Lotus japonicus plant type-related gene LjLAZY3 and its application

Abstract

The present invention discloses a gene LjLAZY3 related to the plant type of Lotus japonicus and its application. The present invention provides a gene LjLAZY3 of the Lotus japonicus LAZY family. By constructing an overexpression vector of the LjLAZY3 gene and transferring the overexpression vector into a wild-type Lotus japonicus through Agrobacterium-mediated genetic transformation, the transgenic Lotus japonicus plant can be significantly promoted to stand upright and reduce the branching angle. The LjLAZY3 gene can be applied to genetic engineering breeding of crops to cultivate upright and densely planted varieties; the LjLAZY3 gene also has important reference significance for the study of photosynthetic utilization efficiency of other crops.

Description

Technical Field

[0001] The invention belongs to the technical field of plant genetic engineering, and specifically relates to a gene LjLAZY3 related to plant type of Lotus japonicus and an application thereof. Background Art

[0002] Lotus corniculatus L. of the genus Lotus in the Leguminosae family is a perennial high-quality forage grass native to the warm regions of Europe and Asia. It is now widely planted as a high-quality forage grass in countries with developed animal husbandry, such as the United States, Canada, and New Zealand. It is also distributed in Inner Mongolia, Northwest China, Southwest China, and South China. It has the advantages of high yield, good quality, tolerance to barrenness, and a long green period. Because it has the characteristics of symbiotic nitrogen fixation with rhizobia, it has broad application prospects in soil improvement and ecological restoration. Therefore, cultivating excellent Lotus corniculatus cultivars so that Lotus corniculatus can be promoted for planting as forage is of great significance to expanding my country's forage cultivars and improving and improving land utilization.

[0003] Ideal plant type is of great significance for utilizing and improving field microclimate, increasing light energy utilization rate and crop yield. For forage plants that mainly produce grass, the selection and breeding of ideal plant type is particularly critical. A good plant type enables the plant to make full use of light energy and soil fertility to obtain higher yields and excellent quality. Forage grasses of different plant types can be suitable for different production needs. Forage grasses with better uprightness have high light energy utilization rate, are suitable for dense planting, have good production performance, accumulate more dry matter, have good ventilation, and are suitable for producing hay. As a leguminous plant that can fix nitrogen, Lotus japonicus not only has strong adaptability, but also has the characteristics of high protein content and good palatability. It is a high-quality forage grass with great potential and can be used as a research object for exploring high-quality plant genetic resources for in-depth research.

[0004] LAZY family genes are widely present in plants. According to their effects on plant phenotypes, they can be divided into two groups: one is the LAZY1 class that positively regulates plant gravitropic responses, and the other is the NGR (Negative Gravitropic Response of Roots 1) class that negatively regulates plant gravitropic responses. LAZY1 class regulates plant type by affecting the asymmetric distribution and accumulation of auxin in leaves and stems; and affects plant phototropism by affecting the asymmetric distribution and accumulation of auxin in the hypocotyl; NGR class participates in regulating the polar transport of auxin in roots and the asymmetric distribution of auxin in the root tip, thereby affecting root gravitropism. LAZY family genes are involved in various aspects of plant growth and development, plant hormone signal transduction, and abiotic stress resistance. Summary of the invention

[0005] The purpose of the present invention is to provide a Lotus japonicus plant type-related gene LjLAZY3 and an application thereof.

[0006] The present invention adopts a method for analyzing the LAZY family genes of Lotus japonicus to screen and obtain the LjLAZY3 gene. The open reading frame sequence of the LjLAZY3 gene is shown in SEQ ID NO.1, and the full length of the reading frame is 792bp, encoding 263 amino acids, and the amino acid sequence is shown in SEQ ID NO.2. By genetically transforming a plasmid that overexpresses the LjLAZY3 gene, the expression level of the LjLAZY3 gene in Lotus japonicus is increased, and the gravitropism phenotype of the plants overexpressing LjLAZY3 is observed, and it is found that overexpressing LjLAZY3 changes the plant type of Lotus japonicus, the plants grow upright, and the branch angle is reduced; the LjLAZY3 gene plays a very important role in regulating the plant type.

[0007] Therefore, the first object of the present invention is to provide a Lotus japonicus LjLAZY3 protein, whose amino acid sequence is shown in SEQ ID NO.2.

[0008] It should be understood that, without affecting the structure and activity of the above-mentioned Lotus japonicus LjLAZY3 protein, those skilled in the art can make various substitutions, additions and / or deletions of one or several amino acids to the amino acid sequence shown in SEQ ID NO.2 to obtain a protein with equivalent function.

[0009] The present invention also provides a Lotus japonicus LjLAZY3 gene, which encodes the Lotus japonicus LjLAZY3 protein.

[0010] Preferably, the nucleotide sequence of the Lotus japonicus LjLAZY3 gene is as shown in SEQ ID NO.1.

[0011] The invention also provides a recombinant expression vector containing the Lotus japonicus LjLAZY3 gene.

[0012] Preferably, the expression vector is pCAMBIA1302 vector.

[0013] The present invention also provides application of the Lotus japonicus LjLAZY3 gene in regulating plant type.

[0014] Preferably, the plant is Lotus japonicus.

[0015] Preferably, the application is to overexpress the Lotus japonicus LjLAZY3 gene in reducing the branching angle of the plant and improving the upright and compact plant type variety.

[0016] Advantages and effects of the present invention:

[0017] 1. The successful cloning of the LAZY3 gene from Lotus japonicus and further proof that LAZY3 from Lotus japonicus is involved in regulating plant morphology are of great significance for a comprehensive understanding of the biological functions of LAZY family genes in plants.

[0018] 2. The LAZY3 gene of Lotus japonicus is overexpressed by a transgenic method, and as the expression of the LJLAZY3 gene increases, the angle of the branches of Lotus japonicus becomes smaller and the plants become upright. Therefore, the present invention can be applied to genetic engineering breeding of crops, cultivate upright crop varieties, improve light energy utilization, and be suitable for dense planting.

[0019] 3. Plant type breeding is one of the important research contents of mechanized production. The results of the present invention show that only LjLAZY3 gene in the LAZY family of Lotus japonicus affects plant type after overexpression, and verify its biological function in regulating plant type, which has important reference significance for obtaining other crops with compact plant type, suitable for dense planting and suitable for mechanized harvesting. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is the map of the pCAMBIA1302 vector used to overexpress the LjLAZY3 gene.

[0021] Figure 2 It is the restriction enzyme digestion detection of the vector overexpressing LjLAZY3 gene; wherein: lane 1 is the result of EcoR I+Spe I digestion; lane 2 is the result of Nco I+Spe I digestion.

[0022] Figure 3 It is a semi-quantitative detection and plant type of wild-type and LjLAZY3-overexpressing Lotus japonicus; Figure A is the semi-quantitative detection of LjLAZY3 gene expression in wild-type and LjLAZY3-overexpressing Lotus japonicus lines; Figure B is a photo of the plant type of wild-type and LjLAZY3-overexpressing Lotus japonicus lines.

[0023] Figure 4 The branch angle statistics of wild-type and LjLAZY3-overexpressing Lotus japonicus lines.

[0024] Figure 5 It is a semi-quantitative detection and plant type of wild-type and overexpressing LAZY family genes in Lotus japonicus. DETAILED DESCRIPTION

[0025] The following examples are provided to further illustrate the present invention, rather than to limit the present invention.

[0026] Example 1: Construction of LjLAZY3 gene overexpression vector and genetic transformation

[0027] 1. Cloning of LjLAZY3

[0028] The overexpression vector used in the present invention is pCAMBIA1302 preserved in this laboratory. The LjLAZY3 fragment ( Figure 1 ), and obtained the overexpression vector of LjLAZY3 fused with GFP. The construction steps of the LjLAZY3 overexpression vector are as follows: using the mixed cDNA of various tissues (roots, stems and leaves) of Lotus japonicus as templates, referring to the requirements of In-Fusion HD Cloning Kits of clontech, the upstream primer was designed according to the sequence of LjLAZY3: 5'-GACTCTTGA CCATGG ATGAAGTTTCTCAGCTGGAT-3' and downstream primer: 5'-TTCTCCTTT ACTAGT TATCTCTAGGACAATATATT-3', after PCR amplification, a fragment containing the LjLAZY3 coding sequence (792bp, the nucleotide sequence is shown in SEQ ID NO.1) was obtained. The PCR amplified fragment was recombined into the pCAMBIA1302 plasmid double-digested with NcoI and SpeI by the In-Fusion method. Reaction system: 2μL (0.02-0.5pmols) and 6μL of the double-digested vector and PCR amplified fragment, 10μL of HB-infusion MasterMix (2X), and ddH2O were added to the total volume of 20μL, and incubated at 50℃ for 20min. Take 10μL of the ligation product and heat-shock transform Escherichia coli DH5α at 42℃, add 1mL LB liquid culture medium, recover at 37℃ for 45min, apply to LB plate containing Kan, and incubate at 37℃ overnight. Pick a single clone and culture it in liquid LB medium containing Kan at 37°C. Use bacterial liquid PCR and sequencing to identify the correct single clone, extract the plasmid of the correct clone, and check for correct restriction enzyme digestion ( Figure 2 ) is the overexpression vector pCAMBIA1302-35S:LjLAZY3-GFP. After sequencing analysis, the accession fragment contains a 792bp reading frame. We named the gene corresponding to the reading frame LjLAZY3. The nucleotide sequence of the open reading frame of the gene is shown in SEQ ID NO.1; the amino acid sequence of the protein encoded by it is shown in SEQ ID NO.2, 263 amino acids.

[0029] 2. Genetic transformation to overexpress the LjLAZY3 gene

[0030] The genetic transformation method mediated by Agrobacterium AGL1 was used to infect the hypocotyls of wild-type Lotus japonicus (MG-20), and the overexpression vector pCAMBIA1302-35S:LjLAZY3-GFP was transformed into wild-type Lotus japonicus. Positive transgenic plants were screened by hygromycin resistance, and positive plants were obtained after PCR identification. Individual plants were harvested, and T3 obtained a homozygous Lotus japonicus strain overexpressing LjLAZY3. Non-transgenic wild-type Lotus japonicus was used as a control.

[0031] Example 2: Comparison of plant types of Lotus japonicus overexpressing LjLAZYs and wild-type Lotus japonicus

[0032] 1. Comparison of plant types between LjLAZY3-overexpressing and wild-type Lotus japonicus

[0033] Wild-type Lotus japonicus seeds (WT) and seeds of homozygous LjLAZY3 overexpression (three strains, OE1, OE2, and OE3) were treated with concentrated sulfuric acid for 10 min, then surface disinfected with 2% sodium hypochlorite, sown on sterilized filter paper, and placed at 22±2°C and 16 h light intensity (60 μmol·m 2 ·s) / 8h dark cycle, and after 3 days, the seedlings with the same growth were selected and moved to a mixed matrix of nutrient soil: vermiculite = 3:1, and the plant phenotypes were observed under the above culture conditions. After 2 months of plant growth, the wild-type strain had creeping stems and branches, while the three strains overexpressing LjLAZY3 all showed upright growth ( Figure 3 We further measured the branching angles of the plants (n=30) and found that overexpression of LjLAZY3 reduced the branching angles of the plants. The branching angles of the two strains with higher expression levels (OE1 and OE3) were reduced by 20% ( Figure 4 ). These results indicate that overexpression of LjLAZY3 makes the plant type of Lotus japonicus more compact, and the extent of the reduction in plant branching angle is positively correlated with the expression level of LjLAZY3 in Lotus japonicus.

[0034] 2. Comparison of plant morphology between overexpressing LjLAZY1, LjLAZY1L, LjLAZY2, LjLAZY3, LjLAZY4, LjLAZY6 and wild-type Lotus japonicus

[0035] Wild-type Lotus japonicus seeds (WT) and homozygous seeds overexpressing other genes of the LjLAZY family (select one representative strain from each of 35S:LjLAZY1 (inserted the nucleotide sequence shown in SEQ ID NO.3 between the CaMV35S promoter and GFP in the pCAMBIA1302 vector), 35S:LjLAZY1L (inserted the nucleotide sequence shown in SEQ ID NO.4), 35S:LjLAZY2 (inserted the nucleotide sequence shown in SEQ ID NO.5), 35S:LjLAZY3 (inserted the nucleotide sequence shown in SEQ ID NO.1), 35S:LjLAZY4 (inserted the nucleotide sequence shown in SEQ ID NO.6) and 35S:LjLAZY6 (inserted the nucleotide sequence shown in SEQ ID NO.7)) were treated with concentrated sulfuric acid for 10 min, surface disinfected with 2% sodium hypochlorite, sown on sterilized filter paper, and placed at 22±2°C under 16 h of light (60 μmol·m 2 ·s) / 8h dark cycle, and after 3 days, the seedlings with the same growth were selected and moved to the mixed matrix of nutrient soil: vermiculite = 3:1, and the plant phenotype was observed under the above culture conditions. After 1 month of plant growth, the stems and branches of the wild-type strain and the overexpression strains of LjLAZY1, LjLAZY1L, LjLAZY2, LjLAZY4, and LjLAZY6 all grew prostrate, and there was no significant change in the branch angle, while the strain overexpressing LjLAZY3 showed upright growth ( Figure 5 ). Therefore, among the six members of the LAZY gene family in Lotus japonicus, only overexpression of LjLAZY3 affected the plant architecture. Sequence Listing <110> South China Botanical Garden, Chinese Academy of Sciences <120> Lotus japonicus plant type-related gene LjLAZY3 and its application <160> 7 <170> SIPOSequenceListing 1.0 <210> 1 <211> 792 <212> DNA <213> Lotus corniculatus L. <400> 1 atgaagtttc tcagctggat gcaaaataaa cttggtggaa agcaagagaa cagaaaatca 60 aacacaaatg cagccccacc tgcatgtcaa gcaaaacaag agcctcgaga agagttcagt 120 180 240 ttcacgcctg aagaaatcgg gaactacaa aaggagttaa caaggctcct gagacgaaaa 300 cccaatgtgg aaaaagaaat ctctgagctt cctcttgaca gatttcttaa ttgcccatca 360 agcttggagg ttgataggag aatcagcaac gcgctttgca gcgaatcaga tgataaagaa 420 gaagacattg agaagacact cagtgtcata cttgatagat gcaaagatgt ttgtgcagag 480 aagagcaaga agtcaattgg gaagaaatct attctttcc tgttgaagaa gatgtttgtt 540 tgtagaagtg ggtttgctcc aacacctagc cttagagata cccttcaaga atcaagaatg 600 gagaagcttt taggcaat gcttcacaag agaatttaca cccaaaaattc ttctcgggca 660 720 ccagaagaga gaaaagatga aggctgtaaa tgggtcaaga ccgactctga atatattgtc 780 ctagatat aa 792 <210> 2 <211> 263 <212> PRT <213> Lotus corniculatus L. (bird's-foot trefoil) <400> 2 Met Lys Phe Leu Ser Trp Met Gln Asn Lys Leu Gly Gly Lys Gln Glu 1 5 10 15 Asn Arg Lys Ser Asn Thr Asn Ala Ala Pro Pro Ala Cys Gln Ala Lys 20 25 30 Gln Glu Pro Arg Glu Glu Phe Ser Asp Trp Pro His Gly Phe Leu Ser 35 40 45 Ile Gly Thr Phe Gly Lys Lys Asp Glu Ile Lys Gln Asn Leu Asp Asp 50 55 60 Gln Asn Thr Gln Asp Gln Asp Pro Ser Ser Ser Glu Glu Ile Ala Asp 65 70 75 80 Phe Thr Pro Glu Glu Ile Gly Lys Leu Gln Lys Glu Leu Thr Arg Leu 85 90 95 Leu Arg Arg Lys Pro Asn Val Glu Lys Glu Ile Ser Glu Leu Pro Leu 100 105 110 Asp Arg Phe Leu Asn Cys Pro Ser Ser Leu Glu Val Asp Arg Arg Ile 115 120 125 Ser Asn Ala Leu Cys Ser Glu Ser Asp Asp Lys Glu Glu Asp Ile Glu 130 135 140 Lys Thr Leu Ser Val Ile Leu Asp Arg Cys Lys Asp Val Cys Ala Glu 145 150 155 160 Lys Ser Lys Lys Ser Ile Gly Lys Lys Ser Ile Ser Phe Leu Leu Lys 165 170 175 Lys Met Phe Val Cys Arg Ser Gly Phe Ala Pro Thr Pro Ser Leu Arg 180 185 190 Asp Thr Leu Gln Glu Ser Arg Met Glu Lys Leu Leu Arg Thr Met Leu 195 200 205 His Lys Arg Ile Tyr Thr Gln Asn Ser Ser Arg Ala Ser Ser Met Lys 210 215 220 Lys Cys Leu Glu Asp Lys Lys Leu Ile Arg Asn Arg Asn Glu Asp Glu 225 230 235 240 Pro Glu Glu Arg Lys Asp Glu Gly Cys Lys Trp Val Lys Thr Asp Ser 245 250 255 Glu Tyr Ile Val Leu Glu Ile 260 <210> 3 <211> 1146 <212> DNA <213> Lotus corniculatus L. <400> 3 atgaagttac ttggttggat gcatagaaag tttcgccaga atagcaatga acctttcaaa 60 gacttggtca ttgggaatgc ttgcaattgt ctttcagggc agccatcact tgatgatgaa 180. 180. 180. 180. 180. 180. 180. 180. 180. 180. 180 agtcttagga aatctttttc tggtgtgga gcaacaggag tagtagtca tgaagactat gagggagaag aaccttcagc tgctatgagt gatctcttcc ctggcttctt ttcaattgga actcttggtt cagaacaagt agttacttct tacccatcaa caccaacatt tgcaatctct gttgagagca taactgaga agagatga gtcactgaga atgatcttaa actcatcaat gatgagctag agaaagtgct aggagctgaa accaaggatg atgttgtcag caatgattca tctagcagaa atagccatgt cagcactggg agaagcagcc atgtcagcac tgggagaagc 540 agccatgtta gcataatac actcagtgga aagccaatag atggtgctgc tgcagaatca aatgggaata aataatgg aactgcaatt tgtccactcc aggaattct ctttggaaca gctattgaat tatctgaaac aacaactaca gtggcaaaga aagaacacag gacttcactt ggggagctgt ttcagagaag caaattagct gaggagaatt atggtgctaa aattgaaaag gaggaaagaa gagctgagag ggaaggagat aaatctgcta tgaacttgat gaaagagaag 840 ctgaaaagaa gaatgttgca tgcttgttct aggaattcta catcaacaag tggtggccct 900 tttgattctg cttcagcaga tacaaaactg aataagattc ttcatatgtt ccgcaagaaa 960 gttcaccctg aaagctcaac agctgcacac aagtcaacta agcacagaaa gaatgagaac 1020 aagaagaaga taatcaacga tggaggccac accaaaggtg acctggggca tccagatgaa 1080 gactcatctg ctatcaggga gcattggatc aaaacagatg cagatgactt agttctagag 1140 ctctga 1146 <210> 4 <211> 969 <212> DNA <213> Lotus corniculatus L. <400> 4 atgcacagaa aatttcgcca gaatagcact gaaccgttta aagatcactt ggttattggg 60 aaggcaggac atgaaattga tgaacgcagt taccatctga aaccgaactt tgggagcaaa 120 catgtccaac aaattgagaa agagcacagc cttagaaaat ctttcagtgg tcaggaagct 180 gcagatgcaa gagtagaaga tgatgagtac gtgtatgagg aagatggtgc tatgtatgag 240 cttttccatg ggttcctcgc aattggaact cttgggcttg ggtgcagtac tgatccatca 300 gcaacaccaa catttggcaa tatctgtgtt gagaatataa ttactgaaaa ggaagatgaa 360 gtgactgaga atgaattgaa gctgatcaat gatgagctcg agaaagttct tggagatgat 420 gagtgtatta attctgccag aaatagccat gtaagtagca gtactgcagg gagaagtagt 480 catggaagca tgatatcatt gagcggaaag ccaatgaata tgatggaagg taattctggt 540 aatattggaa cttcttcaat ttgtccactt cagggttatc tttttggatc agcttccatt 600 gaattaccaa aagtagtacc agcaaagagt aggacatcac ttggggagct atttcagaga 660 agcaaactag cagaccagga gattaatgaa aagggtgata aaatcagtga aaaagaagat 720 aaatctgccc tgcatctggt aagaaagaag ctaaaaaaaa aatattgctc catacttgtt 780 cccgaaactc ctcctcaaat aacggtggac ctgttgattc agctaattca gcagacacaa 840 aattgcgtaa gattctccat atgttccaca ggaaagttca ccctgaaaat tcagcagctg 900 aaccaaagaa tgagtgtaaa aagaaaacaa tcaatgtgga cggaggccat aacaaatttg 960 aagcattag 969 <210> 5 <211> 813 <212> DNA <213> Lotus corniculatus L. <400> 5 atgcagattc ttcattggct ctttaaaagg gcacaagaac aagaaagaaa gagcagcacc 60 ttcagggaag aaatcagagc acaaaatgga agagggaaaa aggttagtct actgaagcat 120 gatggatcat atagaagaac aaagagattc gaatcatgca gcaagaatct gaagttattg 180 gcttttgtgt atagaaaaga cattccgaag gcttgcttct acaaaaccat caacttaaag 240 agattaggaa gctttcatag aaggcatttt ctgatctcta tgaaaatgaa gaaagaagaa 300 tcatcaagag aagaaggggt tgttaccaag acagattcaa cagctcatgt gggaaacaag 360 gttttgccta taactgaggc accattgtca tcctcagctc aaagaaatgg ccaatgtgat 420 gccacagaaa ataaagatca gagcaagggg gataggaaaa agaccatttc gaagatgaaa 480 gagctaatta gatgggctgc tttttccaaa acagagaaag gaggaaaatt ctatggaaag 540 gagattttaa tgttccggag gcgtggaacc ttgaaagcag ttccggatga ggatcaagtt 600 tgcagtgagt cacccaagat cagtttcaga tgggatgtgg aaagctgctc caccacttcc 660 tctgcctact cagctatctc agtagcatcc tcatcaagat ttggacaaaa ccaaattgca 720 acttccacca tatccattcc tcattcagag aatggctgta gaaaaggaag ctggatcaca 780 acagactctg aatttgtggt gctggagcta tga 813 <210> 6 <211> 780 <212> DNA <213> Lotus corniculatus L. <400> 6 atgaaggttt ttctatggaa gaagaacaag cttaatggaa ccaatggcaa gaagaaacca 60 acctcagctt caactactca ttatatgctg catgaatctt gcaagcaaga gttcaatgac 120 tggcctcagg cattccttca cattggaact ttagctaaca gttatgcaga agattcattt 180 ccttttcaga aatacacaaa ggagttcaag ctggaggaag ttgggaaact tcagaatgaa 240 ttgaacatga aattggagga tgaacaagaa gaaaacctca atgatttgat caatgaacat 300 tcaagcttgg aacctgcagc acaagaagtt gagacaattt ctgatgtctc caatatcaaa 360 catggtaatt tctacccagg caacagtcta atacacagca cgggaagaga tctttgttat 420 gatagcaaaa agggtggcat tgggaagaaa tcactttctt tcctttttaa gaagatgttt 480 gcttgcagaa aaggatttca acctactcct tctttcaaag atccactact ctccacagat 540 tccagaatgg agaagattct gagggcaatg cttcagaaaa agatacatcc ccaaggttca 600 tcttcaacaa catttgtaaa gaagtacctg gaaaaggatc aaatgcccca gtttgatgaa 660 gatgataatg atgatgatga tgataatgat gatgatgatg aagaggaacc tactagtgct 720 acaaacaatg gaagtaaatg ggtcaagaca gattctgagt gtatcgttct ggagatataa 780 <210> 7 <211> 735 <212> DNA <213> Lotus corniculatus L. <400> 7 atgcagattc ttcaacggct cactaaaggg gtacatgaac aagacagaga gatctggacc 60 tttagtcctt ccaccaatac agttaccaga gatataaaca tacctaagca tcacagaaca 120 aggagaatag tatggagcaa acttcgatta aagaagttga agctgtttac attcatgtgc 180 ggatcaaaag acattccaaa ggcctgcatt tgcaggaccc ccgacttata cggattagga 240 aacttgagaa attackataaag gacagattct gctgctgcta atgtaggaaa gaaggttttg cccgtaactg aagcatcatt ctctagaaca acagaaagaa atgagtttgt cacagcagaa gctaaggatc aagttaaga ggataaaaag aaagcaattt caagaatgaa agagctactt ggtctggttt ctgctgcca aaaagataa agaggaaat tcaatgaaca aaaagtttcaa 540. acataagga acataaggggc aactccagaa gattatcaac tttgcagtga gtcacccaag atcgaattca catgggatga ggaaattcag tccattactt cttctgtcta ctcagttatc tcatcaatag cttctttgtc taagagtgga caagcccaga ttgcacactc gccaattcat atctcttgta ggaaagaaa ctggatcaca acagactctg aatttgtggt gttggaacta aagacgtgtg tctaa 735

Claims

1. Use of overexpression of the LjLAZY3 gene in Lotus japonicus to reduce the branching angle of the stems and branches of Lotus japonicus and improve Lotus japonicus varieties with upright and compact stems and branches, characterized in that: The nucleotide sequence of the Lotus japonicus LjLAZY3 gene is shown in SEQ ID NO.1.

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