggc atggtttctt gggctaagac atgatagctt ctcagcggat gcgttcttct 2640

acaaacgatt catggctatt ggaactttag ctagtcatgc tcaatttgtt attgccttct 2700

tattttggat ggctgcaaac tgctgcaagg atacagaact acaatcaatc atattggctt 2760

ctccagtaag tctggaatag cacctctaga acccatatgt ttatgagaat ctttatattg 2820

ttggccagaa taaagtgatt ggagcaagtg atcaaatatt tgtattttca acagagcgta 2880

tccttcataa gattattttt tttctaagac tatatcttca ttcatgttca ttcaacacta 2940

gacaacctag agcagaggcc gaaccttcaa gttttatgaa tcatgcagct agtattctcg 3000

ttttggtaaa tgaattgcta tttctgtttg gcagacatga ggtgaggtga tcaatataag 3060

tattaattta agggagatca ttttgagcat gttatgatgg agctctacca tggatacctt 3120

gaagatcatt tcaacctcca caagcttagg tgtgtatttc ctcagacccg atccccctgt 3180

ttcatctgaa cgttctgtcg atccgatcct tcacaaaata ccaatgtttt gatgactgtc 3240

caatcagtca tctgaagtag aacaagacgt gaagcaaaga gcagagctaa tagctttgtg 3300

tcactagcac taatagacta gtagtgcaat ttgcattaga ttttcattac ctgtgggtgg 3360

cacatcatga tgtaaactga atactagatc tttagaggtg caagtggact agtgcatcta 3420

gagtgagatc atgttagtta aaataaacaa agtgaagctc aaaagtagga aaaataaaaa 3480

gatggctacc gtcttttgga atcaccagtt aggaccaaaa cgcatgctac atcaagaaat 3540

actttgtgct aactctccaa aagcagattg tgaagggaga cgtgtccctt tgttctctgc 3600

ctagcctagg cagccatggg ctctcgtttt cgacaagctt tagcccacag gttccactga 3660

attatgcagg acgcatagct gttcctgaat agcacacatt agccatcagt accgttcctt 3720

ttagcggcca tgacggtgag ctttgcatca ggtctttctt tagttgaggt cctctgatag 3780

ttgaccccgt aacggaaaaa agctgagagc cttatttggc aaaacagtgc atgggcagtg 3840

aggcactagg ctctggcgaa ggacaggtgt tttgttttgc accgtctcca gcccactatt 3900

ctgcagggca ggggctgcca gcctcctgct caagtggcct ggtgctgatc tgaaggaagt 3960

ggtgaccagt tctgtactgc tgtgcatcag cagctttcac cactggcaac tcaatcagac 4020

attccctttt cttttgtcat cacatttccc tttcatcagc atcattgtgc acagtaaaca 4080

cacgctacaa cttgcaggtt ttttttttca tatatttcag ggtgcgtgca tgtatcacat 4140

actatgttct tgctttcatg catatctcct ctgagatctg cataatcggc tcgatatggt 4200

tatacaccat tagaattgca tcttctgtag actgtttcaa taggagaatt ggatccagat 4260

ccttgcatat taacctggta cagtgacaaa tcgcacccac acgtgtggcg aaattaagcc 4320

gaaaaaaaat gctacgctca tcacgcactt ctgtttcaga taagttggat ggtggggtta 4380

agatgatgcc cagcgaccac ttctttatct gcagcctgtt cctgtgtgct ggatagagca 4440

gagaccaacc taacctgctt cgccgtaaaa gccaagtggc ctcgccaata attgaggggc 4500

catccaatcc aaaccttctt ttggccgtgc tggcttggcg agccctccac tatgtgtgat 4560

gaagcttctg cttcctgtcg tgcgggagca agcattgcat tttgcacgta atcattgagg 4620

cccatgaata attggataga tcatgctaga aacactgtcg ttttattacg gtattctgag 4680

ctctcatgtg gtcatgtgga gttgtggact ataaaaaaaa caagagggtc aaaagaatac 4740

aacatgcatg cagctggcca tcagcttaag cttgtggttt aggatctaat tctagctata 4800

atctagtgac gttgtggggt ttccttcagg agccttcttg ctttgcactg ctattcctgc 4860

ggtgtgatgc caccgctgat gaactgatgg gcgtcaggcg tcaccgtcac cacacccttt 4920

acatgcttaa tccttaacca ctagtttagc ctctgctact aagtataatg gccggaaatt 4980

cctagcactg gccgggcggt gtggtggccg tccttgtcag cccaggggta gagatgtact 5040

tggccgcata gcgtcaccgc cggtcggccg gtgcgcgcct gactgagacc ggggcgcggc 5100

tatcatcgcg tccgagtaga tcagcacacg ggcttatccc gtatctcgct ggtcagccgc 5160

acgcagcgca ccaagacggc gtcgtaactc gatcaagtgg agatcgaggt ggccatggac 5220

ccacgctctc gtggtcgtcg tttgtcgcgc ctgctgccgt cgccggccgg acggcggccg 5280

cgctgagcag aagagcgaaa cgagcatcgc ggacgcagaa attatgggga cgagaagcag 5340

ctgccgaagc aggagcatga aaaattttca gaggtggcaa ctcttccgtt ggagtccggg 5400

ctcgaattat tcgttccgga gccggattcc agattccagg ttccactcga ggtcgaggcg 5460

acggcatcgt ccagccatca ctggccacgg cagctgccgc tcccgattct cttctctcga 5520

tcatctcgtt gagccgatgc agatgcaccc cgcctagcta ggctcctagg ccgcaccagc 5580

acaggttctc gctcgcatcc cagtaggcaa cctcagctcg ccgtcgatcc gttccttccc 5640

ttgcccgggc cggcggccac cgatcttttc cttcttgtga aaccattgcc gtccggaggt 5700

gacggagacg ccgtcgtgga cgtttagatt tttttgttcc gtgacaggag tgcacggcgt 5760

cttctcggcg cgccatccat catcatgcca tcatttgttg cgtacgttgt cagcctcggc 5820

ggctttgcca aaagtgctcg ccggggttgc ctttcccagc gcctttcctg agcacagctg 5880

cggccgcaat cggagagagc atatttgcct ttgccgctgc gctcattcca ttcggagctc 5940

gaaagcttcg cttcgcctgc ctttctctca tttttttttt ctttcgctct ttaacctggt 6000

tgacacctgc aacggcgctc tgtcgttaac tcacgctgcc tcccccgctg gttgtttttc 6060

tgtttgggta gcatcggcag cgccgcctcc ccgccggagt tcatgacctc tgcgtccgcg 6120

gtgcagttcg cggcggtgcc tgtcagaatg ggggtgtacg agcggccggc gccggcgccc 6180

gtgatgggca tgtggaacag cgatccgttc aaggtcgata gcggccaggc caccagcggc 6240

tccaccgtca tggaagcgga caagaagttc gacaataggg tgaggaagga tgaactcatc 6300

gatcagcttg atttcttgca tggcttctgt gatttgtcca tcgatgggtg gagtagagta 6360

ctgatgttga tgagctttgt gcagctagaa gatgttccac aggtagcact agagccggcg 6420

agaagtacgg atcaggaaac aagcaggccg ccagagaggg tacaattcaa ggatctttca 6480

cttgttttcc tttttgtctt tttgactgcc gcagcatgtt tagtctttct taaatactcc 6540

tacgttacaa cagagaaaag tgatatttta agtcttaact tcgacagtcc atttcatagg 6600

ttatttgatt tgttgttgca agagaaataa aatgctctga atataaacct gaaatgattt 6660

gcctcaggtc ctgagaagac ttgcacagaa cagagaagct gctcggaaga gtcgcctgcg 6720

aaaaaaggta accacaagaa cttcaccaat cttttggtgc tatcatgctt agtgaccttc 6780

tatctcacaa ttggccccga ttttgttggt gtttaatagg cttacatcca gcagctagag 6840

acaagccgga tgaagctggc acaactagag caagagcttc aacgggctcg acaacaggta 6900

caagaaagaa tccgttggtt aaagccatct catcagtccc tgctacattc cctgattcag 6960

tgcattggct gatctgcagg gtgcatatgc aaatgggaac atgggagact cgactctcgg 7020

attcacagga ccaatggatc caggtcaggt tttgcaccta tcattctcaa gaacaaacac 7080

ttgtcgttca gtggcattca gcaaaatgtt cagtggcaga ccctgtgtat ctgacttccg 7140

gcttgcttgt atggttcact tccaacaggt gttgctgggt tcgagataga ctacagcaac 7200

tgggtcgaag agcagaacag gcacacagcc gagctgaggt ctgctctcca ggggcagcca 7260

tcggagctgg agctccgcac gctcgtggag accgggctca acaactacga gcacctcttc 7320

aggatcaagg cgctggccgc gaacgccgac gttttctacg tcatgtccgg catgtggaag 7380

acacccgccg agcggttctt cctgtggatc ggcgggttcc ggccgtcgga agtgctcaag 7440

gtggccggct cagactcact taccgtgcag cttcagacat aaaacaagtg ctgtcctccc 7500

gagagatttt gtctcacacg ttggatcttg ctgttgctgg tgtcactcag atcctaaggc 7560

cgcagctgga gccactgacg gagccgcagc tcatggccgt gggcggcctg cagcacacct 7620

cgacgcaggc cgaggacgcc ctgtcgcagg gcatggagaa gctgcagcag aacctcgccg 7680

agaccctgac ggccgcggcc gacccattcg gactccccga cggctacatg ctgcagatgg 7740

cgaccgccgt ggagaagctg aaagagctcg tcggcttcgt cacccaggta accaaccgac 7800

ccggtcgcca agcaccatcc gcaagtggaa gatcccctgc ttctcacacg ctacccattt 7860

tgcaggcgga ccatctccgg cagacgacga tgcagcaaat gcataagatc ctgacgacgc 7920

ggcaggcggc gaggggcctc ctggcgctcg gcgactactt ccagcgcctc cgcgcgctga 7980

gccatctgtg ggcgacgcgg cgcgaggcag cgattagcta gacgtcggtg gcgaaaaccc 8040

actgaagatg aacgaagacg acgacgaaga gtagtaggtc gttgtaggcc agccgaggac 8100

gaacaagtgg tcgacctcgc tgggattgga tgaagaatgc gaaggcttct cagccagcgg 8160

cgccgccatg cttgccggcg tctctgggga acaggaagtt ggtgtgcgct tgtcaggtgc 8220

agtgatcaat gaataacgaa aattgaggcc tttttctgag acgtgtacat atcttgtttt 8280

ggtatacctg gccggaggcc ttctacagcc gtataattaa ttggtataaa tgatgggttt 8340

tacgagtagt gtttacggta tatatgatgt aacagtatgt aactagtgtc tatccatttc 8400

tcatgagttg ggcatgcagt ccttgtgcca ttgtattgta cggagtatgg aacaagctag 8460

ttcacctgtt gaggaatgag atgcatccgt agcagttc 8498

Claims

1. Use of a protein as shown in a1) or a2) below for modulating plant stress tolerance and/or plant yield-related traits:

the stress tolerance is low nitrogen tolerance;

the plant yield-related trait is dry grain weight and/or number of ears and/or ear length and/or number of grains per ear and/or total number of grains per ear and/or thousand grain weight and/or rice weight and/or straw weight.

2. Use of a biological material related to a protein as defined in claim 1 for modulating stress tolerance and/or plant yield-related traits in a plant;

the biological material is any one of the following B1) -B10):

B1) a nucleic acid molecule encoding the protein of claim 1;

B2) an expression cassette comprising the nucleic acid molecule of B1);

B3) a recombinant vector comprising the nucleic acid molecule of B1);

B4) a recombinant vector comprising the expression cassette of B2);

B5) a recombinant microorganism comprising the nucleic acid molecule of B1);

B6) a recombinant microorganism comprising the expression cassette of B2);

B7) a recombinant microorganism containing the recombinant vector of B3);

B8) a recombinant microorganism containing the recombinant vector of B4);

B9) a transgenic cell line comprising the nucleic acid molecule of B1);

B10) a transgenic cell line comprising the expression cassette of B2);

the stress tolerance is low nitrogen tolerance;

3. Use according to claim 2, characterized in that: B1) the nucleic acid molecule is C1) or C2) as follows:

C1) the coding sequence is a DNA molecule shown in a sequence 1 in a sequence table;

C2) DNA molecule shown in sequence 3 in the sequence table.

4. Use according to any one of claims 1 to 3, characterized in that: the modulation is an increase.

5. Use according to any one of claims 1 to 3, characterized in that: the plant is a monocotyledon or a dicotyledon.

6. Use of a protein according to claim 1 or a biological material according to claim 2 or 3 for the cultivation of transgenic plants with increased stress tolerance and/or yield; the stress tolerance is low nitrogen tolerance;

or, the use of a protein as defined in claim 1 or a biological material as defined in claim 2 or 3 in plant breeding; the breeding aims to breed plants with high stress tolerance and/or high yield; the stress tolerance is low nitrogen tolerance.

7. Use according to claim 6, characterized in that: the plant is a monocotyledon or a dicotyledon.

8. A method for producing a transgenic plant with improved stress tolerance and/or yield, comprising the steps of increasing the content and/or activity of the protein of claim 1 in a recipient plant to obtain a transgenic plant; the transgenic plant has higher stress tolerance and/or higher yield than the recipient plant; the stress tolerance is low nitrogen tolerance;

y2) the transgenic plant has more ears than the recipient plant;

y4) the transgenic plant has more kernels per ear than the recipient plant;

y7) the transgenic plant has a higher straw weight than the recipient plant;

y8) the transgenic plant has a higher rice weight than the recipient plant;

9. The method of claim 8, wherein: the method for increasing the content and/or activity of the protein of claim 1 in a recipient plant is to overexpress the protein of claim 1 in the recipient plant.

10. The method of claim 9, wherein: the method of overexpression, which comprises introducing a gene encoding the protein of claim 1 into a recipient plant.

11. The method according to any one of claims 8-10, wherein: the plant is a monocotyledon or a dicotyledon.