CN110628810B - A kind of method to improve plant photosynthetic efficiency - Google Patents

Abstract

The invention discloses a method for improving plant photosynthetic efficiency. The method comprises the following steps of inhibiting or knocking out a sodium bile co-transporter gene in a plant, and simultaneously overexpressing a glycolate dehydrogenase gene, a glyoxylate carboxylase gene and a Tartrate semialdehyde reductase gene. The invention creatively finds that inhibiting the expression of BASS6 gene in plants, combined with overexpression of glycolate dehydrogenase gene, glyoxylate carboxylase gene and tartrate semialdehyde reductase gene in chloroplast, can significantly reduce photorespiration and improve plant The photosynthetic efficiency increases plant biomass or yield, and more importantly, the drought tolerance of this transgenic plant is significantly higher than that of plants with suppressed PLGG1 gene expression, and the biomass or yield under drought conditions is increased by 3% compared with the non-transgenic control. 45 %.

Description

A kind of method to improve plant photosynthetic efficiency

(1) Technical field

The invention relates to a method for improving the photosynthetic efficiency of plants. The photosynthetic efficiency, biomass or yield of plants can be improved by means of transgenic methods, and plant planting resources can be improved.

(2) Background technology

The increase in the number of human beings and the improvement of living standards require more food and feed to be consumed, which requires more food to be harvested on limited land. Therefore, it is very important to breed new high-yielding plant varieties.

The photosynthesis products of the whole plant are all derived from the enzyme-catalyzed conversion of _CO2 to organic carbon compounds. Ribulose 1,5-diphosphate carboxylase/oxygenase (RubisCO) is a carboxylase in the Calvin-Benson (CB) cycle. Since RubisCO reacts with either _CO or O, _RubisCO reacts with O to produce phosphoglycolic _acid , which enters the photorespiration cycle, which leads to a waste of fixed carbon and nitrogen in plants. Globally, this process re-releases approximately 29GT of fresh assimilated carbon into the atmosphere each year (Anav A, et al. Spatiotemporal patterns of terrestrial grossprimary production: a review. Rev Geophys 2015, 53:785-818.).

In order to reduce the loss caused by photorespiration and improve the photosynthetic efficiency of plants, the commonly used method is to recover CO ₂ in glycolic acid through a new photorespiration branch, so as to achieve the purpose of reducing photorespiration and improving photosynthetic efficiency (Peterhansel C, Blume C, Offermann S. Photorespiratory bypasses: how can they work? [J]. Journal of Experimental Botany, 2013, 64(3):709-715.).

Glycolate dehydrogenase (GDH) can convert glycolate to glyoxylate. Glycolic acid dehydrogenases currently used in plant transgenic research and applications are mainly derived from lower plant Chlamydomonas reinhardtii or Escherichia coli. The glycolate dehydrogenase in green algae is encoded by one gene, while the glycolate dehydrogenase in Bacillus is composed of three subunits, D, E, and F encoded by three genes. It has been reported that after overexpressing the fusion gene of the three subunits of D, E, and F in potatoes, the expression of DEFp fusion protein in the plant increases, and the sugars such as glucose, fructose and sucrose are doubled, and the biomass is also significant. Increase (Nolke G, Houdelet M, Kreuzaler F, et al. The expression of a recombinant glycolate dehydrogenase polyprotein inpotato (Solanum tuberosum) plastids strongly enhances photosynthesis and tuberyield [J]. Plant Biotechnology Journal, 2014, 12(6): 734- 742.). However, the functions and activities of glycolate dehydrogenases derived from Escherichia coli and green algae are significantly different, so the performance in transgenic plants is also very different.

Glyoxylate carboxylase (GCL, glyoxylate carboxylyase) can catalyze the conversion of glyoxylate to tartronic semialdehyde (TS), this enzyme was first cloned from Escherichia coli (Chang YY, WangAY, Cronan Jr JE. 1993. Molecular cloning, DNAsequencing, and biochemical analyses of Escherichia coli glyoxylatecarboligase. An enzyme of the acetohydroxy acidsynthase-pyruvateoxidase family. Journal of Biological Chemistry 268, 3911-3919.). Tartrate semialdehyde reductase (tartronic semialdehydereductase, TSR) can reduce tartar semialdehyde to glyceric acid.

Bile Acid Sodium Symporter (BASS) and plastidal glycolate/glyceratetranslocator 1 (PLGG1) are responsible for transporting chloroplast glycolate to peroxides in photorespiration The key protein of the enzyme body (South P F, Walker B J, Cavanagh AP, et al. Bile Acid Sodium Symporter BASS6Can Transport Glycolate and IsInvolved in Photorespiratory Metabolism in Arabidopsis thaliana [J]. The PlantCell, 2017:tpc.00775.2016.). Both BASS and PLGG1 genes have the function of transporting glycolic acid, but PLGG1 also has the function of transporting glycolic acid and glyceric acid. Previous studies have shown that overexpression of GDH and MS genes in tobacco chloroplasts while inhibiting the expression of PLGG1 gene can significantly increase tobacco biomass (PF South, AP Cavanagh, HW Liu, et al. Synthetic glycolate metabolism pathways stimulate crop growth and productivity in the field, Science, 2019:363(6422):eaat9077.).

Drought tolerance is a very important characteristic of plants. Plants have a certain degree of drought tolerance, which is conducive to their ability to resist drought stress and adapt to different geographical environments. In the context of increasing water stress, it is very important to cultivate new crop varieties with strong drought tolerance.

However, our study found that plants that overexpressed GDH and MS genes in chloroplasts while suppressing the expression of PLGG1 gene had significantly reduced drought tolerance compared with controls. In contrast, plants that overexpressed GDH and MS genes in chloroplasts while suppressing the expression of the BASS6 gene had significantly higher drought tolerance than plants that suppressed the expression of the PLGG1 gene.

(3) Contents of the invention

The object of the present invention is to provide a method for maintaining or improving plant drought tolerance, reducing plant photorespiration, improving plant photosynthetic efficiency, and increasing plant biomass or yield. In order to achieve the above technical purpose, the technical means of the present invention is to inhibit or knock In addition to BASS gene expression in plants, GDH, GCL and TSR genes were overexpressed in chloroplasts.

The technical scheme adopted in the present invention is:

The present invention provides a method for improving plant photosynthetic efficiency. The method comprises the following steps: inhibiting or knocking out sodium bile co-transporter (BASS) gene in plants, and simultaneously overexpressing glycolate dehydrogenase (GDH) gene, glyoxylate Carboxylase (GCL) gene and tartrate semialdehyde reductase (TSR) gene.

Further, the gene encoding the sodium bile co-transporter is derived from plants (Table 1), and its amino acid sequence is shown in one of SEQ ID NO.1, SEQ ID NO.2 or SEQ ID NO.3. When the plant is rice, the amino acid sequence of the BASS6 gene is shown in SEQ ID NO.1; when the plant is soybean, the amino acid sequence of the BASS6 gene is shown in SEQ ID NO.2 and SEQ ID NO. 3 shown.

Further, the method for inhibiting the sodium bile cotransporter gene in the plant is RNA interference method, specifically, introducing a double-stranded RNA nucleotide sequence that forms a hairpin structure targeting the sodium bile cotransporter gene into the plant. Preferably, when the plant is rice, an OsBASS-RNAi sequence targeting the OsBASS gene of rice to form a hairpin structure is introduced, and the nucleotide sequences are respectively SEQ ID NO. 4; when the plant is soybean, the GmBASS gene targeting soybean is introduced to form a hairpin structure. The GmBASS-RNAi sequence of the hairpin structure, the nucleotide sequence is shown in SEQ ID NO.5.

Further, the glycolate dehydrogenase (GDH) gene can be derived from prokaryotes or eukaryotes, including but not limited to those shown in Table 2, and the preferred nucleotide sequence is SEQ ID NO.6 (the amino acid sequence is SEQ ID NO. .7).

Further, the glyoxylate carboxylase (GCL) gene can be derived from prokaryotes or eukaryotes, preferably the nucleotide sequence of the GCL gene is shown in SEQ ID NO.8, and the amino acid sequence is shown in SEQ ID NO.9 .

Further, the tartrate semialdehyde reductase (TSR) gene can be derived from prokaryotes or eukaryotes (Table 2), preferably the nucleotide sequence of the TSR gene is shown in SEQ ID NO.10, and the amino acid sequence is SEQ ID NO. .11 shown.

Table 1: Sodium bile cotransporter (BASS) genes

Numbering source species NCBI Accession Number 1 Arabidopsisthaliana <u>NP</u> 567671 2 Zeamays <u>NP</u> 001158917 3 Sorghumbicolor XP 021308938 4 Oryzasativa <u>XP</u>015612294 5 Glycinemax XP 003538535/XP 003517442

Table 2: Glycolic acid dehydrogenase (GDH) genes from different species

Numbering source species NCBI Accession Number 1 Chlamydomonas reinhardtii <u>XP 001695381.1</u> 2 Volvox carteri f.nagariensis <u>XP002946459.1</u> 3 Gonium pectorale <u>KXZ46746.1</u> 4 Chlamydomonas eustigma <u>GAX77289.1</u> 5 Escherichia coli K-12 <u>NP</u> 417453.1, YP 026191.1, YP 026190.1

The method of the present invention is to construct a T-DNA vector and introduce it into a plant; the T-DNA vector construction method is as follows: the pCambia1300 binary vector containing the glufosinate-resistant bar gene is used as the basic vector, and then glycolic acid is respectively connected to it. Dehydrogenase gene (GDH) expression cassette, glyoxylate carboxylase gene (GCL) expression cassette, tartrate semialdehyde reductase gene (TSR) expression cassette and sodium bile co-transporter gene RNAi expression cassette; the containing The pCambia1300 binary vector of the glufosinate resistance bar gene replaces the original hygromycin resistance gene hptII with the glufosinate resistance gene bar.

In the present invention, the RNAi expression box of BASS gene, the overexpression box of GDH gene, the overexpression box of GCL gene and the overexpression box of TSR gene can be realized by the method of molecular polymerization or hybridization polymerization. Molecular polymerization refers to constructing the BASS gene suppressor expression cassette, GDH, GCL and TSR gene expression cassettes on the T-DNA of the same vector, and transferring the T-DNA into the recipient plant genome by transgenic method, so that In the target plants, the expression of BASS gene was inhibited at the same time, and GDH gene, GCL and TSR gene were overexpressed. Hybrid polymerization refers to obtaining plants containing the BASS gene suppression expression box, GDH overexpression box, GCL overexpression box and TSR overexpression box respectively, and then using traditional breeding methods to carry out the plants containing one or two of the above expression boxes respectively. Crossed to obtain transgenic plants containing the above three expression cassettes at the same time.

The signal peptide that mediates the overexpression of GDH, GCL and TSR genes in the chloroplast of the present invention is derived from the plant RuBisCO small subunit (RbcS) or the phosphoglucomutase transit peptide sequence, preferably the amino acid sequence of the chloroplast signal peptide is as SEQ ID NO.12 Or as shown in SEQ ID NO. 13, the chloroplast signal peptide sequence is fused to the N-terminus of GDH, GCL and TSR proteins. The promoter that mediates the overexpression of GDH, GCL and TSR in the present invention can be derived from eukaryotes or prokaryotes, or can be obtained by artificial synthesis, and can be a constitutive promoter or a specific promoter. The promoters include p35S (NCBIACCESSION: MG719235 REGION: 848-1628), maize UBI promoter (NCBI ACCESSION: KR297238 REGION: 4879-6876) and rice Actin1 promoter (NCBI ACCESSION: AY452735 REGION: 2428-3797).

The terminators of the present invention can be derived from eukaryotes or prokaryotes, and can also be obtained by artificial synthesis. Preferably, the terminators are ter1 (NCBI ACCESSION: KJ716235 REGION: 3962-4158) and ter2 (NCBI ACCESSION: MG733984 REGION: 2092-2314).

The plants in the present invention are C3 plants, which refer to plants in which the initial product of CO ₂ assimilation is the three-carbon compound 3-phosphoglycerate in the photosynthetic carbon cycle, mainly including rice and soybean.

The present invention provides a method for significantly increasing plant biomass or yield and maintaining or improving its drought tolerance by increasing plant photosynthetic efficiency, by performing RNA interference on plant BASS gene, combined with overexpressing glycolic acid in plant chloroplasts Dehydrogenase gene (GDH), glyoxylate carboxylase gene (GCL), and tartrate semialdehyde reductase gene (TSR), which convert glycolic acid to glyoxylic acid and further to tartaric semialdehyde in the chloroplast, Tartar semialdehyde is converted into glyceric acid under the action of tartar semialdehyde reductase, so as to reduce photorespiration and improve photosynthetic efficiency and yield. RNA interference of BASS gene in rice and soybean chloroplasts and overexpression of glycolate dehydrogenase gene, glyoxylate carboxylase gene and tartrate semialdehyde reductase gene increased rice yield by 3%-45%, soybean Yield was increased by 3%-45%, and drought tolerance was comparable to, or better than, non-transgenic controls.

Compared with the prior art, the beneficial effects of the present invention are mainly reflected in:

The present invention creatively finds that inhibiting the expression of BASS6 gene in plants, combined with overexpression of glycolate dehydrogenase gene (GDH), glyoxylate carboxylase gene (GCL) and tartrate semialdehyde reductase gene (TSR) in chloroplasts , can significantly reduce photorespiration, increase the photosynthetic efficiency of plants, and increase plant biomass or yield. More importantly, the drought tolerance of this transgenic plant is significantly higher than that of plants that inhibit PLGG1 gene expression, and the biomass or yield under drought conditions 3%-45% increase over non-transgenic controls.

(4) Specific implementations

The present invention is further described below in conjunction with specific embodiment, but the protection scope of the present invention is not limited to this:

Embodiment 1, the construction of carrier

The GDH genes of the present invention can be derived from prokaryotes or eukaryotes. The GDH genes provided by the present invention include but are not limited to the genes shown in Table 2. In order to construct the transformation vector, the E. coli-derived GDH gene and the corresponding terminator sequence were artificially synthesized, including the chloroplast signal peptide, the GDH coding gene and the terminator. The nucleotide sequence is shown in SEQID NO. ' ends are provided with BamHI and KpnI sites, respectively. The artificially synthesized GCL gene includes a chloroplast signal peptide, a GCL coding gene and a terminator, the nucleotide sequence is shown in SEQ ID NO. 8, and the 5' end and the 3' end are respectively provided with BamHI and HindIII sites. The artificially synthesized TSR gene includes a chloroplast signal peptide, a TSR coding gene and a terminator, the nucleotide sequence is shown in SEQ ID NO. 10, and the 5' end and the 3' end are respectively provided with BamHI and EcoRI sites.

In order to realize the expression inhibition or knockout of the BASS gene, the present invention provides the BASS gene of the plant itself; when the plant is rice, the amino acid sequence of the BASS6 gene is shown in SEQ ID NO. 1; when the plant is rice In the case of soybean, the amino acid sequence of the BASS6 gene is shown in SEQ ID NO.2 and SEQ ID NO.3. In order to construct the BASS gene interference expression cassette, the OsBASS-RNAi and GmBASS-RNAi sequences targeting the rice OsBASS gene and soybean GmBASS gene, which can form hairpin structures, were artificially synthesized, as shown in SEQ ID NO.4 and SEQ ID NO.5. Show. As controls, Os PGGL1-RNAi and Gm PGGL1-RNAi sequences targeting rice OsPGGL1 gene and soybean GmPGGL1 gene that can form hairpin structures were synthesized, as shown in SEQ ID NO.14 and SEQ ID NO.15. Terminator ter1 is added to the 3' ends of the above sequences respectively, and finally constitutes OsBASS-RNAi-ter, GmBASS-RNAi-ter, Os PGGL1-RNAi-ter, Gm PGGL1-RNAi-ter. The 5' and 3' ends are provided with BglII and HindIII sites, respectively.

At the same time, the maize Ubi promoter sequence, the rice Actin promoter and the 35S promoter sequence of cauliflower mosaic virus (CaMV) were synthesized. EcoRI and BamHI sites are set at the 5' and 3' ends of the Ubi promoter, respectively, EcoRI and BamHI sites are set at the 5' and 3' ends of the Actin promoter, respectively, and the 5' and 3' ends of the 35S promoter are respectively set KpnI and BamHI sites are provided. At the same time, a Ubi promoter with HindIII and BamHI sites at the 5' and 3' ends was synthesized to mediate the transcription of RNAi sequences.

In order to construct a binary vector that can be used to transform plants by Agrobacterium, the commercial binary vector pCambia1300 was used as the basis, and the previous hptII (hygromycin resistance) gene was replaced by a glufosinate-resistant bar through the XhoI restriction site. Gene (NCBI ACCESSIONp:MG719235 REGION:287-835), the substituted vector was named pCambia1300-bar.

The Ubi promoter and GDH gene were ligated into the pCambia1300-bar vector through EcoRI and KpnI sites to obtain the transition vector pCambia1300-bar-GDH. Then, the 35S promoter and GCL gene were ligated into the transition vector pCambia1300-bar-GDH through KpnI and HindIII sites to obtain the transition vector pCambia1300-bar-GDH-GCL. Then, pCambia1300-bar-GDH-GCL was single digested with EcoRI, and then ligated with the Actin promoter digested with EcoRI and BamHI and the TSR gene digested with BamHI and EcoRI to obtain the over-vector pCambia1300-bar-GDH- GCL-TSR. Finally, pCambia1300-bar-GDH-GCL-TSR was single digested by HindIII, then OsBASS-RNAi-ter or GmBASS-RNAi-ter digested with BglII and HindIII and Ubi digested with BamHI and HindIII were digested The promoters were ligated to construct final vectors, named pCambia1300-bar-GDH-GCL-TSR-OsBASS-RNAi (GGTOsBi) and pCambia1300-bar-GDH-GCL-TSR-GmBASS-RNAi (GGTGmBi), respectively.

As a control, vectors containing the PGGL1 gene expression cassette were constructed in the same way and named as pCambia1300-bar-GDH-GCL-TSR-OsPGGL1-RNAi (GGTOsPi) and pCambia1300-bar-GDH-GCL-TSR-GmPGGL1-RNAi, respectively (GGTGmPi).

Finally, the T-DNA plasmid was transferred into Agrobacterium LB4404 by electroporation, and positive clones were screened out by YEP solid medium containing 15 μg/ml tetracycline and 50 μg/mL kanamycin, and the bacteria were preserved for the next step. plant transformation.

Example 2, rice transformation

The method of obtaining transgenic rice is to use the existing technology (Lu Xiongbin, Gong Zuxun (1998) Life Science 10: 125-131; Liu Fan et al. (2003) Molecular Plant Breeding 1: 108-115). The mature and plump "Xiushui-134" seeds were selected and shelled to induce callus as the transformation material. Take the Agrobacterium plates constructed in Example 1 containing the plasmids pCambia1300-bar-GDH-GCL-TSR-OsBASS-RNAi (GGTOsBi) and pCambia1300-bar-GDH-GCL-TSR-OsPGGL1-RNAi (GGTOsPi) respectively. Pick a single colony to inoculate and prepare for transformation with Agrobacterium. Put the callus to be transformed into the Agrobacterium liquid with an OD of about 0.6 (the preparation of the Agrobacterium liquid: inoculate the Agrobacterium into the culture medium, and cultivate to an OD of about 0.6; the medium consists of 3 g/LK ₂ HPO ₄ , 1 g/L NaH ₂ PO ₄ , 1 g/L NH ₄ Cl, 0.3 g/L MgSO ₄ ·7H ₂ O, 0.15 g/L KCl, 0.01 g/L CaCl ₂ , 0.0025 g/L FeSO ₄ ·7H ₂ O, 5g/L sucrose, 20mg/L acetosyringone, the solvent is water, pH=5.8), let Agrobacterium bind to the surface of the callus, and then transfer the callus to the co-cultivation medium (MS+2mg/ L 2,4-D+30g/L glucose+30g/L sucrose+3g/L agar (sigma 7921)+20mg/L acetosyringone), co-cultured for 2-3 days. The transformed callus was washed with sterile water and transferred to screening medium (MS+2mg/L 2,4-D(2,4-dichlorophenoxyacetic acid)+30g/L sucrose+3g/L agar (sigma 7921) + 20 mg/L acetosyringone + 2 mM glyphosate (Sigma)), screened and cultured for two months (one intermediate passage). After selection, the callus with good growth vigor was transferred to pre-differentiation medium (MS+0.1g/L inositol+5mg/L ABA (abscisic acid)+1mg/L NAA (naphthalene acetic acid)+5mg/L 6-BA (6-benzylaminoadenine)+20g/L sorbitol+30g/L sucrose+2.5g/L phytogel (gelrite)) for about 20 days, and then the pre-differentiated callus was transferred to differentiated On the medium, 14 hours a day of light differentiation and germination. After 2-3 weeks, the resistant regenerated plants were transferred to rooting medium (1/2MS+0.2mg/L NAA+20g/L sucrose+2.5g/L gelrite) for strong seedlings and rooting, and finally the regenerated plants were washed off the agar Transplant in the greenhouse, select transgenic lines with high yield, large seeds or high biomass that can increase rice yield, and cultivate new varieties. Transgenic rice plants containing the above transformation vectors were obtained respectively.

Example 3. Soybean Transformation

The steps used here to obtain transgenic soybeans were derived from existing techniques (Deng et al., 1998, PlantPhysiology Communications 34:381-387; Ma et al., 2008, Scientia Agricultura Sinica 41:661-668; Zhou et al., 2001 , Journal of Northeast Agricultural University 32:313-319). Select healthy, plump and mature soybeans of "Tianlong No. 1", sterilize them with 80% ethanol for 2 minutes, rinse with sterile water, and place them in a desiccator filled with chlorine gas (produced by the reaction of 50ml NaClO and 2ml concentrated HCl) Sterilize for 4-6 hours. The sterilized soybeans were sown into B5 medium in an ultra-clean workbench, and cultivated at 25°C for 5 days, while the optical density was at the level of 90-150 μmol photons/m ² ·s. When the cotyledons turn green and burst the seed coat, sterile sprouts will grow. The sprouts with the hypocotyls removed were cut in half in length so that both explants had cotyledons and epicotyls. Explants are cut at approximately 7-8 points at the junction of cotyledons and epicotyls and can be used as target tissue for infection.

The monoclonal Agrobacterium containing the vectors pCambia1300-bar-GDH-GCL-TSR-GmBASS-RNAi (GGTGmBi) and pCambia1300-bar-GDH-GCL-TSR-GmPGGL1-RNAi (GGTGmPi) constructed by Example 1 were separated Cultivated for use. The prepared explants were immersed in the Agrobacterium suspension (prepared by the method of Example 2) and co-cultured for about 30 minutes. Then, the excess cell suspension on the infected tissue was absorbed with absorbent paper, and then transferred to 1/10B5 co-culture medium for 3-5 days in the dark at 25°C.

The co-cultured plant tissues were washed with B5 liquid medium to remove excess Agrobacterium, and then placed in B5 solid medium for 5 days at 25°C and allowed to germinate. The induced embryonic tissue was transferred to B5 selection medium containing 0.1 mM glyphosate, and incubated in the light at 25°C for 4 weeks, during which the medium was changed every two weeks. The screened embryo tissues were then transferred to solid medium and cultured at 25°C until they grew into seedlings. Subsequently, the transgenic plantlets were transferred to 1/2B5 medium for rooting induction. Finally, the grown plantlets were washed to remove the agar and planted in the greenhouse.

Example 4: Identification of transgenic rice

Transgenic rice plants of vectors pCambia1300-bar-GDH-GCL-TSR-OsBASS-RNAi(GGTOsBi) and pCambia1300-bar-GDH-GCL-TSR-OsPGGL1-RNAi(GGTOsPi) were obtained in Example 2, respectively. The above-mentioned transgenic plants showed increased biomass and yield compared with non-transgenic controls, and GGTOsBi plants showed the greatest increase in biomass or yield. In order to further identify the performance changes of GGTOsBi transgenic plants, we evaluated and compared the biomass and seed yield of the above transgenic plants, and the results are shown in Table 3. Under normal conditions, the biomass or yield of GGTOsBi transgenic plants and GGTOsPi transgenic plants were significantly increased compared with non-transgenic controls, with an increase of 5%-45%; but under drought conditions (Table 5), GGTOsBi transgenic plants were significantly higher than those of non-transgenic controls. The control still had a significant increase in biomass or yield, while the GGTOsPi transgenic plants did not have a significant increase in biomass or yield compared to the non-transgenic controls.

Table 3 Comparison of biomass and yield

Example 5: Identification of Transgenic Soybeans

The transgenic soybean plants of the vectors pCambia1300-bar-GDH-GCL-TSR-GmBASS-RNAi (GGTGmBi) and pCambia1300-bar-GDH-GCL-TSR-GmPGGL1-RNAi (GGTGmPi) were obtained in Example 3, respectively. The above transgenic plants showed increased biomass and yield compared to non-transgenic controls, and the transgenic plants showed the greatest increase in biomass or yield. In order to further identify the performance changes of the transgenic plants, we evaluated the biomass and seed yield of the above-mentioned transgenic plants, and the results are shown in Table 4. Under normal conditions, the biomass or yield of GGTGmBi transgenic plants and GGTGmPi transgenic plants were significantly increased compared with non-transgenic controls by 5%-45%; Controls still had significant increases in biomass or yield, while GGTGmPi transgenic plants did not have significant increases in biomass or yield compared to non-transgenic controls.

Table 4 Comparison of biomass and yield

Table 5. Index grades based on farmland and crop drought morphology (GB/T 32136-2015)

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<213> Unknown

<400> 3

Met Ile Ser Ser Gly Leu Lys Pro Lys His Phe Asn Asn Val His Ser

1 5 10 15

Leu Phe Asn Leu Ser Lys Ser Gln Gln Pro Pro Asn Pro Ile Ile Val

20 25 30

Pro Cys Cys Arg Thr Asn Thr Asn Asn Asn Ile Ser Ser Pro Phe Ser

35 40 45

Ile Arg Phe Asn Ser Pro Phe Pro Tyr Arg Ser Pro Lys Ile Pro Leu

50 55 60

Lys Cys Ala Pro Leu His Ser Ser Asp Ser Leu Pro Pro Asp Pro Ser

65 70 75 80

Ser Ala Ser Thr Gln Met Glu Gln Asn Ser Met Ser Ile Leu Glu Ile

85 90 95

Leu Lys Gln Ser Asn Ser Tyr Leu Pro His Ala Leu Ile Ala Ser Ile

100 105 110

Leu Leu Ala Leu Ile Tyr Pro Arg Ser Leu Thr Trp Phe Thr Ser Arg

115 120 125

Phe Tyr Ala Pro Ala Leu Gly Phe Leu Met Phe Ala Val Gly Val Asn

130 135 140

Ser Asn Glu Asn Asp Phe Leu Glu Ala Phe Lys Arg Pro Ala Glu Ile

145 150 155 160

Val Thr Gly Tyr Phe Gly Gln Phe Ala Val Lys Pro Leu Leu Gly Tyr

165 170 175

Leu Phe Cys Met Ile Ala Val Thr Val Leu Gly Leu Pro Thr Thr Val

180 185 190

Gly Ala Gly Ile Val Leu Val Ala Cys Val Ser Gly Ala Gln Leu Ser

195 200 205

Ser Tyr Ala Thr Phe Leu Thr Asp Pro Gln Met Ala Pro Leu Ser Ile

210 215 220

Val Met Thr Ser Leu Ser Thr Ala Ser Ala Val Phe Val Thr Pro Leu

225 230 235 240

Leu Leu Leu Leu Leu Ile Gly Lys Lys Leu Pro Ile Asp Val Lys Gly

245 250 255

Met Val Tyr Asn Ile Thr Gln Ile Val Val Val Pro Ile Ala Ala Gly

260 265 270

Leu Leu Leu Asn Arg Phe Phe Pro Arg Ile Cys Asn Val Ile Arg Pro

275 280 285

Phe Leu Pro Pro Leu Ser Val Leu Val Ala Ser Ile Cys Ala Gly Ala

290 295 300

Pro Leu Ala Leu Asn Val Glu Thr Met Lys Ser Pro Leu Gly Val Ala

305 310 315 320

Ile Leu Leu Leu Val Val Ala Phe His Leu Ser Ser Phe Ile Ala Gly

325 330 335

Tyr Ile Leu Ser Gly Phe Val Phe Arg Asp Ser Leu Asp Val Lys Ala

340 345 350

Leu Gln Arg Thr Ile Ser Phe Glu Thr Gly Met Gln Ser Ser Leu Leu

355 360 365

Ala Leu Ala Leu Ala Asn Lys Phe Phe Glu Asp Pro Lys Val Ala Ile

370 375 380

Pro Pro Ala Ile Ser Thr Ser Ile Met Ser Leu Met Gly Phe Val Leu

385 390 395 400

Val Leu Ile Trp Thr Arg Arg Gly Lys Ser Glu Ile Lys Asn Ser Ser

405 410 415

<210> 4

<211> 856

<212> DNA

<213> Unknown

<400> 4

gcttttgatg gaaagacaga catcataccg aatttataaa aggaaaagaa ataaattcaa 60

aactttacat tttttatgcc accaaccaaa ggtgaatcaa agatatgaac aagagtttct 120

taagactatt agcccccccc cccccccccc aacgacctcc aactccaatc ctccttaatc 180

gccaacccac acagctataa aaaggggata tttcagatcg gatcaagcag agcacctacg 240

ccgtgaaaac ggcggcgaga ccgcctgggg aggagccaga cggggcagtc gccggccggt 300

gggcagatgg tggacgccgg cgatgtgggg ccgcaccagg acggcggcgt tgggggccat 360

tcgagcgccg gcgaccgcga gggtgggtgg gttttggttt cagagtttca gagctgatga 420

cgcaacgcag cgaaagagac gattcagatt tcagtgagaa gttgggagtt tcgacaagga 480

acgaacaatc agtcgaatgg cccccaacgc cgccgtcctg gtgcggcccc acatcgccgg 540

cgtccaccat ctgcccaccg gccggcgact gccccgtctg gctcctcccc aggcggtctc 600

gccgccgttt tcacggcgta ggtgctctgc ttgatccgat ctgaaatatc ccctttttat 660

agctgtgtgg gttggcgatt aaggaggatt ggagttggag gtcgttgggg gggggggggg 720

ggggctaata gtcttaagaa actcttgttc atatctttga ttcacctttg gttggtggca 780

taaaaaatgt aaagttttga atttatttct tttcctttta taaattcggt atgatgtctg 840

tctttccatc aaaagt 856

<210> 5

<211> 771

<212> DNA

<213> Unknown

<400> 5

aagaggccag cagaaattgt cactggttat tttggccagt ttgctgtgaa gcctcttctt 60

ggatatctgt tttgcatgat tgcagtaact gttttaggcc taccaacaac agtaggcgca 120

ggaattgtat tggtggcttg tgttagtggt gctcagcttt caagttatgc tactttcctg 180

actgatccac aaatggcacc tttaagcata gttatgacat cactgtccac tgcttctgca 240

gtttttgtca cgccactctt attactgttg ctcattggga agaaattgcc ttcatagtct 300

caacatttaa ggcaagtggc gctccggcac agatagatgc caccagtaca gatagcggag 360

gcaaaaatgg tcgaataaca ttacaaatac gaggaaagaa tcgatttaga agcaggccag 420

ctgcaatagg cacaaccaca atctgtgtaa tgttatacac cattcctttt acatctatag 480

gcaatttctt cccaatgagc aacagtaata agagtggcgt gacaaaaact gcagaagcag 540

tggacagtga tgtcataact atgcttaaag gtgccatttg tggatcagtc aggaaagtag 600

cataacttga aagctgagca ccactaacac aagccaccaa tacaattcct gcgcctactg 660

ttgttggtag gcctaaaaca gttactgcaa tcatgcaaaa cagatatcca agaagaggct 720

tcacagcaaa ctggccaaaa taaccagtga caatttctgc tggcctcttt a 771

<210> 6

<211> 3668

<212> DNA

<213> Unknown

<400> 6

ggatccaaca atggccccgt ccgtgatggc ctcctccgcc accaccgtgg ccccgttcca 60

gggcctcaag tccaccgccg gcatgccggt ggcccgccgc tccggcaact cctccttcgg 120

caacgtgtcc aacggcggcc gcatccgctg catgccgcgc ggccagggca agcgcctcgc 180

ccagctcctc ggcgcccagc tcaagcagta cgccgccgag gtgcgcggca tctccaccgc 240

cggcggcgcc tcccgcggcg gcgcccgcgg cccggcctcc ccgtcctccc tcgagcagca 300

gacccgccag gtggcccagg tggccgtgca gcagtccacc cagcaggccg tgaaggtggt 360

ggtgccggcc atcaaggtgg acctcgtggg cgccgtgtcc tccgtgtccg agtccgacaa 420

ggtggagccg ggcgtgttca agaacgtgga cggccaccgc ttcgaggacg gccgctacgc 480

cgccttcgtg gaggagatca ccaagttcat cccgaaggag cgccagtact ccgacccggt 540

gcgcaccttc gcctacggca ccgacgcctc cttctaccgc ctcaacccga agctcgtggt 600

gaaggtgcac aacgaggacg aggtgcgccg catcatgccg atcgccgagc gcctccaggt 660

gccgatcacc ttccgcgccg ccggcacctc cctctccggc caggccatca ccgactccgt 720

gctcatcaag ctctcccaca ccggcaagaa cttccgcaac ttcaccgtgc acggcgacgg 780

ctccgtgatc accgtggagc cgggcctcat cggcggcgag gtgaaccgca tcctcgccgc 840

ccaccagaag aagaacaagc tcccgatcca gtacaagatc ggcccggacc cgtcctccat 900

cgactcctgc atgatcggcg gcatcgtgtc caacaactcc tccggcatgt gctgcggcgt 960

gtcccagaac acctaccaca ccctcaagga catgcgcgtg gtgttcgtgg acggcaccgt 1020

gctcgacacc gccgacccga actcctgcac cgccttcatg aagtcccacc gctccctcgt 1080

ggacggcgtg gtgtccctcg cccgccgcgt gcaggccgac aaggagctca ccgccctcat 1140

ccgccgcaag ttcgccatca agtgcaccac cggctactcc ctcaacgccc tcgtggactt 1200

cccggtggac aacccgatcg agatcatcaa gcacctcatc atcggctccg agggcaccct 1260

cggcttcgtg tcccgcgcca cctacaacac cgtgccggag tggccgaaca aggcctccgc 1320

cttcatcgtg ttcccggacg tgcgcgccgc ctgcaccggc gcctccgtgc tccgcaacga 1380

gacctccgtg gacgccgtgg agctcttcga ccgcgcctcc ctccgcgagt gcgagaacaa 1440

cgaggacatg atgcgcctcg tgccggacat caagggctgc gacccgatgg ccgccgccct 1500

cctcatcgag tgccgcggcc aggacgaggc cgccctccag tcccgcatcg aggaggtggt 1560

gcgcgtgctc accgccgccg gcctcccgtt cggcgccaag gccgcccagc cgatggccat 1620

cgacgcctac ccgttccacc acgaccagaa gaacgccaag gtgttctggg acgtgcgccg 1680

cggcctcatc ccgatcgtgg gcgccgcccg cgagccgggc acctccatgc tcatcgagga 1740

cgtggcctgc ccggtggaca agctcgccga catgatgatc gacctcatcg acatgttcca 1800

gcgccacggc taccacgacg cctcctgctt cggccacgcc ctcgagggca acctccacct 1860

cgtgttctcc cagggcttcc gcaacaagga ggaggtgcag cgcttctccg acatgatgga 1920

ggagatgtgc cacctcgtgg ccaccaagca ctccggctcc ctcaagggcg agcacggcac 1980

cggccgcaac gtggccccgt tcgtggagat ggagtggggc aacaaggcct acgagctcat 2040

gtgggagctc aaggccctct tcgacccgtc ccacaccctc aacccgggcg tgatcctcaa 2100

ccgcgaccag gacgcccaca tcaagttcct caagccgtcc ccggccgcct ccccgatcgt 2160

gaaccgctgc atcgagtgcg gcttctgcga gtccaactgc ccgtcccgcg acatcaccct 2220

caccccgcgc cagcgcatct ccgtgtaccg cgagatgtac cgcctcaagc agctcggccc 2280

gggcgcctcc gaggaggaga agaagcagct cgccgccatg tcctcctcct acgcctacga 2340

cggcgagcag acctgcgccg ccgacggcat gtgccaggag aagtgcccgg tgaagatcaa 2400

caccggcgac ctcatcaagt ccatgcgcgc cgagcacatg aaggaggaga agaccgcctc 2460

cggcatggcc gactggctcg ccgccaactt cggcgtgatc aactccaacg tgccgcgctt 2520

cctcaacatc gtgaacgcca tgcactccgt ggtgggctcc gccccgctct ccgccatctc 2580

ccgcgccctc aacgccgcca ccaaccactt cgtgccggtg tggaacccgt acatgccgaa 2640

gggcgccgcc ccgctcaagg tgccggcccc gccggccccg gccgccgccg aggcctccgg 2700

catcccgcgc aaggtggtgt acatgccgtc ctgcgtgacc cgcatgatgg gcccggccgc 2760

ctccgacacc gagaccgccg ccgtgcacga gaaggtgatg tccctcttcg gcaaggccgg 2820

ctacgaggtg atcatcccgg agggcgtggc ctcccagtgc tgcggcatga tgttcaactc 2880

ccgcggcttc aaggacgccg ccgcctccaa gggcgccgag ctcgaggccg ccctcctcaa 2940

ggcctccgac aacggcaaga tcccgatcgt gatcgacacc tccccgtgcc tcgcccaggt 3000

gaagtcccaa atctccgagc cgtccctccg cttcgccctc tacgagccgg tggagttcat 3060

ccgccacttc ctcgtggaca agctcgagtg gaagaaggtg cgcgaccagg tggccatcca 3120

cgtgccgtgc tcctccaaga agatgggcat cgaggagtcc ttcgccaagc tcgccggcct 3180

ctgcgccaac gaggtggtgc cgtccggcat cccgtgctgc ggcatggccg gcgaccgcgg 3240

catgcgcttc ccggagctca ccggcgcctc cctccagcac ctcaacctcc cgaagacctg 3300

caaggacggc tactccacct cccgcacctg cgagatgtcc ctctccaacc acgccggcat 3360

caacttccgc ggcctcgtgt acctcgtgga cgaggccacc gccccgaaga agcaggccgc 3420

cgccgccaag accgcctaag tagatgccga ccggatctgt cgatcgacaa gctcgagttt 3480

ctccataata atgtgtgagt agttcccaga taagggaatt agggttccta tagggtttcg 3540

ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta 3600

tcaataaaat ttctaattcc taaaaccaaa atccagtact aaaatccaga tcccccgaat 3660

taaagctt 3668

<210> 7

<211> 1136

<212> PRT

<213> Unknown

<400> 7

Met Ala Ser Ser Met Leu Ser Ser Ala Thr Met Val Ala Ser Pro Ala

1 5 10 15

Gln Ala Thr Met Val Ala Pro Phe Asn Gly Leu Lys Ser Ser Ala Ala

20 25 30

Phe Pro Ala Thr Arg Lys Ala Asn Gly Gly Pro Arg Gly Gln Gly Lys

35 40 45

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

50 55 60

Val Arg Gly Ile Ser Thr Ala Gly Gly Ala Ser Arg Gly Gly Ala Arg

65 70 75 80

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

85 90 95

Gln Val Ala Val Gln Gln Ser Thr Gln Gln Ala Val Lys Val Val Val

100 105 110

Pro Ala Ile Lys Val Asp Leu Val Gly Ala Val Ser Ser Val Ser Glu

115 120 125

Ser Asp Lys Val Glu Pro Gly Val Phe Lys Asn Val Asp Gly His Arg

130 135 140

Phe Glu Asp Gly Arg Tyr Ala Ala Phe Val Glu Glu Ile Thr Lys Phe

145 150 155 160

Ile Pro Lys Glu Arg Gln Tyr Ser Asp Pro Val Arg Thr Phe Ala Tyr

165 170 175

Gly Thr Asp Ala Ser Phe Tyr Arg Leu Asn Pro Lys Leu Val Val Lys

180 185 190

Val His Asn Glu Asp Glu Val Arg Arg Ile Met Pro Ile Ala Glu Arg

195 200 205

Leu Gln Val Pro Ile Thr Phe Arg Ala Ala Gly Thr Ser Leu Ser Gly

210 215 220

Gln Ala Ile Thr Asp Ser Val Leu Ile Lys Leu Ser His Thr Gly Lys

225 230 235 240

Asn Phe Arg Asn Phe Thr Val His Gly Asp Gly Ser Val Ile Thr Val

245 250 255

Glu Pro Gly Leu Ile Gly Gly Glu Val Asn Arg Ile Leu Ala Ala His

260 265 270

Gln Lys Lys Asn Lys Leu Pro Ile Gln Tyr Lys Ile Gly Pro Asp Pro

275 280 285

Ser Ser Ile Asp Ser Cys Met Ile Gly Gly Ile Val Ser Asn Asn Ser

290 295 300

Ser Gly Met Cys Cys Gly Val Ser Gln Asn Thr Tyr His Thr Leu Lys

305 310 315 320

Asp Met Arg Val Val Phe Val Asp Gly Thr Val Leu Asp Thr Ala Asp

325 330 335

Pro Asn Ser Cys Thr Ala Phe Met Lys Ser His Arg Ser Leu Val Asp

340 345 350

Gly Val Val Ser Leu Ala Arg Arg Val Gln Ala Asp Lys Glu Leu Thr

355 360 365

Ala Leu Ile Arg Arg Lys Phe Ala Ile Lys Cys Thr Thr Gly Tyr Ser

370 375 380

Leu Asn Ala Leu Val Asp Phe Pro Val Asp Asn Pro Ile Glu Ile Ile

385 390 395 400

Lys His Leu Ile Ile Gly Ser Glu Gly Thr Leu Gly Phe Val Ser Arg

405 410 415

Ala Thr Tyr Asn Thr Val Pro Glu Trp Pro Asn Lys Ala Ser Ala Phe

420 425 430

Ile Val Phe Pro Asp Val Arg Ala Ala Cys Thr Gly Ala Ser Val Leu

435 440 445

Arg Asn Glu Thr Ser Val Asp Ala Val Glu Leu Phe Asp Arg Ala Ser

450 455 460

Leu Arg Glu Cys Glu Asn Asn Glu Asp Met Met Arg Leu Val Pro Asp

465 470 475 480

Ile Lys Gly Cys Asp Pro Met Ala Ala Ala Leu Leu Ile Glu Cys Arg

485 490 495

Gly Gln Asp Glu Ala Ala Leu Gln Ser Arg Ile Glu Glu Val Val Arg

500 505 510

Val Leu Thr Ala Ala Gly Leu Pro Phe Gly Ala Lys Ala Ala Gln Pro

515 520 525

Met Ala Ile Asp Ala Tyr Pro Phe His His Asp Gln Lys Asn Ala Lys

530 535 540

Val Phe Trp Asp Val Arg Arg Gly Leu Ile Pro Ile Val Gly Ala Ala

545 550 555 560

Arg Glu Pro Gly Thr Ser Met Leu Ile Glu Asp Val Ala Cys Pro Val

565 570 575

Asp Lys Leu Ala Asp Met Met Ile Asp Leu Ile Asp Met Phe Gln Arg

580 585 590

His Gly Tyr His Asp Ala Ser Cys Phe Gly His Ala Leu Glu Gly Asn

595 600 605

Leu His Leu Val Phe Ser Gln Gly Phe Arg Asn Lys Glu Glu Val Gln

610 615 620

Arg Phe Ser Asp Met Met Glu Glu Met Cys His Leu Val Ala Thr Lys

625 630 635 640

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

645 650 655

Pro Phe Val Glu Met Glu Trp Gly Asn Lys Ala Tyr Glu Leu Met Trp

660 665 670

Glu Leu Lys Ala Leu Phe Asp Pro Ser His Thr Leu Asn Pro Gly Val

675 680 685

Ile Leu Asn Arg Asp Gln Asp Ala His Ile Lys Phe Leu Lys Pro Ser

690 695 700

Pro Ala Ala Ser Pro Ile Val Asn Arg Cys Ile Glu Cys Gly Phe Cys

705 710 715 720

Glu Ser Asn Cys Pro Ser Arg Asp Ile Thr Leu Thr Pro Arg Gln Arg

725 730 735

Ile Ser Val Tyr Arg Glu Met Tyr Arg Leu Lys Gln Leu Gly Pro Gly

740 745 750

Ala Ser Glu Glu Glu Lys Lys Gln Leu Ala Ala Met Ser Ser Ser Tyr

755 760 765

Ala Tyr Asp Gly Glu Gln Thr Cys Ala Ala Asp Gly Met Cys Gln Glu

770 775 780

Lys Cys Pro Val Lys Ile Asn Thr Gly Asp Leu Ile Lys Ser Met Arg

785 790 795 800

Ala Glu His Met Lys Glu Glu Lys Thr Ala Ser Gly Met Ala Asp Trp

805 810 815

Leu Ala Ala Asn Phe Gly Val Ile Asn Ser Asn Val Pro Arg Phe Leu

820 825 830

Asn Ile Val Asn Ala Met His Ser Val Val Gly Ser Ala Pro Leu Ser

835 840 845

Ala Ile Ser Arg Ala Leu Asn Ala Ala Thr Asn His Phe Val Pro Val

850 855 860

Trp Asn Pro Tyr Met Pro Lys Gly Ala Ala Pro Leu Lys Val Pro Ala

865 870 875 880

Pro Pro Ala Pro Ala Ala Ala Glu Ala Ser Gly Ile Pro Arg Lys Val

885 890 895

Val Tyr Met Pro Ser Cys Val Thr Arg Met Met Gly Pro Ala Ala Ser

900 905 910

Asp Thr Glu Thr Ala Ala Val His Glu Lys Val Met Ser Leu Phe Gly

915 920 925

Lys Ala Gly Tyr Glu Val Ile Ile Pro Glu Gly Val Ala Ser Gln Cys

930 935 940

Cys Gly Met Met Phe Asn Ser Arg Gly Phe Lys Asp Ala Ala Ala Ser

945 950 955 960

Lys Gly Ala Glu Leu Glu Ala Ala Leu Leu Lys Ala Ser Asp Asn Gly

965 970 975

Lys Ile Pro Ile Val Ile Asp Thr Ser Pro Cys Leu Ala Gln Val Lys

980 985 990

Ser Gln Ile Ser Glu Pro Ser Leu Arg Phe Ala Leu Tyr Glu Pro Val

995 1000 1005

Glu Phe Ile Arg His Phe Leu Val Asp Lys Leu Glu Trp Lys Lys Val

1010 1015 1020

Arg Asp Gln Val Ala Ile His Val Pro Cys Ser Ser Lys Lys Met Gly

1025 1030 1035 1040

Ile Glu Glu Ser Phe Ala Lys Leu Ala Gly Leu Cys Ala Asn Glu Val

1045 1050 1055

Val Pro Ser Gly Ile Pro Cys Cys Gly Met Ala Gly Asp Arg Gly Met

1060 1065 1070

Arg Phe Pro Glu Leu Thr Gly Ala Ser Leu Gln His Leu Asn Leu Pro

1075 1080 1085

Lys Thr Cys Lys Asp Gly Tyr Ser Thr Ser Arg Thr Cys Glu Met Ser

1090 1095 1100

Leu Ser Asn His Ala Gly Ile Asn Phe Arg Gly Leu Val Tyr Leu Val

1105 1110 1115 1120

Asp Glu Ala Thr Ala Pro Lys Lys Gln Ala Ala Ala Ala Lys Thr Ala

1125 1130 1135

<210> 8

<211> 2135

<212> DNA

<213> Unknown

<400> 8

ggatccaaca atggccccgt ccgtgatggc ctcctccgcc accaccgtgg ccccgttcca 60

gggcctcaag tccaccgccg gcatgccggt ggcccgccgc tccggcaact cctccttcgg 120

caacgtgtcc aacggcggcc gcatccgctg catggccaag atgcgcgccg tggacgccgc 180

catgtacgtg ctcgagaagg agggcatcac caccgccttc ggcgtgccgg gcgccgccat 240

caacccgttc tactccgcca tgcgcaagca cggcggcatc cgccacatcc tcgcccgcca 300

cgtggagggc gcctcccaca tggccgaggg ctacacccgc gccaccgccg gcaacatcgg 360

cgtgtgcctc ggcacctccg gcccggccgg caccgacatg atcaccgccc tctactccgc 420

ctccgccgac tccatcccga tcctctgcat caccggccag gccccgcgcg cccgcctcca 480

caaggaggac ttccaggccg tggacatcga ggccatcgcc aagccggtgt ccaagatggc 540

cgtgaccgtg cgcgaggccg ccctcgtgcc gcgcgtgctc cagcaggcct tccacctcat 600

gcgctccggc cgcccgggcc cggtgctcgt ggacctcccg ttcgacgtgc aggtggccga 660

gatcgagttc gacccggaca tgtacgagcc gctcccggtg tacaagccgg ccgcctcccg 720

catgcagatc gagaaggccg tggagatgct catccaggcc gagcgcccgg tgatcgtggc 780

cggcggcggc gtgatcaacg ccgacgccgc cgccctcctc cagcagttcg ccgagctcac 840

ctccgtgccg gtgatcccga ccctcatggg ctggggctgc atcccggacg accacgagct 900

catggccggc atggtgggcc tccagaccgc ccaccgctac ggcaacgcca ccctcctcgc 960

ctccgacatg gtgttcggca tcggcaaccg cttcgccaac cgccacaccg gctccgtgga 1020

gaagtacacc gagggccgca agatcgtgca catcgacatc gagccgaccc agatcggccg 1080

cgtgctctgc ccggacctcg gcatcgtgtc cgacgccaag gccgccctca ccctcctcgt 1140

ggaggtggcc caggagatgc agaaggccgg ccgcctcccg tgccgcaagg agtgggtggc 1200

cgactgccag cagcgcaagc gcaccctcct ccgcaagacc cacttcgaca acgtgccggt 1260

gaagccgcag cgcgtgtacg aggagatgaa caaggccttc ggccgcgacg tgtgctacgt 1320

gaccaccatc ggcctctccc agatcgccgc cgcccagatg ctccacgtgt tcaaggaccg 1380

ccactggatc aactgcggcc aggccggccc gctcggctgg accatcccgg ccgccctcgg 1440

cgtgtgcgcc gccgacccga agcgcaacgt ggtggccatc tccggcgact tcgacttcca 1500

gttcctcatc gaggagctcg ccgtgggcgc ccagttcaac atcccgtaca tccacgtgct 1560

cgtgaacaac gcctacctcg gcctcatccg ccagtcccag cgcgccttcg acatggacta 1620

ctgcgtgcag ctcgccttcg agaacatcaa ctcctccgag gtgaacggct acggcgtgga 1680

ccacgtgaag gtggccgagg gcctcggctg caaggccatc cgcgtgttca agccggagga 1740

catcgccccg gccttcgagc aggccaaggc cctcatggcc cagtaccgcg tgccggtggt 1800

ggtggaggtg atcctcgagc gcgtgaccaa catctccatg ggctccgagc tcgacaacgt 1860

gatggagttc gaggacatcg ccgacaacgc cgccgacgcc ccgaccgaga cctgcttcat 1920

gcactacgag taagagctct agatcgttct gcacaaagtg gagtagtcag tcatcgatca 1980

ggaaccagac accagacttt tattcataca gtgaagtgaa gtgaagtgca gtgcagtgag 2040

ttgctggttt ttgtacaact tagtatgtat ttgtatttgt aaaatacttc tatcaataaa 2100

atttctaatt cctaaaacca aaatccagga agctt 2135

<210> 9

<211> 593

<212> PRT

<213> Unknown

<400> 9

Met Ala Lys Met Arg Ala Val Asp Ala Ala Met Tyr Val Leu Glu Lys

1 5 10 15

Glu Gly Ile Thr Thr Ala Phe Gly Val Pro Gly Ala Ala Ile Asn Pro

20 25 30

Phe Tyr Ser Ala Met Arg Lys His Gly Gly Ile Arg His Ile Leu Ala

35 40 45

Arg His Val Glu Gly Ala Ser His Met Ala Glu Gly Tyr Thr Arg Ala

50 55 60

Thr Ala Gly Asn Ile Gly Val Cys Leu Gly Thr Ser Gly Pro Ala Gly

65 70 75 80

Thr Asp Met Ile Thr Ala Leu Tyr Ser Ala Ser Ala Asp Ser Ile Pro

85 90 95

Ile Leu Cys Ile Thr Gly Gln Ala Pro Arg Ala Arg Leu His Lys Glu

100 105 110

Asp Phe Gln Ala Val Asp Ile Glu Ala Ile Ala Lys Pro Val Ser Lys

115 120 125

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

130 135 140

Gln Ala Phe His Leu Met Arg Ser Gly Arg Pro Gly Pro Val Leu Val

145 150 155 160

Asp Leu Pro Phe Asp Val Gln Val Ala Glu Ile Glu Phe Asp Pro Asp

165 170 175

Met Tyr Glu Pro Leu Pro Val Tyr Lys Pro Ala Ala Ser Arg Met Gln

180 185 190

Ile Glu Lys Ala Val Glu Met Leu Ile Gln Ala Glu Arg Pro Val Ile

195 200 205

Val Ala Gly Gly Gly Val Ile Asn Ala Asp Ala Ala Ala Leu Leu Gln

210 215 220

Gln Phe Ala Glu Leu Thr Ser Val Pro Val Ile Pro Thr Leu Met Gly

225 230 235 240

Trp Gly Cys Ile Pro Asp Asp His Glu Leu Met Ala Gly Met Val Gly

245 250 255

Leu Gln Thr Ala His Arg Tyr Gly Asn Ala Thr Leu Leu Ala Ser Asp

260 265 270

Met Val Phe Gly Ile Gly Asn Arg Phe Ala Asn Arg His Thr Gly Ser

275 280 285

Val Glu Lys Tyr Thr Glu Gly Arg Lys Ile Val His Ile Asp Ile Glu

290 295 300

Pro Thr Gln Ile Gly Arg Val Leu Cys Pro Asp Leu Gly Ile Val Ser

305 310 315 320

Asp Ala Lys Ala Ala Leu Thr Leu Leu Val Glu Val Ala Gln Glu Met

325 330 335

Gln Lys Ala Gly Arg Leu Pro Cys Arg Lys Glu Trp Val Ala Asp Cys

340 345 350

Gln Gln Arg Lys Arg Thr Leu Leu Arg Lys Thr His Phe Asp Asn Val

355 360 365

Pro Val Lys Pro Gln Arg Val Tyr Glu Glu Met Asn Lys Ala Phe Gly

370 375 380

Arg Asp Val Cys Tyr Val Thr Thr Ile Gly Leu Ser Gln Ile Ala Ala

385 390 395 400

Ala Gln Met Leu His Val Phe Lys Asp Arg His Trp Ile Asn Cys Gly

405 410 415

Gln Ala Gly Pro Leu Gly Trp Thr Ile Pro Ala Ala Leu Gly Val Cys

420 425 430

Ala Ala Asp Pro Lys Arg Asn Val Val Ala Ile Ser Gly Asp Phe Asp

435 440 445

Phe Gln Phe Leu Ile Glu Glu Leu Ala Val Gly Ala Gln Phe Asn Ile

450 455 460

Pro Tyr Ile His Val Leu Val Asn Asn Ala Tyr Leu Gly Leu Ile Arg

465 470 475 480

Gln Ser Gln Arg Ala Phe Asp Met Asp Tyr Cys Val Gln Leu Ala Phe

485 490 495

Glu Asn Ile Asn Ser Ser Glu Val Asn Gly Tyr Gly Val Asp His Val

500 505 510

Lys Val Ala Glu Gly Leu Gly Cys Lys Ala Ile Arg Val Phe Lys Pro

515 520 525

Glu Asp Ile Ala Pro Ala Phe Glu Gln Ala Lys Ala Leu Met Ala Gln

530 535 540

Tyr Arg Val Pro Val Val Val Glu Val Ile Leu Glu Arg Val Thr Asn

545 550 555 560

Ile Ser Met Gly Ser Glu Leu Asp Asn Val Met Glu Phe Glu Asp Ile

565 570 575

Ala Asp Asn Ala Ala Asp Ala Pro Thr Glu Thr Cys Phe Met His Tyr

580 585 590

Glu

<210> 10

<211> 1232

<212> DNA

<213> Unknown

<400> 10

ggatccaaca atggccccgt ccgtgatggc ctcctccgcc accaccgtgg ccccgttcca 60

gggcctcaag tccaccgccg gcatgccggt ggcccgccgc tccggcaact cctccttcgg 120

caacgtgtcc aacggcggcc gcatccgctg catgaagctc ggcttcatcg gcctcggcat 180

catgggcacc ccgatggcca tcaacctcgc ccgcgccggc caccagctcc acgtgaccac 240

catcggcccg gtggccgacg agctcctctc cctcggcgcc gtgaacgtgg acaccgcccg 300

ccaggtgacc gaggccgccg acatcatctt catcatggtg ccggacaccc cgcaggtgga 360

ggaggtgctc ttcggcgaga acggctgcac caaggcctcc ctcaagggca agaccatcgt 420

ggacatgtcc tccatctccc cgatcgagac caagcgcttc gcccgccagg tgaacgagct 480

cggcggcgac tacctcgacg ccccggtgtc cggcggcgag atcggcgccc gcgagggcac 540

cctctccatc atggtgggcg gcgacgaggc cgtgttcgag cgcgtgaagc cgctcttcga 600

gctcctcggc aagaacatca ccctcgtggg cggcaacggc gacggccaga cctgcaaggt 660

ggccaaccag atcatcgtgg ccctcaacat cgaggccgtg tccgaggccc tcctcttcgc 720

ctccaaggcc ggcgccgacc cggtgcgcgt gcgccaggcc ctcatgggcg gcttcgcctc 780

ctcccgcatc ctcgaggtgc acggcgagcg catgatcaag cgcaccttca acccgggctt 840

caagatcgcc ctccaccaga aggacctcaa cctcgccctc cagtccgcca aggccctcgc 900

cctcaacctc ccgaacaccg ccacctgcca ggagctcttc aacacctgcg ccgccaacgg 960

cggctcccag ctcgaccact ccgccctcgt gcaggccctc gagctcatgg ccaaccacaa 1020

gctcgcctaa gagctctaga tcgttctgca caaagtggag tagtcagtca tcgatcagga 1080

accagacacc agacttttat tcatacagtg aagtgaagtg aagtgcagtg cagtgagttg 1140

ctggtttttg tacaacttag tatgtatttg tatttgtaaa atacttctat caataaaatt 1200

tctaattcct aaaaccaaaa tccagggaat tc 1232

<210> 11

<211> 292

<212> PRT

<213> Unknown

<400> 11

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

1 5 10 15

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

20 25 30

Pro Val Ala Asp Glu Leu Leu Ser Leu Gly Ala Val Asn Val Asp Thr

35 40 45

Ala Arg Gln Val Thr Glu Ala Ala Asp Ile Ile Phe Ile Met Val Pro

50 55 60

Asp Thr Pro Gln Val Glu Glu Val Leu Phe Gly Glu Asn Gly Cys Thr

65 70 75 80

Lys Ala Ser Leu Lys Gly Lys Thr Ile Val Asp Met Ser Ser Ile Ser

85 90 95

Pro Ile Glu Thr Lys Arg Phe Ala Arg Gln Val Asn Glu Leu Gly Gly

100 105 110

Asp Tyr Leu Asp Ala Pro Val Ser Gly Gly Glu Ile Gly Ala Arg Glu

115 120 125

Gly Thr Leu Ser Ile Met Val Gly Gly Asp Glu Ala Val Phe Glu Arg

130 135 140

Val Lys Pro Leu Phe Glu Leu Leu Gly Lys Asn Ile Thr Leu Val Gly

145 150 155 160

Gly Asn Gly Asp Gly Gln Thr Cys Lys Val Ala Asn Gln Ile Ile Val

165 170 175

Ala Leu Asn Ile Glu Ala Val Ser Glu Ala Leu Leu Phe Ala Ser Lys

180 185 190

Ala Gly Ala Asp Pro Val Arg Val Arg Gln Ala Leu Met Gly Gly Phe

195 200 205

Ala Ser Ser Arg Ile Leu Glu Val His Gly Glu Arg Met Ile Lys Arg

210 215 220

Thr Phe Asn Pro Gly Phe Lys Ile Ala Leu His Gln Lys Asp Leu Asn

225 230 235 240

Leu Ala Leu Gln Ser Ala Lys Ala Leu Ala Leu Asn Leu Pro Asn Thr

245 250 255

Ala Thr Cys Gln Glu Leu Phe Asn Thr Cys Ala Ala Asn Gly Gly Ser

260 265 270

Gln Leu Asp His Ser Ala Leu Val Gln Ala Leu Glu Leu Met Ala Asn

275 280 285

His Lys Leu Ala

290

<210> 12

<211> 47

<212> PRT

<213> Unknown

<400> 12

Met Ala Pro Ser Val Met Ala Ser Ser Ala Thr Thr Val Ala Pro Phe

1 5 10 15

Gln Gly Leu Lys Ser Thr Ala Gly Met Pro Val Ala Arg Arg Ser Gly

20 25 30

Asn Ser Ser Phe Gly Asn Val Ser Asn Gly Gly Arg Ile Arg Cys

35 40 45

<210> 13

<211> 42

<212> PRT

<213> Unknown

<400> 13

Met Ala Ser Ser Met Leu Ser Ser Ala Thr Met Val Ala Ser Pro Ala

1 5 10 15

Gln Ala Thr Met Val Ala Pro Phe Asn Gly Leu Lys Ser Ser Ala Ala

20 25 30

Phe Pro Ala Thr Arg Lys Ala Asn Gly Gly

35 40

<210> 14

<211> 838

<212> DNA

<213> Unknown

<400> 14

cgatcggcag gtcatgcgaa atcgcgacga ggctgcgtgc attttgactg attgacgact 60

cacgctgggt aggcccgagg gagaggtggc gcccgcgcct cctcctcctc ctccggcggc 120

ggcggcggcg tggatcgggg cgatgaggcg gtggcgagga ttggcgggag ccatggagat 180

tgccgccgct tggcgggagg aggaggagga ggaggagggg ttgcgtcggc atcggcgggg 240

aggaagcgtg cggaggcggg gcggcgacgt ggcggtggcg gagggcgaaa ggcggcagcg 300

atggctgctg cgtagcgagg caatcatgca gggggaggat gatgatgagg tcgctgccat 360

ctcttctctt ctcttctctt cttctccttc tcctttggcc agcgagagag cagtggcagt 420

gacagtggat gagaagggag ctgggagcag tggcagaggc caggtggaag agaggagatg 480

gcagcgacct catcatcatc ctccccctgc atgattgcct cgctacgcag cagccatcgc 540

tgccgccttt cgccctccgc caccgccacg tcgccgcccc gcctccgcac gcttcctccc 600

cgccgatgcc gacgcaaccc ctcctcctcc tcctcctcct cccgccaagc ggcggcaatc 660

tccatggctc ccgccaatcc tcgccaccgc ctcatcgccc cgatccacgc cgccgccgcc 720

gccggaggag gaggaggagg cgcgggcgcc acctctccct cgggcctacc cagcgtgagt 780

cgtcaatcag tcaaaatgca cgcagcctcg tcgcgatttc gcatgacctg ccgatcgg 838

<210> 15

<211> 491

<212> DNA

<213> Unknown

<400> 15

gccgtgacat aaatgacagt ctcagccaac aattgcaagc aaactttgtc taacagcagg 60

aatcgagcaa agaatggatc caaatatacc attcagagca taagcaatgg cacaaaatgg 120

gagagcctcg ggttccttgg cagacaaagc tgctgttcca agcccgtggg cactgaaaaa 180

taaaatttag gtcaagagag taagtaaata accatgcacg cacaaagaaa atgtcattca 240

actattattg catgtttggt atagattaat ttttgagaaa taatcactta ttttacagaa 300

actaatttta tttttcagtg cccacgggct tggaacagca gctttgtctg ccaaggaacc 360

cgaggctctc ccattttgtg ccattgctta tgctctgaat ggtatatttg gatccattct 420

ttgctcgatt cctgctgtta gacaaagttt gcttgcaatt gttggctgag actgtcattt 480

atgtcacggc c 491

Claims (10)

1. A method for improving the photosynthetic efficiency of a plant, characterized in that the method comprises: inhibiting or knocking out sodium bile cotransporter gene in plants, and simultaneously over-expressing glycollic acid dehydrogenase gene, glyoxylate carboxylase gene and tartrate semialdehyde reductase gene.

2. The method of claim 1, wherein the amino acid sequence of the sodium bile acid cotransporter is as set forth in one of SEQ ID No.1, SEQ ID No.2 or SEQ ID No. 3.

3. The method of claim 1, wherein the method of inhibiting a sodium bile acid cotransporter gene in a plant comprises: introducing into a plant a double-stranded RNA nucleotide sequence that forms a hairpin structure of a targeted bile acid sodium cotransporter gene.

4. The method of claim 3, wherein the double stranded RNA has the nucleotide sequence set forth in SEQ ID No.4 and SEQ ID No. 5.

5. The method according to claim 1, wherein the nucleotide sequence of the ethanol dehydrogenase gene is as shown in SEQ ID No. 6.

6. The method according to claim 1, wherein the nucleotide sequence of the glyoxylate carboxylase gene is as set forth in SEQ ID No. 8.

7. The method of claim 1, wherein the tartrate semialdehyde reductase gene has the nucleotide sequence set forth in SEQ ID No. 10.

8. The method of claim 3, wherein said method is performed by constructing a T-DNA vector, and introducing the vector into a plant; the construction method of the T-DNA vector comprises the following steps: taking pCambia1300 binary vector containing glufosinate-ammonium-resistant bar gene as a basic vector, and respectively connecting a glycollic acid dehydrogenase gene expression frame, a glyoxylate carboxylase gene expression frame, a tartrate semialdehyde reductase gene expression frame and a bile acid sodium cotransporter gene RNAi expression frame.

9. The method of claim 8, wherein the promoters used for the expression cassette construction include the maize Ubi promoter sequence, the rice Actin promoter, and the 35S promoter of cauliflower mosaic virus CaMV; the terminator is the terminator ter.

10. The method of claim 8, wherein said plant is rice or soybean.