CN111944772A

CN111944772A - Eggplant cryptochrome blue light inhibitor SmBIC1 protein and coding gene

Info

Publication number: CN111944772A
Application number: CN202010763196.5A
Authority: CN
Inventors: 刘杨; 何永军; 陈火英; 张信童
Original assignee: Shanghai Jiaotong University
Current assignee: Shanghai Jiaotong University
Priority date: 2020-07-31
Filing date: 2020-07-31
Publication date: 2020-11-17
Anticipated expiration: 2040-07-31
Also published as: CN111944772B

Abstract

The invention discloses eggplant cryptochrome blue light inhibitor SmBIC1 protein and a coding gene, wherein cDNA of the gene has a nucleotide sequence shown in SEQ ID NO. 1. The amino acid sequence of the gene code is shown in SEQ ID NO. 2. The gene participates in regulating and controlling anthocyanin synthesis and hypocotyl elongation of a photosensitive eggplant, has expression in roots, stems, leaves, flowers, fruit peels, sepals and fruit pulp, and has an expression level remarkably higher than that of other tissues in anthocyanin-enriched parts. The method provides a theoretical basis for improving the quality of eggplants by utilizing a genetic engineering technology in the future, and has great application value.

Description

Eggplant cryptochrome blue light inhibitor SmBIC1 protein and coding gene

Technical Field

The invention belongs to the technical field of biology, relates to key enzymes in an eggplant optical signal path and coding genes thereof, and particularly relates to an eggplant cryptochrome blue light inhibitor SmBIC1(blue light inhibitor 1 of cryptochromes) protein and a coding gene thereof.

Background

BIC1 is an cryptochrome blue light inhibitor in plants, and can interact with blue light receptors CRY1 and CRY2 to regulate and control plant photomorphogenesis. It is found that under dark conditions, the CRYs exists as an inactive monomer, and light changes the CRYs into an active dimer, thereby triggering a series of photomorphogenesis, while the BIC1 protein can maintain the photosensitivity of cells by inhibiting the dimerization of the CRYs, thereby participating in the regulation of photomorphogenesis of plants.

BIC1 can regulate plant photomorphogenesis and transcription factors in anthocyanin synthesis process, such as HY5, TT8, MYB1, LZF1, HFR1, CO, etc. BIC1 regulates elongation of hypocotyl of Arabidopsis seedlings by inhibiting HY5, LZF1 and HFR1, regulates accumulation of eggplant anthocyanin by inhibiting HY5, TT8 and MYB1, and regulates photoperiod flowering of Arabidopsis by regulating protein stability of CO.

BIC1 regulates the photomorphogenesis of plants, how does light regulate BIC 1? Research shows that the transcription of BIC1 is not only regulated by blue light, red light and far-red light can promote the expression of BIC1, and red light, far-red light receptor and blue light receptor can regulate the expression of HY5 through COP, so the light is supposed to regulate the transcription of BIC1 through HY 5.

At present, the BIC1 gene was cloned only in Arabidopsis thaliana and functionally analyzed. Eggplants are not the most important vegetable crops rich in anthocyanin, but the related research is relatively lagged. At present, no literature report related to eggplant SmBIC1 gene and protein coded by the gene is available.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to fill the blank of eggplant SmBIC1 gene and provides eggplant cryptochrome blue light inhibitor SmBIC1 protein and a coding gene. The invention provides a cDNA and an amino acid sequence of eggplant SmBIC 1; further, the invention provides an expression pattern of eggplant SmBIC1 gene in different tissues and organs. The invention also provides a result of the phenotype change after SmBIC1 is transferred into arabidopsis and eggplant.

The purpose of the invention is realized by the following technical scheme:

in a first aspect, the invention provides an eggplant cryptochrome blue light inhibitor SmBIC1 protein, which comprises an amino acid sequence shown as SEQ ID No. 2.

In a second aspect, the invention provides a gene for coding eggplant cryptochrome blue light inhibitor SmBIC1 protein, wherein the cDNA sequence of the gene comprises:

(a) a base sequence shown as 1st to 420 th positions in SEQ ID NO. 1; or

(b) A base sequence having at least 70% homology with the base sequences represented by the 1st to 420 th positions represented by SEQ ID NO. 1; or

(c) A base sequence capable of hybridizing with the base sequences shown in 1st to 420 th positions of SEQ ID NO. 1.

As an embodiment of the invention, the cDNA sequence comprises deletion, insertion and/or substitution of 1-90 nucleotides in the nucleic acid sequence shown in the 1 st-420 th position of SEQ ID NO.1, and a sequence formed by adding 60 nucleotides below at the 5 'and/or 3' end.

In a third aspect, the invention provides a primer pair for amplifying the gene of the eggplant cryptochrome blue light inhibitor SmBIC1 protein, wherein the base sequences of the primer pair are shown as SEQ ID No.3 and SEQ ID No. 4.

In a fourth aspect, the invention provides a primer pair for gene fluorescence quantitative PCR analysis of coding eggplant cryptochrome blue light inhibitor SmBIC1 protein, wherein the base sequence of the primer pair is shown as SEQ ID NO.5 and SEQ ID NO. 6.

In a fifth aspect, the invention provides application of the gene for coding eggplant cryptochrome blue light inhibitor SmBIC1 protein in regulating and controlling hypocotyl elongation of plants through genetic engineering.

In a sixth aspect, the invention provides application of the gene for coding eggplant cryptochrome blue light inhibitor SmBIC1 protein in genetic engineering regulation and control of plant anthocyanin synthesis.

As an embodiment of the present invention, the plant includes arabidopsis thaliana, eggplant.

In the present invention, the term "coding sequence of SmBIC1 gene" refers to the nucleotide sequence from position 1 to 420 shown in SEQ ID NO. 1. The term also includes variants of the sequence shown in SEQ ID No.1 which encode a protein having the same function as native eggplant SmBIC 1. These variants include (but are not limited to): usually 1-90 nucleotides, and within 60 nucleotides at the 5 'and/or 3' end.

In the present invention, the expression pattern of the eggplant SmBIC1 gene product, i.e. the presence, absence and amount of mRNA transcript of eggplant SmBIC1 gene in cells, can be analyzed by a real-time fluorescent quantitative PCR method.

In addition, the eggplant SmBIC1 nucleotide sequence and the amino acid sequence can be used for screening eggplant SmBIC1 related homologous genes or homologous proteins on the basis of nucleic acid homology or expressed protein homology.

To obtain a lattice of genes related to eggplant SmBIC1, an eggplant cDNA library is screened using DNA probes under low stringency conditions³²P is obtained by radioactive labeling all or part of eggplant SmBIC 1. A suitable cDNA library for screening is a library from eggplant. Methods for constructing cDNA libraries from cells or tissues of interest are well known in the field of molecular biology. In addition, many such cDNA libraries are also commercially available, for example, from Clontech, Stratagene, Palo Alto, Calif. This screening method allows to identify the nucleotide sequences of the gene family associated with eggplant SmBIC 1.

The full-length eggplant SmBIC1 related nucleotide sequence or a fragment thereof can be obtained by a PCR amplification method, a recombination method or an artificial synthesis method. For PCR amplification, primers can be designed based on the nucleotide sequences disclosed herein, particularly open reading frame sequences, and the sequences can be amplified using commercially available cDNA libraries or cDNA libraries prepared by conventional methods known to those skilled in the art as templates. When the sequence is long, two or more PCR amplifications are often required, and then the amplified fragments are spliced together in the correct order.

When the sequence of interest is obtained, the sequence of interest can be obtained in large quantities by recombinant methods. This is usually done by cloning it into a vector, transferring it into a cell, and isolating the relevant sequence from the propagated host cell by conventional methods.

Furthermore, mutations can also be introduced into the protein sequences of the invention by chemical synthesis.

Light signals can regulate many secondary metabolic pathways, such as anthocyanin synthesis. Eggplants are widely planted vegetable crops, and the peel of purple eggplants contains rich anthocyanin. Recent research results show that the antioxidant health-care effect of purple eggplant is the best in common vegetable crops.

Compared with the prior art, the invention has the following beneficial effects:

1. the SmBIC1 gene is over-expressed in wild arabidopsis thaliana and eggplants, so that the elongation of hypocotyls of the arabidopsis thaliana and the eggplants can be promoted, and the synthesis of anthocyanin is inhibited;

2. aiming at the current situation that the research foundation of eggplants is weak at present, the key gene SmBIC1 in an optical signal path is cloned, so that a theoretical basis is provided for improving the plant quality by utilizing a genetic engineering technology and obtaining a medicine or food with high oxidation resistance in the future, and the method has great application value.

Drawings

Other features, objects and advantages of the invention will become more apparent upon reading of the detailed description of non-limiting embodiments with reference to the following drawings:

FIG. 1 shows the results of amino acid sequence comparison (FASTA) of eggplant SmBIC1 protein of the present invention and Arabidopsis BIC1 protein, wherein the same amino acids are marked in black between the two sequences and the CID domain interacting with the blue light receptor CRY is marked by the horizontal line;

FIG. 2 shows the expression of the eggplant SmBIC1 gene in different tissues;

FIG. 3 shows RT-PCR detection of SmBIC1 transgenic Arabidopsis plants;

FIG. 4 shows phenotypic changes of wild type Arabidopsis seedlings transformed with SmBIC1 gene;

FIG. 5 shows the hypocotyl change of SmBIC1 transgenic wild type Arabidopsis seedlings;

FIG. 6 shows the change of anthocyanin synthesis in wild type Arabidopsis seedlings transformed with SmBIC1 gene;

FIG. 7 shows the phenotypic changes of eggplant seedlings transformed with SmBIC1 gene;

FIG. 8 shows the hypocotyl changes of eggplant seedlings transformed with SmBIC1 gene;

FIG. 9 shows the change of anthocyanin synthesis in eggplant seedlings transformed with SmBIC1 gene;

FIG. 10 shows the changes of the major structural genes for anthocyanin synthesis in eggplant seedlings transformed with SmBIC1 gene.

Detailed Description

The present invention will be described in detail with reference to specific examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that variations and modifications can be made by persons skilled in the art without departing from the spirit of the invention. All falling within the scope of the present invention.

Experimental procedures without specific conditions noted in the following examples, generally followed by conventional conditions, such as molecular cloning in Sambrook et al: the conditions described in the Laboratory Manual (New York: Cold Spring Harbor Laboratory Press,1989), or according to the manufacturer's recommendations.

Example 1 cloning of eggplant SmBIC1 Gene

1. Obtaining plant material

The plant material used in the experiment is eggplant excellent germplasm resource 'blue mountain standing grain solanum torvum'. The experimental material was cultivated in an artificial plastic greenhouse in a Min-river space breeding base in Shanghai. And (4) seedling culture, growth and fruit setting under natural conditions. Collecting leaf of eggplant for extracting RNA.

Extraction of RNA

Total RNA was extracted by TRIzol (TRIzol is available from Biotechnology engineering, Shanghai, Ltd.). The integrity of the RNA was verified by electrophoresis on formaldehyde denaturing gel and the purity and concentration of the RNA was determined on a Spectrophotometer (Thermo Scientific NanODROP1000 Spectrophotometer).

3. Full Length cloning of Gene

Based on eggplant genome database (http:// eggplant.kazusa.or. jp /), the homologous sequence of SmBIC1 is found by utilizing the sequence of Arabidopsis AtBIC1 gene, and a specific primer is designed. Reverse transcription of the extracted RNA (Prime Script II 1st Strand cDNA Synthesis Kit: Takara Bio Inc.) was performed using the first Strand cDNA as a template and a primer

F1(SEQ ID NO.3)：5′-ATGTCAACTATGGATGATGGAGATA-3′

R1(SEQ ID NO.4)：5′-CTATCCACTTTTGATCTCTAGCATT-3′

And carrying out PCR, recovering and connecting the amplified product with an expected length to a Blunt Simple Vector (Beijing all-purpose gold biotechnology limited) Vector, transforming escherichia coli DH5 alpha, screening positive clones by using colony PCR, and sending the positive clones to Shanghai Biotechnology limited for sequencing to obtain a cDNA sequence SEQ ID NO. 1.

Example 2 sequence information of eggplant SmBIC1 Gene

The full-length CDS open reading frame sequence of the novel eggplant SmBIC1 gene is 420bp, and the detailed sequence is shown in SEQ ID NO. 1. Deducing the amino acid sequence of eggplant SmBIC1 according to the CDS open reading frame sequence, wherein the amino acid sequence has 139 amino acid residues in total, the molecular weight is 15.77KDa, the theoretical isoelectric point (pI) is 4.606, and the detailed sequence is shown in a sequence shown in SEQ ID NO. 2.

By comparing the amino acid sequence of the eggplant SmBIC1 encoding protein with the amino acid sequence of the Arabidopsis AtBIC1 protein, it was found that eggplant SmBIC1 and Arabidopsis AtBIC1 also have CID domain that interacts with the blue light receptor CRY as shown in FIG. 1 (SmBIC 1: the amino acid sequence of eggplant; AtBIC 1: the amino acid sequence of Arabidopsis AtBIC 1; CID domain: the domain that interacts with CRY). Thus, the eggplant SmBIC1 gene has a similar structural domain with the Arabidopsis AtBIC1 gene.

Example 3 expression of eggplant SmBIC1 Gene in different tissues

1. Obtaining plant material

Collecting root, stem, leaf, flower, sepal, peel and pulp of eggplant at the mature period of fruit, wrapping the sample with aluminum-platinum paper, immediately putting into liquid nitrogen, and storing in a-80 deg.C ultra-low temperature refrigerator for later use.

Extraction of RNA

Total RNA was extracted by TRIzol (TRIzol is available from Biotechnology engineering, Shanghai, Ltd.). Integrity was checked by normal agarose gel electrophoresis (gel concentration 1.2%; 0.5 × TBE electrophoresis buffer; 150v, 15 min). The maximum rRNA brightness in the electrophoretic band should be 1.5-2.0 times the brightness of the second rRNA, otherwise it represents degradation of the rRNA sample. RNA of better purity, A₂₆₀/A₂₈₀And A₂₆₀/A₂₃₀About 2.0 or so. OD was measured with a spectrophotometer and RNA content was calculated.

Obtaining of cDNA

500ng of total RNA was used as a template in accordance with TaKaRa PrimeScript, Takara Shuzo Co., Ltd^TMThe RT reagent Kit Perfect Real Time Kit operation instruction carries out reverse transcription to obtain cDNA for later use.

4. Specific primers are designed to carry out real-time fluorescent quantitative PCR analysis on the expression quantity of the genes in different tissues. Designing a specific primer for quantitative analysis of SmBIC1 gene in Real-time PCR by utilizing primer design software according to the obtained eggplant SmBIC1 gene sequence,

SmBIC1-F(SEQ ID NO.5)：5′-GATGAAGGCAAATCAAGGTGAATGT-3′

SmBIC1-R(SEQ ID NO.6)：5′-CTGTTTCATCTTCATCATCCACCAT-3′

the reference gene is Actin (GU984779.1), and the primer is

SmACTIN-F(SEQ ID NO.7)：5′-GTCGGAATGGGACAGAAGGATG-3′

SmACTIN-R(SEQ ID NO.8)：5′-GTGCCTCAGTCAGGAGAACAGGGT-3′

5. A standard curve of the target gene and the reference gene is prepared. Using EASY Dilution (provided by a kit) to carry out gradient Dilution on a standard cDNA solution, then respectively using the diluted cDNA solution as a template, carrying out Real-time PCR amplification by using specific primers of a target gene and an internal reference gene, and drawing a dissolution curve and a standard curve after the reaction is finished. Analyzing the dissolution curve, judging whether the dissolution curves of the target gene and the internal reference gene obtain a single peak or not, and judging whether a single PCR amplification product can be obtained by using the primer or not. The appropriate dilution of the template cDNA was determined by standard curve.

6. And (3) carrying out real-time fluorescence quantitative analysis on the target gene in the sample to be detected. The first strand of the synthesized cDNA is taken as a template, the specific primers of the target gene and the reference gene are respectively used for amplification to carry out fluorescent quantitative analysis, the Real-time PCR reaction is carried out on an FTC-3000 Real-time quantitative instrument, and the reaction system is 20 mu L. The reaction was performed in three steps, denaturation at 95 ℃ for 1min, followed by 40 cycles: 30s at 95 ℃; 30s at 58 ℃; 45s at 72 ℃. After each amplification, a melting curve was made to examine whether the amplified product was specifically produced.

7. By using 2^-△△CtThe method is used for relative quantitative analysis. The results show that the SmBIC1 gene is expressed in the root, stem, leaf, flower, sepal, peel and pulp of eggplant, wherein the expression level is highest in the flower and is next to the leaf, and the expression level is lower in the pulp, which indicates that the SmBIC1 gene is higher in the anthocyanin synthesis part (figure 2).

Example 4 functional verification of eggplant SmBIC1 Gene transfer to Arabidopsis thaliana

The transformation vector used was PHB, and the Arabidopsis material was a wild-type (Columbia) plant. Culturing Agrobacterium containing SmBIC1-PHB to OD 0.8-2.0, centrifuging at 6000rpm for 5min, discarding supernatant, resuspending thallus precipitate with MS liquid (purchased from Shanghai Shaoxing Biotech Co., Ltd.), and adjusting OD₆₀₀0.8-1.2, infecting arabidopsis inflorescence for 10-60 sec, performing dark culture for 12h, performing normal culture, and collecting mature seeds.

Collected T₀Transgenic seeds are laid on a plate containing 50mg/L hygromycin resistance 1/2MS (purchased from Shanghai Shaoxing Biotechnology Co., Ltd.), vernalized for 3d at 4 ℃, cultured for 6-10d in an illumination incubator, selected from robust seedlings with long roots and true leaves, transplanted to a soil pot, and cultured. Collected T₁The seeds were further plated on 1/2MS plates containing 50mg/L hygromycin resistance, and resistance was selected: collection of T-reactive strains with a non-resistance ratio of 3:1₂And (5) seed generation. T is₂Pure and resistant plants were selected with a 100% survival rate on 50mg/L hygromycin-resistant plates. The expression level of SmBIC1 was detected by Real-time PCR on the obtained strain (FIG. 3). The Arabidopsis actin (NM-179953) primers used were as follows:

SEQ ID NO.9：5′-GTCTGGATTGGAGGGTC-3′

SEQ ID NO.10：5′-TGAGAAATGGTCGGAAA-3′

the positive transformants obtained by the screening were cultured together with the wild type in 8h of dark/16 h of light. The results show that the BIC1-OE transgenic plant has a longer hypocotyl compared with the wild type (FIGS. 4 and 5) and the anthocyanin content of the hypocotyl is also obviously lower than that of the wild type (FIGS. 4 and 6), which indicates that the SmBIC1 gene has the functions of promoting the elongation of the hypocotyl and inhibiting the synthesis of anthocyanin.

Example 5 functional verification of eggplant SmBIC1 Gene transfer to Arabidopsis thaliana

The transformation vector is PHB, and the eggplant material is blue mountain standing grain solanum torvum. Culturing Agrobacterium containing SmBIC1-PHB to OD 0.8-2.0, centrifuging at 6000rpm for 5min, discarding supernatant, resuspending thallus precipitate with MS liquid, adjusting OD₆₀₀And (3) infecting the explants which are pre-cultured for 2d for 15min, and co-culturing (dark culture) for 2d after absorbing surface water. Washing explant with sterile water for 4 times, soaking in 300mg/L Cb carbenicillin solution (CAS: 4800-94-6, Shanghai Biotech Co., Ltd.) for 30min, and inoculating to first recovery medium (MS + zeatin (2.0 mg. L.))^-1) + carbenicillin (300 mg. L)^-1) + agar powder (7.0 g.L)^-1) On the surface, screening culture (MS + zeatin (2.0 mg. L.) was performed after 7 days of culture^-1) + carbenicillin (300 mg. L)^-1) + hygromycin (50 mg. L)^-1) + agar powder (7.0 g.. timesL^-1)). 14d subculture (MS + zeatin (2.0 mg. L.)^-1) + carbenicillin (300 mg. L)^-1) + hygromycin (50 mg. L)^-1) + agar powder (7.0 g.L)^-1)1 time, screening and culturing (MS + zeatin (2.0 mg. L))^-1) + carbenicillin (300 mg. L)^-1) + hygromycin (50 mg. L)^-1) + agar powder (7.0 g.L)^-1)42 d) when the explant forms adventitious buds and has true leaves, the base thereof is excised and transferred to a rooting medium (MS + auxin (0.1 mg. L.)^-1) + agar powder (7.0 g.L)^-1)). And (3) opening the bottle mouth for 2d when the main root of the regenerated plant grows to 3-5cm, washing the root, transplanting the plant into a nutrition pot, placing the plant into a culture box, and transferring to a greenhouse for culture after the plant grows well.

The positive plants successfully transformed were identified by PCR and were selfed three generations consecutively to obtain homozygous transgenic lines, and the expression level of SmBIC1 was detected by Real-time qPCR on the obtained homozygous lines (FIG. 10). Eggplant actin (GU984779.1) primers used were as follows:

SEQ ID NO.9：5′-GTCGGAATGGGACAGAAGGATG-3′

SEQ ID NO.10：5′-GTGCCTCAGTCAGGAGAACAGGGT-3′

homozygous positive transformants were incubated with wild type for 14 days in 8h dark/16 h white light. The result shows that the BIC1-OE transgenic plant has a longer hypocotyl (FIGS. 7 and 8) compared with the wild type, the anthocyanin content of the hypocotyl is also obviously lower than that of the wild type (FIGS. 7 and 9), and meanwhile, the expression quantity of major structural genes SmCHS, SmDFR and SmANS for anthocyanin synthesis is detected by adopting Real-time qPCR (polymerase chain reaction), and the expression quantity of the structural genes in the BIC1-OE transgenic plant is found to be obviously lower than that of the wild type (FIG. 10), which shows that the SmBIC1 gene has the functions of promoting elongation of the hypocotyl and inhibiting anthocyanin synthesis.

The SmCHS primers for synthesizing the structural gene of the eggplant anthocyanin are as follows:

SEQ ID NO.11：5′-GGGAACAGTACTCCGGCTAGCC-3′

SEQ ID NO.12：5′-AACACCTGAAATTGGGTCTGAACCA-3′

the primers for synthesizing the structural gene SmDFR of the eggplant anthocyanin are as follows:

SEQ ID NO.13：5′-CATTGAGACTTGCCGACAGA-3′

SEQ ID NO.14：5′-ATTCTCCTTGCCACTTGCAT-3′

the SmANS primers for the anthocyanin synthesis structural gene of the eggplant are as follows:

SEQ ID NO.15：5′-CTCGATTCCCACCTCGGACCTT-3′

SEQ ID NO.16：5′-TCAGCTGCAGCGTCCTGTTTGT-3′

the foregoing description of specific embodiments of the present invention has been presented. It is to be understood that the present invention is not limited to the specific embodiments described above, and that various changes and modifications may be made by one skilled in the art within the scope of the appended claims without departing from the spirit of the invention.

Sequence listing

<110> Shanghai university of transportation

<120> eggplant cryptochrome blue light inhibitor SmBIC1 protein and coding gene

<130> DD09801

<160> 16

<170> SIPOSequenceListing 1.0

<210> 1

<211> 420

<212> DNA

<213> eggplant (Solanum Melongena L.)

<400> 1

atgtcaacta tggatgatgg agataaaata gccgaatttt gtgactccgc gacgatgaag 60

gcaaatcaag gtgaatatga atatcaagat tctatcaaga actttggatc attgtcatta 120

atggtggatg atgaagatga aacaggcgat ctttatgttg aatcggagaa attagggtta 180

tcggggcgtg agaagttgaa gaggcattgg agagaagttg gagatagggt ttttgtgcca 240

gaaagatggg agcatgaggg ttctttgaag gaatggatgg attgttcttc ttttgataaa 300

atactagctc caaaggaatt aaaatcagct cgagaagcct tgatgtctca aggaaaacgt 360

gtacgttcaa gttcaggttc agattcaact tcaagaatgc tagagatcaa aagtggatag 420

<210> 2

<211> 139

<212> PRT

<213> eggplant (Solanum Melongena L.)

<400> 2

Met Ser Thr Met Asp Asp Gly Asp Lys Ile Ala Glu Phe Cys Asp Ser

1 5 10 15

Ala Thr Met Lys Ala Asn Gln Gly Glu Tyr Glu Tyr Gln Asp Ser Ile

20 25 30

Lys Asn Phe Gly Ser Leu Ser Leu Met Val Asp Asp Glu Asp Glu Thr

35 40 45

Gly Asp Leu Tyr Val Glu Ser Glu Lys Leu Gly Leu Ser Gly Arg Glu

50 55 60

Lys Leu Lys Arg His Trp Arg Glu Val Gly Asp Arg Val Phe Val Pro

65 70 75 80

Glu Arg Trp Glu His Glu Gly Ser Leu Lys Glu Trp Met Asp Cys Ser

85 90 95

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

100 105 110

Ala Leu Met Ser Gln Gly Lys Arg Val Arg Ser Ser Ser Gly Ser Asp

115 120 125

Ser Thr Ser Arg Met Leu Glu Ile Lys Ser Gly

130 135

<210> 3

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 3

atgtcaacta tggatgatgg agata 25

<210> 4

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 4

ctatccactt ttgatctcta gcatt 25

<210> 5

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 5

gatgaaggca aatcaaggtg aatgt 25

<210> 6

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 6

ctgtttcatc ttcatcatcc accat 25

<210> 7

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 7

gtcggaatgg gacagaagga tg 22

<210> 8

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 8

gtgcctcagt caggagaaca gggt 24

<210> 9

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 9

gtcggaatgg gacagaagga tg 22

<210> 10

<211> 24

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 10

gtgcctcagt caggagaaca gggt 24

<210> 11

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 11

gggaacagta ctccggctag cc 22

<210> 12

<211> 25

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 12

aacacctgaa attgggtctg aacca 25

<210> 13

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 13

cattgagact tgccgacaga 20

<210> 14

<211> 20

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 14

attctccttg ccacttgcat 20

<210> 15

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 15

ctcgattccc acctcggacc tt 22

<210> 16

<211> 22

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 16

tcagctgcag cgtcctgttt gt 22

Claims

1. Eggplant cryptochrome blue light inhibitor SmBIC1 protein, which is characterized by comprising an amino acid sequence shown as SEQ ID NO. 2.

2. A gene encoding the eggplant cryptochrome blue light inhibitor SmBIC1 protein as claimed in claim 1, wherein the nucleotide sequence of the gene comprises:

(a) a base sequence shown as 1st to 420 th positions in SEQ ID NO. 1; or

3. The nucleotide sequence of a gene encoding eggplant cryptochrome blue light inhibitor SmBIC1 protein as claimed in claim 2, which comprises deletions, insertions and/or substitutions of 1-90 nucleotides in the base sequence shown in 1 st-420 th position of SEQ ID NO.1, and a sequence formed by adding 60 or less nucleotides at the 5 'and/or 3' end.

4. A primer pair for amplifying the gene encoding eggplant cryptochrome blue light inhibitor SmBIC1 protein as claimed in claim 2, wherein the base sequences of the primer pair are shown as SEQ ID No.3 and SEQ ID No. 4.

5. A primer pair for the fluorescent quantitative PCR analysis of the gene encoding eggplant cryptochrome blue light inhibitor SmBIC1 protein as claimed in claim 2, wherein the base sequences of the primer pair are shown as SEQ ID No.5 and SEQ ID No. 6.

6. Use of the gene encoding eggplant cryptochrome blue light inhibitor SmBIC1 protein as claimed in claim 2 in genetically engineering plants to regulate hypocotyl elongation.

7. Use according to claim 6, wherein said plant comprises Arabidopsis thaliana, eggplant.

8. Use of the gene encoding eggplant cryptochrome blue light inhibitor SmBIC1 protein as claimed in claim 2 in genetic engineering to regulate plant anthocyanin synthesis.

9. Use according to claim 8, wherein said plant comprises Arabidopsis thaliana, eggplant.