Application of maize ZmGOLS2 and ZmRAFS genes to improve grain yield of crops

Abstract

The application discloses corn ZmGOLS2 and ZmRAFS genes for improving crop grain yield, and also discloses the application of superimposed use of the corn ZmGOLS2 and ZmRAFS genes for improving corn harvest index. The application proves that the simultaneous overexpression of the corn ZmGOLS2 and ZmRAFS genes in corn changes the source-sink relationship, improves the harvest index, and improves the corn grain yield.

Description

Technical Field

[0001] This invention belongs to the field of bioengineering technology. Specifically, it relates to the application of two genes that increase the raffinose content of maize seeds, namely the superposition of the maize ZmGOLS2 and ZmRAFS genes, to improve maize grain yield. Background Technology

[0002] Inositol galactoside synthase (GOLS2) and raffinose synthase (RAFS) are key enzymes responsible for raffinose synthesis. In 2016, Gu reported that overexpression of maize ZmGOLS2 in Arabidopsis improved the drought, salt, and heat resistance of Arabidopsis seedlings. This paper only addressed the biomass of Arabidopsis plants under stress conditions (Gu et al., 2016, ZmGOLS2, a target of transcription factor ZmDREB2A, offers similar protection against abiotic stress as ZmDREB2A. PlantMolecular Biology. 90:157-170). In 2023, Liu reported that overexpression of the raffinose synthase (RAFS) gene in maize improved the biomass of maize plants under drought conditions (Liu et al., 2023, Raffinose positively regulates maize drought tolerance by reducing leaf transpiration. The Plant Journal 114:55-67). Summary of the Invention

[0003] The inventors discovered that simultaneously increasing the expression levels of two genes, ZmGOLS2 and ZmRAFS, in crops improved crop harvest index and grain yield.

[0004] Based on this, the present invention provides the application of the corn inositol galactoside synthase encoding gene ZmGOLS2 and the corn raffinose synthase encoding gene ZmRAFS for improving crop grain yield and / or harvest index. The nucleotide sequence of the corn inositol galactoside synthase encoding gene is shown in SEQ ID NO:1; the nucleotide sequence of the corn raffinose synthase encoding gene is shown in SEQ ID NO:3; or, the amino acid sequence of the corn inositol galactoside synthase is shown in SEQ ID NO:2; the amino acid sequence of the corn raffinose synthase is shown in SEQ ID NO:4.

[0005] Furthermore, this invention improves crop grain yield and / or harvest index by simultaneously increasing the expression levels of the gene encoding corn inositol galactoside synthase and the gene encoding corn raffinose synthase.

[0006] Furthermore, the gene encoding zeatinol galactosidase ZmGOLS2 and the gene encoding zeatinol raffinose synthase ZmRAFS are used to improve grain yield and / or harvest index under normal crop growth conditions.

[0007] This invention also provides a crop breeding method. The method includes high expression of the zeatinositol galactoside synthase encoding gene ZmGOLS2 and the zeatinositol raffinose synthase encoding gene ZmRAFS in the crop. Further, the method includes obtaining transgenic plants by high expression of the zeatinositol galactoside synthase encoding gene ZmGOLS2 and the zeatinositol raffinose synthase encoding gene ZmRAFS in maize, and then backcrossing the transgenic plants to obtain improved plants.

[0008] This invention demonstrates that simultaneous overexpression of the maize ZmGOLS2 and ZmRAFS genes in crops such as maize can improve the maize harvest index and increase crop grain yield without increasing total biomass. Attached Figure Description

[0009] Figure 1 This is a schematic diagram of the pTF-ZmGOLS2-ZmRAFS vector structure.

[0010] Figure 2 The results of the identification of ZmGOLS2 / ZmRAFS overexpression strains 31 (#1 and #2) were as follows: (A) PCR identification of the insertion of Bar, ZmGOLS2, and ZmRAFS genes; (B) Western blot identification of the protein expression levels of ZmGOLS2 and ZmRAFS; (C) HPLC identification of the content of inositol galactoside in leaves; (D) HPLC identification of the content of raffinose in leaves.

[0011] Figure 3 The process and results of backcrossing and breeding of transgenic hybrid 31 into commercial maize inbred line Chang 7-2, molecular identification, and preparation of hybrid Zhengdan 958 are as follows: (A) Schematic diagram of backcrossing and breeding of transgenic hybrid 31 into commercial maize inbred line Chang 7-2 and preparation of hybrid Zhengdan 958; (B) PCR identification of the insertion of Bar, ZmGOLS2, and ZmRAFS genes; (C) Western blot identification of the protein expression levels of ZmGOLS2 and ZmRAFS; (D) HPLC identification of the content of inositol galactoside in leaves; (E) HPLC identification of the content of raffinose in leaves.

[0012] Figure 4 The results show the biomass comparison between the improved variety (improved Zhengdan 958) and the control variety (Zhengdan 958) under irrigation, water control, and drought conditions.

[0013] Figure 5 This is a comparison of the harvest index of the improved variety and the control variety.

[0014] Figure 6 Comparison of grain yield between improved varieties and control varieties. Detailed Implementation

[0015] Unless otherwise specified, the scientific and technical terms used in this article are intended for understanding by those skilled in the art.

[0016] The normal growth conditions described in this invention refer to conditions such as irrigation, light, and temperature that are suitable for crop growth.

[0017] The biomass mentioned in this invention refers to the weight of the aboveground straw (including ears of grain) of the crop; the ratio of ear weight to total biomass is the harvest index; the ears of grain are threshed and the grain weight is taken as the grain yield.

[0018] The crops described in this invention include common economic crops such as corn, wheat, soybeans, rice, and sorghum.

[0019] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0020] Example 1: In this invention, the coding regions of the maize ZmGOLS2 and ZmRAFS genes were cloned using PCR, and expression frameworks of the two genes were constructed on the same vector.

[0021] The vector construction method is as follows: Using plasmid vector PHP69219 (Lowe, K. et al, Morphogenic Regulators Baby boom and Wuschel Improve Monocot Transformation. Plant Cell 2016, 28(9), 1998-2015.) DNA as a template, maize Ubiquitin promoter (Ubi) was amplified by PCR using primers (pF: 5'-TTCGAGCTCGGTACCAGCTTGCATGCCTGC-3' and pR: 5'-TGGGAGCCAGGATCCCGAGTTGAGCCATGGTCTAGAGTCGACCTGCAG-3'). The PCR product was then ligated into pCSGFPBT-ZmRAFS vector (Li, T, et al, Regulation of Seed Vigor by Manipulation of Raffinose Family Oligosaccharides in Maize and Arabidopsis thaliana. Molecular Biology). Plant, 2017, 10(12), 1540-1555.) Replace the 35S promoter to obtain the pCSGFPBT-Ubi:ZmRAFS vector.

[0022] Subsequently, using pCSGFPBT-Ubi:ZmRAFS vector DNA as a template, the Ubi-ZmRAFS-PolyA expression framework was amplified and ligated into the pTF101.1 vector by PCR using primers 5'-GACTCTAGAGGATCCGCATGCCTGCAGTGCAGCGT-3' and 5'-GGTACCCGGGGATCCGTCACTGGATTTTGGTTTTAG-3' (Zheng, Y, et al, Expression of the Arabidopsis thaliana Histone Gene AtHTA1 Enhances Rice Transformation Efficiency. Molecular Plant, 2009, 2(4), 832-837). The Ubi-ZmGOLS2-PolyA amplification (primers: 5'-GACTCTAGAGGATCCGCATGCCTGCAGTGCAGCGT-3' and 5'-GGTACCCGGGGATCCGTCACTGGATTTTGGTTTTAG-3') was ligated into the pTF101.1-Ubi-ZmRAFS vector using the same strategy, ultimately obtaining the pTF101.1-Ubi-ZmRAFS+Ubi-ZmGOLS2 vector, the structure of which is shown below. Figure 1 As shown.

[0023] Example 2: Genetic transformation of maize

[0024] Genetic transformation of maize was performed according to the published method (Ishida Y, Hiei Y, Komari T (2007) Agrobacterium-mediated transformation of maize. Nat Protoc 2(7):1614-1621.). The recipient material was the immature embryo of maize inbred line 31 (Zong31), and transgenic lines overexpressing ZmGOLS2 and ZmRAFS were obtained.

[0025] Example 3: Backcrossing, molecular identification, and hybridization of transgenic inbred line 31 to commercial maize inbred line Chang 7-2

[0026] See Figure 3As shown in (A), in this embodiment, seeds of the transgenic 31 inbred line overexpressing ZmGOLS2 and ZmRAFS and the maize Chang7-2 inbred line were simultaneously sown in the field. During the flowering period, the female and male ears were bagged separately, and pollen from the transgenic 31 was sprinkled onto the silks of the female ears of Chang7-2 before bagging. After maturity, the seeds were harvested. Subsequently, backcrossing was performed for 6 generations. Specifically, the harvested Chang7-2 and transgenic 31 materials were backcrossed into Chang7-2 for 6 consecutive generations. In the 6th generation, self-pollination was performed, and homozygous transgenic improved Chang7-2 was selected as the male parent and crossed with the non-transgenic Zheng58 as the female parent to obtain the improved Zhengdan 958 hybrid (improved). The unimproved Zhengdan 958 hybrid was obtained by crossing the female parent Zheng58 and the male parent Chang7-2 (control).

[0027] Example 4: Molecular identification of transgenic lines

[0028] First, the transgenic plants obtained in Example 2, the improved plants obtained in Example 3, and the unimproved plants were identified at the genomic level by PCR. Then, the expression levels of ZmGOLS2 and ZmRAFS proteins in the transgenic plants obtained in Example 2, the improved plants obtained in Example 3, and the unimproved plants were identified by Western blot.

[0029] The PCR amplification program was as follows: 95℃ pre-denaturation for 5 min; 95℃ denaturation for 30 s, 60℃ annealing for 30 s, 72℃ extension for 30 s, 35 cycles; 72℃ final extension for 8 min. Primers 5'-ATGGAACAAGGGCAGAAGATT-3' and 5'-GCACCATCGTCAACCACTAC-3' were used to identify the Bar gene, primers 5'-TTTTAGCCCTGCCTTCATAC-3' and 5'-TCCACGAACTCCCAGATG-3' were used to identify the ZmGOLS2 gene, and primers 5'-TTTTAGCCCTGCCTTCATAC-3' and 5'-CTTGAAACGGAAGATGCT-3' were used to identify the ZmRAFS gene.

[0030] The results are as follows Figure 2 A and Figure 3 As shown in B, transgenic maize lines that simultaneously overexpress the ZmGOLS2 and ZmRAFS genes were successfully obtained.

[0031] The Western blot procedure follows the existing technique described below (Gu, L., et al., ZmGOLS2, a target of transcription factor ZmDREB2A, offers similar protection against abiotic stress as ZmDREB2A. Plant Mol Biol, 2016, 90(1-2): p. 157-70.).

[0032] The ZmGOLS2 and ZmRAFS primary antibodies were prepared by our research group for immunizing rabbits at a dilution of 1:5000; the secondary antibody (goat anti-rabbit) was purchased from Kangwei Company and diluted 1:10000. Results are as follows... Figure 2 B and Figure 3 As shown in C, the obtained transgenic lines are able to express ZmGOLS2 and ZmRAFS proteins.

[0033] Example 5: Identification of free sugar components in leaves of transgenic lines

[0034] The inositol galactoside and raffinose content in the leaves of the transgenic plants obtained in Example 2, the improved plants obtained in Example 3, and the unimproved plants were determined. The specific methods are as follows: 0.5 g of leaves from each line were collected, ground into powder using liquid nitrogen, and then 5 mL of 80% ethanol (containing 200 μg / mL lactose) was added. Grinding continued until homogenized. The homogenate was poured into a 10 mL centrifuge tube, and 2 mL of 80% ethanol (containing 200 μg / mL lactose) was added to the mortar. The mortar was cleaned, and the samples were combined in the centrifuge tube. The sample was heated in an 80°C water bath for 30 min, centrifuged at 12000×g for 20 min at room temperature, and the supernatant was transferred to another clean centrifuge tube. The ethanol was evaporated by heating at 95°C. The sample was then frozen at -80°C for 24 hours and vacuum dried. The sample was resuspended in HPLC-grade double-distilled water and centrifuged at 12000×g for 20 min. The supernatant was frozen at -20°C and filtered through a 0.22 μm filter before determination. The HPLC detector was a Waters 2424 ELSD, and the column was a Waters Xbrige amino column. The sample loading volume was 5 μL.

[0035] The results are as follows Figure 2 C, D and Figure 3 As shown in D and E, the contents of inositol galactoside and raffinose were significantly increased in the obtained transgenic lines.

[0036] Example 6: Normal planting, water control, and drought treatment of transgenic maize in the field

[0037] The improved material (improved Zhengdan 958) and the control material (unimproved Zhengdan 958) were planted in the field with a row spacing of 55 cm and a plant spacing of 22 cm, and double seeds were sown. At the same time, each material was divided into three groups: one group was watered throughout the entire growth period (normal irrigation); one group was watered once before flowering and once after flowering for two weeks (water control); and one group was treated with drought and did not receive water (drought).

[0038] Harvest the above-ground corn stalks (including ears), and the weight collected is the biomass. Then, remove the ears and weigh them. The ratio of ear weight to total biomass is the harvest index. Thresh the ears and weigh the kernels; this is the yield. Results are as follows: Figure 4 , Figure 5 and Figure 6 As shown.

[0039] The above description is merely a preferred embodiment of the present invention, and the scope of protection of the present invention is not limited thereto. Any simple changes or equivalent substitutions of the technical solutions that can be obviously obtained by those skilled in the art within the scope of the technology disclosed in the present invention shall fall within the scope of protection of the present invention.

[0040] SEQ ID NO:1

[0041] ZmGOLS2 gene encoding zeatinositol galactoside synthase

[0042] ATGGCTCCCGAGCTGATGACAGCCAAGATGACCGCCAAAGCCGCGGCGGCGGCGGCGGCGGTGAAGCC

[0043] TGCCACGAGGGCGTACGTGACGTTCCTTGCGGGCGACGGCGACTACTGGAAGGGCGTGGTGGGGCTGG

[0044] CCAAAGGCCTGCGCAAGGTCCGCTCGGCCTACCCCCTGGTGGTGGCGGTGCTGCCCGACGTGCCCGAG

[0045] TCCCACCGCCGCATCCTCGTCTCGCAGGGCTGCGTCGTCCGCGAGATCGAGCCCGTGTACCCGCCTGA

[0046] GAACCAGACGCAGTTCGCCATGGCGTACTACGTCATCAACTACTCCAAGCTCCGCATCTGGGAGTTCG

[0047] TGGAGTACGAGAGGATGGTGTACCTGGACGCGGACATCCAGGTGTTCGAGAACATCGACGGCCTGTTC

[0048] GAGCTCGAGAAGGGGTACTTCTACGCGGTGATGGACTGCTTCTGCGAGAAGACGTGGAGCCACACCCC

[0049] GCAGTACAGGATCGGCTACTGCCAGCAGTGCCCGGACAAGGTGGCGTGGCCAGCGGCGACCGCCGAGC

[0050] TGGGCCCTCCGCCGTCGCTCTACTTCAACGCCGGCATGTTCGTGCACGAGCCCAGCGTGGCCACCGCC

[0051] AAGGCGCTCCTCGACACCCTCCGCGTGACTCCGCCCACCCCCTTCGCAGAGCAGGACTTCCTGAACAT

[0052] GTTCTTCAGGGATCAGTACAGGCCGATCCCCAACGTGTACAACCTCGTGCTGGCCATGCTCTGGAGGC

[0053] ACCCCGAGAACGTGCAGCTGGAGAAGGTGAAGGTCGTGCACTACTGCGCGGCGGGGTCCAAGCCGTGG

[0054] AGGTTCACGGGCAAGGAGGCGAACATGGACAGGGAGGACATCAACGCCCTGGTGAACAAGTGGTGGGA

[0055] CATCTACAACGACGAGACGCTGGACTTGAAGGGCCTGCCCTCGCTCTCGCCCGACGACGACGACGAGG

[0056] TGGAGGCGGTGGCCAAGAAGCCGCTTCGCGCGGCCCTGGCGGAGGCCGGCACGGTCAAGTACGTCACC

[0057] GCGCCCTCGGCCGCGTGA

[0058] SEQ ID NO:2

[0059] Maize galactinol synthase

[0060] MAPELMTAKMTAKAAAAAAAVKPATRAYVTFLAGDGDYWKGVVGLAKGLRKVRSAYPLVVAVLPDVPE

[0061] SHRRILVSQGCVVREIEPVYPPENQTQFAMAYYVINYSKLRIWEFVEYERMVYLDADIQVFENIDGLF

[0062] ELEKGYFYAVMDCFCEKTWSHTPQYRIGYCQQCPDKVAWPAATAELGPPPSLYFNAGMFVHEPSVATA

[0063] KALLDTLRVTPPTPFAEQDFLNMFFRDQYRPIPNVYNLVLAMLWRHPENVQLEKVKVVHYCAAGSKPW

[0064] RFTGKEANMDREDINALVNKWWDIYNDETLDLKGLPSLSPDDDDEVEAVAKKPLRAALAEAGTVKYVT

[0065] APSAA

[0066] SEQ ID NO:3

[0067] Maize raffinose synthase encoding gene ZmRAFS

[0068] ATGGCTCCCAACCTCAGCAAGAAGACGCCTGCTGGCCTCCTCGGCGACGAGGTGGCCCCGGTGGAGGG

[0069] ACTCAAGCCGTCGCGGTTCACCCTTAAGGGCAAGGACCTGGCCGTGGACGGGCACCCGGTCCTGCTGG

[0070] ACGTGCCGGCCAACATCCGTCTCACCCCGGCGTCGACGCTCGTGCCCGCCGCGGACGTCCCCGCAGCG

[0071] GGCGGCGGCAGCTTCCTCGGCTTCGACGCGGCGGCGGCCGAGAGCCGGCACGTGGTGCCCGTCGGAAA

[0072] GCTCCGTGACATTCGGTTCATGAGCATCTTCCGTTTCAAGGTGTGGTGGACGACGCACTGGGTGGGGG

[0073] ACAGCGGCAGGGACGTGGAGAACGAGACGCAGATGATGGTGCTCGACCGCTCCGCCGGCGAGCCCGGC

[0074] GGCGGCGGCCGACCCTACGTGCTGCTGCTCCCCATCATCGAGGGCTCGTTCCGGGCCTGCCTCGAGGC

[0075] CGGGAAGGTGGAAGACTACGTGGACCTGTGCGTGGAGAGCGGGTCGTCGGCGGTGCGCGGCGCCGCGT

[0076] TCCGGAGCTCGCTGTACCTGCACGCGGGCGACGACCCGTTCGAGCTCGTCGCGGACGCCGTCAGGGTG

[0077] GTCCGTGCGCACCTGGGCACGTTCCGGACCATGGAGGAGAAGACGCCGCCGCCGATCGTGGACAAGTT

[0078] CGGGTGGTGCACGTGGGACGCCTTCTACCTCAAGGTGCACCCGGAGGGCGTGTGGGAGGGCGTGCGCC

[0079] GCCTGGCGGAGGGCGGCTGCCCGCCGGGGCTGGTGCTCATCGACGACGGCTGGCAGTCCATCTGCCAC

[0080] GACGAGGACGACCCGAACAGCGGCGAGGAGGGCATGAACCGCACCTCCGCCGGCGAGCAGATGCCCTG

[0081] CCGCCTCATCAAGTTCCAGGAGAACCACAAGTTCAGGGAGTACAAGCAGGGCGGGATGGGCGCGTTCG

[0082] TGCGGGAGATGAAGGCGGCGTTCCCCACCGTGGAGCAGGTGTACGTGTGGCACGCGCTGTGCGGGTAC

[0083] TGGGGCGGCCTCCGCCCCGGCGCGCCCGGCCTGCCGCCCGCCAAGGTGGTGGCGCCCAAGCTCTCCCC

[0084] CGGCCTGCAGCGCACCATGGAGGACCTCGCCGTCGACAAGATCGTCAACAACGGTGTCGGCCTCGTCG

[0085] ACCCCAAGCGCGCGCACGAGCTCTACGATGGTTTGCACTCCCACCTCCAGGCCTCCGGCATCGACGGC

[0086] GTCAAGGTCGACGTCATTCACTTGCTGGAGATGCTGTGCGAGGAGTACGGCGGCCGTGTCGAGCTGGC

[0087] CAAGGCCTACTTCGCCGGGCTGACGGCGTCGGTGCGGCGGCACTTCGGCGGCAACGGCGTGATCGCGA

[0088] GCATGGAGCACTGCAACGACTTCATGCTGCTGGGCACGGAGGCGGTGGCGCTGGGCCGCGTGGGCGAC

[0089] GACTTCTGGTGCACGGACCCCTCCGGCGACCCCAACGGCACCTTCTGGCTGCAGGGGTGCCACATGGT

[0090] GCACTGCGCCTACAACTCGCTGTGGATGGGCAACTTCATCCACCCGGACTGGGACATGTTCCAGTCCA

[0091] CGCACCCCTGCGCCGCCTTCCACGCCGCGTCCCGCGCCATCTCCGGCGGGCCCATCTACGTCAGCGAC

[0092] TCGGTGGGGCAGCACGACTTCGCGCTGCTCCGCCGCCTGGCGCTCCCCGACGGCACCGTCCTCCGGTG

[0093] CGAGGGCCACGCGCTGCCCACGCGCGACTGCCTCTTCGCCGACCCGCTCCACGACGGCCGGACCGTGC

[0094] TCAAGATCTGGAACGTGAACCGCTTCGCCGGCGTCGTCGGCGCCTTCAACTGCCAGGGCGGCGGGTGG

[0095] AGCCCCGAGGCGCGGCGGAACAAGTGCTTCTCGGAGTTCTCCGTGCCCCTGGCCGCGCGCGCCTCGCC

[0096] GTCCGACGTCGAATGGAAGAGCGGCAAAGCGGGACCAGGCGTCAGCGTCAAGGGCGTCTCCCAGTTCG

[0097] CCGTGTACGCGGTCGAGGCCAGGACGCTGCAGCTGCTGCGCCCCGACGAGGGCGTCGACCTCACGCTG

[0098] CAGCCCTTCACCTACGAGCTCTTCGTCGTCGCCCCCGTGCGCGTCATCTCGCACGAGCGGGCCATCAA

[0099] GTTCGCGCCCATCGGACTCGCCAACATGCTCAACACCGCCGGCGCCGTGCAGGCGTTCGAGGCCAAGA

[0100] AAGATGCTAGCGGCGTCACGGCAGAGGTGTTCGTGAAGGGCGCAGGGGAGCTGGTGGCGTACTCGTCG

[0101] GCGACGCCCAGGCTCTGCAAGGTGAACGGCGACGAGGCCGAGTTCACGTACAAGGACGGCGTGGTCAC

[0102] CGTCGACGTGCCGTGGTCGGGGTCGTCGTCGAAGCTGTGTCGCGTCCAGTACGTCTACTGA

[0103] SEQ ID NO:4

[0104] Corn raffinose synthase

[0105] MAPNLSKKTPAGLLGDEVAPVEGLKPSRFTLKGKDLAVDGHPVLLDVPANIRLTPASTLVPAADVPAA

[0106] GGGSFLGFDAAAAESRHVVPVGKLRDIRFMSIFRFKVWWTTHWVGDSGRDVENETQMMVLDRSAGEPG

[0107] GGGRPYVLLLPIIEGSFRACLEAGKVEDYVDLCVESGSSAVRGAAFRSSLYLHAGDDPFELVADAVRV

[0108] VRAHLGTFRTMEEKTPPPIVDKFGWCTWDAFYLKVHPEGVWEGVRRLAEGGCPPGLVLIDDGWQSICH

[0109] DEDDPNSGEEGMNRTSAGEQMPCRLIKFQENHKFREYKQGGMGAFVREMKAAFPTVEQVYVWHALCGY

[0110] WGGLRPGAPGLPPAKVVAPKLSPGLQRTMEDLAVDKIVNNGVGLVDPKRAHELYDGLHSHLQASGIDG

[0111] VKVDVIHLLEMLCEEYGGRVELAKAYFAGLTASVRRHFGGNGVIASMEHCNDFMLLGTEAVALGRVGD

[0112] DFWCTDPSGDPNGTFWLQGCHMVHCAYNSLWMGNFIHPDWDMFQSTHPCAAFHAASRAISGGPIYVSD

[0113] SVGQHDFALLRRLALPDGTVLRCEGHALPTRDCLFADPLHDGRTVLKIWNVNRFAGVVGAFNCQGGGW

[0114] SPEARRNKCFSEFSVPLAARASPSDVEWKSGKAGPGVSVKGVSQFAVYAVEARTLQLLRPDEGVDLTL

[0115] QFPTYELFVVAPVRVISHERAIKFAPIGLANMLNTAGAVQAFEAKKDASGVTAEVFVKGAGELVAYSS

[0116] ATPRLCKVNGDEAEFTYKDGVVTVDVPWSGSSSKLCRVQYVY

Claims

1. Overexpression of the gene encoding zeatinositol galactoside synthase ZmGOLS2 and the gene encoding maize raffinose synthase ZmRAFS For applications to increase corn kernel yield, the nucleotide sequence of the corn inositol galactoside synthase encoding gene is shown in SEQ ID NO:1; the nucleotide sequence of the corn raffinose synthase encoding gene is shown in SEQ ID NO:

3.

2. Overexpression of the gene encoding zeatinositol galactoside synthase ZmGOLS2 and the gene encoding maize raffinose synthase ZmRAFS For applications to increase corn kernel yield, the amino acid sequence of the corn inositol galactoside synthase is shown in SEQ ID NO:2; the amino acid sequence of the corn raffinose synthase is shown in SEQ ID NO:

4.

3. The application according to claim 1 or 2, characterized in that, Increase maize grain yield by simultaneously increasing the expression levels of the gene encoding maize inositol galactoside synthase and the gene encoding maize raffinose synthase.

4. Overexpression of the gene encoding zeatinositol galactoside synthase ZmGOLS2 and the gene encoding maize raffinose synthase ZmRAFS For applications aimed at improving the corn harvest index, the nucleotide sequence of the corn inositol galactoside synthase encoding gene is shown in SEQ ID NO:1; the nucleotide sequence of the corn raffinose synthase encoding gene is shown in SEQ ID NO:

3.

5. Overexpression of the gene encoding zeatinositol galactoside synthase ZmGOLS2 and the gene encoding maize raffinose synthase ZmRAFS For applications aimed at improving the corn harvest index, the amino acid sequence of the corn inositol galactoside synthase is shown in SEQ ID NO:2; the amino acid sequence of the corn raffinose synthase is shown in SEQ ID NO:

4.

6. The application according to claim 4 or 5, characterized in that, The maize harvest index can be improved by simultaneously increasing the expression levels of the maize inositol galactoside synthase encoding gene and the maize raffinose synthase encoding gene.

7. The application according to claim 4 or 5, characterized in that, Gene encoding zeatinol galactoside synthase ZmGOLS2 and the gene encoding maize raffinose synthase ZmRAFS Applications to improve the harvesting of maize under normal growing conditions.

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