Application of pumpkin CmoEXPA23 gene in regulating fruit flesh thickness of crops

Abstract

The application discloses application of a pumpkin CmoEXPA23 gene in regulation of pulp thickness of crops, a nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and an amino acid sequence of a coded protein is shown in SEQ ID NO:2. It is found that the pumpkin CmoEXPA23 gene has a biological function of positively regulating pulp cell development in a fruit expansion process, can be used as an important gene resource for regulating pulp thickness of crops, and is expected to improve fruit traits and promote a molecular breeding process of crops.

Description

Technical Field

[0001] This invention belongs to the field of agricultural biotechnology, specifically to the field of crop genetics and breeding technology, and more specifically, to the application of the pumpkin CmoEXPA23 gene in regulating crop flesh thickness. Background Technology

[0002] Fruit flesh development is crucial for fleshy vegetable crops. As a globally important cucurbitaceous crop, pumpkin's fruit flesh development is closely related to its yield and final marketability. While pumpkin resources are abundant, significant differences exist in fruit flesh development among different resources.

[0003] Different expansin genes have different functions in plants, and there are no reports of their role in regulating early fruit pulp development. Summary of the Invention

[0004] Based on this, the purpose of the present invention is to provide an expansin gene that regulates the thickness of crop pulp.

[0005] The specific technical solutions for achieving the above-mentioned objectives are as follows.

[0006] In a first aspect, the present invention provides the application of the pumpkin CmoEXPA23 gene in regulating the thickness of crop flesh, wherein the nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

[0007] In a second aspect, the present invention provides the application of a protein encoded by the pumpkin CmoEXPA23 gene in regulating crop flesh thickness, wherein the amino acid sequence of the protein is shown in SEQ ID NO:2, and the crop is a cucurbitaceous crop or a solanaceous crop.

[0008] In a third aspect, the present invention provides the application of an overexpression vector of the pumpkin CmoEXPA23 gene in promoting increased fruit flesh thickness in crops, wherein the nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

[0009] In a fourth aspect, the present invention provides the application of engineered bacteria transformed with an overexpression vector of the pumpkin CmoEXPA23 gene in promoting increased fruit flesh thickness in crops, wherein the nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

[0010] In a fifth aspect, the present invention provides a biological agent that promotes the increase of crop flesh thickness, the active ingredient of which includes an overexpression vector of the pumpkin CmoEXPA23 gene or an engineered bacterium transformed with an overexpression vector of the pumpkin CmoEXPA23 gene, wherein the nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

[0011] In a sixth aspect, the present invention provides a method for promoting increased fruit flesh thickness in crops, comprising the following steps: increasing the expression level of the pumpkin CmoEXPA23 gene in the crop, wherein the nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

[0012] The inventors of this invention have discovered that the CmoEXPA23 gene in pumpkin has a positive regulatory function on the development of fruit pulp cells and increases the thickness of fruit pulp during the fruit enlargement process. It can serve as an important gene resource for regulating the thickness of crop fruit pulp and is expected to improve fruit traits and promote the process of molecular breeding of crops. Attached Figure Description

[0013] Figure 1 Fruit growth and development (0–40 days) of high-generation pumpkin inbred lines H (thick flesh) and B (thin flesh). A: Longitudinal section of fruit; B: Flesh thickness; C: Flesh cell size; D: Paraffin section of H flesh; E: Paraffin section of B flesh. * and ** in the figure indicate significant (p < 0.05) and highly significant (p < 0.01), respectively.

[0014] Figure 2 Identification of the expansion protein gene family in Chinese pumpkin (Cmo), and phylogenetic diagrams constructed from expansion proteins in species such as Arabidopsis thaliana (At), rice (Ost), Capsella bursa-pastoris (Cgr), potato (St), and cucumber (Cs).

[0015] Figure 3 The results of WGCNA analysis were conducted on the pulp transcriptome data of thick-fleshed pumpkin material H and thin-fleshed pumpkin material B at different developmental stages, combined with pulp development traits.

[0016] Figure 4 This is a chromosomal map showing the distribution of the expansion protein gene in Chinese pumpkin.

[0017] Figure 5 This presents the results of gene expression analysis of the expansion protein family in Chinese pumpkin. A: Tissue expression analysis; B: Expression analysis in the flesh of thick-fleshed pumpkin material H and thin-fleshed pumpkin material B at different developmental stages.

[0018] Figure 6Subcellular localization results for CmoEXPA23 protein.

[0019] Figure 7 This study investigated the application of the CmoEXPA23 gene in regulating tomato flesh thickness. The data includes: A: CmoEXPA23 overexpression validation; B: flowering plants; C: fruiting plants; D: longitudinal section of tomato fruit; E: longitudinal diameter of fruit; F: transverse diameter of fruit; G: longitudinal flesh thickness; H: transverse flesh thickness; I: fruit firmness at green ripening stage; J: fruit firmness at red ripening stage; K: paraffin section of tomato flesh tissue. WT: wild-type tomato; OE: CmoEXPA23 overexpressing tomato lines. ** indicates highly significant differences (p < 0.01). Detailed Implementation

[0020] To facilitate understanding of the present invention, a more complete description will be provided below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0021] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. The term "and / or" as used in this invention includes any and all combinations of one or more of the associated listed items.

[0022] Unless otherwise specified, all examples were performed under standard experimental conditions, such as those described in Sambrook et al., Molecular Cloning: a Laboratory Manual (Sambrook J & Russell DW, 2013), or as recommended by the manufacturer.

[0023] In some embodiments of the present invention, the application of the pumpkin CmoEXPA23 gene in regulating crop flesh thickness is disclosed, wherein the nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

[0024] In one embodiment, the cucurbitaceous crop includes pumpkin, and the solanaceous crop includes tomato.

[0025] In other embodiments of the present invention, the application of a protein encoded by the pumpkin CmoEXPA23 gene in regulating crop flesh thickness is disclosed, wherein the amino acid sequence of the protein is shown in SEQ ID NO:2, and the crop is a cucurbitaceous crop or a solanaceous crop.

[0026] In one embodiment, the cucurbitaceous crop includes pumpkin, and the solanaceous crop includes tomato.

[0027] In other embodiments of the present invention, the application of the overexpression vector of the pumpkin CmoEXPA23 gene in promoting the increase of crop flesh thickness is disclosed, wherein the nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

[0028] In one embodiment, the cucurbitaceous crop includes pumpkin, and the solanaceous crop includes tomato.

[0029] In other embodiments of the present invention, the application of engineered bacteria transformed with an overexpression vector of the pumpkin CmoEXPA23 gene in promoting increased fruit flesh thickness in crops is disclosed, wherein the nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

[0030] In one embodiment, the engineered bacteria is Escherichia coli or Agrobacterium.

[0031] In one embodiment, the cucurbitaceous crop includes pumpkin, and the solanaceous crop includes tomato.

[0032] In other embodiments of the present invention, a biological agent that promotes the increase of crop flesh thickness is disclosed, the active ingredient of which includes an overexpression vector of the pumpkin CmoEXPA23 gene or an engineered bacterium transformed with an overexpression vector of the pumpkin CmoEXPA23 gene, the nucleotide sequence of the pumpkin CmoEXPA23 gene being shown in SEQ ID NO:1, and the crop being a cucurbitaceous crop or a solanaceous crop.

[0033] In one embodiment, the cucurbitaceous crop includes pumpkin, and the solanaceous crop includes tomato.

[0034] In some other embodiments of the present invention, a method for promoting increased fruit flesh thickness in crops is disclosed, comprising the following steps: increasing the expression level of the pumpkin CmoEXPA23 gene in the crop, wherein the nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

[0035] In one embodiment, the cucurbitaceous crop includes pumpkin, and the solanaceous crop includes tomato.

[0036] The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.

[0037] Example 1: Discovery of candidate genes regulating pumpkin flesh thickness

[0038] Includes the following steps:

[0039] 1. Analysis of two high-generation pumpkin inbred lines, H (thick flesh) and B (thin flesh), with significant differences in flesh thickness (resources collected or created by the applicant's research group), revealed that their growth and development cycles (0-40 days after flowering) were similar, and their fruit sizes were also similar. Figure 1 (A and B in the text). The period from 0 to 20 days after flowering is the main period for pumpkin fruit enlargement and flesh development; during this time, the flesh thickness changes relatively little. Figure 1 (B in the text). The pulp cells rapidly swell 5-10 days after flowering. Figure 1 (C in the text), and the pulp cells of H are significantly larger than those of B ( Figure 1 The differences in cell size (D and E) suggest that the difference in cell size is an important factor leading to the difference in flesh thickness between the two fruits.

[0040] 2. To identify genes related to fruit pulp development, based on the two functional domains of the expansion protein family, DPBB_1 (PF03330) and pollen_allerg_1 (PF01357), a total of 54 expansion protein genes (CmoEXP) were identified from the Chinese pumpkin genome. Figure 2 Further comparison with the amino acid sequences of dilatants from species such as Arabidopsis thaliana, rice, and cucumber revealed that 54 dilatants in pumpkin were divided into four subfamilies: 32 CmoEXPA (α-dilatants), 9 CmoEXLA (dilatant-like A), 5 CmoEXPB (β-dilatants), and 8 CmoEXLB (dilatant-like B). EXPA is the largest dilatant subfamily in pumpkin, accounting for approximately 60%.

[0041] 3. Based on the combined flesh transcriptome data of thick-fleshed material H and thin-fleshed material B at different developmental stages, and combined with flesh development traits, WGCNA analysis was conducted, and the candidate gene for flesh thickness, CmoEXPA23, was screened in the grey60 module. Figure 3 Furthermore, CmoEXPA23 is the only expansion protein gene on chromosome 13 of Chinese pumpkin ( ), Figure 4 ).

[0042] 4. Tissue expression analysis revealed that CmoEXPA23 had a high expression level in fruit, significantly higher than other expansion protein genes. Figure 5 (A) CmoEXPA23 expression remained elevated during fruit enlargement, exhibited high expression during the critical period of pulp development (5-15 days after flowering), and significantly decreased during the later stages of fruit development to maturity. Figure 5Of all the expansion protein genes, CmoEXPA23 had the highest expression level in the pulp, and its expression changes were correlated with the trend of pulp cell enlargement and pulp thickening. Furthermore, its expression was significantly higher in the thick-fleshed pumpkin material H (with larger cells) than in the thin-fleshed pumpkin material B (with smaller cells). Figure 1 , Figure 5 (B in the middle).

[0043] The results of this embodiment suggest that the CmoEXPA23 gene (whose nucleotide sequence is shown in SEQ ID NO:1 and the amino acid sequence of the protein it encodes is shown in SEQ ID NO:2) may regulate pulp thickness.

[0044] SEQ ID NO:1 (containing the 2000 bp upstream promoter sequence of ATG and the 780 bp CDS sequence)

[0045] gttttagaaagaaaacaaataaataaattatttccatgaaaaataaaaaatagggcagctaattaatg ccattgtatattacccaaaaaaaaagggcaaaataaataattgattttctgattctaattattccccaaccccaac tactgcattgtctgctggggtaacaggtttttatcctctgtggaacgccacacagcttttatagtgtaccttcctt gtgagccccattcttatcctcaccacctaattcccttccatcccagccgtccatttcttttgctgtttttttgtaa taattaaaaattttgaattggattaaattgtacccttatttttaggttagtcgggttaaatatcagctctcgaacc gacttgattttttcagttttatatgtatatatgtatcaatattatattttaaaatatttaatacgttcataaactt ctaaatatatatttaataagtccaatactctcaattctatatctaataagtcgtttaaagattttattaacataat cgttaatatctcattttaatttaatattagattacgagagctacgtttaaataggttaaatcatgatttattatat tagagataggagaaattgacctaaggtaaagaaaaactacaataactttatggttaaaattgaattttttactttt tattaaaaaaaccaattatctaaatatattttttaatctgagtgaaataatttgtacttgaaaaaatatggatgaa aataggtaaaaaggggaaaaaagaaagaaggttaaaatggtaaaagatcaagaacaatagttaccctgtgtgacaa agcgcgttgacacgtaagagagagaacgtggaagcttgaccatatatagctggattccatttcatttcatatttac aaatattattttaaactattttttttttcacattatatatatattttttaattttttcgaccaatttgaatctctc gtttcaaaatttaagtgattatatatatatatatatatatatatatatatatatatatatatatatataaaacata ttattggtgtatttttaaatttatctaaaattaaaatattaacataaatataataacactcgtaataattttatac tcgactctaaaggataaatatttatacatacgaaaatatttgtcaaatactaattttgtcatgttgaaaatttctt agacctattttattgtgatatatgtaattcacgtgaaagatttaaatcgctacaaatatcgattgaatctattgtt taaatttaaaatatttattggataaaaaaattaaaggctaaattaaaaataaaaattataaattaaaaaaaaaaaa aaaaaatatatatatatatatagatatatatatatattctcaacccacccctagcagatattgtattctttagttt atcctcacaattataacggtgtttcattatcctccccaaccgatgtgggatcttacatcctcgcttgcacttgttc ctttctccaattaatgtgggatcccctccaaatccacccccttcaaggctcagcgtccctataggcacaccgcctc gtgaccacccttttggggctcaatcttcttactggcatatcgcccgatgtctggctctgacaccatttgtaacggt ccaagtttatcgctagcagatattgtcttatttgaacttttcgttttggagttacctcaaagtttttaaaatgagt aggctagacacaagtttacagggtgtttcattctccttaatttttttttttaaaatatccttaatttttttaaaaa tattcattaaaaccgttatttaaaaaaaaaaaaaaaaatttctagaactgttttcacacacaggcacacaaaagat tcgacattaaatgaggggcgtaattgtaaattcacacaaaaagacctccccgtcctcctcctccgttaagtactcc tcccattccctcttccagattgaaccaaaaaaATGGCTAATCCCGCCGTTCTCCTCTGCCTTGTGTCTCTCATCTCAGCAATGTGGACGGCAGATGCTAGAATTCCCGGCGCCTACTCCGGTGGCCCATGGCAGACCGCTCACGCCACCTTCTACGGCGGCTCCGACGCATCGGGCACAATGGGTGGGGCTTGTGGGTATGGGAATTTGTACAGCCAAGGGTACGGGGTGAACACGGCGGCGCTGAGTACGGCGCTATTCAACAATGGGCTGAGCTGTGGCGCGTGTTTCGAGATCAAATGTGCGAATGACCCGAGATGGTGCCATTCGGGTAGCCCGTCGATTGTCATAACGGCCACCAACTTCTGCCCGCCCAACTTTGCTCTGCCCAGTGACAATGGCGGCTGGTGTAACCCTCCTCGCCCCCATTTCGATTTGGCCATGCCTATGTTTCTCAAGATCGCTGAATATCGCGCCGGCATCGTCCCCGTCGCCTTCCGCCGGGTGCCATGCCGGAAGCAAGGGGGGATCCGGTTCACGGTCCACGGGTTCAGGTACTTCAACTTGGTTTTAATCACGAACGTCGCGGGTGCAGGGGATATCGTGAAGGTGAGCGTGAAGGGCACGCGAACCGGGTGGATGAGCATGAGCCGGAACTGGGGTCAAAACTGGCAGTCAAACGCCGTGTTGGTGGGGCAGGCACTGTCTTTTAGAGTCACAGGCAGTGACAGACGGACATCCACCACGTACAACGTGGCGCCAGCTGATTGGCAGTTTGGTCAGACCTTCATCGGCAAGAATTTTAGGGTTTGA

[0046] SEQ ID NO:2 (259 aa)

[0047] MANPAVLLCLVSLISAMWTADARIPGAYSGGPWQTAHATFYGGSDASGTMGGACGYGNLYSQGYGVNTAALSTALFNNGLSCGACFEIKCANDPRWCHSGSPSIVITATNFCPPNFALPSDNGGWCNPP RPHFDLAMPMFLKIAEYRAGIVPVAFRRVPCRKQGGIRFTVHGFRYFNLVLITNVAGAGDIVKVSVKGTRTGWMSMSRNWGQNWQSNAVLVGQALSFRVTGSDRRTSTTYNVAPADWQFGQTFIGKNFRV

[0048] Example 2 Subcellular localization of CmoEXPA23 protein

[0049] Cellular localization of CmoEXPA23 protein was predicted using the Plant-mPLoc website (http: / / www.csbio.sjtu.edu.cn / bioinf / plant-multi / ), revealing its location within the cell wall. This embodiment performed subcellular localization analysis, specifically including the following steps:

[0050] 1. The full-length coding sequence of the CmoEXPA23 gene was cloned, and based on this, the pBWA(V)HS-CmoEXPA23-GFP fusion expression vector was further constructed.

[0051] The full-length coding sequence of NAA60 was cloned into the target vector pBWA(V)HS-mKATE (vector and gene were purchased from Boyuan Biotechnology, Wuhan). mKATE is a red fluorescent protein, which was fused with NAA60 as a membrane localization marker.

[0052] 2. The two constructed vectors were transformed into Agrobacterium tumefaciens GV3101 strain, respectively. After culturing on YEB medium, the bacterial cells were collected, and the two bacterial solutions were mixed at a 1:1 ratio and injected into the leaves of 4-week-old Nicotiana benthamiana. Three days later, the fluorescence signal was observed using a laser confocal scanning microscope (Nikon, Japan).

[0053] 3. A 20% sucrose solution was slowly injected into leaves of *Nicotiana benthamiana* that had already shown fluorescence to induce plasmolysis. The sucrose-treated leaf segments were placed on glass slides and kept moist with the 20% sucrose solution. After incubation at room temperature for 20 min, coverslips were applied, and the localization of the green fluorescent CmoEXPA23-GFP and the red fluorescent plasma membrane marker was observed under a confocal scanning microscope (Nikon, Japan). The results showed that after plasmolysis treatment, the co-expressed plasma membrane marker NAA60-mKATE (red fluorescence) contracted synchronously with the protoplast, while CmoEXPA23-GFP (green fluorescence) remained in the cell wall. Figure 6 ).

[0054] The dilatation protein is a secreted protein that is synthesized intracellularly and transported to the cell wall to exert its function. The results of this embodiment show that the cell wall localization of the CmoEXPA23 protein is consistent with the properties of this family.

[0055] Example 3: CmoEXPA23 gene regulates tomato flesh thickness

[0056] The CmoEXPA23 open reading frame (ORF) without a stop codon was amplified from the flesh of the thick-fleshed squash material H and then inserted into the overexpression vector BG Plant-GFP (BioGround Crop). The recombinant plasmid was transformed into Escherichia coli for validation. Subsequently, the plasmid was transformed into Agrobacterium tumefaciens GV3101 and transfected into Solanum lycopersicum microTom via Agrobacterium-mediated transformation to obtain plants (OE-14 and OE-15) overexpressing the CmoEXPA23 gene, which were then compared with wild-type (WT).

[0057] Wild-type tomatoes (WT) and CmoEXPA23 overexpression lines (OE) began to bear fruit at 45 days of growth, but the fruits of the CmoEXPA23 overexpression lines matured a week later than those of the wild-type. The fruits at the top of the wild-type tomatoes and CmoEXPA23 overexpression plants were completely red and white-green, respectively. Figure 7 AC in the middle.

[0058] Further statistical analysis of the fruit phenotype at maturity yielded the following results: Figure 7 In the D~H region, compared to the wild type, the longitudinal diameter of the fruit in the CmoEXPA23 overexpression lines was slightly smaller. Figure 7 In the E), the transverse diameter did not change significantly ( Figure 7 The F in the middle). However, the longitudinal pulp thickness of the fruit of the CmoEXPA23 overexpression line ( Figure 7 (G in the middle) and transverse flesh thickness ( Figure 7 The H in the sample increased significantly, by approximately 30% and 40%, respectively.

[0059] The determination of tomato fruit firmness revealed no significant difference between the CmoEXPA23 overexpression lines and wild-type tomatoes. Figure 7 (I and J in the text).

[0060] To further compare the differences in tomato pulp cells between the CmoEXPA23 overexpression line and the wild-type, paraffin sections and microstructures of pulp were taken at the color-breaking stage. The results showed that the pulp cells of the CmoEXPA23 overexpression line were significantly larger than those of the wild-type. Figure 7 (K in the middle).

[0061] These results from this embodiment demonstrate that CmoEXPA23 can promote cell enlargement and increase tomato pulp thickness. The CmoEXPA23 gene has a positive regulatory function on pulp cell development during fruit enlargement and can serve as an important gene resource for regulating crop pulp thickness, potentially improving fruit traits.

[0062] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0063] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. The application of the pumpkin CmoEXPA23 gene in regulating crop flesh thickness, characterized by, The nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

2. The application of the protein encoded by the pumpkin CmoEXPA23 gene in regulating crop fruit flesh thickness, characterized by: The amino acid sequence of the protein is shown in SEQ ID NO:2, and the crop is a cucurbitaceous crop or a solanaceous crop.

3. The application of the overexpression vector of the pumpkin CmoEXPA23 gene in promoting increased fruit flesh thickness, characterized by: The nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

4. The application of engineered bacteria transformed with an overexpression vector of the pumpkin CmoEXPA23 gene in promoting increased fruit flesh thickness in crops, characterized in that... The nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

5. The application according to claim 4, characterized in that, The engineered bacteria are Escherichia coli or Agrobacterium.

6. The application according to any one of claims 1 to 5, characterized in that, The cucurbitaceous crops include pumpkin, and the solanaceous crops include tomato.

7. A biological agent that promotes increased fruit flesh thickness in crops, characterized in that, Its active ingredients include an overexpression vector of the pumpkin CmoEXPA23 gene or an engineered bacterium transformed with an overexpression vector of the pumpkin CmoEXPA23 gene. The nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:

1. The crop is a cucurbitaceous crop or a solanaceous crop.

8. The biological agent according to claim 7, characterized in that, The cucurbitaceous crops include pumpkin, and the solanaceous crops include tomato.

9. A method for promoting increased fruit flesh thickness in crops, characterized in that, Includes the following steps: To increase the expression level of the pumpkin CmoEXPA23 gene in crops, wherein the nucleotide sequence of the pumpkin CmoEXPA23 gene is shown in SEQ ID NO:1, and the crop is a cucurbitaceous crop or a solanaceous crop.

10. The method according to claim 9, characterized in that, The cucurbitaceous crops include pumpkin, and the solanaceous crops include tomato.

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