FaCP5L1 and FaCP5L3 genes and their application in regulating fruit ripening and postharvest preservation quality.

Abstract

The FaCP5L1 and FaCP5L3 genes and application thereof in regulating fruit ripening and postharvest quality belong to the field of agricultural biological genetic engineering technology. It is found for the first time that the cruciferin coding genes FaCP5L1 and FaCP5L3 can directly participate in the regulation of strawberry ripening and postharvest quality, including but not limited to the regulation of abscisic acid (ABA) content, ripening process, soluble solid content, hardness, color, anthocyanin content. The expression of FaCP5L1 and FaCP5L3 can promote the ripening of strawberry fruit and the formation of postharvest quality; the expression reduction can slow down the ripening of strawberry fruit and the formation of postharvest quality. Accordingly, the ripening of strawberry fruit and the postharvest quality traits can be regulated by reasonably utilizing the genes, and the ripening and postharvest quality related breeding can be guided. The present application provides a theoretical basis and technical support for regulating the ripening of strawberry fruit and the postharvest quality traits by using genetic engineering technology, and has important application value.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of agricultural bioengineering technology, and particularly relates to FaCP5L1 and FaCP5L3 genes and their application in regulating fruit ripening and postharvest quality. BACKGROUND

[0002] Strawberry (Fragaria x ananassa Duch.) is one of the most important berry crops in the world due to its bright color, unique aroma and rich nutrition. The maturity and quality of strawberry fruits, including color, soluble solids content and firmness, are important factors for consumer selection and market demand. Therefore, the regulation of strawberry ripening and postharvest quality has important research significance and practical application value.

[0003] Cruciferin PGCRURSE5-like (CP5L) is a type of seed storage protein. Previous studies have shown that this type of protein accumulates in large quantities during seed development and provides amino acid nutrition as the main nitrogen and sulfur source for seed germination and early seedling growth. There is no literature report that CP5L is directly involved in fruit quality regulation. In the early stage of the present application, it was found that the CP5L homologous protein coding genes FaCP5L1 and FaCP5L3 in strawberry fruits are regulated by the DNA methylation maintenance pathway core protein FaCMT3 and are involved in the regulation of strawberry fruit ripening and postharvest quality. The present application takes FaCP5L1 and FaCP5L3 in strawberry as the research object and finds that they can positively regulate strawberry ripening and postharvest quality formation.

[0004] Genetic engineering methods such as gene editing or transgenic technology can regulate the expression of FaCP5L1 and FaCP5L3, which may provide an effective molecular tool for strawberry quality improvement. Based on the regulatory role of FaCP5L1 and FaCP5L3 genes and their encoded proteins, the present application proposes a new method for regulating strawberry fruit ripening and postharvest, which aims to control the ripening of strawberry fruits and the formation of postharvest quality by regulating the expression of cruciferin protein coding genes FaCP5L1 and FaCP5L3. This method not only helps to enhance the market competitiveness of strawberry, but also provides an innovative genetic engineering solution for realizing the quality upgrading and variety improvement of strawberry. SUMMARY

[0005] In view of the problems existing in the prior art, the purpose of the present application is to design a technical solution of FaCP5L1 and FaCP5L3 genes and their application in regulating fruit ripening and postharvest quality.

[0006] The present application specifically adopts the following technical solutions to achieve the above purposes:

[0007] The application provides a strawberry fruit ripening and postharvest quality regulation gene, which comprises a FaCP5L1 gene and / or a FaCP5L3 gene, wherein the FaCP5L1 gene comprises the following nucleotide sequences (A1), (A2), (A3) or (A4):

[0008] (A1) the nucleotide sequence shown in SEQ ID NO. 1;

[0009] (A2) a nucleotide sequence obtained by substituting, deleting and / or adding one or more bases of the nucleotide sequence shown in SEQ ID NO. 1, and having the same function;

[0010] (A3) a nucleotide sequence with more than 70% identity to the nucleotide sequence shown in SEQ ID NO. 1, and having the same function;

[0011] (A4) a nucleotide sequence obtained by connecting a tag coding sequence to the 5' end and / or 3' end of any one of the sequences defined in (A1) to (A3);

[0012] The FaCP5L3 gene comprises the following nucleotide sequences (a1), (a2), (a3) or (a4):

[0013] (a1) the nucleotide sequence shown in SEQ ID NO. 2;

[0014] (a2) a nucleotide sequence obtained by substituting, deleting and / or adding one or more bases of the nucleotide sequence shown in SEQ ID NO. 2, and having the same function;

[0015] (a3) a nucleotide sequence with more than 70% identity to the nucleotide sequence shown in SEQ ID NO. 2, and having the same function;

[0016] (a4) a nucleotide sequence obtained by connecting a tag coding sequence to the 5' end and / or 3' end of any one of the sequences defined in (a1) to (a3).

[0017] The application provides a protein encoded by the above-mentioned strawberry fruit ripening and postharvest quality regulation gene, which comprises the following amino acid sequences (B1), (B2), (B3) or (B4):

[0018] (B1) the amino acid sequence shown in SEQ ID NO. 5;

[0019] (B2) a protein obtained by substituting, deleting and / or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID NO. 5, and having the same function;

[0020] (B3) An amino acid sequence that is more than 70% identical to the amino acid sequence shown in SEQ ID NO.5 and has the same function;

[0021] (B4) The amino acid sequence obtained by attaching a protein tag to the N-terminus and / or C-terminus of any of the proteins defined in (B1) to (B3).

[0022] A third aspect of this invention provides a protein encoding the aforementioned strawberry fruit ripening and postharvest preservation quality regulation gene, the protein containing the amino acid sequence described in (b1), (b2), (b3), or (b4):

[0023] (b1) The amino acid sequence as shown in SEQ ID NO. 6;

[0024] (b2) Proteins with the same function but with one or more amino acid residues substituted and / or deleted and / or added, as shown in SEQ ID NO.6;

[0025] (b3) An amino acid sequence that is more than 70% identical to the amino acid sequence shown in SEQ ID NO.6 and has the same function;

[0026] (b4) The amino acid sequence obtained by attaching a protein tag to the N-terminus and / or C-terminus of any of the proteins defined in (b1) to (b3).

[0027] The fourth aspect of this invention provides the application of the above-mentioned gene in regulating strawberry fruit ripening and postharvest preservation quality.

[0028] Furthermore, the regulation of strawberry fruit ripening and post-harvest preservation quality specifically includes at least one of the following:

[0029] (1) Regulating the abscisic acid content in strawberry fruits;

[0030] (2) Regulating the ripening process of strawberry fruits;

[0031] (3) Regulate the soluble solids content of strawberry fruit;

[0032] (4) Regulate the firmness of strawberry fruits;

[0033] (5) Regulating the color of strawberry fruits;

[0034] (6) Regulate the anthocyanin content of strawberry fruit.

[0035] The fifth aspect of this invention provides the application of the above-mentioned gene as a target in the breeding process related to strawberry fruit ripening and postharvest preservation quality.

[0036] The sixth aspect of this invention provides a method for positively regulating strawberry fruit ripening and postharvest preservation quality, comprising:

[0037] Increase the expression level of FaCP5L1 or FaCP5L3 genes in strawberries; or

[0038] To increase the activity of proteins encoded by the FaCP5L1 or FaCP5L3 genes in strawberries; or

[0039] Increase the content of proteins encoded by the FaCP5L1 or FaCP5L3 genes in strawberries.

[0040] The seventh aspect of this invention provides a method for negatively regulating strawberry fruit ripening and postharvest preservation quality, comprising:

[0041] Knock out the FaCP5L1 or FaCP5L3 gene in strawberries; or

[0042] Silent FaCP5L1 or FaCP5L3 genes in strawberries; or

[0043] Reduce the activity of proteins encoded by the FaCP5L1 or FaCP5L3 genes in strawberries; or

[0044] Reduce the content of proteins encoded by the FaCP5L1 or FaCP5L3 genes in strawberries.

[0045] The beneficial effects of this invention are:

[0046] This invention is the first to discover that FaCP5L1 and FaCP5L3 can positively regulate the ripening and postharvest ripening rate of strawberry fruits, affecting characteristics such as ABA content, color accumulation, anthocyanin content, firmness, and soluble solids content. Inhibiting FaCP5L1 and FaCP5L3 expression suppresses the strawberry ripening process; promoting FaCP5L1 and FaCP5L3 expression accelerates the ripening process. By rationally utilizing these genes, the ripening process of strawberry fruits and the formation of postharvest preservation quality can be regulated, providing important theoretical basis and technical means for the regulation of strawberry quality and preservation quality and for molecular breeding, thus possessing significant application value. Attached Figure Description

[0047] Figure 1 Phylogenetic analysis of strawberry FaCP5L homologous proteins with carrot (RsCP5L) and potato (StCP5L).

[0048] Figure 2 FaCP5L1 and FaCP5L3 overexpression vector backbones.

[0049] Figure 3 The relative expression levels of FaCP5L1 and FaCP5L3 in fruits overexpressing FaCP5L1 and FaCP5L3.

[0050] Figure 4FaCP5L1 and FaCP5L3 silencing vector backbone.

[0051] Figure 5 Relative expression levels of FaCP5L1 and FaCP5L3 in FaCP5L1 and FaCP5L3 silenced fruits.

[0052] Figure 6 ABA content (a) of fruits transformed by overexpression and silencing of FaCP5L1 and FaCP5L3; soluble solids content (b); firmness (c); color chromaticity angle (d); anthocyanin content (e). Detailed Implementation

[0053] The present invention will be further illustrated below with reference to specific embodiments, but these are not intended to limit the scope of the invention. Unless otherwise specified, the reagents, consumables, etc., used in the embodiments are all commercially available, and the embodiments are all conducted according to conventional experimental methods or the manufacturer's instructions. In the quantitative experiments in the following embodiments, three replicate experiments were performed, and the results were averaged.

[0054] Example 1: Phylogenetic analysis of the strawberry CP5L gene

[0055] Using the CDS sequences of CP5L (RsCP5L in carrot (Raphanus sativus) and StCP5L in potato (Solanum tuberosum)) with high-level annotation in the Uniprot protein database and prior research in the literature, a local BLAST was performed on the octoploid strawberry genome “Royal Royce 1” using TBtools, with a p-value threshold set to 1×e. -100 The amino acid sequences corresponding to the candidate FaCP5L-CDS were compared with those of SrCP5L and StCP5L using MEGA 7.0 software via ClustalW. Neighbor-Joining phylogenetic analysis was then performed, and the bootstrap method was used to obtain the phylogenetic tree. The No. of bootstrap replications parameter was set to 1,000, the image mode was selected as Displayonly topology, and other parameters were left at their default settings. The results are shown below. Figure 1 .

[0056] The analysis results show that this invention identified four possible CP5L homologs in strawberry (Fxa2Ag102170, Fxa2Bg202008, Fxa2Cg202928, and Fxa2Dg201871). Notably, the mRNA length of Fxa2Ag102170 is 7,204 bp, while the mRNA lengths of the other homologs are only 1,926–1,943 bp. Furthermore, in the original annotation of the IGV genome visualization based on RNA-seq abundance, it was found that the original annotation of the Fxa2Ag102170 mRNA contained nine exons, while the other homologs each had only four exons; and no reads were aligned to the fifth exon of the Fxa2Ag102170 mRNA. Based on these significant differences compared to the other homologs, this invention concludes that the original annotation of the Fxa2Ag102170 gene may contain certain errors. To this end, the present invention conducted a 3' RACE experiment to detect the true sequence of the 3' end of the mRNA in strawberry. The results showed that, unlike the original annotation, a TAA codon corresponding to a triplet codon position actually exists at the end of the 4th exon of the Fxa2Ag102170 mRNA. The first four exons constitute an independent CP5L transcript (CDS sequence is sequence 1 in this invention). The 5th exon is presumed to be a false positive annotation, and the last four exons also constitute an independent transcript, which is annotated as seed storage protein CRA1 in the nr protein assay. Based on this, the present invention partially corrects the annotation of the Fxa2Ag102170 gene and its corresponding mRNA, and provides the CDS sequence of FaCP5L1 (as shown in SEQ ID NO.1), the encoded protein of FaCP5L1 as shown in SEQ ID NO.5; and provides the CDS sequence of FaCP5L3 (as shown in SEQ ID NO.2), the encoded protein of FaCP5L3 as shown in SEQ ID NO.6.

[0057] Example 2: Cloning and overexpression of strawberry FaCP5L1& and FaCP5L3 gene CDS in fruit

[0058] Cloning and vector construction: Primers (F-ATGGCCAGGTCTTTAGCTCTCT; R-TTAAGCACTAGCCCTCTCGGAC) were designed to clone the CDS sequences of FaCP5Ls. Sequencing was performed to distinguish between FaCP5L1-CDS and FaCP5L3-CDS, and the cloned CDS sequences were then ligated into the pAEnTopo vector. The pAEnTopo vector containing FaCP5L1 & 3-CDS sequences was then... Figure 2The recombinant plasmid pCambia-FlagN, which is resistant to kanamycin, was subjected to a displacement reaction (incubated at 50°C for 1 h) to obtain pCambia-FaCP5L1&3 recombinant plasmid (where the Flag tag is 2×Flag). This plasmid was transformed into competent DH5α Escherichia coli cells. Single colonies were picked from resistant solid LB medium containing 50 mg / mL kanamycin for colony PCR screening. Positive clones were then sequenced for verification. After successful alignment, the recombinant plasmid was extracted.

[0059] Recombinant plasmid transformation: The successfully ligated pCambia-FaCP5L1&3 plasmid (and the pCambia empty vector without FaCP5L1&3-CDS) were transformed into GV3101 Agrobacterium competent cells. The transformed bacterial culture was plated on LB agar containing 50 μg / mL kanamycin and 25 μg / mL rifampin antibiotics and incubated upside down at 28°C for 2–3 days. Single colonies were picked and positive clones were screened by colony PCR for sequencing verification. After successful sequencing, a portion of the corresponding bacterial culture was mixed with 50% glycerol at a 1:1 ratio and stored at -80°C.

[0060] Infecting strawberry fruits: Resuspend the recombinant plasmid Agrobacterium to OD600 of 0.8–1.0 using a pre-prepared infection solution (1% sucrose, 10 mM MES, 10 mM MgCl2, 150 μM acetylsylcholine, pH = 5.6), and then let it stand at room temperature for 2 hours. Inject the solution into strawberry fruits from the green to white ripe stage until the infection solution overflows onto the fruit surface. Take samples and flash-freeze them in liquid nitrogen 4 days later.

[0061] Real-time quantitative PCR (qRT-PCR) detection: Total RNA was extracted from empty vector and overexpression fruits. The quality-tested RNA solution was reverse transcribed into cDNA in a one-step process, simultaneously removing genomic gDNA and synthesizing first-strand cDNA. Based on the experimental design and calculations, a 15 μL reaction system was prepared in a 200 μL 96-well PCR plate, including SYBR qPCR Master Mix, ultrapure water, the cDNA obtained in the previous step, and upstream and downstream primer pairs. The qRT-PCR experiment was performed using the standard three-step ROX dye qRT-PCR mode. FaCP5L1 primers: upstream primer 5'-3'cttgccgatgctctcaacatcga; downstream primer 5'-3'tgtcgtccttgttgttgtctttcc. FaCP5L3 primers: upstream primer 5'-3'ctcgccgatgctctcaacattga; downstream primer 5'-3'cttgttgtccttgaagttgtcttt. Internal reference FaUBQ primers: upstream primer 5'-3'cagaccagcagaggcttatctt; downstream primer 5'-3'ttctggatattgtagtctgctaggg.

[0062] The results are as follows Figure 3 The display shows that FaCP5L1 ( Figure 3 a) and FaCP5L3 ( Figure 3 b) The overexpression fruit model was successfully constructed, with the average overexpression fold reaching more than 25 times that of the empty vector control group, and significantly higher than that of the empty vector control group, indicating that the overexpression effect of the target gene is outstanding.

[0063] Example 3: Strawberry FaCP5L1& and FaCP5L3 gene silencing experiment

[0064] Vector construction: First, primers were designed (FaCP5L1 upstream primer 5'-3'AATTGGACGCCAACAAGGCCAA; FaCP5L1 downstream primer 5'-3'TTGTTGTTGTCTTTCCTCCCTCATT; FaCP5L3 upstream primer 5'-3'AAAGTGCAACCCATCAAGGAAGGT; FaCP5L3 downstream primer 5'-3'CTTGTTGTCCTTGAAGTTGTCTTT). An RNAi fragment for RNA interference (RNAi) was cloned into the pAEnTopo vector ligated with FaCP5L1&3-CDS and ligated into the pAEnTopo vector. The pAEnTopo vector ligated with the RNAi fragment was then subjected to a displacement reaction with the pCambia-RNAi vector containing kanamycin resistance (incubated at 50℃ for 1 h) to obtain the pCambia-RNAi-FaCP5L1&3 fragment recombinant plasmid. Figure 4 The cells were transformed into competent DH5α *E. coli* cells. Single colonies were picked from LB broth containing 50 mg / mL kanamycin and colony PCR was performed to screen for positive clones. Sequencing verification was then performed (colony PCR and sequencing primers: upstream primer 5'-3'gtcgaacatgaataaacaaggt; downstream primer 5'-3'aatcatcgcaagaccggc). After successful alignment, the recombinant plasmid was extracted. The sequence of FaCP5L1 RNAi is shown in SEQ ID NO.3, and the sequence of FaCP5L3 RNAi is shown in SEQ ID NO.4.

[0065] Transformation with recombinant plasmid: Transform the successfully ligated pCambia-RNAi-FaCP5L1&3fragment plasmid ( Figure 4 (The pCambia-RNAi empty vector without RNAi fragments) was transformed into GV3101 Agrobacterium competent cells. The transformed bacterial culture was plated on LB agar containing 50 μg / mL kanamycin and 25 μg / mL rifampicin and incubated upside down at 28°C for 2–3 days. Single colonies were picked and positive clones were screened by colony PCR (colony PCR and sequencing primers: upstream primer 5'-3'gtcgaacatgaataaacaaggt; downstream primer 5'-3'aatcatcgcaagaccggc) for sequencing verification. After successful sequencing, a portion of the corresponding bacterial culture was mixed with 50% glycerol at a 1:1 ratio and stored at -80°C.

[0066] The infection of strawberry fruits and the qRT-PCR process were consistent with those in Example 2.

[0067] The results are as follows Figure 5 The display shows that FaCP5L1 (Figure 5 a) and FaCP5L3 ( Figure 5 b) The RNAi-based silent fruit model was successfully constructed, with the average expression level being less than 25% of the empty vector control group, and significantly lower than that of the empty vector control group, indicating that the silencing effect of the target gene is outstanding.

[0068] Example 4: Effects of strawberry FaCP5L1& and FaCP5L3 gene transformation on ripening and quality traits

[0069] Abscisic acid (ABA) content detection: 0.20 g of strawberry powder pre-ground in liquid nitrogen was extracted using 1.5 mL of buffer solution containing methanol, ethyl acetate, and formic acid (50:50:1). The mixture was sonicated and vortexed for 10 min each, and incubated overnight at 4 °C. Subsequently, it was centrifuged at 14,000 rpm for 15 min at 4 °C, and the supernatant was stored at 4 °C. The precipitate was then subjected to the same extraction process twice more to obtain second and third supernatants. These were combined with the first supernatant and filtered through a 0.22 μm filter to obtain the test solution.

[0070] The analyte was detected using a UPLC-qTOF-MS / MS system equipped with a PDA detector and a Vanguard ACQUITY UPLC BEH C18 column (2.1 × 10 mm, 1.7 μm, Waters Corporation, Milford, USA) with a pre-column. The mobile phase was methanol:acetonitrile:0.1% formic acid solution = 20:10:70, the flow rate was 0.25 mL / min, the injection volume was 10 μL, positive ion MRM mode was used, the ion source voltage was +3.0 kV, the ion source temperature was 500 °C, and the curtain gas (nitrogen) pressure was 35 psi. ABA was identified and quantified using the 2H6-ABA internal standard and the corresponding external standard curve, based on peak area and retention time.

[0071] Total soluble solids (TSS) determination: Total soluble solids were determined using an ATAGO PAL-BX portable refractometer (Japan). Four strawberry fruits were collected, diced, and flash-frozen in liquid nitrogen before extraction. The juice was extracted by squeezing the juice tightly through four layers of gauze, and 200 μL was collected for TSS determination.

[0072] Hardness determination: 9. Fruit hardness was determined using a TA-XT2i texture analyzer. Three points were evenly selected on the equatorial plane of the strawberry fruit for measurement. A TA / 2 stainless steel probe was used for the puncture test, with a testing speed of 0.5 mm / s and a testing depth of 6 mm. The peak value of a single probe test was recorded as the fruit hardness.

[0073] Hue angle measurement: Color was measured using a colorimeter. Four points were evenly selected on the equatorial ring of the fruit for sampling. The L* and a* color space parameters were set and recorded during color measurement. The hue angle was then calculated as follows: Hue angle = arctan(b* / a*).

[0074] Anthocyanin content detection: For crude extraction, 1.0 g of strawberry powder pre-ground with liquid nitrogen was first extracted in an ultrasonic bath with 8 mL of acetone / water (80 / 20, v / v) solution containing HCl (0.1 / 10, v / v) for 15 min, and then centrifuged (12000×g, 4℃, 10 min). The process was repeated twice, and the supernatants obtained were combined and diluted to 25 mL to obtain crude extract for subsequent detection.

[0075] To determine the anthocyanin content, the crude extract was diluted 5-fold with a 70 / 30 / 1 (v / v / v) ethanol / water / HCl solution. After standing for 5 minutes, the absorbance was measured at 540 nm using an ELISA reader. A standard curve for pelargonidin-3-O-glucoside was established to quantify the total anthocyanin level, which was ultimately expressed as pelargonidin-3-O-glucoside equivalents.

[0076] Figure 6 The results showed that the FaCP5L1 and FaCP5L3 genes play an important role in the regulation of strawberry fruit ripening and quality. Figure 6 As can be seen, overexpression significantly increased ABA accumulation, while silencing significantly reduced ABA content, indicating that these two genes have a positive regulatory effect on ABA accumulation, and ABA is a key plant hormone that promotes strawberry ripening. Figure 6 b shows that overexpression significantly increased TSS content, indicating that it promoted sugar accumulation, while silencing reduced TSS, reflecting the regulation of fruit sweetness by the target gene. Figure 6 In c, gene overexpression significantly reduced fruit firmness, while silencing led to a significant increase in firmness. This indicates that FaCP5L1 and FaCP5L3 have a negative regulatory effect on cell wall stability or related metabolism. Figure 6 The results showed that overexpression led to a decrease in the hue angle, while gene silencing increased the hue angle, demonstrating the regulation of fruit color. Figure 6The results showed that overexpression significantly increased anthocyanin content, while gene silencing significantly decreased it, further demonstrating the key roles of FaCP5L1 and FaCP5L3 in fruit color formation. Overall, significant changes in the expression levels of FaCP5L1 and FaCP5L3 can regulate multiple quality traits of strawberry fruits, including ABA content, color, anthocyanin content, firmness, and soluble solids content; they can also significantly regulate the fruit ripening process, a conclusion also reached in postharvest experiments. In summary, FaCP5L1 and FaCP5L3 can be applied to the regulation of strawberry fruit ripening and postharvest preservation quality, as well as related breeding work.

[0077] The specific sequences involved in this invention are as follows:

[0078] FaCP5L1 CDS sequence (SEQ ID NO.1)

[0079]

[0080]

[0081] FaCP5L1 RNAi fragment (502bp) (SEQ ID NO.3)

[0082] AATTGGACGCCAACAAGGCCAACAGGGACAACAAGAGAGCTTCGAACAGCTAGACCGACACCAGAAAGTGCGACCCGTCAAGGAAGGTGACGTTATCGCCGTCCCCGCCGGAGTCACAACCTGGTCCTACAACGACGGTGACCAATCTCTTGTTTTCGTCTGTCTTCAAGACACCAGCAACCTTCACAACCAGCTTGACCAAATCCCAAGGCGATTCTATCTTGCTGGTAACCCACAAGATGAGTTCACCCAACAAGGCCAAAGCCGACATTCGATCGCGAGACAACAGGGAAAGATCAGACACCAACAACAGGGAAACAACAACAACAACGTCTTTGCCGGCTTCGACACCAGACTCCTTGCCGATGCTCTCAACATCGACCAACAGACCGCCCAACAGCTCCAGGGACAGAACGACAACAGGCCACAGATCGTTAGGGTTCAGGGACGCCTCGACTTCGTCCAGCCCCCAGAGTCAATGAGGGAGGAAAGACAACAACAA;

[0083] FaCP5L3 RNAi fragment (448bp) (SEQ ID NO.4)

[0084] AAAGTGCAACCCATCAAGGAAGGTGACGTTATCGCCGTCCCAGCCGGAGTCACAACCTGGTCCTACAACGACGGTGACCAATCTCTTGTTTTCGTCTGTCTTCAAGACACCAGCAACCTTCACAACCAACTTGACCAAATCCCAAGGCGATTCTATCTTGCTGGTAACCCACAAGATGAGTTCACCCAACAAGGCCAAAGCCGACATTCGATCGCGAGACAACAGGGAAAGATCAGACGCCAACAACAGGGAAACAACAACAACAACGTCTTTGCCGGCTTCGACACCAGACTCCTCGCCGATGCTCTCAACATTGACCAACAGACAGCCCAACAGCTCCAGGGACAGAACGACAACAGGCCACAGATCGTTAGGGTTCAGGGACGCCTCGACTTCGTCCAGCCCCCAGAGTCAATGAGGGAGGAAAGACAACTTCAAGGACAACAAG;

[0085] Amino acid sequence of FaCP5L1 (SEQ ID NO.5)

[0086] MARSLALSLCLLVLFHGCSAYRQQSPSSQQNECMLNQIQAREPDNRIETEAGRIESWDYNQDDFQCAGVAVQRITVERNGLHLPFYTNSPRLLYVVQGRGQLGMVIPGCPETFEETEQSYEEQLQGSQQQGQGQQGRQQIGRQQGQQGQQESFEQLDRHQKVRPVKEGDVIAVPAGVTTWSYNDGDQSLVFVCLQDTSNLHNQLDQIPRRFYLAGNPQDEFTQQGQSRHSIARQQGKIRHQQQGNNNNNVFAGFDTRLLADALNIDQQTAQQLQGQNDNRPQIVRVQGRLDFVQPPESMREERQQQGRQGQNGFEEVFCHMKMKQNIGKPSMADVFSPQAGRISVLNGLSMPVLRHLRLSAERGFFYNNAIYSPHWNLNAHEVYYVIRGSARVQVVNDNGETILDDQVRQGQLFIVPQNHAVLQKAMSNGYEYIAFKTQDNAIINTLAGRTSVLRALPDVVLANAYQMDRQQARNLKYNRQETVALSSSRSFQSERASA*;

[0087] Amino acid sequence of FaCP5L3 (SEQ ID NO.6)

[0088] MARSLALSLCLLVLFHGCVAYRQQSPSSQQNECMLNQIQAREPDNRIEETEAGGIESWDYNQDDFQCAGVAVQRITVERNGLHLPFYTNTPRLLYVVQGRGQLGMVIPGCPETFEETEQSYEEQLQG SQQQQQGQQGRQQIERRQGQQGQQESFEQLDRHQKVQPIKEGDVIAVPAGVTTWSYNDGDQSLVFVCLQDTSNLHNQLDQIPRRFYLAGNPQDEFTQQGQSRHSIARQQGKIRRQQQGNNNNNVFA GFDTRLLADALNIDQQTAQQLQGQNDNRPQIVRVQGRLDFVQPPESMREERQLQGQQGRQGQNGFEEVFCHMKMKQNIGKPSMADVFSPQAGRISVLNGLSMPVLRHLRLSAERGFFYNNAIYSPH WNLNAHEVYYVIRGSARVQVVNDNGETILDDQVRQGQLFIVPQNHAVLQKAMSNGYEYIAFKTQDNAMINTLAGRTSVLRSLPDVVLANAYQMDRQQARNLKYNRQETVALSSSRSSFQSERASA*.

[0089] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention are within the scope of protection claimed by the present invention.

Claims

1. A gene for regulating ripening and postharvest quality of strawberry fruits, characterized in that, The FaCP5L1 gene is a nucleotide sequence as shown in SEQ ID NO. 1; and the FaCP5L3 gene is a nucleotide sequence as shown in SEQ ID NO.

2.

2. The protein encoded by the gene for controlling ripening and postharvest quality of strawberry fruits according to claim 1, characterized by, The coding protein is a coding protein of FaCP5L1, and the coding protein sequence of FaCP5L1 is an amino acid sequence as shown in SEQ ID NO.

5.

3. The protein encoded by the gene for controlling ripening and postharvest quality of strawberry fruits according to claim 1, characterized by, The coding protein is a coding protein of FaCP5L3, and the coding protein sequence of FaCP5L3 is an amino acid sequence as shown in SEQ ID NO.

6.

4. The gene of claim 1 is applied to regulate strawberry fruit ripening and postharvest quality.

5. The use according to claim 4, wherein the compound is ###0002### The regulation of strawberry fruit ripening and postharvest quality specifically includes at least one of the following: (1) regulating the content of abscisic acid in strawberry fruit; (2) regulating the ripening process of strawberry fruit; (3) regulating the soluble solids content of strawberry fruit; (4) regulating the firmness of strawberry fruit; (5) regulating the color of strawberry fruit; (6) regulating the anthocyanin content of strawberry fruit.

6. The gene of claim 1 is applied as a target in the breeding process related to strawberry fruit ripening and postharvest quality.

7. A method of positively regulating fruit ripening and postharvest quality in strawberry, characterized by, including: increasing the expression level of FaCP5L1 or FaCP5L3 gene in strawberry; or increasing the activity of the coding protein of FaCP5L1 or FaCP5L3 gene in strawberry; or increasing the content of the coding protein of FaCP5L1 or FaCP5L3 gene in strawberry.

8. A method for negatively regulating fruit ripening and postharvest quality in strawberry, characterized by, including: knocking out the FaCP5L1 or FaCP5L3 gene in strawberry; or silencing the FaCP5L1 or FaCP5L3 gene in strawberry; or decreasing the activity of the coding protein of FaCP5L1 or FaCP5L3 gene in strawberry; or decreasing the content of the coding protein of FaCP5L1 or FaCP5L3 gene in strawberry.

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