Application of AhMAGL1b gene in regulation and control of plant leaf shape

Abstract

The invention discloses application of an AhMAGL1b gene in regulation and control of plant leaf shapes, and belongs to the field of plant leaf shape improvement. The nucleotide sequence of the AhMAGL1b gene is as shown in SEQ ID NO. 1 (sequence identifier number 1). According to the invention, an AhMAGL1b gene is obtained through cloning, and the AhMAGL1b gene is used for coding an AhMAGL1b protein. Through verification, the AhMAGL1b gene is overexpressed in arabidopsis thaliana, so that leaves of the arabidopsis thaliana can be curled; and furthermore, the curling of the overexpressed plant leaf is caused by polarity change. Meanwhile, overexpression of the AhMAGL1b changes the oil content, the sugar content and the fatty acid component of plant leaves and seeds. In conclusion, new gene resources and theoretical guidance are provided for peanut leaf improvement and molecular design breeding, ideal characters of peanut leaves are favorably molded, and the method has important application value.

Description

Technical Field

[0001] The present invention relates to the field of plant leaf shape improvement, and in particular to the application of the AhMAGL1b gene in regulating plant leaf shape. Background Art

[0002] The material basis of economic crop yields primarily comes from photosynthesis. The photosynthetic productivity of a crop population is primarily determined by factors such as leaf area, the duration of a specific leaf area, and the net assimilation rate per unit leaf area. Leaves are the primary photosynthetic organ of a crop, and leaf shape traits, such as leaf length, leaf width, leaf length-to-width ratio (leaf shape index), leaf area, and leaf angle, directly influence the canopy structure, leaf area index, and light energy use efficiency of a crop population. In rice breeding, improving plant and leaf morphology to achieve an ideal plant type in hybrid rice is considered the best approach to achieving super-high yields. In wheat, an appropriate flag leaf size is a key factor in high yields. During the reproductive growth period, a leaf area index of 3.5 to 4.0 is essential for achieving maximum yield potential in soybeans. These breeding practices demonstrate that crop leaf morphology is closely related to yield, making improving leaf morphology an effective means of increasing crop yields.

[0003] Peanut (Arachis hypogaea) is an important oilseed and cash crop, and increasing peanut yield per unit area has always been a key goal of peanut breeding. Despite significant progress in peanut breeding, increasing peanut yield per unit area continues to be increasingly challenging. Over the past five years, the average annual growth rate of peanut yield has dropped to less than 0.5%. Relying on traditional breeding methods to increase yields has become increasingly difficult, and peanut yield improvement has reached a bottleneck. Peanut leaf morphology is diverse, including 15 types, including elliptical, subcircular, and lanceolate. In recent years, EMS mutagenesis has created material with curly leaves. This rich resource of leaf shapes provides a foundation for improving peanut leaf shape. However, the discovery of genes regulating peanut leaf shape is currently very limited, hindering the improvement of peanut leaf shape. Summary of the Invention

[0004] The purpose of the present invention is to provide an application of the AhMAGL1b gene in regulating plant leaf shape to solve the problems existing in the above-mentioned prior art. The AhMAGL1b gene positively regulates plant leaf shape.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides the use of the AhMAGL1b gene in any of the following:

[0007] (1) Application in regulating plant leaf shape;

[0008] (2) Application in the preparation of products for regulating plant leaf shape;

[0009] (3) Application in plant leaf shape improvement and breeding;

[0010] (4) Application in cultivating plants with curled leaves;

[0011] The nucleotide sequence of the AhMAGL1b gene is shown in SEQ ID NO.1.

[0012] The present invention also provides the use of the AhMAGL1b protein encoded by the AhMAGL1b gene in any of the following:

[0013] (1) Application in regulating plant leaf shape;

[0014] (2) Application in the preparation of products for regulating plant leaf shape;

[0015] (3) Application in plant leaf shape improvement and breeding;

[0016] (4) Application in cultivating plants with curled leaves;

[0017] The nucleotide sequence of the AhMAGL1b gene is shown in SEQ ID NO.1; the amino acid sequence of the AhMAGL1b protein is shown in SEQ ID NO.2.

[0018] The present invention also provides use of a recombinant vector containing the AhMAGL1b gene in any of the following:

[0019] (1) Application in regulating plant leaf shape;

[0020] (2) Application in the preparation of products for regulating plant leaf shape;

[0021] (3) Application in plant leaf shape improvement and breeding;

[0022] (4) Application in cultivating plants with curled leaves;

[0023] The nucleotide sequence of the AhMAGL1b gene is shown in SEQ ID NO.1.

[0024] The present invention also provides the use of a host bacterium containing the recombinant vector in any of the following:

[0025] (1) Application in regulating plant leaf shape;

[0026] (2) Application in the preparation of products for regulating plant leaf shape;

[0027] (3) Application in plant leaf shape improvement and breeding;

[0028] (4) Application in cultivating plants with curled leaves.

[0029] Optionally, the regulation refers to changing the shape of plant leaves into curled ones.

[0030] Optionally, the plant comprises Arabidopsis thaliana or peanut.

[0031] The present invention also provides a method for regulating plant leaf shape, comprising the steps of overexpressing the AhMAGL1b gene in the plant to increase the expression level of the AhMAGL1b gene to regulate the plant leaf shape;

[0032] The nucleotide sequence of the AhMAGL1b gene is shown in SEQ ID No. 1.

[0033] Optionally, the plant comprises Arabidopsis thaliana or peanut.

[0034] The present invention also provides a method for cultivating plants with curled leaves, comprising the steps of overexpressing the AhMAGL1b gene in the plant, increasing the expression level of the AhMAGL1b gene, and obtaining the plant with curled leaves;

[0035] The nucleotide sequence of the AhMAGL1b gene is shown in SEQ ID No. 1.

[0036] Optionally, the plant comprises Arabidopsis thaliana or peanut.

[0037] The present invention discloses the following technical effects:

[0038] The present invention cloned the AhMAGL1b gene, encoding the AhMAGL1b protein, from the cultivated peanut variety "Yuhua 9326." Overexpression of the AhMAGL1b gene in Arabidopsis thaliana has been shown to cause leaf curling. The authors further discovered that leaf curling in overexpressing plants is caused by changes in polarity. Furthermore, the present invention found that overexpression of AhMAGL1b altered the oil, sugar, and fatty acid composition of leaves and seeds.

[0039] In summary, the present invention provides new genetic resources and theoretical guidance for peanut leaf improvement and molecular design breeding, helps to shape the ideal traits of peanut leaves, and has important application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0041] Figure 1Identification of AhMAGL1b in transgenic Arabidopsis overexpressing AhMAGL1b; A: Schematic diagram of the structure of the AhMAGL1b-containing vector 1300-pUBQ10-MCS-G10evo; B: Expression level of AhMAGL1b in transgenic Arabidopsis;

[0042] Figure 2 Phenotypic analysis of AhMAGL1b overexpressing transgenic lines; AD: Phenotypes of wild type and overexpressing transgenic lines (OE1, OE4, OE7) 20, 30, 50, and 70 days after sowing, scale bar = 4 cm; E: Plant height 30-83 days after sowing; F: Number of siliques per plant 30-83 days after sowing; G: Aboveground fresh weight 30 days after sowing; H: 1000-grain weight at seed maturity;

[0043] Figure 3 The morphological basis of AhMAGL1b overexpressing transgenic leaves; AC: Leaf phenotype, SPAD, and net photosynthetic rate (Pn) of the wild type and overexpressing transgenic lines (OE1, OE4, and OE7) 30 days after sowing, scale bar = 2 cm; D: Paraffin section of leaves, scale bar = 200 μm; E: Paraffin section of leaf veins, scale bar = 20 μm; F: Scanning electron microscopy image;

[0044] Figure 4 Analysis of oil content and fatty acid composition in leaves and seeds of transgenic Arabidopsis thaliana overexpressing AhMAGL1b; A: leaf oil content; B: leaf fatty acid composition; C: seed oil content; D: seed fatty acid composition. DETAILED DESCRIPTION

[0045] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0046] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. The intermediate value within any stated value or stated range, and each smaller range between any other stated value or intermediate value within the stated range, is also encompassed within the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0047] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0048] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be exemplary only.

[0049] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0050] Example 1 AhMAGL1b gene cloning and overexpression vector construction

[0051] 1. Experimental Materials

[0052] The cultivated peanut variety is "Yuhua 9326".

[0053] 2. Experimental methods

[0054] 2.1 Extraction of peanut total RNA and synthesis of first-strand cDNA

[0055] Using peanut leaves as the material, RNA was extracted using TransZol Up Plus RNA Kit (ER501, Transgene). The specific steps are as shown in the instructions. After RNA extraction, the concentration was measured and detected using 1% agarose gel electrophoresis. First Strand cDNA Synthesis SuperMix for qPCR (gDNA digester plus) (11141ES60, Yisheng) reverse transcription kit was used to synthesize the first strand cDNA. 1 μg RNA and 3 μL 5× gDNA digester Mix were added to an RNase-free centrifuge tube. RNase-free ddH2O was added to make up to 15 μL and the mixture was reacted at 42°C for 2 min. After the reaction was completed, 5 μL of III SuperMix plus, gently pipette to mix. The reaction program is: 25℃ for 5 min, 55℃ for 15 min, and 85℃ for 5 min.

[0056] 2.2 Cloning of the AhMAGL1b gene

[0057] The CDS sequence of AhMAGL1b was amplified by PCR using peanut cDNA as a template using a high-fidelity DNA polymerase. Primers used were FP: 5'-AACTGCAGATGGCAGCAGAGAAAGCGGA-3' (SEQ ID NO. 3) and RP: 5'-TCCCCCGGGTTATTTATTTCCATACCTAT-3' (SEQ ID NO. 4). The reaction system consisted of 1 μL of cDNA, 1 μL each of PrimerFP and Primer RP, 4 μL of 10 mM dNTPs, 8 μL of 5× buffer, 1 μL of Takara high-fidelity enzyme Primer Star, and ddH2O to a final volume of 40 μL. The target fragment was amplified according to the protocol. The nucleotide sequence of the AhMAGL1b gene CDS and the amino acid sequence of the AhMAGL1b protein are shown in SEQ ID NOs. 1-2.

[0058] SEQ ID NO.1 (AhMAGL1b gene CDS nucleotide sequence):

[0059] atggcagcagagaaagcggaggtaccccctaacttctggggccacatgcccgaggaggagtactacacctcccagggggtgcgaaacaccaaatcattcttcgacacgccacacggcaaaatcttcacgcagagcttcctcccattggaccttcaacctaacgaagtcaaagccaccgtcttcatgacccacggctacggctccgacaccggttggctcttccagaagatctgcattaacttcgccacctggggctacgccgtcttcgccgccgatctcctaggccacggccgctccgatggcctccgctgctacctcggcgacatggacaaagtggcatccgcctcgctctccttcttcctccacgtccgccgcagtgagccgtacaaggatctcccggcgttcctcttcggtgagtccatgggtggtctcgcaaccctgcttatgtactttaaatcagaacccgatacgtggacgggcctgatgttctctgcaccgcttttcgtgatccccgaggatatgaagcccagcaggcttcatctgttcgcgtatgggcttcttttcggatgggccgacacgtgggccgctatgcctgacaacaagatggttgggaaggcaatcagggatcccgagaagctgaagatcatagcgtcaaacccgaggaggtacacgggtccacctagggttgggaccatgagggagctccttagggttacccagtacgtgcaggataacttctccaaggtaacggcgccgtttctcacagtgcatggcacctctgatggcgttacgtgtccgtcttcatccaagttgctgtatgagaaggcttcgagtgaggacaagtcgttgaagctgtacgaagggatgtaccattctttgattcaaggggagcctgacgagagtgctaaccttgtattgagtgatatgagggaatggattgatcagagggttcataggtatggaaataaataa;

[0060] SEQ ID NO.2 (AhMAGL1b protein amino acid sequence):

[0061] MAAEKAEVPPNFWGHMPEEEYYTSQGVRNTKSFFDTPHGKIFTQSFLPLDLQPNEVKATVFMTHGYGSDTGWLFQKICINFATWGYAVFAADLLGHGRSDGLRCYLGDMDKVASASLSFFLHVRRSEPYKDLPAFLFGESMGGLATLLMYFKSEPDTWTGLM FSAPLFVIPEDMKPSRLHLFAYGLLFGWADTWAAMPDNKMVGKAIRDPEKLKIIIASNPRRYTGPPRVGTMRELLRVTQYVQDNFSKVTAPFLTVHGTSDGVTCPSSSKLLYEKASSEDKSLKLYEGMYHSLIQGEPDESANLVLSDMREWIDQRVHRYGNK.

[0062] 2.3 Overexpression vector construction

[0063] The 1300-35S-g10evo vector (kindly donated by Professor Zhicheng Shen of Zhejiang University, and disclosed in the article "Transfer of g10evo gene to glyphosate-resistant soybean" by Tan Miaomiao) was modified. The pUBQ10 and MCS from the pSY06 vector (kindly donated by Professor Shengjun Feng of Zhejiang Agricultural and Forestry University, and disclosed in the article "Construction of mimicry156 vector for silencing miR156 expression and transformation of tobacco" by Feng Shengjun et al.) were cleaved using restriction endonucleases HindIII and EcoRI. The 1300-35S-g10evo vector was then digested with the same endonucleases, and the pUBQ10 and MCS gene fragments were ligated into the 1300-35S-g10evo vector using T4 ligase. The pUBQ10 (long) promoter of Arabidopsis thaliana (SEQ ID NO.5), replaced the pUBQ10 promoter on the vector, and sequenced to obtain the 1300-pUBQ10-MCS-G10evo overexpression vector. The AhMAGL1b CDS sequence was ligated between the PstI and SmaI restriction sites of the 1300-pUBQ10-MCS-G10evo overexpression vector to obtain the pUBQ10-AhMAGL1b-G10evo vector ( Figure 1 Middle A).

[0064] SEQ ID NO.5 (pUBQ10 (long) promoter sequence of Arabidopsis thaliana):

[0065]

[0066] Example 2 Genetic transformation and positive identification of Arabidopsis thaliana

[0067] 1. Genetic transformation of Arabidopsis thaliana

[0068] The pUBQ10-AhMAGL1b-G10evo plasmid was transformed into Agrobacterium tumefaciens GV3101 and infected with Arabidopsis thaliana by the inflorescence dip method. T1 transgenic Arabidopsis plants, screened for glyphosate and identified by PCR, were cultured normally. Each individual plant was considered a transgenic line, and the harvested seeds were T2 transgenic Arabidopsis seeds. The number and ratio of resistant to non-resistant plants were statistically analyzed. The chi-square test was used to determine whether the phenotypic distribution of the screened T2 lines conformed to the theoretical ratio of 3:1 according to Mendel's law of genetic segregation. Homozygous T3 transgenic lines were obtained. Real-time fluorescence quantitative PCR was used to determine the expression of the exogenous AhMAGL1b gene in the screened T3 pUBQ10:AhMAGL1b lines.

[0069] 2. Real-time fluorescence quantitative PCR analysis of AhMAGL1b gene

[0070] Total RNA was extracted from two-week-old wild-type and transgenic Arabidopsis leaves using the TransZol Up Plus RNA Kit (ER501, Transgene). cDNA was synthesized using the 1stStrand cDNASynthesis SuperMix. Real-time fluorescence quantitative PCR analysis was performed using Hieff qPCR SYBR green Master Mix (11201ES03 / 08 / 60, Yisheng). The primers for AhMAGL1b and AtActin2 were 5'-GACCTTGTTGTTCAGGATTG-3' (SEQ ID NO. 6) and 5'-AGTGAGGATCTGAGGAATTG-3' (SEQ ID NO. 7), and 5'-GTCGTACAACCGGTATTGTGCT-3' (SEQ ID NO. 8) and 5'-TGTCTCTTACAATTTCCCGCTCT-3' (SEQ ID NO. 9), respectively. The following reaction mixture was prepared in a Hard-Shell Low-Profile Skirted 96-Well PCR reaction plate: 10 μL qPCR SYBR Green Master Mix, 0.4 μL 10 μM Primer FP, 0.4 μL 10 μM Primer RP, 9.2 μL cDNA template (diluted 5-fold). Place the PCR reaction plate in a fluorescent quantitative PCR instrument for amplification (two-step method). Set the reaction program as shown in Table 1 below.

[0071] The results are as follows Figure 1 As shown in Figure B, all homozygous transgenic lines overexpressed AhMAGL1b compared to WT. Three lines with high AhMAGL1b expression (OE1, OE4, and OE7) were selected for subsequent experiments.

[0072] Table 1 Reaction procedure

[0073]

[0074] 3. Growth and development analysis of AhMAGL1b overexpressing plants and determination of corresponding indicators

[0075] 3.1 Phenotypic observation and indicator determination

[0076] Phenotypes of the T3 transgenic lines and wild-type plants during the vegetative and reproductive stages were observed, including leaf size, plant height, flowering time, and number of pods per plant. Growth rate and days to maturity were calculated. 30 days after sowing, the SPAD of the third leaf below the growing point was measured using a chlorophyll meter (SPAD-502Plus, Japan). Plant height (cm) was recorded 50 days after sowing. Thousand-grain weight was measured after drying at maturity.

[0077] 3.2 Net photosynthetic rate determination

[0078] Plants with good growth were selected and tested at the beginning of flowering. Six seedlings were tested for each strain, and each test was repeated at least three times. The LI-6800 portable photosynthetic system (LI-COR, USA) was used for the test, along with a CO2 automatic injection system and a red and blue light source leaf chamber 6800-02. The CO2 concentration during the test was 400 μmol·mol -1 , the temperature was 28 °C, and the light intensity was set to 1000 μmol m -2 ·s -1 .

[0079] 3.3 Paraffin sections

[0080] Take the transgenic and wild-type leaves at a distance of about 0.5 cm from the edge and 0.25 cm 2Add the leaves to the pre-cooled FAA fixative, vacuumize to sink the leaves to the bottom, and fix the sample for 2 days. Transfer the sample to a vial for paraffin sectioning. First, use an ethanol solution of the corresponding concentration to wash away the formaldehyde and acetic acid in the fixative. Fix with 70% FAA, first wash away the fixative with 70% ethanol, changing three times for 30 minutes each time. Then, soak in 70%, 85%, 95%, and anhydrous ethanol for 1 hour for gradient dehydration. Xylene ethanol solution 20%, 40%, 60%, and 85%, and xylene soak for 3 hours for gradient transparency. Gradually add paraffin and gradually increase the temperature to completely replace the xylene with liquid paraffin. Embed the sample and trim the wax block. Use an automatic microtome to cut the sample to a thickness of 20μm and then spread it. Finally, dewax and rehydrate, stain with 0.5% toluidine blue solution, and observe and photograph under a microscope.

[0081] 3.4 Scanning electron microscopy

[0082] Transgenic and wild-type leaves were fixed in FAA fixative for at least 2 hours. Dehydration was then performed using a gradient of 70%, 80%, 95%, and anhydrous ethanol, with each step lasting 2 hours. The volume of the dehydrating agent was three times greater than the material. Critical point drying was performed. Finally, the samples were fixed to a copper table with conductive adhesive and adjusted to a suitable position for observation. The samples were then sprayed with gold and observed under a scanning electron microscope.

[0083] 4. Results

[0084] 4.1 Overexpression of AhMAGL1b causes Arabidopsis leaves to curl and shrink, growth rate to slow, and seed set rate to decrease

[0085] Under normal and identical culture conditions, the growth and development of the transgenic lines overexpressing AhMAGL1b were delayed and the plants were short ( Figure 2 ). 20 days after sowing, the transgenic plants showed that their leaves were obviously smaller than those of the wild type ( Figure 2 Middle A), the leaves began to curl about 30 days after sowing ( Figure 2 B), and the fresh weight of leaves was significantly lower than that of wild type ( Figure 2 Middle G). 50 days after sowing, the height of transgenic plants was significantly lower than that of wild type ( Figure 2 70 days after sowing, the seeds of wild-type plants were basically mature, while the transgenic seeds were only partially mature ( Figure 2 Agronomic trait analysis showed that the bolting time of the overexpression lines was generally delayed by 3-5 days compared with the wild type ( Figure 2 The transgenic plants were significantly taller than the wild type, the number of pods per plant was significantly reduced, and the seeds took longer to mature ( Figure 2 Middle F), except for the OE7 line, there was no significant change in the 1000-grain weight of seeds at maturity ( Figure 2These results indicate that overexpression of AhMAGL1b causes Arabidopsis leaves to curl and shrink, leading to reduced plant height and seed setting rate.

[0086] 4.2 Overexpression of AhMAGL1b causes partial changes in leaf polarity

[0087] The most obvious phenotypic difference between AhMAGL1b overexpressing transgenic plants and WT plants is the change in leaf curling, which becomes more and more obvious as the leaves mature ( Figure 3 In the middle (A), OE1 is the most curly. In terms of overall plant shape, OE strains are more compact, while WT leaves are slender and appear loose. In the late stage of plant growth, OE will form a nearly spherical plant shape, while WT will form an elliptical cylindrical plant shape. The chlorophyll content of OE increases ( Figure 3 Middle B), the net photosynthetic rate decreased ( Figure 3 To determine whether OE curling is due to changes in leaf polarity, the present invention observed the morphological characteristics of mesophyll cells and epidermal cells of OE and WT using tissue sections and scanning electron microscopy. In wild-type leaves, the adaxial surface is densely packed with palisade cells, while the abaxial surface is loosely packed with spongy cells. However, cross-sections of OE1 leaves clearly show a greater degree of curling ( Figure 3 Middle D), the polarity of the mesophyll cells of the entire leaf is partially lost, there is no obvious distinction between the palisade tissue and the spongy tissue, and the leaf thickness becomes smaller, and the xylem is arranged more densely ( Figure 3 Scanning electron microscopy showed that the leaf epidermal cells of the wild type were smooth and resembled puzzle pieces, while the leaf epidermal cells of OE1 had unclear boundaries, smaller cells, and increased numbers ( Figure 3 Middle F).

[0088] The researchers also found that overexpressing AhMAGL1b altered the oil, sugar, and fatty acid composition of leaves and seeds. Lipids were extracted from leaves of transgenic lines grown for four weeks, methylated, and analyzed by gas chromatography. Compared to the wild type, overexpressing AhMAGL1b significantly increased the total fatty acid content in Arabidopsis leaves. The fatty acid content in leaves of OE1, OE4, and OE7 increased by 37.60%, 34.28%, and 27.67%, respectively. Figure 4 Overexpression of AhMAGL1b altered the fatty acid composition in leaves, with a significant increase in C16:0 content and a significant decrease in C16:3 and C18:3 contents ( Figure 4The content of saturated fatty acid components in the leaves of the transgenic lines was significantly increased, while the content of unsaturated fatty acid components was significantly decreased. The oil content and fatty acid composition of T4 seeds were analyzed. The results showed that compared with wild-type plants, overexpression of AhMAGL1b significantly reduced the oil content of Arabidopsis seeds. The oil content of OE1, OE4 and OE7 seeds decreased by 26.52%, 29.13% and 24.22%, respectively. Figure 4 Among them, the proportion of C18:3 and C20:1 decreased significantly; the proportion of C18:2 increased significantly ( Figure 4 Furthermore, overexpression of AhMAGL1b significantly increased the contents of soluble sugars and starch in leaves and seeds, with little change in protein content (Table 2). These results suggest that AhMAGL1b may be involved in carbohydrate redistribution.

[0089] Table 2 Soluble sugar, starch and protein contents in leaves and seeds of AhMAGL1b overexpressing Arabidopsis

[0090]

[0091]

[0092] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.

Claims

1. Use of the AhMAGL1b gene in any of the following: (1) Application in regulating plant leaf shape; (2) Application in the preparation of products for regulating plant leaf shape; (3) Application in plant leaf shape improvement and breeding; (4) Application in cultivating plants with curled leaves; The nucleotide sequence of the AhMAGL1b gene is shown in SEQ ID NO.

1.

2. Use of the AhMAGL1b protein encoded by the AhMAGL1b gene in any of the following: (1) Application in regulating plant leaf shape; (2) Application in the preparation of products for regulating plant leaf shape; (3) Application in plant leaf shape improvement and breeding; (4) Application in cultivating plants with curled leaves; The nucleotide sequence of the AhMAGL1b gene is shown in SEQ ID NO.1; the amino acid sequence of the AhMAGL1b protein is shown in SEQ ID NO.

2.

3. Use of a recombinant vector containing the AhMAGL1b gene in any of the following: (1) Application in regulating plant leaf shape; (2) Application in the preparation of products for regulating plant leaf shape; (3) Application in plant leaf shape improvement and breeding; (4) Application in cultivating plants with curled leaves; The nucleotide sequence of the AhMAGL1b gene is shown in SEQ ID NO.

1.

4. Use of a host bacterium containing the recombinant vector according to claim 3 in any of the following: (1) Application in regulating plant leaf shape; (2) Application in the preparation of products for regulating plant leaf shape; (3) Application in plant leaf shape improvement and breeding; (4) Application in cultivating plants with curled leaves.

5. The use according to any one of claims 1 to 4, characterized in that The regulation refers to changing the shape of plant leaves into curled ones.

6. The use according to any one of claims 1 to 4, characterized in that The plant includes Arabidopsis thaliana or peanut.

7. A method for regulating plant leaf shape, characterized in that: The method comprises the steps of overexpressing the AhMAGL1b gene in the plant to increase the expression level of the AhMAGL1b gene to regulate the leaf shape of the plant; The nucleotide sequence of the AhMAGL1b gene is shown in SEQ ID NO.

1.

8. The method according to claim 7, wherein The plant includes Arabidopsis thaliana or peanut.

9. A method for cultivating a plant with curled leaves, characterized in that: The method comprises the steps of overexpressing the AhMAGL1b gene in a plant, increasing the expression level of the AhMAGL1b gene, and obtaining a plant with curled leaves; The nucleotide sequence of the AhMAGL1b gene is shown in SEQ ID NO.

1.

10. The method according to claim 9, wherein The plant includes Arabidopsis thaliana or peanut.

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