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Article

Genome-Wide Analysis of the Serine Carboxypeptidase-like (SCPL) Protein Family of Bitter Gourd and Functional Validation of McSCPL22 in Fusarium oxysporum f. sp. Momordicae (FOM) Resistance

by
Feng Guan
1,2,3,
Xuetong Yang
1,2,3,
Bo Shi
1,2,3,
Kai Wang
1,2,3,
Jingyun Zhang
1,2,3,
Yuanyuan Xie
1,2,3 and
Xinjian Wan
1,2,3,*
1
Institute of Vegetables and Flowers, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China
2
Jiangxi Key Laboratory of Horticultural Crops (Fruit, Vegetable & Tea) Breeding, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China
3
Jiangxi Engineering Research Center of Vegetable Molecular Breeding, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2024, 25(21), 11816; https://doi.org/10.3390/ijms252111816
Submission received: 10 October 2024 / Revised: 31 October 2024 / Accepted: 1 November 2024 / Published: 3 November 2024
(This article belongs to the Section Molecular Genetics and Genomics)
Browse Figures
Versions Notes

Abstract

:
Bitter gourd is increasingly being recognized for its value as a vegetable and medicinal use, but the molecular mechanisms of pathogen resistance remain relatively poorly understood. The serine carboxypeptidase-like (SCPL) protein family plays a key role in plant growth, pathogen defense, and so on. However, a comprehensive identification and functional characterization of the SCPL gene family has yet to be conducted in bitter melon. In this study, 32 SCPL genes were identified in bitter gourd and divided into three classes. The number of SCPL genes contained in the three clusters was 7, 7, and 18, respectively. Most SCPL gene promoters contain cis-acting elements with light, hormone, and stress responses. The RNA sequencing data showed that the expression of several SCPL genes changed significantly after pathogen infection. In particular, expression of the McSCPL4, 10, 17, 22, and 25 genes increased substantially in the resistant varieties after infection, and their expression levels were higher than those in the susceptible varieties. These results suggested that genes such as McSCPL4, 10, 17, 22, and 25 may play a significant role in conferring resistance to fungal infections. Moreover, the expression levels of the McSCPL10, 17, 22, 23, and 25 genes were likewise significantly changed after being induced by salicylic acid (SA) and jasmonic acid (JA). In situ hybridization showed that McSCPL22 was expressed in the vascular tissues of infected plants, which largely overlapped with the location of Fusarium oxysporum f. sp. Momordicae (FOM) infection and the site of hydrogen peroxide production. Our results showed that McSCPL22 may be involved in the regulation of the SA and JA pathways and enhance resistance to FOM in bitter gourd plants. This is the first study to perform SCPL gene family analysis in bitter gourd. McSCPL22 may have the potential to enhance FOM resistance in bitter gourd, and further investigation into its function is warranted. The results of this study may enhance the yield and molecular breeding of bitter gourd.

1. Introduction

Momordica charantia L. is a member of the Cucurbitaceae family; we commonly call it bitter gourd or bitter melon [1]. Bitter gourd has important nutritional and medical value and is used for medicinal purposes such as treating cancer, inflammation, and high cholesterol [2,3]. The production of bitter gourd is affected by some insects, such as the gourd fruit fly [4], and is also susceptible to various fungal infections, such as Fusarium oxysporum f. sp. momordicae (FOM), which results in reduced yields [5,6]. To reduce the damage caused by pests and diseases, researchers aim to identify genetic resources with pest and disease resistance. In our previous studies, the resistant and susceptible varieties of bitter gourd were researched by transcriptome and metabolome methods for screening the candidate genes [7,8].
In plants, serine carboxypeptidase-like proteins (SCPLs) belong to the serine carboxypeptidase family. They have a highly conserved α/β hydrolase fold, which is characterized by a central β-sheet surrounded by α-helices on either side [9,10]. SCPLs exhibit acyltransferase activity rather than peptidase activity. Typically, SCPLs contain a conserved amino acid structure of serine, aspartate, and histidine (Ser-Asp-His) with a typical PF00450 domain [11]. Previous research has demonstrated that SCPLs have complex roles in plant growth and resistance to various stresses [12,13,14,15].
In particular, SCPLs were found to play a significant role in plant defense mechanisms. In soybeans, at least six GmSCPLs were found to respond to various adverse stresses [16]. In cotton, GhSCPL42 may increase the resistance of cotton to Verticillium wilt through the salicylic acid (SA) and jasmonic acid (JA) pathways [17]. The rice OsSCPL1 gene was observed to regulate the defense responses of the plant against the bacterial pathogen Pseudomonas syringae pv. tomato and the fungal pathogen Alternaria brassicicola [14]. Oat SCPL1 was found to be essential for antimicrobial synthesis and disease resistance [15].
SCPLs are widely distributed in various plants, and SCPL families have been successfully characterized in several species, with results showing that there may be 54 SCPL family members in Arabidopsis thaliana [18,19], 71 members in rice (Oryza sativa L. ssp. japonica) [20], 57 members in poplar (Populus tomentosa) [21], 47 members in the tea plant (Camellia sinensis) [22], 209 members in wheat (Triticum aestivum) [23], 117 in Brassica napus [24], and 73 members in soybean (Glycine max) [16]. Moreover, Fu et al. identified 891 SCPL genes from 14 species [25]. Currently, the SCPL gene family has not been researched in bitter gourd.
In this study, genome-wide characterization of the SCPL gene family was performed using the available bitter gourd reference genome. The expression patterns of SCPLs were investigated using available transcriptome data and were detected in the treatment plant with SA and JA. Subsequently, genes that may be related to fungal resistance were screened. These results will provide a reference for further systematic studies on the functions of the SCPL genes in bitter gourd and additional genetic resources to enrich resistance breeding in bitter gourd.

2. Results

2.1. Identification of the Bitter Gourd SCPL Gene Family

We identified 32 McSCPL genes in bitter gourd and confirmed them in the CDD and SMART databases. All McSCPLs can be found in conserved domains in the SMART database. The 32 McSCPL genes were named McSCPL1–32 (Table 1). The coding protein length ranged from 268 to 1457 amino acids. The molecular weight ranged from 31.0 to 161.6 kDa, and their isoelectric point was 4.85–8.8. Among them, 28 proteins were localized in the vacuole, while McSCPL26 was found in the peroxisome. Additionally, McSCPL9, McSCPL11, and McSCPL12 were observed in both the vacuole and peroxisome (Table 1).

2.2. Evolutionary Relationship of SCPL Genes

The predicted SCPL protein sequences from M. charantia and A. thaliana were used to construct a phylogenetic tree. The SCPL proteins clustered into three subgroups: green, blue, and red (Figure 1A). There were 18, 7, and 7 SCPL proteins in the green, blue, and red subgroups of M. charantia, respectively. In the green, blue, and red subgroups, there were 26, 22, and 6 SCPL proteins of A. thaliana, respectively (Figure 1A). The SCPL genes in the green and red subgroups of M. charantia were similar to those of A. thaliana, but in the blue subgroup, M. charantia had only one-third of the number of genes that A. thaliana had.
The multiple sequence alignment shows that McSCPLs and AtSCPLs have a typical Ser-Asp-His conserved domain and two typical G-x-S-x-G motifs. The Ser was surrounded by a Gly structure (Figure 1B).

2.3. Evolutionary Tree, Sequence Structure Analysis of SCPL Genes

A phylogenetic tree of McSCPL proteins showed that all members were divided into three subgroups, which were consistent with the findings in other known plants (Figure 2A). Among them, all members contained motif 3 and motif 5, but the red subgroup did not contain motif 6, and the blue and green subgroups exhibited similar motif characteristics (Figure 2B, Supplementary Figure S1). The green subgroup contains 8–10 exons, the red subgroup members have 9–13 exons, except for McSCPL25, which only has two exons, and the blue subgroup members have 8–15 exons, which have the greatest range of exon numbers (Figure 2C).

2.4. The Cis-Acting Elements of the SCPL Genes Promoter

To predict the possible regulatory factors of the McSCPL family members, the upstream promoter sequences 1500 bp before the start codon of the McSCPL family members were analyzed. The McSCPL9 and McSCPL26 promotor sequences cannot be obtained because the genome sequences are incomplete. The results revealed that the promoters of these genes are mainly cis-elements, including light, hormone, and stress responses. The promoters of most of the gene family members contain light response-related elements, such as Box-4 and G-box; methyl jasmonate-related cis-elements CGTCA-motifs and TGACG-motifs; and SA-related cis-element TCA; abscisic acid-related cis-element ABRE; and defense and stress response-related cis-elements of MYB binding (Figure 3A). In addition, the specific type and quantity of cis-elements present within each gene promoter exhibited variability. This suggests that most of these family members may be regulated by abscisic acid, methyl jasmonate, and SA and are closely associated with biotic and abiotic stress responses. These genes may be regulated by auxin (McSCPL7, McSCPL10, McSCPL14, McSCPL16, McSCPL19, McSCPL22, McSCPL25, McSCPL28, and McSCPL31 containing the TGA-element) or gibberellin (eight genes contained at least one cis-element of the GARE-motif, or P-box, or TATC-box) (Figure 3B).

2.5. Tissue-Specific Expression Profiles of SCPL Genes

The expression patterns of McSCPL genes were calculated based on previous report data [7]. The expression of the McSCPL1, McSCPL5, McSCPL7, McSCPL8, McSCPL12, McSCPL14, McSCPL15, McSCPL20, McSCPL24, McSCPL26, and McSCPL28 genes showed a trend of upregulation after inoculation with FOM both in resistant and susceptible varieties. The expression of McSCPL3, McSCPL6, McSCPL11, McSCPL16, McSCPL18, McSCPL27, and McSCPL31 genes showed a trend of downregulation after inoculation with FOM, both in resistant and susceptible varieties. A comprehensive evaluation of the expression level of the McSCPL2, McSCPL4, McSCPL9, McSCPL10, McSCPL17, McSCPL22, McSCPL23, McSCPL25, McSCPL29, McSCPL30, and McSCPL32 genes revealed that they were higher in the resistant variety than in the susceptible variety at multiple time points in both mock and treated samples (Figure 4A).
In the tissues, McSCPL26 and McSCPL29 showed higher expression in the ovary tissue; McSCPL2, McSCPL12, and McSCPL17 were highly expressed in the flowers; and McSCPL5, McSCPL9, McSCPL18-20, and McSCPL32 were highly expressed in the stem; McSCPL1, McSCPL10, McSCPL11, McSCPL13-16, McSCPL21, McSCPL22, McSCPL24, McSCPL25, McSCPL30, and McSCPL31 were highly expressed in the leaf (Figure 4B).

2.6. Expression Analysis of McSCPLs Response to SA and JA Treatment

SA and JA have important functions in disease resistance. To confirm that McSCPLs are regulated by SA and JA, the results demonstrated that MCSCPL10 and MCSCPL15 had a similar expression change to the control after SA treatment. The expression of McSCPL10 was higher than the control group at three and nine hours, and the expression of McSCPL15 was lower than the control group at nine hours (Figure 5A,B). SA promotes the expression of McSCPL17, McSCPL23, and McSCPL25. Their expression levels were significantly higher at three hours after treatment than in the control group (Figure 5C,E,F). The expression of McSCPL22 was markedly elevated in the control group three hours after treatment, subsequently declining (Figure 5D), indicating that SA affects the expression level of McSPLC22.
After JA treatment, the expression level of McSCPL10, 15, 17, 22, 23, and 25 was elevated at three hours. This elevation was followed by a gradual decrease (Figure 6A–F). This suggests that JA may promote the expression of multiple McSCPLs.

2.7. FOM Infection Promoted the Accumulation of Hydrogen Peroxide

In plants, fungal infection frequently results in the generation of reactive oxygen species. We examined the infected bitter gourd stems and showed that hydrogen peroxide (brown parts) was produced in the stems of both resistant (BK0604) and susceptible (LK0901) varieties (Figure 7A). However, the accumulation of hydrogen peroxide in the susceptible varieties was higher than in the resistant varieties, which suggests that resistant varieties have an inhibitory effect on the accumulation of hydrogen peroxide. The staining of the sites where hydrogen peroxide was produced mainly focused on the locations where the vascular bundles were located, including the xylem and phloem (Figure 7B).

2.8. McSCPL22 May Respond to FOM Infection

From the quantitative PCR results, it can be seen that there is a large difference in the expression of McSCPL22 at 3 h after SA and JA treatments compared with the control group. Subcellular localization studies revealed that McSCPL22 is mainly localized to the cell membrane (Figure 8A).
By in situ hybridization, we found that after FOM inoculation, McSCPL22 was mainly detected in the vascular bundle portion of the plant, in addition to some expression in the cortical thin-walled tissue of the stem (Figure 8B). Interestingly, the hybridization of the infected tissues by the nucleic acid probe for FOM showed that the distribution location of FOM was also mainly concentrated in the vascular bundle part of the plant, which was the same site of McSCPL22 expression (Figure 8C).

3. Discussion

Bitter gourd is increasingly becoming a valuable food and medical resource, but it is relatively understudied for stress tolerance, with members of the SCPL gene family showing tolerance to biotic and abiotic stresses in several species [10,15,23,26,27]. The SCPL gene family is characterized and reported in multiple species [16,17,18,20,21,22,23,24,25]; however, this is the first time that the SCPL gene family has been identified in bitter gourd. In this study, we searched the bitter gourd genome with A. thaliana SCPL family members using BLASTP and simultaneously used HMMER based on the SCPL gene model (PF00450). We found that the 32 members of the bitter gourd theoretical isoelectric point varied widely (4.85–8.8), but most of the isoelectric points were less than 7 (approximately 65%) and shared similar features with other species.
As we know, Arabidopsis has 54 SCPL family members, which is close to twice the number of family members in bitter gourd, but in terms of the evolutionary relationship, we found that the number of SCPLs in the red subfamily was similar in bitter gourd and Arabidopsis, which are 7 and 6, respectively. In the blue cluster subfamily, the number of members in bitter gourd is 7, but the number of members in Arabidopsis is 22, which is close to three times that in bitter gourd. In Arabidopsis, the functions of the members of this subfamily are relatively well studied, such as At5G09640 (sinapoylglucose choline sinapoyltransferases) [28], AT2G22990 (sinapoylglucose malate sinapoyltransferase) [29], AT2G23000 (sinapoylglucose anthocyanin sinapoyltransferase) [30], and AT2G23010 and AT2G22980 (sinapoylglucose sinapoyltransferase) [30]. In addition, the functions of the members of this subfamily in Arabidopsis mainly regulated the formation of a variety of secondary metabolites, whereas the relatively small number of members of this subfamily in bitter gourd may imply that the corresponding secondary metabolites may be lacking in bitter gourd. In addition, Mugford et al. found that AsSCPL1, a gene with functions in the synthesis of antimicrobial compounds and disease resistance, belongs to the blue cluster subfamily. This finding suggests that members of this subfamily may have evolved resistance-related functions [15]. The green cluster subfamily of bitter gourd has 18 SCPL members, whereas Arabidopsis has 26 members; the ratio of subfamily genes is similar to the overall number.
Because of the rich functionality of SCPL family members, the results of analyzing their upstream promoter elements revealed that most of the members contain multiple types of elements, including light, hormone, and stress responses (Figure 3). These results suggest that these members are related to the corresponding regulatory pathways. Combined with previously reported transcriptome data [7], it was shown that the expression of most of the genes changed significantly after FOM infection, especially in the resistant varieties, where the expression level was significantly elevated, such as McSCPL4, McSCPL22, McSCPL25, and McSCPL10 (Figure 4A). We also found several hormone-related regulatory elements in their promoter sequences. In a previous report, cotton GhSCPL42 improved FOM resistance through the SA and JA pathways [15]. We found significant changes in the expression level of multiple SCPL members in bitter gourd after SA and JA treatment (Figure 5 and Figure 6). These results indicate that McSCPL10, McSCPL22, and McSCPL25 might be involved in bitter gourd resistance to FOM via the SA and JA regulatory pathways. Interestingly, McSCPL22 was significantly downregulated after treatment with SA and upregulated after treatment with JA, which varied dramatically and could serve as a genetic resource to consider in subsequent studies.
To preliminarily determine the function of McSCPL22, we found it mainly localized to the cell membrane, which differed from the predicted result in the vacuole (Figure 8A). Following inoculation with FOM, the susceptible varieties produced more hydrogen peroxide, mainly concentrated within the vascular bundles (Figure 7). This observation is similar to the FOM distribution after infection (Figure 8C). Interestingly, we also found that the expression of McSCPL22 in the infected susceptible varieties was also concentrated in the vascular bundles of the plants (Figure 8B). This result suggests that the site fungal infection and the expression of the McSCPL22 gene may overlap. The expression level of McSCPL22 was markedly increased in the resistant varieties after FOM inoculation, suggesting a potential relation between the stress response and elevated resistance of bitter gourd.
This study identified and reported the SCPL gene family of bitter melon for the first time. Furthermore, the evolutionary relationships, gene structures, conserved motif features, and expression patterns of the SCPL family in bitter melon were then systematically analyzed and compared. It was found that most of the McSCPLs might be associated with light, hormone, and stress responses. The results analyzed by gene expression patterns and in situ hybridization showed that McSCPL22 may respond to FOM infection in bitter melon via the SA and JA regulatory pathways. The results presented herein provide a foundation for further investigation into the functions of McSCPLs and the molecular mechanism of McSCPL22 in fungal resistance, as well as the potential use of McSCPL22 in the breeding of highly resistant bitter melon varieties.

4. Methods

4.1. Plant Material

This research used two bitter gourd varieties; BK0604 was highly resistant to Fusarium wilt and selected and bred from a local variety in Jiangxi Province by the study team. LK0901 was highly susceptible to Fusarium wilt collected from Taiwan Province in 2004 and was bred by our team [7]. Bitter gourd seeds were soaked in water for 17 h and germinated in a controlled environment at 32 °C. After germination, the seedlings were transplanted into pots with the seedling substrate (Pindstrup, Ryomgård, Danmark) for further experimentation.

4.2. Identification of SCPL Genes

The genome data of bitter gourd were downloaded from the National Center for Biotechnology Information (NCBI) database (Assembly version: GCF_001995035.1; PRJNA397875) [31]. Arabidopsis SCPL gene data were acquired from the TAIR database (https://www.arabidopsis.org, accessed on 30 September 2023). The SCPL gene model PF00450 was obtained from the Pfam website (http://pfam.xfam.org, accessed on 30 September 2023). The PF00450 data were queried in encoding protein sequence data of bitter gourd using HMMER software (v3.0). The results were further verified through Pfam, NCBI CDD, and SMART databases [32,33,34,35].

4.3. Phylogenetic Analysis of the SCPL Gene Family

The ClustalW program facilitated multiple alignments of the SCPL protein sequences from M. charantia and Arabidopsis. A phylogenetic tree was constructed utilizing the MEGA7 software (v7) through the neighbor-joining method, employing the Poisson correction model with 1000 bootstrap test replications [36].

4.4. Structure Analysis of the SCPL Gene Family

The gene structure file was submitted to the GSDS 2.0 server to obtain the diagram [37]. The chromosome location of the McSCPL genes was retrieved from M. charantia genome data. We used the MEME website (https://meme-suite.org/meme/, accessed on 10 July 2023) to identify conserved protein motifs using default parameters. The Plant-mPLoc website (http://www.csbio.sjtu.edu.cn/bioinf/Cell-PLoc-2/, accessed on 20 June 2023) was used to predict the subcellular location of McSCPLs. The molecular weight and isoelectric point of the McSCPL proteins were predicted using Expasy. Then, the cis-acting elements of the 1500 bp promoter sequences of each McSCPL gene were analyzed via the PlantCARE website (http://bioinformatics.psb.ugent.be/webtools/plantcare/html/ , accessed on 10 July 2023).

4.5. Expression Pattern of SCPL Genes Based on RNA Sequencing Data

RNA sequencing (RNA-seq) data of the tissue samples (ovary, male, flower, leaf, stem, root) were obtained from the website at http://www.ncbi.nlm.nih.gov/sra/ through the accession numbers SRR13308550–SRR13308554 (PRJNA687997) (accessed on 5 October 2023). The data related to the fungal infection assays were obtained from the following source: http://www.ncbi.nlm.nih.gov/bioproject/PRJNA825666 (accessed on 5 October 2023) [7]. The fragments per kilobase of transcript per million mapped reads (FPKM) values for each gene in all samples were calculated according to our previous report [7].

4.6. Fungal Infection Assay and Treatment with SA and JA

The FOM strain was collected from the experimental base of the Institute of Vegetables and Flowers at Jiangxi Academy of Agricultural Sciences [38]. The FOM was cultured and collected according to previous methods [7]. Liquid potato dextrose agar medium at 25 °C with shaking at 150 rpm for 5–7 days. The conidia with 1 × 106 CFU/mL concentration were used to infect the bitter gourd root. Bitter gourd seedlings were washed and dipped in spore suspension. These seedlings were then replanted into a sterile growing substrate and positioned in an artificial climate chamber set at 28 °C with a 12/12 h light/dark cycle and 70% relative humidity [8]. Seedling leaves were sprayed with SA (1.5 mmol/L) and JA (100 µmol/L) on the leaves at 0, 3, 6, and 9 days after inoculation, respectively [7,8,14]. The control group was treated with distilled water. Then, samples were collected at 2-day intervals after the treatments. Three independent experiments and at least six plants for each experiment were performed.

4.7. Gene Expression Analysis by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR)

Total RNA was extracted, and cDNA was synthesized using RNA Isolation Reagent and cDNA Synthesis Kit (GeneCopoeia, China), respectively. The primers were designed on the Primer3 website (Supplementary Table S1). McActin7 was used as the internal control. qRT-PCR was performed according to previous methods [7]. The 2−△△Ct method was used to assess gene expression level. Three independent experiments and three technical replicates for each experiment were performed. The obtained data were subjected to unpaired two-tailed Student’s t-tests using GraphPad Prism software (version 8).

4.8. Subcellular Localization of the McSCPL22 Protein

McSCPL22 was synthesized into the pR101-35S-GFP vector, then transformed into Agrobacterium tumefaciens strain GV3101. Agrobacterium containing pCambia1300-35S-PM-mcherry and the pR101-35S-McSCPL22-GFP vector were injected into the 2–4-week-old Nicotiana benthamiana leaves. The plants were darkened for 2–3 days. The leaves were cut to observe the fluorescence using a confocal microscope (Leica SP8, Leica, Japan).

4.9. Analysis of Hydrogen Peroxide Accumulation

The plant stems were collected after 12 days of the fungal infection assay. H2O2 content was detected using 3,3′-diaminobenzidine (DAB) staining (Servicebio, Wuhan, China). The stems of BK0604 (R) and LK0901 (S) plants were dipped in 1 mg/mL DAB solution (pH 3.8) for 24 h. Thereafter, the stem sections were boiled in 95% ethanol for 10 min, and the image was captured using Nikon Eclipse CI microscope (Nikon, Japan) [39].

4.10. Toluidine Blue Staining

The collected stems were cut into sections and placed into a dewaxing transparent liquid (20 min) (Servicebio, Wuhan, China), anhydrous ethanol (5 min), and 75% alcohol (5 min). The plant tissue sections were stained with toluidine blue dye for approximately 2 min, according to the depth of tissue coloring, and then dried in an oven at 60 °C after washing with water. Finally, the sections were treated with xylene and imaged by Nikon Eclipse E100 (Nikon, Japan) [40,41].

4.11. In Situ Hybridization Experiment

This experiment was performed by using standard in situ hybridization and NBT histochemistry techniques [42]. In brief, the tissues of LK0901(S) were placed in situ hybridization fixative (Servicebio, Wuhan, China) overnight at 4 °C, then sliced into 6 μm sections using a microtome. After dewaxing and dehydration, the hybridization solution containing the probes was added as previously described (Supplementary Table S1) [43,44]. Imaging was performed using a Nikon Eclipse CI microscope (Nikon, Japan).

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/ijms252111816/s1.

Author Contributions

F.G. designed the study and wrote the manuscript. X.Y. and B.S. carried out bioinformatic analyses. K.W., J.Z. and Y.X. carried out the experiments. X.W., resources, conceptualization, funding acquisition, project administration. All authors have read and agreed to the published version of the manuscript.

Funding

This work was supported by the Basic Research Project of Jiangxi Academy of Agricultural Sciences (JXSNKYJCRC202313), China Agriculture Research System of MOF, and MARA (CARS-23).

Institutional Review Board Statement

The collection of resistant genotype BK0604 and susceptible genotype LK0901 plant materials used in the experiment in this article was provided by Jiangxi Academy of Agricultural Sciences and permitted by Prof. Xinjian Wan, who is responsible for the preservation and management of bitter gourd germplasm resources.

Informed Consent Statement

Not applicable.

Data Availability Statement

The RNA-seq datasets of the tissue samples (ovary, male, flower, leaf, stem, root) are obtained from the SRA database (http://www.ncbi.nlm.nih.gov/sra/, accessed on 5 October 2023) through the accession numbers SRR13308550–SRR13308554 (PRJNA687997). The RNA-seq datasets of the fungal infection assays are available in the NCBI repository (http://www.ncbi.nlm.nih.gov/bioproject/PRJNA825666, accessed on 5 October 2023). The genome data of bitter gourd are available in the NCBI database (Assembly version: GCF_001995035.1; PRJNA397875).

Acknowledgments

The authors thank Minshan Sun and Henan Assist Research Biotechnology Co., Ltd. (Zhengzhou, China) for their help with the bioinformatics analysis.

Conflicts of Interest

The authors declare no competing interests.

Abbreviations

SCPL: serine carboxypeptidase-like; FOM: Fusarium oxysporum f. sp. Momordicae; SA: salicylic acid; JA: jasmonic acid; RNA-seq: RNA sequencing; FPKM: fragments per kilobase of transcript per million mapped reads; qRT-PCR: real-time quantitative reverse transcription polymerase chain reaction; SE: standard errors.

References

  1. Behera, T.K.; Behera, S.; Bharathi, L.K.; John, K.J.; Staub, J.E. Bitter Gourd: Botany, Horticulture, Breeding; John Wiley & Sons, Inc.: Hoboken, NJ, USA, 2010. [Google Scholar]
  2. Dandawate, P.R.; Subramaniam, D.; Padhye, S.B.; Anant, S. Bitter melon: A panacea for inflammation and cancer. Chin. J. Nat. Med. 2016, 14, 81–100. [Google Scholar] [CrossRef] [PubMed]
  3. Tan, S.P.; Kha, T.C.; Parks, S.E.; Roach, P.D. Bitter melon (Momordica charantia L.) bioactive composition and health benefits: A review. Food Rev. Int. 2015, 32, 181–202. [Google Scholar] [CrossRef]
  4. Dhillon, M.K.; Singh, R.; Naresh, J.S.; Sharma, H.C. The melon fruit fly, Bactrocera cucurbitae: A review of its biology and management. J. Insect Sci. (Online) 2005, 5, 40. [Google Scholar] [CrossRef] [PubMed]
  5. Tian, Y.H.; Hou, Y.Y.; Peng, C.Y.; Wang, Y.Y.; He, B.L.; Gao, K.X. Genetic diversity and phylogenetic analysis of Fusarium oxysporium strains isolated from the Cucurbitaceae hosts revealed by SRAPs. Ying Yong Sheng Tai Xue Bao J. Appl. Ecol. 2017, 28, 947–956. [Google Scholar]
  6. Li, X.F.; Tian, Y.H.; Peng, H.Y.; He, B.L.; Gao, K.X. Isolation, screening and identification of anantagonistic actinomycetes to control Fusarium wilt of Momordica charantia. Ying Yong Sheng Tai Xue Bao J. Appl. Ecol. 2020, 31, 3869–3879. [Google Scholar]
  7. Guan, F.; Shi, B.; Zhang, J.; Wan, X. Transcriptome analysis provides insights into lignin synthesis and MAPK signaling pathway that strengthen the resistance of bitter gourd (Momordica charantia) to Fusarium wilt. Genomics 2023, 115, 110538. [Google Scholar] [CrossRef]
  8. Guan, F.; Shi, B.; Zhang, J.; Wan, X. Metabolome Revealed the Potential Mechanism of Fusarium Wilt Resistance in Bitter Gourd (Momordica charantia) Based on Liquid Chromatography with Mass Spectrometry. Plant Dis. 2024, 108, 920–929. [Google Scholar] [CrossRef]
  9. Milkowski, C.; Strack, D. Serine carboxypeptidase-like acyltransferases. Phytochemistry 2004, 65, 517–524. [Google Scholar] [CrossRef]
  10. Mugford, S.T.; Milkowski, C. Serine carboxypeptidase-like acyltransferases from plants. Nat. Prod. Biosynth. Microorg. Plants Pt B 2012, 516, 279–297. [Google Scholar]
  11. Agarwal, V.; Tikhonov, A.; Metlitskaya, A.; Severinov, K.; Nair, S.K. Structure and function of a serine carboxypeptidase adapted for degradation of the protein synthesis antibiotic microcin C7. Proc. Natl. Acad. Sci. USA 2012, 109, 4425–4430. [Google Scholar] [CrossRef]
  12. Ciarkowska, A.; Ostrowski, M.; Starzynska, E.; Jakubowska, A. Plant SCPL acyltransferases: Multiplicity of enzymes with various functions in secondary metabolism. Phytochem. Rev. 2019, 18, 303–316. [Google Scholar] [CrossRef]
  13. Remington, S.J. Serine carboxypeptidases: A new and versatile family of enzymes. Curr. Opin. Biotechnol. 1993, 4, 462–468. [Google Scholar] [CrossRef] [PubMed]
  14. Liu, H.; Wang, X.; Zhang, H.; Yang, Y.; Ge, X.; Song, F. A rice serine carboxypeptidase-like gene OsBISCPL1 is involved in regulation of defense responses against biotic and oxidative stress. Gene 2008, 420, 57–65. [Google Scholar] [CrossRef] [PubMed]
  15. Mugford, S.T.; Qi, X.; Bakht, S.; Hill, L.; Wegel, E.; Hughes, R.K.; Papadopoulou, K.; Melton, R.; Philo, M.; Sainsbury, F.; et al. A serine carboxypeptidase-like acyltransferase is required for synthesis of antimicrobial compounds and disease resistance in oats. Plant Cell 2009, 21, 2473–2484. [Google Scholar] [CrossRef]
  16. He, L.; Liu, Q.N.; Han, S.J. Genome-wide analysis of serine carboxypeptidase-like genes in soybean and their roles in stress resistance. Int. J. Mol. Sci. 2024, 25, 6712. [Google Scholar] [CrossRef]
  17. Wang, Y.; Zhao, J.; Deng, X.; Wang, P.; Geng, S.; Gao, W.; Guo, P.; Chen, Q.; Li, C.; Qu, Y. Genome-wide analysis of serine carboxypeptidase-like protein (SCPL) family and functional validation of Gh_SCPL42 unchromosome conferring cotton Verticillium der Verticillium wilt stress in Gossypium hirsutum. BMC Plant Biol. 2022, 22, 421. [Google Scholar] [CrossRef]
  18. Feng, Y.; Liu, Q.P.; Jia, J.; Xue, Q.Z. Genomic analysis of serine carboxypeptidase-like protein family of Arabidopsis thaliana. Yi Chuan Xue Bao Acta Genet. Sin. 2005, 32, 864–873. [Google Scholar]
  19. Fraser, C.M.; Rider, L.W.; Chapple, C. An expression and bioinformatics analysis of the Arabidopsis serine carboxypeptidase-like gene family. Plant Physiol. 2005, 138, 1136–1148. [Google Scholar] [CrossRef]
  20. Feng, Y.; Xue, Q. The serine carboxypeptidase like gene family of rice (Oryza sativa L. ssp. japonica). Funct. Integr. Genom. 2006, 6, 14–24. [Google Scholar] [CrossRef]
  21. Zhu, D.Y.; Chu, W.Y.; Wang, Y.J.; Yan, H.W.; Chen, Z.; Xiang, Y. Genome-wide identification, classification and expression analysis of the serine carboxypeptidase-like protein family in poplar. Physiol. Plant. 2018, 162, 333–352. [Google Scholar] [CrossRef]
  22. Ahmad, M.Z.; Li, P.; She, G.; Xia, E.; Benedito, V.A.; Wan, X.C.; Zhao, J. Genome-wide analysis of serine carboxypeptidase-like acyltransferase gene family for evolution and characterization of enzymes involved in the biosynthesis of galloylated catechins in the tea plant (Camellia sinensis). Front. Plant Sci. 2020, 11, 848. [Google Scholar] [CrossRef] [PubMed]
  23. Xu, X.M.; Zhang, L.L.; Zhao, W.; Fu, L.; Han, Y.X.; Wang, K.K.; Yan, L.Y.; Li, Y.; Zhang, X.-H.; Min, D.-H. Genome-wide analysis of the serine carboxypeptidase-like protein family in Triticum aestivum reveals TaSCPL184-6D is involved in abiotic stress response. BMC Genom. 2021, 22, 350. [Google Scholar] [CrossRef] [PubMed]
  24. Liu, Y.L.; Ce, F.; Tang, H.; Tian, G.F.; Yang, L.; Qian, W.; Dong, H.L. Genome-wide analysis of the serine carboxypeptidase-like (SCPL) proteins in Brassica napus L. Plant Physiol. Biochem. 2022, 186, 310–321. [Google Scholar] [CrossRef] [PubMed]
  25. Fu, G.Y.; Chen, B.J.; Pei, X.X.; Wang, X.Y.; Wang, X.; Nazir, M.F.; Wang, J.J.; Zhang, X.; Xing, A.; Pan, Z.; et al. Genome-wide analysis of the serine carboxypeptidase-like protein family reveals Ga09G1039 is involved in fiber elongation in cotton. Plant Physiol. Biochem. 2023, 201, 107759. [Google Scholar] [CrossRef] [PubMed]
  26. Moura, D.S.; Bergey, D.R.; Ryan, C.A. Characterization and localization of a wound-inducible type I serine-carboxypeptidase from leaves of tomato plants (Lycopersicon esculentum Mill.). Planta 2001, 212, 222–230. [Google Scholar] [CrossRef]
  27. Sheahan, J.J. Sinapate esters provide greater UV-B attenuation than flavonoids in Arabidopsis thaliana (Brassicaceae). Am. J. Bot. 1996, 83, 679–686. [Google Scholar] [CrossRef]
  28. Shirley, A.M.; McMichael, C.M.; Chapple, C. The sng2 mutant of Arabidopsis is defective in the gene encoding the serine carboxypeptidase-like protein sinapoylglucose:choline sinapoyltransferase. Plant J. 2001, 28, 83–94. [Google Scholar] [CrossRef]
  29. Lehfeldt, C.; Shirley, A.M.; Meyer, K.; Ruegger, M.O.; Cusumano, J.C.; Viitanen, P.V.; Strack, D.; Chapple, C. Cloning of the SNG1 gene of Arabidopsis reveals a role for a serine carboxypeptidase-like protein as an acyltransferase in secondary metabolism. Plant Cell 2000, 12, 1295–1306. [Google Scholar] [CrossRef]
  30. Fraser, C.M.; Thompson, M.G.; Shirley, A.M.; Ralph, J.; Schoenherr, J.A.; Sinlapadech, T.; Hall, M.C.; Chapple, C. Related Arabidopsis serine carboxypeptidase-like sinapoylglucose acyltransferases display distinct but overlapping substrate specificities. Plant Physiol. 2007, 144, 1986–1999. [Google Scholar] [CrossRef]
  31. Urasaki, N.; Takagi, H.; Natsume, S.; Uemura, A.; Taniai, N.; Miyagi, N.; Fukushima, M.; Suzuki, S.; Tarora, K.; Tamaki, M.; et al. Draft genome sequence of bitter gourd (Momordica charantia), a vegetable and medicinal plant in tropical and subtropical regions. DNA Res. 2017, 24, 51–58. [Google Scholar] [CrossRef]
  32. Letunic, I.; Bork, P. 20 years of the SMART protein domain annotation resource. Nucleic Acids Res. 2018, 46, D493–D496. [Google Scholar] [CrossRef] [PubMed]
  33. Letunic, I.; Doerks, T.; Bork, P. SMART: Recent updates, new developments and status in 2015. Nucleic Acids Res. 2015, 43, D257–D260. [Google Scholar] [CrossRef] [PubMed]
  34. Finn, R.D.; Mistry, J.; Tate, J.; Coggill, P.; Heger, A.; Pollington, J.E.; Gavin, O.L.; Gunasekaran, P.; Ceric, G.; Forslund, K.; et al. The Pfam protein families database. Nucleic Acids Res. 2010, 38, D211–D222. [Google Scholar] [CrossRef] [PubMed]
  35. Mistry, J.; Chuguransky, S.; Williams, L.; Qureshi, M.; Salazar, G.A.; Sonnhammer, E.L.L.; Tosatto, S.C.E.; Paladin, L.; Raj, S.; Richardson, L.J.; et al. Pfam: The protein families database in 2021. Nucleic Acids Res. 2021, 49, D412–D419. [Google Scholar] [CrossRef] [PubMed]
  36. Kumar, S.; Stecher, G.; Tamura, K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol. Biol. Evol. 2016, 33, 1870–1874. [Google Scholar] [CrossRef]
  37. Hu, B.; Jin, J.; Guo, A.Y.; Zhang, H.; Luo, J.; Gao, G. GSDS 2.0: An upgraded gene feature visualization server. Bioinformatics 2015, 31, 1296–1297. [Google Scholar] [CrossRef]
  38. Guan, F.; Zhang, J.Y.; Shi, B.; Wan, X.J.; Huang, C.L. Identification and fungicides screening of bitter gourd fusarium wilt pathogen. Acta Agric. Jiangxi 2021, 33, 68–72. [Google Scholar]
  39. Li, H.; Wang, X.M.; Chen, L.; Ahammed, G.J.; Xia, X.J.; Shi, K.; Considine, M.J.; Yu, J.Q.; Zhou, Y.H. Growth temperature-induced changes in biomass accumulation, photosynthesis and glutathione redox homeostasis as influenced by hydrogen peroxide in cucumber. Plant Physiol. Biochem. PPB 2013, 71, 1–10. [Google Scholar] [CrossRef]
  40. Meenakshi, G.; Solomon, J.J.; Sulochana, C.B. Toluidine blue metachromasia; a means for farly detection of plant virus infection. Stain. Technol. 2009, 47, 267–268. [Google Scholar] [CrossRef]
  41. Sakai, W.S. Simple method for differential staining of paraffin embedded plant material using toluidine blue O. Stain technology 2009, 48, 247–249. [Google Scholar] [CrossRef]
  42. Trinh, L.A.; McCutchen, M.D.; Bonner-Fraser, M.; Fraser, S.E.; Bumm, L.A.; McCauley, D.W. Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain. BioTechniques 2018, 42, 756–759. [Google Scholar] [CrossRef] [PubMed]
  43. Hanna, W.M.; Kwok, K. Chromogenic hybridization: A viable alternative to fluorescence hybridization in the HER2 testing algorithm. Mod. Pathol. 2006, 19, 481–487. [Google Scholar] [CrossRef] [PubMed]
  44. Todorović-Raković, N. Detection of c-myc amplification in formalin-fixed paraffin-embedded tumor tissue by chromogenic in situ hybridization (CISH). Methods Mol. Biol. 2013, 1012, 249–254. [Google Scholar] [PubMed]
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Figure 1. Evolutionary relationships of SCPL proteins from M. charantia and A. thaliana. (A) The evolutionary history analysis. (B) The main motif logo of SCPL. The characters show the consensus sequence. G, glycine; S, serine; x, any amino acid.
Figure 1. Evolutionary relationships of SCPL proteins from M. charantia and A. thaliana. (A) The evolutionary history analysis. (B) The main motif logo of SCPL. The characters show the consensus sequence. G, glycine; S, serine; x, any amino acid.
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Figure 2. Gene structure of McSCPLs. (A) Phylogenetic tree of McSCPL-encoding proteins. (B) The conserved motif distribution in the McSCPL protein sequence. The character sequence of each motif is shown in Supplementary Figure S1. (C) Genomic structure diagram of McSCPLs in bitter gourd. Exons are represented with green boxes; introns are shown with lines.
Figure 2. Gene structure of McSCPLs. (A) Phylogenetic tree of McSCPL-encoding proteins. (B) The conserved motif distribution in the McSCPL protein sequence. The character sequence of each motif is shown in Supplementary Figure S1. (C) Genomic structure diagram of McSCPLs in bitter gourd. Exons are represented with green boxes; introns are shown with lines.
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Figure 3. cis-element distribution of McSCPL genes. (A) Location of the cis-elements. (B) Quantitative analysis of the cis-elements.
Figure 3. cis-element distribution of McSCPL genes. (A) Location of the cis-elements. (B) Quantitative analysis of the cis-elements.
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Figure 4. Expression pattern of McSCPLs. (A) Expression patterns after being treated by FOM. RM, mock root tissues of the resistant variety (BK0604); SM, mock root tissues of the susceptible variety (LK0901). Samples of the inoculated roots were collected at 6 days (RI6 and SI6), 12 days (RI12 and SI12), and 15 days (RI15). (B) Expression patterns in tissues of bitter gourd cultivar Dali-11. The data are log2-transformed and represented by the color bar.
Figure 4. Expression pattern of McSCPLs. (A) Expression patterns after being treated by FOM. RM, mock root tissues of the resistant variety (BK0604); SM, mock root tissues of the susceptible variety (LK0901). Samples of the inoculated roots were collected at 6 days (RI6 and SI6), 12 days (RI12 and SI12), and 15 days (RI15). (B) Expression patterns in tissues of bitter gourd cultivar Dali-11. The data are log2-transformed and represented by the color bar.
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Figure 5. Expression of McSCPLs in response to SA treatment. (A) Expression level of McSCPL10. (B) Expression level of McSCPL15. (C) Expression level of McSCPL17. (D) Expression level of McSCPL22. (E) Expression level of McSCPL23. (F) Expression level of McSCPL25. The bar showed standard errors (SE). ns: no significance; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 (Student’s t-test).
Figure 5. Expression of McSCPLs in response to SA treatment. (A) Expression level of McSCPL10. (B) Expression level of McSCPL15. (C) Expression level of McSCPL17. (D) Expression level of McSCPL22. (E) Expression level of McSCPL23. (F) Expression level of McSCPL25. The bar showed standard errors (SE). ns: no significance; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 (Student’s t-test).
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Figure 6. Analysis of the expression of certain McSCPLs in response to JA treatment. (A) Expression level of McSCPL10. (B) Expression level of McSCPL15. (C) Expression level of McSCPL17. (D) Expression level of McSCPL22. (E) Expression level of McSCPL23. (F) Expression level of McSCPL25. The bar showed standard errors (SE). ns: non significance; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 (Student’s t-test).
Figure 6. Analysis of the expression of certain McSCPLs in response to JA treatment. (A) Expression level of McSCPL10. (B) Expression level of McSCPL15. (C) Expression level of McSCPL17. (D) Expression level of McSCPL22. (E) Expression level of McSCPL23. (F) Expression level of McSCPL25. The bar showed standard errors (SE). ns: non significance; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 (Student’s t-test).
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Figure 7. Detection of hydrogen peroxide accumulation. (A) Staining results of hydrogen peroxide in the stems after FOM infection. (B) Methylamine blue staining of stems. BK0604, resistant variety; LK0901, susceptible variety.
Figure 7. Detection of hydrogen peroxide accumulation. (A) Staining results of hydrogen peroxide in the stems after FOM infection. (B) Methylamine blue staining of stems. BK0604, resistant variety; LK0901, susceptible variety.
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Figure 8. The expression of McSCPL22. (A) Subcellular localization of McSCPL22. (B) In situ hybridization of the McSCPL22 gene. (C) In situ hybridization of the FOM probe. The red arrow shows the expression site.
Figure 8. The expression of McSCPL22. (A) Subcellular localization of McSCPL22. (B) In situ hybridization of the McSCPL22 gene. (C) In situ hybridization of the FOM probe. The red arrow shows the expression site.
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Table 1. List of bitter gourd SCPL genes.
Table 1. List of bitter gourd SCPL genes.
NCBI_IDRenamedCDDSMARTLengthMW (Da)pISubcellular Localization
LOC111013428McSCPL1++56963,021.24.87Vac
LOC111007166McSCPL2++43348,972.98.8Vac
LOC111010259McSCPL3++45651,938.37.22Vac
LOC111010625McSCPL4++48254,264.15.53Vac
LOC111014585McSCPL5++48054,343.48.31Vac
LOC111020681McSCPL6++47252,963.47.09Vac
LOC111022214McSCPL7++47052,931.48.41Vac
LOC111014800McSCPL8++51257,242.17.25Vac
LOC111018253McSCPL9++50556,220.96.32Pero
LOC111011999McSCPL10++48053,752.86.24Vac
LOC111006031McSCPL11++50957,031.15.2Pero, Vac
LOC111020491McSCPL12++50656,443.75.44Pero, Vac
LOC111021720McSCPL13++47753,267.26.94Vac
LOC111020263McSCPL14++47954,015.86.79Vac
LOC111024700McSCPL15++47153,692.95.09Vac
LOC111024586McSCPL16++49855,155.16.67Vac
LOC111007100McSCPL17++46752,517.27.05Vac
LOC111023593McSCPL18++46551,808.36.7Vac
LOC111021347McSCPL19++48053,215.96.79Vac
LOC111025862McSCPL20 +47053,137.96.1Vac
LOC111025866McSCPL21 +46351,972.66.98Vac
LOC111019830McSCPL22++49855,001.18.58Vac
LOC111015180McSCPL23++39544,625.36.85Vac
LOC111025759McSCPL24 +49856,128.56.97Vac
LOC111018332McSCPL25 +45650,580.46.42Vac
LOC111013427McSCPL26++36640,877.84.85Pero
LOC111020504McSCPL27 +46752,041.86.79Vac
LOC111011240McSCPL28 +46251,115.85.79Vac
LOC111011335McSCPL29 +43348,114.76.27Vac
LOC111011336McSCPL30 +43347,813.47.86Vac
LOC111019252McSCPL31 +1457161,625.47.24Vac
LOC111008438McSCPL32 +26831,091.98.34Vac
+: Confirmed in according databases. Vac: Vacuole; Pero: Peroxisome.
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Guan, F.; Yang, X.; Shi, B.; Wang, K.; Zhang, J.; Xie, Y.; Wan, X. Genome-Wide Analysis of the Serine Carboxypeptidase-like (SCPL) Protein Family of Bitter Gourd and Functional Validation of McSCPL22 in Fusarium oxysporum f. sp. Momordicae (FOM) Resistance. Int. J. Mol. Sci. 2024, 25, 11816. https://doi.org/10.3390/ijms252111816

AMA Style

Guan F, Yang X, Shi B, Wang K, Zhang J, Xie Y, Wan X. Genome-Wide Analysis of the Serine Carboxypeptidase-like (SCPL) Protein Family of Bitter Gourd and Functional Validation of McSCPL22 in Fusarium oxysporum f. sp. Momordicae (FOM) Resistance. International Journal of Molecular Sciences. 2024; 25(21):11816. https://doi.org/10.3390/ijms252111816

Chicago/Turabian Style

Guan, Feng, Xuetong Yang, Bo Shi, Kai Wang, Jingyun Zhang, Yuanyuan Xie, and Xinjian Wan. 2024. "Genome-Wide Analysis of the Serine Carboxypeptidase-like (SCPL) Protein Family of Bitter Gourd and Functional Validation of McSCPL22 in Fusarium oxysporum f. sp. Momordicae (FOM) Resistance" International Journal of Molecular Sciences 25, no. 21: 11816. https://doi.org/10.3390/ijms252111816

APA Style

Guan, F., Yang, X., Shi, B., Wang, K., Zhang, J., Xie, Y., & Wan, X. (2024). Genome-Wide Analysis of the Serine Carboxypeptidase-like (SCPL) Protein Family of Bitter Gourd and Functional Validation of McSCPL22 in Fusarium oxysporum f. sp. Momordicae (FOM) Resistance. International Journal of Molecular Sciences, 25(21), 11816. https://doi.org/10.3390/ijms252111816

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