CN111518183B

CN111518183B - Application of SiMYB61 protein and related biomaterial thereof in regulation and control of plant stress resistance

Info

Publication number: CN111518183B
Application number: CN202010401862.0A
Authority: CN
Inventors: 陈明; 孙黛珍; 马有志; 黎毛毛; 张玥玮; 唐文思; 周永斌; 徐兆师; 陈隽
Original assignee: Shanxi Agricultural University; Institute of Crop Sciences of Chinese Academy of Agricultural Sciences
Current assignee: Shanxi Agricultural University; Institute of Crop Sciences of Chinese Academy of Agricultural Sciences
Priority date: 2020-05-13
Filing date: 2020-05-13
Publication date: 2022-08-23
Anticipated expiration: 2040-05-13
Also published as: CN111518183A

Abstract

The invention discloses an application of SiMYB61 protein and related biological materials thereof in regulation and control of plant stress resistance. The invention provides a new gene for rice stress genetic engineering, and simultaneously introduces japonica rice fine varieties through genetic transformation to culture new stress-resistant transgenic varieties, so that the stress resistance of rice can be effectively improved, the influence of stress on rice production is reduced, and important economic and social benefits are achieved.

Description

Application of SiMYB61 protein and related biomaterial thereof in regulation and control of plant stress resistance

Technical Field

The invention relates to an application of SiMYB61 protein and related biological materials thereof in regulation and control of plant stress resistance.

Background

China is one of countries with the lowest grain input and output of unit chemical fertilizers in the world, the utilization rate of nitrogen fertilizers is only 30-35% (45% in developed countries), and the utilization rate of phosphate fertilizers is only 10-20%. According to the current consumption, for example, 10 percent of nitrogen fertilizer and 20 percent of phosphate fertilizer are saved, 241 million yuan of capital can be saved every year. The low-efficiency utilization of nitrogen and phosphorus fertilizers enables agricultural non-point source pollution to become the most important factor for water system eutrophication, soil acidification and heavy metal pollution, and threatens ecological safety and sustainable development. Therefore, the cultivation of new crop varieties with high nutrient utilization efficiency is urgently needed in China, so that the utilization efficiency of nitrogen and phosphorus fertilizers in China is greatly improved.

The cultivation of new stress-resistant rice varieties improves the stress resistance of the rice varieties, and is an important measure for improving the yield level of the rice under the conditions of reduced fertilizer application amount and drought and water shortage.

Disclosure of Invention

The invention aims to provide SiMYB61 protein related to plant stress resistance, a related biological material and application thereof.

In a first aspect, the present invention first protects the use of a SiMYB61 protein or related biomaterial as follows (a1) and/or (a 2):

(a1) regulating and controlling plant yield-related traits;

(a2) regulating and controlling the stress resistance of plants;

the related biological material is a nucleic acid molecule capable of expressing the SiMYB61 protein or an expression cassette, a recombinant vector, a recombinant bacterium or a transgenic cell line containing the nucleic acid molecule;

the SiMYB61 protein is any one of the following proteins:

(A1) protein with an amino acid sequence of SEQ ID No. 4;

(A2) protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues to the amino acid sequence shown in SEQ ID No.4 and has the same function;

(A3) a protein having 99% or more, 95% or more, 90% or more, 85% or more, or 80% or more identity to the amino acid sequence defined in any one of (A1) to (A2) and having the same function;

(A4) a fusion protein obtained by attaching a protein tag to the N-terminus and/or C-terminus of the protein defined in any one of (A1) to (A3).

The protein can be artificially synthesized, or can be obtained by synthesizing the coding gene and then carrying out biological expression.

In the above protein, the tag may be a Flag tag, a His tag, an MBP tag, an HA tag, a myc tag, a GST tag, and/or a SUMO tag, etc.

The nucleic acid molecule may specifically be a gene encoding a SiMYB61 protein. The encoding gene of the SiMYB61 protein is a DNA molecule described in any one of the following items:

(B1) DNA molecule shown in SEQ ID No. 1;

(B2) DNA molecule shown in SEQ ID No. 2;

(B3) a DNA molecule shown as SEQ ID No. 3;

(B4) a DNA molecule that hybridizes under stringent conditions to the DNA molecule defined in (B1) or (B2) or (B3) and encodes the SiMYB61 protein;

(B3) a DNA molecule which has more than 99%, more than 95%, more than 90%, more than 85% or more than 80% of identity with the DNA sequence defined in (B1) or (B2) or (B3) and encodes the SiMYB61 protein.

The stringent conditions may beThe method comprises the following steps: 50 ℃ in 7% Sodium Dodecyl Sulfate (SDS), 0.5M NaPO ₄ Hybridization with 1mM EDTA, rinsing in 2 XSSC, 0.1% SDS at 50 ℃; also can be: 50 ℃ in 7% SDS, 0.5M NaPO ₄ Hybridization with 1mM EDTA, rinsing at 50 ℃ in 1 XSSC, 0.1% SDS; also can be: 50 ℃ in 7% SDS, 0.5M NaPO ₄ Hybridization with 1mM EDTA, rinsing in 0.5 XSSC, 0.1% SDS at 50 ℃; also can be: 50 ℃ in 7% SDS, 0.5M NaPO ₄ Hybridization with 1mM EDTA, rinsing in 0.1 XSSC, 0.1% SDS at 50 ℃; also can be: 50 ℃ in 7% SDS, 0.5M NaPO ₄ Hybridization with a mixed solution of 1mM EDTA, rinsing in 0.1 XSSC, 0.1% SDS at 65 ℃; can also be: in a solution of 6 XSSC, 0.5% SDS at 65 ℃ and then washed once with each of 2 XSSC, 0.1% SDS and 1 XSSC, 0.1% SDS.

The expression cassette can be specifically an expression cassette consisting of a ubiquitin constitutive promoter, the coding gene of the SiMYB61 protein and a terminator nos 3'. The expression cassette can be obtained by double enzyme digestion of a recombinant vector by Hind III and EcoRI. The recombinant vector can be specifically a recombinant vector obtained by cloning SEQ ID NO.3 into BamHI and SpeI sites of the vector LP 0471118-Bar-ubi-EDLL.

The recombinant strain can be obtained by introducing the expression cassette or the recombinant vector into an agrobacterium strain. The agrobacterium strain may specifically be agrobacterium strain EHA 105.

In said use, said plant yield-related traits comprise ear number, ear length, grain per ear and/or biomass; the biomass comprises a straw weight and/or a rice weight.

The plant stress resistance is the resistance of a plant to low nitrogen stress.

The modulation is a forward modulation.

In a second aspect, the invention features the use of a SiMYB61 protein or a related biomaterial thereof in plant breeding;

the SiMYB61 protein is as shown previously.

In the application, the breeding aims to breed plants with high yield and/or high stress resistance. The high yield can be embodied in particular as a high number of ears and/or a high ear length and/or a high number of grains per ear and/or a high biomass. The biomass comprises the weight of rice straw and/or the weight of rice. The stress tolerance is in particular resistance to low nitrogen stress. The high resistance to low nitrogen stress is reflected by high yield and/or plant height under low nitrogen stress. The high resistance to low nitrogen stress is reflected by a high nitrogen content.

In a third aspect, the present invention provides a method for increasing plant yield and/or increasing plant stress tolerance, comprising the step of increasing the expression level and/or activity of a SiMYB61 protein in a recipient plant.

The yield is specifically the ear number, ear length, grain number per ear, grain weight and/or biomass.

The biomass comprises the weight of rice straw and/or the weight of rice.

The stress tolerance is in particular resistance to low nitrogen stress.

The improved stress tolerance can be embodied as an increased yield and/or an increased plant height under low nitrogen stress.

The high resistance to low nitrogen stress is reflected by a high nitrogen content.

The SiMYB61 protein is as shown above.

In a fourth aspect, the present invention provides a method of growing a transgenic plant, comprising the steps of: introducing a nucleic acid molecule capable of expressing a SiMYB61 protein into a recipient plant to obtain a transgenic plant with increased SiMYB61 protein expression; the transgenic plants have increased yield and/or stress resistance as compared to the recipient plant.

The yield is particularly the number of ears, ear length, grain number per ear, grain weight and/or biomass.

The biomass comprises the weight of rice straw and/or the weight of rice.

The stress tolerance is in particular resistance to low nitrogen stress.

The "introducing into a recipient plant a nucleic acid molecule capable of expressing a SiMYB61 protein" is effected by introducing into the recipient plant an expression cassette comprising a gene encoding the SiMYB61 protein.

The encoding gene of the SiMYB61 protein is shown in the specification.

The number of grains per spike is the number of solid grains per spike and/or the total grains per spike.

Any one of the plants is a dicotyledonous plant or a monocotyledonous plant;

further, the monocotyledon is a gramineous plant;

further, the gramineous plant is rice or millet.

The rice can be rice variety Kitaake.

The millet can be Yu Gu I.

The invention provides a new gene for rice stress genetic engineering, and simultaneously introduces japonica rice fine varieties through genetic transformation to culture new stress-resistant transgenic varieties, so that the stress resistance of rice can be effectively improved, the influence of stress on rice production is reduced, and important economic and social benefits are achieved.

Drawings

FIG. 1 shows SSR detection results of stress-resistant rice transformed with SiMYB61 gene. Marker: DL1000 Marker; negative control: kitaake; positive control: the plant expression vector psSiMYB 61.

Fig. 2 shows the statistical result of 18-year field test data.

FIG. 3 is a comparison of transgenic plants with wild type phenotype.

FIG. 4 shows the statistical results of the test data under the condition of normal field treatment for 19 years.

FIG. 5 is a statistical result of detection data under 19-year field low nitrogen stress conditions.

FIG. 6 shows the statistical results of 19 years field straw and rice weight measurement data.

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.

LP0471118-Bar-ubi-EDLL vector: are described in the literature: ning bud, Wang Shuang, ju Peng Gao, Bai Xin Xuan, Ge Lin Hao, Qixin, Jiangqin, Sun Xijun, Chenming, Sun Dazhen, over-expression millet SiANT1 has influence on rice salt tolerance [ J ]. Chinese agricultural science, 2018,51(10): 1830) 1841.

Yugu I: are described in the literature: ning bud, Wang Shuang, ju Peng lifting, Bai Xin Xuan, Ge Lin Hao, Qixin, Jiangqin, Sun Shi Jun, Chenming, Sun Dazhen, overexpression of millet SiANT1 on rice salt tolerance [ J ]. Chinese agricultural science, 2018,51(10):1830 one 1841.; the public is available from the institute of crop science, academy of agricultural sciences, china.

bar gene expression vector pSBAR: are described in the literature: obtaining drought resistant transgenic wheat [ D ] using an improved minimum expression box technique, university of inner mongolia agriculture, 2012; the public is available from the institute of crop science, academy of agricultural sciences, china.

Agrobacterium strain EHA 105: beijing Ongke New Biotechnology Ltd.

Rice variety Kitaake: are described in the literature: the influence of over-expressed millet SiANT1 on the salt tolerance of rice [ J ] Chinese agricultural science, 2018,51(10): 1830-; the public is available from the institute of crop science, academy of agricultural sciences, china.

Example 1 acquisition of SimYB61 protein and Gene encoding the same

A stress resistance related gene is cloned from the first confirmation of the stress resistance millet and is named as SiMYB61 gene, the genome sequence of the gene is shown as SEQ ID NO.1, the transcription sequence is shown as SEQ ID NO.2, and the CDS is shown as SEQ ID NO. 3. The protein (SiMYB61 protein) coded by the SiMYB61 gene is shown as SEQ ID NO. 4.

Example 2, SiMYB61 protein and application of encoding gene thereof in rice breeding

Preparation of linear minimal expression cassette

1. Obtaining of plant expression vector psSiMYB61

Extracting total RNA of Yugu No. one, and reverse transcribing into cDNA. The cDNA was used as a template, PCR amplification was carried out using primers F and R, and the CDS sequence of SiMYB61 (SEQ ID NO: 3) was cloned into the BamHI and SpeI sites of vector LP0471118-Bar-ubi-EDLL using the seamless cloning kit (cat # 639649) from Clotech according to the procedures described in the specification to obtain plant expression vector psSiMYB61 (which was sequence-verified).

And (3) primer F: 5'-AGACCGATCTGGATCATGGGGAAGCACTCCTGC-3', respectively;

and (3) primer R: 5'-CGATCGATCCACTAGCTAGATATTCTCGAAAGACAAGGACATTCTCTG-3' is added.

2. Preparation of Linear minimal expression cassette

Adopting HindIII and EcoRI to carry out double enzyme digestion on the plant expression vector psSiMYB61 obtained in the step 1, recovering a fragment of about 3500bp to obtain a SiMYB61 gene linear minimum expression frame, wherein the linear minimum expression frame consists of a ubiquitin constitutive promoter (1800bp), a SiMYB61 gene (1275bp) and a terminator nos 3' (300 bp). The minimal expression frame transformation method can remove the vector skeleton sequence, has no ampicillin resistance gene, has no obvious difference between the transgenic rice and the receptor, has no pathogenicity, and reduces the safety risk of inserting the plant genome exogenous fragment.

Preparing a marker gene expression frame by using a bar gene as a marker gene: adopting Hind III enzyme to cut the bar gene expression vector pSBAR, and recovering the enzyme cutting product, namely a marker gene expression frame; the marker gene expression frame consists of a maize ubiquitin promoter, a marker gene bar and a nos terminator.

Second, transformation of rice

1. And (3) transforming the SiMYB61 gene linear minimum expression frame and the marker gene expression frame prepared in the step one into an agrobacterium tumefaciens strain EHA105 to obtain the recombinant agrobacterium tumefaciens.

2. Soaking immature embryos of a rice variety Kitaake 12-14 days after pollination in 70% ethanol for 1 minute, then disinfecting the immature embryos with 10% sodium hypochlorite for 15 minutes, washing the immature embryos with sterile water for 3-5 times, taking out the immature embryos on an ultra-clean bench, and inoculating the immature embryos to SD (secure digital) ₂ The young embryo callus is induced on a culture medium (MS basic culture medium (containing no vitamin), 2 mg/L2, 4-D +1mg/LVB1+150mg/LAsn asparagine +30g/L sucrose +2.4g/L plant gel, pH is 5.8) for 7 days, and then the induced callus is transferred to a hypertonic culture medium (MS basic culture medium, 5 mg/L2, 4-D +0.4mol/L mannitol +3g/L plant gel) to be subjected to hypertonic treatment for 4-6 hours.

3. After step 2, the callus was infected with Agrobacterium and soaked in OD ₆₀₀ And oscillating and infecting the recombinant agrobacterium tumefaciens liquid of 0.8 for about 30min at 180 r/min.

4. After the step 3 is finished, continuously culturing the callus on a hypertonic culture medium for 16-18 hours, and then transferring the infected callus to SD ₂ Dark culture was performed on the medium for two weeks.

5. After the step 4 is completed, transferring the callus onto a selection culture medium containing 2-3mg/L of herbicide Bialaphos (MS basic culture medium (containing no vitamin) +2 mg/L2, 4-D +1mg/LVB1+150mg/LAsn asparagine +30g/L sucrose +2.4g/L plant gel +2-3mg/L herbicide Bialaphos, pH is 5.8) to perform callus screening, differentiation and seedling strengthening, so as to obtain a T0 generation transgenic rice plant.

Identification of transgenic positive rice

Extracting DNA of T0 rice leaves to be detected by adopting an SDS method, designing a primer for amplifying a partial sequence according to a SiMYB61 gene sequence by taking the DNA as a template to carry out PCR amplification, carrying out PCR amplification by adopting a primer F and a primer R, detecting a PCR amplification product by 0.8% agarose gel, and carrying out ultraviolet photographing. Replacing T0 generation rice with Kitaake rice variety to carry out the operation as negative control; the plant expression vector psSiMYB61 was used as a positive control.

F：5’-TGGCAACTTGAGCCATCTCC-3’；

R：5’-GTTGATGTTGAGAGAGTT-3’。

The PCR reaction system is shown in Table 1. And (3) PCR reaction conditions: denaturation at 94 deg.C for 5 min; 50sec at 94 ℃, 50sec at 62 ℃, 1min at 72 ℃ and 35 cycles; extension for 10min at 72 ℃.

TABLE 1

Composition (A)	Dosage (ul)
		(Takara)10xPCRbuffer(Mg ²⁺ Plus)	2.50
(Takara)25mMMg ²⁺	0.05
		(Takara)2.5mMdNTPMixture	2.00
Primer F	0.80
		Primer R	0.80
(Takara)r-TaqDNAPloymerase(5U/ul)	0.25
		Form panel	1.00
ddH ₂ O	17.6
		Total of	25

The results are shown in FIG. 1. 5 positive plants are obtained through PCR detection, and the positive rate is 2%. Through greenhouse generation adding, screening and identification, SiMYB 61-transgenic rice homozygous strains are selected and selected, and are named as OE21 and OE44 in sequence for the following experiments.

Fourth, phenotype detection of transgenic rice

1. 2018 field phenotype detection

And (3) the plant to be detected: rice variety kitaake (wt), transgenic line OE21, transgenic line OE 44.

Test site: transgenic test base of rice institute of agricultural science institute of Gao Jiangxi province of Jiangxi province.

The seedling bed management is the same as the field production (no nitrogen fertilizer is added), and each test material can grow normally.

8 plants are harvested from each material, and the biological yield and the rice yield of each material are determined in three times.

The N content of the paddy and the N content of the straw of the reference material are respectively measured by a Kjeldahl method. The detection method is specifically described in the references "comparison of total nitrogen in plants by Yangyi, Chua-Shunlin. flow analysis and Kjeldahl method [ J ]. proceedings of the Seiki institute of America, 2016,32(08): 51-54" and "Dahonglin, Wu Xiaojun. determination of nitrogen content in dried samples of plants by Kjeldahl method [ J ]. proceedings of the Jiangsu institute of agriculture, 1995(03): 70").

3 plants of each material are respectively taken for indoor seed test, and the number of the single plant effective spikes (more than 5 single spikes are effective spikes), the solid grains per spike, the total grains per spike, the seed setting rate and the like are inspected.

The results are shown in FIG. 2. In FIG. 2, the nitrogen contents of rice and straw are expressed in g/8, and the nitrogen contents are expressed by the weight of 8 rice/straw. The unit of the total nitrogen content of the straws is percentage (%), which means the nitrogen content of the straws/the weight of the 8 straws.

The results show that the dry grain weight and the dry straw weight of the transgenic rice line are higher than those of the control rice variety Kitaake under the treatment without applying nitrogen fertilizer. The effective spike number, the solid grain number and the total grain number of each spike of the single plant are all higher than those of the control. The N content of the rice and the N content of the straw are higher than those of the control.

2. 2019 field data

And (3) the plant to be detected: rice variety kitaake (wt), transgenic line OE 44.

Test site: transgenic test base of rice institute of agricultural science institute of high-safety Jiangxi province in Jiangxi province.

The management of the rice seedling bed is the same as the production of the field. Each test material was able to grow normally.

The test sets two treatments of applying 12 kilograms of pure nitrogen and not applying nitrogen fertilizer per mu, and each treatment is set for two times.

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. Other fertilizer and water management in the field is the same as local production.

20 plants of each material are harvested for each treatment, and the weight of the straw and the weight of the paddy are respectively measured for each material after three times of repetition.

Each material was harvested 8 plants per treatment, and the biological yield and rice yield of each material were determined in triplicate.

And 3 plants of each material are respectively taken for indoor seed test in each treatment, and the number of the effective ears of each plant, the solid grains of each ear, the total grains of each ear and the like are inspected.

The phenotypic observations are shown in figure 3.

The statistical results are shown in fig. 4 (normal treatment), fig. 5 (low nitrogen treatment) and fig. 6.

The results show that under the condition of normal nitrogen fertilizer application, the biomass (straw weight and rice weight), the ear length and the total grain per ear of the transgenic plants are obviously higher than those of the receptor plants. Under the low nitrogen stress, the biomass (the weight of the straws and the weight of the paddy) of the transgenic plants, the plant height, the number of ears, the length of the ears, the solid grains per ear and the total grains per ear are also higher than those of the receptor plants.

Sequence listing

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

SHANXI AGRICULTURAL University

Application of SiMYB61 protein and related biological material thereof in regulation and control of plant stress resistance

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actcaagtgg cctgacctgc tccatggaac cttcacagat acaccagcaa ccatgcagaa 1500

tcttagccaa tcactgtatg aagatgtggt caaagccgag aaccaattca acatggaggg 1560

cctctgtgca gcttggtctc aaaatttgca gccacagcaa catctgcagg tagcatcaga 1620

tttgtatgac aaggatttgc agagaatgtc cttgtctttc gagaatatct aggcaacatg 1680

tttcagcatg catggaacag aacaatagca gaaagctgag ttaatgaagc ctagaaaaca 1740

caacatttct tcacagattt atttgcaggc tatgtggagg tgtttgcgta ggtacattct 1800

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gaaaaaaagg agacaaagta tgaatttata ttacagcata atgtatagcc tagagaaaag 1920

tatgtctcga agtttacttt cctttcactt aaagttgctg tctttatctg tacatgcatg 1980

cagctcgact ttttgtatac gtgagtcatg gttgatcttt ttctaccaaa taaatgaggc 2040

aaggatgtgg aacactg 2057

<210> 3

<211> 1275

<212> DNA

<213> millet (Setaria italica)

<400> 3

atggggaagc actcctgctg ttacaagcag aagctgagga aggggctctg gtctcctgag 60

gaagacgaga agctcatgaa ccacataacc aagcatgggc atggctgctg gagcactgtt 120

ccaaagcttg cagggcttca aaggtgtggc aagagctgca ggctgaggtg gataaattac 180

ctgaggcctg accttaaaag aggtgcattc tctcaggagg aggaagacct tatcattgaa 240

cttcatgctg tcttgggcaa caggtggtct cagattgcaa cacggttgcc tggaagaact 300

gataacgaga tcaagaatct ctggaattca agcatcaaga agaagctccg gcagaaaggc 360

atcgacccca acacccacaa gccccttgct gaggttgatc gcaaaatagc tccaacaatc 420

agtactgaga gaacctccga gtccagcgat attgaccctt caagtggtgg tgcacttggc 480

aacttgagcc atctcctcag tgagacagca caatcaccag agctgctgcc agtgctcggt 540

aagcatcgca aagaaactac tagtttggca catctaaggg tgccatcgaa ggagctattc 600

cttgaccagc ttgtttctgg taatgataac ctccccagct gccgctcaac aggcccaatt 660

ccaaatttcc ctttccagca gttgatgtgt tacagcaatg aatttggcgg caagcatgga 720

ggcagcacga atccactctg gtttaaccag aatgagtcaa gctgcagcac catttccact 780

gtgatgccac cagtttcgcc atcaactctc tcaacatcaa caggactcaa taggtcaccg 840

gacaatccac actctggagg tactggcatt cagagtaacc aattctactg ggacaccact 900

aatcctagca gcagtagcag taaaggaagc agtggaagca atagcttggg atttgagctg 960

caaagcacaa gctcaattct ggagaatagt atcttcccat ggacagattt atcgccagat 1020

aaaaatagcc acctagagga agaactcaag tggcctgacc tgctccatgg aaccttcaca 1080

gatacaccag caaccatgca gaatcttagc caatcactgt atgaagatgt ggtcaaagcc 1140

gagaaccaat tcaacatgga gggcctctgt gcagcttggt ctcaaaattt gcagccacag 1200

caacatctgc aggtagcatc agatttgtat gacaaggatt tgcagagaat gtccttgtct 1260

ttcgagaata tctag 1275

<210> 4

<211> 424

<212> PRT

<213> millet (Setaria italica)

<400> 4

Met Gly Lys His Ser Cys Cys Tyr Lys Gln Lys Leu Arg Lys Gly Leu

1 5 10 15

Trp Ser Pro Glu Glu Asp Glu Lys Leu Met Asn His Ile Thr Lys His

20 25 30

Gly His Gly Cys Trp Ser Thr Val Pro Lys Leu Ala Gly Leu Gln Arg

35 40 45

Cys Gly Lys Ser Cys Arg Leu Arg Trp Ile Asn Tyr Leu Arg Pro Asp

50 55 60

Leu Lys Arg Gly Ala Phe Ser Gln Glu Glu Glu Asp Leu Ile Ile Glu

65 70 75 80

Leu His Ala Val Leu Gly Asn Arg Trp Ser Gln Ile Ala Thr Arg Leu

85 90 95

Pro Gly Arg Thr Asp Asn Glu Ile Lys Asn Leu Trp Asn Ser Ser Ile

100 105 110

Lys Lys Lys Leu Arg Gln Lys Gly Ile Asp Pro Asn Thr His Lys Pro

115 120 125

Leu Ala Glu Val Asp Arg Lys Ile Ala Pro Thr Ile Ser Thr Glu Arg

130 135 140

Thr Ser Glu Ser Ser Asp Ile Asp Pro Ser Ser Gly Gly Ala Leu Gly

145 150 155 160

Asn Leu Ser His Leu Leu Ser Glu Thr Ala Gln Ser Pro Glu Leu Leu

165 170 175

Pro Val Leu Gly Lys His Arg Lys Glu Thr Thr Ser Leu Ala His Leu

180 185 190

Arg Val Pro Ser Lys Glu Leu Phe Leu Asp Gln Leu Val Ser Gly Asn

195 200 205

Asp Asn Leu Pro Ser Cys Arg Ser Thr Gly Pro Ile Pro Asn Phe Pro

210 215 220

Phe Gln Gln Leu Met Cys Tyr Ser Asn Glu Phe Gly Gly Lys His Gly

225 230 235 240

Gly Ser Thr Asn Pro Leu Trp Phe Asn Gln Asn Glu Ser Ser Cys Ser

245 250 255

Thr Ile Ser Thr Val Met Pro Pro Val Ser Pro Ser Thr Leu Ser Thr

260 265 270

Ser Thr Gly Leu Asn Arg Ser Pro Asp Asn Pro His Ser Gly Gly Thr

275 280 285

Gly Ile Gln Ser Asn Gln Phe Tyr Trp Asp Thr Thr Asn Pro Ser Ser

290 295 300

Ser Ser Ser Lys Gly Ser Ser Gly Ser Asn Ser Leu Gly Phe Glu Leu

305 310 315 320

Gln Ser Thr Ser Ser Ile Leu Glu Asn Ser Ile Phe Pro Trp Thr Asp

325 330 335

Leu Ser Pro Asp Lys Asn Ser His Leu Glu Glu Glu Leu Lys Trp Pro

340 345 350

Asp Leu Leu His Gly Thr Phe Thr Asp Thr Pro Ala Thr Met Gln Asn

355 360 365

Leu Ser Gln Ser Leu Tyr Glu Asp Val Val Lys Ala Glu Asn Gln Phe

370 375 380

Asn Met Glu Gly Leu Cys Ala Ala Trp Ser Gln Asn Leu Gln Pro Gln

385 390 395 400

Gln His Leu Gln Val Ala Ser Asp Leu Tyr Asp Lys Asp Leu Gln Arg

405 410 415

Met Ser Leu Ser Phe Glu Asn Ile

420

Claims

Use of a SiMYB61 protein or related biomaterial thereof in (a1) and/or (a2) as follows:

(a1) improving plant yield-related traits;

(a2) the stress resistance of the plants is improved;

the amino acid sequence of the SiMYB61 protein is shown in SEQ ID No. 4;

the related biological material is a nucleic acid molecule capable of expressing the SiMYB61 protein or an expression cassette, a recombinant vector, a recombinant bacterium or a transgenic cell line containing the nucleic acid molecule;

the plant yield-related traits are ear number, ear length and/or grain per ear;

the plant stress resistance is the resistance of the plant to low nitrogen stress;

the plant is rice or millet.
The application of SiMYB61 protein or related biological materials thereof in breeding plants with high yield and/or high stress resistance;

the amino acid sequence of the SiMYB61 protein is shown as SEQ ID No. 4;

the related biological material is a nucleic acid molecule capable of expressing the SiMYB61 protein or an expression cassette, a recombinant vector, a recombinant bacterium or a transgenic cell line containing the nucleic acid molecule;

the high yield is represented by a large number of spikes and/or a high spike length and/or a large number of grains per spike;

the stress resistance is resistance to low nitrogen stress;

the plant is rice or millet.
3. A method for increasing yield-related traits in plants and/or increasing stress tolerance in plants, comprising the step of increasing expression level and/or activity of a SiMYB61 protein in a recipient plant;

the amino acid sequence of the SiMYB61 protein is shown in SEQ ID No. 4;

the yield-related traits are ear number, ear length and/or grain number per ear;

the stress tolerance is resistance to low nitrogen stress;

the plant is rice or millet.
4. A method of growing a transgenic plant comprising the steps of: introducing a nucleic acid molecule capable of expressing a SiMYB61 protein into a recipient plant to obtain a transgenic plant with increased SiMYB61 protein expression; the transgenic plant has increased yield-related traits and/or stress resistance as compared to the recipient plant;

the amino acid sequence of the SiMYB61 protein is shown as SEQ ID No. 4;

the yield is the ear number, the ear length and/or the grain number per ear;

the stress resistance is resistance to low nitrogen stress;

the plant is rice or millet.
5. The method of claim 4, wherein: the step of introducing into a recipient plant a nucleic acid molecule capable of expressing a SiMYB61 protein comprises introducing into the recipient plant an expression cassette comprising a gene encoding the SiMYB61 protein.
6. The method of claim 5, wherein:

the encoding gene of the SiMYB61 protein is a DNA molecule described in any one of the following items:

(B1) DNA molecule shown in SEQ ID No. 2;

(B2) a DNA molecule shown as SEQ ID No. 3.