LjCYCD2 gene and application thereof in regulating and controlling glandular hair density of plant leaves

Abstract

The invention discloses an LjCYCD2 gene and application thereof in regulation and control of plant leaf glandular hair density, and belongs to the technical field of gene engineering. The amino acid sequence coded by the LjCYCD2 gene is as shown in SEQ ID NO: 2, and the nucleotide sequence of the LjCYCD2 gene is as shown in SEQ ID NO: 1. The gene is expressed in the honeysuckle leaf, the expression quantity is gradually reduced along with the maturity of the leaf, and after the LjCYCD2 gene of the honeysuckle leaf is silenced by VIGS, the gland hair density of the honeysuckle leaf is obviously reduced. The honeysuckle LjCYCD2 gene is used for constructing a plant overexpression vector and transforming tobacco, and the number of glandular hairs of leaves of the obtained transgenic tobacco is obviously increased. It can be seen that the honeysuckle LjCYCD2 gene is related to plant leaf glandular hair development and has important application value in regulation and control of plant leaf surface glandular hair density.

Description

Technical Field

[0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a LjCYCD2 gene and an application thereof in regulating the density of glandular hairs on plant leaves. Background Art

[0002] Glandular hairs are usually distributed on the surface of plant organs such as stems, leaves, flowers, and fruits. They can not only perform physical barrier functions like non-glandular hairs, but also secrete a variety of secondary metabolites such as terpenes, alkaloids, phenols, and sterols. Studies have shown that glandular hairs can regulate the response of plants to abiotic stresses, help plants resist low temperatures, and improve tolerance in heavy metal-contaminated soils; they can also resist damage from animals. For example, the glandular hairs of Solanaceae plants can produce acyl sugar metabolites with insecticidal activity. Metabolites such as δ-elemene and β-caryophyllene produced in the glandular hairs of tomatoes (Lycopersicon esculentum) not only improve the tolerance of tomatoes to red spider mites, but also have a repellent effect on a variety of herbivores. The glandular hairs of many medicinal plants are also the site where their medicinal ingredients are produced. Artemisinin is a sesquiterpene lactone produced in the glandular hairs of Artemisia annua, which can be used as an antimalarial drug; the shield-shaped glandular hairs on the surface of the Chinese medicine Nepeta cataria contain a large amount of volatile oil, which is its main active ingredient.

[0003] Glandular hairs are an exocrine structure formed by cell division of the primitive cells of the epidermis. In the early stages of glandular hair development, some primitive cells in the epidermis receive cell division signals that can form glandular hairs, and then divide through a strictly regulated cell cycle. The number, direction, and degree of cell extension of these cell divisions will lead to the formation of different types of glandular hairs. The density and size of glandular hairs are important factors in regulating the content of metabolites, so increasing the density of glandular hairs is considered an effective way to increase the content of secondary metabolites.

[0004] Cell cycle regulation plays a key role in plant cell differentiation, leading to the formation of unicellular or multicellular glandular hairs. Studies have shown that cell cycle-related genes play an important role in the early stages of glandular hair development, but these studies have mostly focused on unicellular glandular hairs. For example, in Arabidopsis thaliana, SIM can effectively promote cell division and cell differentiation of trichomes. There are few studies on the molecular regulatory mechanisms of multicellular glandular hair development. In a study of Artemisia annua multicellular glandular hairs (Artemisia annua glandular hairs are composed of 10 cells, namely two basal cells, two stalk cells, four subapical cells and two apical cells), it was found that overexpression of TLR1 and TLR2 can significantly reduce the density of glandular hairs and the content of artemisinin. The CsMYB6 gene in cucumber (Cucumis sativus) plays an important role in regulating the development of glandular hairs in cucumber fruits, inhibiting the formation of glandular hairs on the surface of cucumber fruits by forming a CsMyb6-CsTRY complex.

[0005] Honeysuckle (Lonicera japonica) is a twining shrub of the genus Lonicera in the family Caprifoliaceae. Its flowers are called honeysuckle and its stems and leaves are called honeysuckle vines. It is a traditional Chinese herbal medicine rich in organic acids, triterpenes, flavonoids and other active ingredients, and has good therapeutic effects in treating colds, headaches, fevers and rashes. Studying the genes related to the development of honeysuckle glandular hairs can provide further basis for the development and molecular regulation of multicellular glandular hairs and provide reference for the cultivation of high-quality honeysuckle varieties. Summary of the invention

[0006] In view of the problems existing in the prior art, the object of the present invention is to provide a LjCYCD2 gene and its application in regulating the density of glandular hairs on plant leaves.

[0007] In order to achieve the above object, the present invention adopts the following technical solution:

[0008] The LjCYCD2 gene, the amino acid sequence it encodes is shown in SEQ ID NO:2, and the nucleic acid sequence thereof is shown in SEQ ID NO:1.

[0009] The application of the above-mentioned LjCYCD2 gene in regulating the density of glandular hairs on plant leaves.

[0010] A method for regulating the density of glandular hairs on plant leaves, comprising silencing a LjCYCD2 gene in the plant to reduce the density of glandular hairs on the plant leaves; or overexpressing the LjCYCD2 gene in the plant to increase the density of glandular hairs on the plant leaves; the amino acid sequence of the LjCYCD2 gene is shown in SEQ ID NO: 2.

[0011] Based on the above scheme, the nucleic acid sequence of the LjCYCD2 gene is shown in SEQ ID NO:1.

[0012] On the basis of the above scheme, a silencing vector or an overexpression vector containing the LjCYCD2 gene is constructed and transformed into a plant body, so that the LjCYCD2 gene in the plant body is silenced or overexpressed, thereby reducing or increasing the density of glandular hairs on plant leaves.

[0013] On the basis of the above scheme, the method of transforming the plant body is one of the following: Agrobacterium-mediated method, gene gun method, electric shock method, PEG method, and liposome method.

[0014] Based on the above scheme, the Agrobacterium is Agrobacterium tumefaciens or Agrobacterium rhizogenes.

[0015] Based on the above scheme, the plant is honeysuckle or tobacco.

[0016] The application of the LjCYCD2 gene in the preparation of products for regulating the density of glandular hairs on plant leaves.

[0017] A product for regulating the density of glandular hairs on plant leaves is a recombinant expression vector, an expression cassette, a recombinant bacterium, a recombinant virus or a transgenic cell line containing a LjCYCD2 gene sequence; the nucleic acid sequence of the LjCYCD2 gene is shown in SEQ ID NO: 1.

[0018] Advantages of the technical solution of the present invention

[0019] The invention clones and obtains the LjCYCD2 gene of honeysuckle, which is expressed in all leaves of honeysuckle, and the expression amount gradually decreases with the maturity of the leaves. After silencing the LjCYCD2 gene of honeysuckle leaves by VIGS, the density of glandular hairs of the leaves of honeysuckle is significantly reduced. The LjCYCD2 gene of honeysuckle is constructed into a plant overexpression vector, and tobacco is transformed, and the number of glandular hairs of the leaves of the obtained transgenic tobacco is significantly increased. It can be seen that the LjCYCD2 gene of honeysuckle is related to the development of glandular hairs on plant leaves, and has important application value in regulating the density of glandular hairs on the surface of plant leaves. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 is the relative expression level of LjCYCD2 gene in different leaves of Honeysuckle;

[0021] Figure 2 The relative expression level of LjCYCD2 gene in the first pair of leaves of Lonicera japonica after gene silencing was analyzed by qRT-PCR (error bars indicate SD; asterisks indicate significant differences, one-way ANOVA, *P<0.1, **P<0.01, ***P<0.001, ****P<0.0001);

[0022] Figure 3 The phenotype and glandular hair density of LjCYCD2 gene in honeysuckle leaves after silencing by VIGS (A is TRV1-TRV2, B is TRV2 / LjCYCD2, and C is the change of glandular hair density after different treatments);

[0023] Figure 4 qRT-PCR analysis of the relative expression level of LjCYCD2 gene in transgenic tobacco leaves after LjCYCD2 gene overexpression;

[0024] Figure 5 The phenotype and density of glandular hairs in transgenic tobacco leaves (A is pSuper1300, B is OE-LjCYCD2, and C is the change in glandular hair density after different treatments). DETAILED DESCRIPTION

[0025] The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those of ordinary skill in the art. The present invention will be further described in detail below in conjunction with specific examples and with reference to data. The following examples are intended to illustrate the present invention and are not intended to limit the scope of the present invention in any way.

[0026] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The experimental materials, reagents, drugs, etc. used in the following examples, unless otherwise specified, can all be purchased through general channels.

[0027] In the following embodiments,

[0028] Honeysuckle "Beihua No. 1": obtained from Linyi, Shandong; Nicotiana benthamiana: provided by the Plant Genetics and Development Laboratory of Qingdao Agricultural University; pSuper1300 vector, TRV2 vector: provided by the Plant Genetics and Development Laboratory of Qingdao Agricultural University; P19, TRV1: provided by the Plant Genetics and Development Laboratory of Qingdao Agricultural University; Escherichia coli DH5α competent cells, Agrobacterium tumefaciens EHA105, Agrobacterium tumefaciens GV3101: Shanghai Weidi Biotechnology Co., Ltd.

[0029] Example 1

[0030] Cloning of LjCYCD2 gene

[0031] Total RNA was extracted from the leaves of Lonicera japonica 'Beihua No. 1' and reverse transcribed into cDNA as a template. PCR amplification was performed using specific primers of the LjCYCD2 gene. After sequencing, the nucleic acid sequence of the LjCYCD2 gene was obtained as shown in SEQ ID NO: 1, and the amino acid sequence encoded by it was shown in SEQ ID NO: 2.

[0032] SEQ ID NO:1(5’→3’)

[0033]

[0034] SEQ ID NO:2:

[0035] MAVTPTSPSSKSISTSQNNKNKKQKPHTKIKKKNNKNKNKNDDHNPTPIINPINPQQLPPSKPVKRLKSPGVRVIGARIYDSEHGKTCHQCRQKTMDFVVSCKSQTQNKNNKQCTFRFCHKCLLNRYGEKAEELAELVDWICPKCRGICNCSFCMKKRGFHPTGILVHTAKANGFSSVSDMLLIDNSAKAETEEGSPSKKRAAKSENGDLELKRSRKDDNKEQQSRGIIKDAGASSKKATALAKEPAVGPEKAQKNVTGLDGNSDMKSQPESLKTKTNKKDTKNTNLEGLRESNEIDDSSETSMNNGAPAENITTKNDIELIKTNSEALMPNKVSTRSIKIEDNESEVPSNINVNTTAAKSRIVDEFDQFTKTEEVQGKDSVAEIPLPQGTELTNVANIDMPAEDVGHALQFLEFCEAFGKVLDLKKGQPEMLLRDLACDRRSRRKDESSVVQFHIKLLSMIQETSGKEFSSSKATVRNPWLEGLRKCISESQDPSKESLLGFLDVGVDGYEKLDTSKKLRLLTFLCDEALGTPVLRSWIDEQNSEYVKEVKKAKETILPAQKEKEKNMKRKVQNEVARAIIMNNGVPLSIAEHENLVFKIRAEAAQTLAETLEEIEMLTKTLPKETPRSDAVRSEPILFDASGRIFWKLRGYDGSSDILLQDIGSGELVTCKDRWFAYDVQEKEMVEKYLSSKRNL

[0036] The specific primer sequences of the LjCYCD2 gene are as follows:

[0037] Forward primer: 5’-atacaccaaatcgactctagaATGGCTGTAACTCCTACATCTCCTT-3’ (SEQ IDNO:3);

[0038] Reverse primer: 5′-gcccttgctcaccatggtaccTCAAAGATTCCTTTTTGATGAAAGAT-3′ (SEQ ID NO: 4);

[0039] PCR reaction system: PrimeSTARMax DNA Polymerase (2×) 12.5μL, Primer F / R (10μM) 1μL; cDNA 1μL; RNase Free Water 9.5μL.

[0040] The PCR reaction procedure is:

[0041]

[0042] Example 2

[0043] Expression analysis of LjCYCD2 gene

[0044] The first, second, third and fourth pairs of leaves of Honeysuckle "Beihua No. 1" were taken, frozen in liquid nitrogen and stored at -80°C. According to the instructions of the kit of Accurate Biotechnology (Hunan) Co., Ltd., the total RNA of the leaves was extracted, reverse transcribed into cDNA as a template, and qRT-PCR was performed using the qRT-PCR specific primers of the LjCYCD2 gene, and the Lonja.ACT2 / 7 gene was used as the internal reference gene.

[0045] The specific primer sequences for qRT-PCR of LjCYCD2 gene are as follows:

[0046] LjCYCD2-F: 5'-TCTGAGCATGGCAAGACCTG-3' (SEQ ID NO: 5);

[0047] LjCYCD2-R: 5'-TAGGGTGAAAACCCCGCTTC-3' (SEQ ID NO: 6);

[0048] The specific primer sequences for qRT-PCR of Lonja.ACT2 / 7 gene are as follows:

[0049] Lonja.ACT2 / 7-F: 5'-CCCTAAAGCCAACAGAGAGAAG-3' (SEQ ID NO:7);

[0050] Lonja.ACT2 / 7-R: 5'-CGACCACTAGCATACAGAGAAAG-3' (SEQ ID NO: 8);

[0051] The reaction system of qRT-PCR was: DNA Polymerase (2×) 10 μL; Primer F / R (10 μM) 0.8 μL; cDNA 1 μL; RNase Free Water 7.4 μL.

[0052] The reaction conditions for qRT-PCR were:

[0053]

[0054] qRT-PCR results Figure 1 As shown in the figure, the density of glandular hairs on honeysuckle leaves gradually decreases with the maturity of the leaves. From the first pair of leaves to the fourth pair of leaves, the maturity of the leaves gradually increases, and the density of glandular hairs gradually decreases. Figure 1 It can be seen that the LjCYCD2 gene is expressed in the first to fourth pairs of leaves of Honeysuckle, and its relative expression level decreases with the maturity of the leaves, which is consistent with the changing trend of the number of glandular hairs on different leaves. Therefore, the LjCYCD2 gene is related to the development of honeysuckle glandular hairs.

[0055] Example 3

[0056] Construction of LjCYCD2 gene silencing vector

[0057] According to the sequences on both sides of the XbaI and KpnI restriction sites in the TRV2 vector and the requirements for homologous recombination, primers for the LjCYCD2 specific sequence were designed and synthesized using CE Design V1.04 software. The primer sequences are as follows:

[0058] TRV2 / LjCYCD2-F:5'-aaggttaccgaattctctagaGACGACCACAATCCAACTCCG-3'(SEQID

[0059] NO:9)

[0060] TRV2 / LjCYCD2-R:5'-gagacgcgtgagctcggtaccTATCCAATCAACCAATTCGGC-3'(SEQID NO:10)

[0061] The RNA of leaves of four different leaf positions of Lonicera japonica treated with quick freezing was extracted using the kit instructions of Accurate Biotechnology (Hunan) Co., Ltd., and reverse transcribed into cDNA using PrimerScriptTM RT Reagent Kit. After equal amounts were mixed as templates, the specific primers mentioned above were used to amplify the specific fragment of LjCYCD2. The amplified target fragment was detected by electrophoresis, and the bright bands with the correct position were selected for The electrophoresis product was purified using the Gel DNA Extraction Mini Kit to obtain the target fragment.

[0062] The TRV2 vector was double-digested with QuickCut restriction enzyme to obtain a linearized vector.

[0063] After adding the target fragment and linearized vector as required for the recombination reaction, add homologous recombination ligase, and perform the recombination reaction at 50°C for 1 hour to obtain the recombinant plasmid.

[0064] The recombinant plasmid was transformed into Escherichia coli DH5α competent cells, and the positive clones were picked out. After PCR detection, sequencing was performed. The one with correct sequencing was the successfully constructed LjCYCD2 gene silencing vector (TRV2 / LjCYCD2).

[0065] Example 4

[0066] Effects of silencing LjCYCD2 gene on the density of glandular trichomes in leaves of Honeysuckle

[0067] The silencing vector TRV2 / LjCYCD2 of Example 3 was transformed into Agrobacterium tumefaciens EHA105, and TRV1 and TRV2 were respectively transferred into Agrobacterium tumefaciens EHA105. The three bacterial solutions containing the recombinant plasmids TRV1, TRV2, and TRV2 / LjCYCD2 were placed in a shaker at 28°C, 200rpm, and cultured overnight; the next day, the three bacterial solutions were shaken for a second time to about OD600 = 1.0-1.2; then the bacteria were collected, and an equal volume of infection solution was prepared (1ml MES, 40μLAS, and 1ml MgCl2 were added to every 100mlH2O), and the bacteria were resuspended to OD600 = 1.0-1.2; the bacterial solution containing the TRV1 plasmid and the bacterial solution containing the TRV2 plasmid were mixed at a volume ratio of 1:1, and the bacterial solution containing the TRV1 plasmid and the bacterial solution containing the TRV2 / LjCYCD2 recombinant plasmid were mixed at a volume ratio of 1:1; after mixing, the mixture was left to stand in the dark for 3 hours before infection.

[0068] The first pair of leaves of honeysuckle branches were selected and infected by high-pressure suction using a vacuum pump. During the suction infection, the young branches were immersed in the infection liquid, and the air was pumped for 4-5 minutes, kept for 20 minutes, and slowly released for 3 hours, and repeated twice; then it was dark treated at room temperature for 3 days and cultured normally. The empty vector TRV2 was used as the control (CK), and the relative expression of the LjCYCD2 gene in the first pair of honeysuckle leaves was determined by qRT-PCR 14 days after VIGS transformation, and the difference in glandular hair density was observed.

[0069] The relative expression results of LjCYCD2 gene in the first pair of leaves of Honeysuckle after silencing are as follows Figure 2As shown, the relative expression level of the LjCYCD2 gene in the first pair of leaves of Honeysuckle decreased significantly after silencing.

[0070] After LjCYCD2 gene was silenced, the density of glandular hairs on honeysuckle leaves changed. Figure 3 Under the microscope, we can see that the glandular hairs at the base of the main vein of the control leaf are denser ( Figure 3 Middle A), after TRV2 / LjCYCD2 was silenced, the density of glandular hairs on the lower surface of honeysuckle leaves decreased significantly ( Figure 3 Middle B), compared with the control, the density of glandular hairs decreased by 2 times ( Figure 3 C), which is consistent with the results of gene expression, further indicating that this gene is a positive regulatory gene for the development of honeysuckle glandular hairs.

[0071] Example 5

[0072] Construction of LjCYCD2 gene overexpression vector

[0073] According to the gene sequence of LjCYCD2 of honeysuckle and the sequences on both sides of the Xbal and Kpnl restriction sites in the pSuper1300 vector and the requirements for homologous recombination, primers of the LjCYCD2 specific sequence were designed and synthesized using CE Design V1.04 software. The primer sequences are as follows:

[0074] pSuper1300-LjCYCD2-F: 5'-atacaccaaatcgactctagaATGGCTGTAACTCCTACATCTCCTT-3' (SEQ ID NO: 11);

[0075] pSuper1300-LjCYCD2-R: 5'-gcccttgctcaccatggtaccTCAAAGATTCCTTTTTTGATGAAAGAT-3' (SEQ ID NO: 12);

[0076] The RNA of leaves of four different leaf positions of honeysuckle treated by quick freezing was extracted using the kit instructions of Accurate Biotechnology (Hunan) Co., Ltd., and reverse transcribed into cDNA using PrimerScriptTM RT Reagent Kit. After that, equal amounts were mixed as templates to amplify the specific fragment of LjCYCD2. The amplified target fragment was detected by electrophoresis, and the bright bands with the correct position were selected for The electrophoresis product was purified using the Gel DNA Extraction Mini Kit to obtain the target fragment.

[0077] The pSuper1300 vector was double-digested with QuickCut restriction enzyme to obtain a linearized vector.

[0078] After adding the target fragment and linearized vector as required for the recombination reaction, add homologous recombination ligase, and perform the recombination reaction at 50°C for 1 hour to obtain the recombinant plasmid.

[0079] The recombinant plasmid was transformed into E. coli DH5α competent cells, and the positive clones were picked out. After PCR detection, sequencing was performed. The one with correct sequencing was the successfully constructed overexpression vector pSuper1300-LjCYCD2.

[0080] Example 6

[0081] Application of overexpression of LjCYCD2 gene in increasing the density of glandular hairs on plant leaves

[0082] The overexpression vector pSuper1300-LjCYCD2 of Example 5 was transformed into Agrobacterium tumefaciens GV3101, and the P19 and pSuper1300 empty vectors were respectively transferred into Agrobacterium tumefaciens GV3101; the three bacteria containing P19, pSuper1300 empty vector, and pSuper1300-LjCYCD2 recombinant vector were activated, and the bacterial solution was expanded and shaken to about OD600 = 0.6-0.8 on the second day; then the bacteria were collected and an equal volume of infection solution was prepared (1 ml MES, 40 μL HCl were added to every 100 ml H2O) AS, 1mlMgCl2), resuspend the bacteria to OD600 = 0.6-0.8; mix the bacterial solution containing P19 plasmid and the bacterial solution containing pSuper1300 plasmid at a volume ratio of 1:1, and mix the bacterial solution containing pSuper1300 plasmid and the bacterial solution containing pSuper1300-LjCYCD2 recombinant plasmid at a volume ratio of 1:1; after mixing, let it stand for 3 hours in the dark before infection.

[0083] First, the leaves of sterile tobacco seedlings were placed in MS medium for pre-culture, and then the pre-cultured tobacco leaves were completely immersed in the infection solution. After infection for 8-10 minutes, they were inoculated on the co-culture medium and inverted in the dark at 28°C for 2-4 days; then differentiation culture, bud proliferation culture, rooting culture, and transplanting were carried out. The DNA of the leaves of empty vector and transgenic tobacco were extracted for identification, and then RNA was extracted to detect the expression level, and the transgenic strains with high expression levels were identified. The empty vector pSuper1300 was used as the control (CK), and T1 generation tobacco seeds were collected. After T2 generation culture, the leaves of tobacco plants with consistent growth were taken to observe the difference in glandular hair density.

[0084] The relative expression results of LjCYCD2 gene in tobacco overexpressing LjCYCD2 gene are as follows Figure 4As shown, compared with the control, the relative expression level of LjCYCD2 gene increased significantly.

[0085] After LjCYCD2 gene overexpression, transgenic tobacco leaves with basically the same growth potential were taken and the density of glandular hairs was observed under a fluorescence microscope, as shown in the figure ( Figure 5 ). Compared with the control group pSuper1300, it can be seen that the density of glandular hairs in the LjCYCD2 overexpression strain is significantly increased. Therefore, overexpressing the LjCYCD2 gene in plants can increase the number of glandular hairs on plant leaves.

[0086] The above is only a preferred embodiment of the present invention, and does not limit the present invention in other forms. Any technician familiar with the profession may use the above disclosed technical content to change or modify it into an equivalent embodiment with equivalent changes. However, any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention without departing from the technical solution of the present invention still belongs to the protection scope of the technical solution of the present invention.

Claims

1. LjCYCD2 gene, characterized in that The amino acid sequence encoded by it is shown in SEQ ID NO:

2.

2. The LjCYCD2 gene according to claim 1, characterized in that Its nucleic acid sequence is shown in SEQ ID NO:

1.

3. Use of the LjCYCD2 gene according to claim 1 or 2 in regulating the density of glandular hairs on plant leaves.

4. A method for regulating the density of glandular hairs on plant leaves, characterized in that: The density of glandular hairs on plant leaves is reduced by silencing the LjCYCD2 gene in the plant; or the density of glandular hairs on plant leaves is increased by overexpressing the LjCYCD2 gene in the plant; the amino acid sequence of the LjCYCD2 gene is shown in SEQ ID NO:

2.

5. The method for regulating the density of glandular hairs on plant leaves according to claim 4, characterized in that: The nucleic acid sequence of the LjCYCD2 gene is shown in SEQ ID NO:

1.

6. The method for regulating the density of glandular hairs on plant leaves according to claim 4, characterized in that: A silencing vector or an overexpression vector containing the LjCYCD2 gene is constructed and transformed into a plant body, so that the LjCYCD2 gene in the plant body is silenced or overexpressed, thereby reducing or increasing the density of glandular hairs on plant leaves.

7. The method for regulating the density of glandular hairs on plant leaves according to claim 6, characterized in that: The method of transforming the plant body is one of Agrobacterium-mediated method, gene gun method, electric shock method, PEG method and liposome method.

8. The method for regulating the density of glandular hairs on plant leaves according to claim 7, characterized in that: The Agrobacterium is Agrobacterium tumefaciens or Agrobacterium rhizogenes.

9. Use of the LjCYCD2 gene according to claim 1 or 2 in preparing a product for regulating the density of glandular hairs on plant leaves.

10. A product for regulating the density of glandular hairs on plant leaves, characterized in that: It is a recombinant expression vector, expression cassette, recombinant bacteria, recombinant virus or transgenic cell line containing the LjCYCD2 gene sequence; the nucleic acid sequence of the LjCYCD2 gene is shown in SEQ ID NO:1.

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