Proteins containing steroidogenic acute regulatory protein-related lipid transfer (START) domain exist widely in the biological world, and are generally divided into 2 classes, one containing both the START domain and other domains, and the other containing the START domain only. In Arabidopsis thaliana, there are 35 proteins containing START domain, 8 of them only have START domain, and this protein subfamily containing START domain only provides a good research material for the biological function of START domain. This study analyzed the tissue expression characteristics of the 8 members by qPCR and bioinformatics method. The results showed that 1st, 3rd, 4th, and 5th chromosome each had 2 of the 8 members, and the size of the START domain and the amino acid sequence of the 8 members were not exactly the same. The 8 members had different tissue expression. In order to clarify the biological function of START domain, the gene overexpression plants and T-DNA insertion mutants of the 8 members were used as materials, the phenotypes of seedlings, leaves and pods were analyzed. Gene overexpression plants and T-DNA insertion mutants of subfamily members containing only the START domain showed asymmetric leaf growth, curling and arrested development. At the same time, the pod was no longer in whorl growth, the pod initiation became disordered, and there was a continuous growth on the same side of the stem. The above results indicate that the START domain family plays an important role in plant morphogenesis. The present study provides basic material for elucidating the molecular regulating mechanism of START domain in plant growth and development.

The storability of rice (Oryza sativa) seeds is an important guarantee for food security and the safety of seed use. It is the basis of genetic improvement of rice seed storability to explore QTL loci and screen excellent storability haplotypes. In this study, 431 core collections from 3 000 rice (3K rice) resources in the world were used as research materials, which were sown in Changsha, Hunan in 2019 and in Sanya, Hainan in 2020, making the maturity period of each trial material basically consistent. The harvested seeds were identified for storability after natural storage and artificial aging treatment. Genome wide association study (GWAS) was performed on rice seed storability based on genotypic data. Using P<1×10^-4 as the threshold, 14 QTL for seed storability were repeatedly detected, which were distributed on chromosome 2, 3, 5, 8, 9, 10, 11 and 12. The explained phenotypic variation ranged from 5% to 24%, and qSS3-2 and qSS5-2 were new QTL loci for seed storability. A total of 17 candidate genes related to rice seed storage tolerance were mined in these 14 QTL intervals, and haplotype analysis of the candidate genes in the qSS5-2 interval yielded 8 rice seed storability haplotypes. The results of this study provides theoretical reference and important gene resources for genetic improvement of rice seed storability.

Zinc is a trace element necessary for plant growth and development. The lacking or presence of excessive zinc have severe impacts on growth of plants. Zinc transporter plays an important role in regulating zinc homeostasis in plants. In this study, the potato (Solanum tuberosum) zinc transporter gene StZnT2 (GenBank No. Xm_006358632.2) was cloned from the in vitro seedlings of potato cultivar 'Atlantic'. The results showed that the gene was 1 294 bp in length and 1 011 bp in CDS region, encoding 336 amino acids. Protein molecular weight was 36 682.78 kD. Theoretical isoelectric point was 5.63. StZnT2 was a hydrophobic transmembrane protein with a typical ZIP conserved domain. StZnT2 was located on the cell membrane. Comparative analysis of homologous proteins showed that the proteins encoded by StZnT2 gene were most similar to S. pennellii, S. lycopersicum and S. commersonii. Potato plants were stunted and root elongation was inhibited under zinc deficiency and zinc toxicity. The expression of genes under different zinc concentration gradient stress were analyzed by qPCR. The results showed that the expression level of StZnT2 was up-regulated under zinc deficiency and excessive zinc stress, and the expression level was the highest in roots. These results provide theoretical basis for further elucidating the function of StZnT2 gene in potato.

GGB (geranyl geranyltransferase beta) gene encodes type Ⅰ protein geranyltransferase, which is involved in protein isoprene modification and plays an important regulatory role in plant response to stress. In order to explore the biological function of GhGGB gene in cotton (Gossypium hirsutum), the plant complementary expression vector pBI101-AtGGBP-GhGGB-AtGGBT was constructed and transformed into ggb mutant of Arabidopsis thaliana, and GhGGB transgenic lines were screened. Phenotypic, physiological and biochemical indexes of GhGGB transgenic lines, ggb mutant and wild type Arabidopsis Col-0 under natural drought stress were detected. The results showed that with wild-type Arabidopsis Col-0 as the control, the sensitivity of transgenic GhGGB plants to drought stress was significantly enhanced, while the ggb mutant plants showed stronger drought resistance. The determination results of physiological indexes of each component were consistent with the phenotype, indicating that GhGGB gene complemented the functional deficiency of ggb mutant in Arabidopsis and reduced drought tolerance. The results of this study preliminarily showed that GhGGB was a negative regulatory gene for drought resistance, which provides a new gene selection for cotton drought-resistant germplasm cultivation.

Cajanus cajan is a medicinal and dietary woody legume with rich active substances and high economic value. Abiotic and biotic stresses, including drought and diseases, are important factors in the reduction of yield and quality of C. cajan; therefore, mining key C. cajan resistance genes and analyzing their functions are the focus of C. cajan breeding research. As an important member of the MYB (v-myb avian myeloblastosis viral) family of transcription factors, MYB-related (MYB-R) subfamily genes are thought to play an important role in adversity stress. In this study, 48 CcMYB-R (CcMYB-related) family genes (CcMYB-R1~CcMYB-R48) containing conserved structural domains of the MYB-R subfamily were screened by bioinformatic methods using Arabidopsis thaliana MYB-R subfamily genes for sequence alignment in the C. cajan genome. Analysis of transcriptome data revealed that CcMYB-R48 was significantly up-regulated and highly expressed under abscisic acid (ABA), Methyl Jasmonate (MeJA) stress treatments, suggested that it might be play an important function in stress response and hormone induction in C. cajan. Cloning and analysis of the CcMYB-R48 revealed that it had a typical MYB functional domain with high homology to Arabidopsis thaliana AT1G19000 and AT1G74840. The semi-quantitative RT-PCR (SqRT-PCR) revealed that CcMYB-R48 was significantly up-regulated in expression under both 6 and 12 h of drought stress, whereas no difference was observed under 6 and 12 h of high-temperature stress, suggested its possible involved in drought resistance in C. cajan. CcMYB-R48 overexpression vector was constructed and overexpressed CcMYB-R48 in C. cajan plants were established using the C. cajan instantaneous conversion system. The results showed that the overexpression plants had lower mortality of 19.44% under drought conditions, and their electrical conductivity, malondialdehyde (MDA) content and peroxidase (POD) content were significantly lower than those of the control plants, indicated that CcMYB-R48 could enhance the drought resistance of C. cajan. This study is important to further investigate the CcMYB-R subfamily genes of C. cajan and their drought resistance mechanism and to carry out molecular resistance breeding work.

Golden2-like (G2-like) transcription factors play important roles during chloroplast development, chlorophyll accumulation, photosynthesis and senescence in plants. In order to explore the role of G2-like transcription factor during grape (Vitis vinifra) development, bioinformatics method was used to identifiy G2-like family transcription factor in grape, on the basis of phylogenetic, conserved domain and synteny analysis, expression of G2-like in different organs and during rachis browning was quantified. A total of 43 G2-like (VvG1~VvG43) transcription factors uneven distributed in 17 chromosomes were identified in grape genome, and were classified into 6 groups based on phylogeny analysis with Arabidopsis thaliana. 34 pairs of synteny between A. thaliana and grape, 5 segmental duplication, 1 tandem duplication and conserved Myb DNA-binding domain were found in grape. qPCR results revealed that VvG1, VvG28, VvG43, VvG2, VvG14, VvG6, VvG24 showed positive correlation with chlorophyll content variation during rachis browning of grape, but VvG27 showed negative expression pattern. This results indicated G2-like genes played the important role in tissue development and rachis browning of grape, which provides a reference basis for further function analysis of G2-like genes.

Calmodulin (CaMs) are the primary calcium-binding proteins in plants which control diverse cellular processes, including plant development and hormonal response. In this study, a genome-wide analysis conducted in Rosa roxburghii and identifified 2 RrCaM-encoding genes named RrCaM1 and RrCaM2. The RrCaM1 contains 3 introns while the RrCaM2 gene has no introns. Both of the RrCaMs contained 4 typical EF-hands. The RrCaM1 and RrCaM2 genes were located on the 3^rd and 4^th chromosome, repectively. The phylogenetic relationship and synteny analyses showed that RrCaM1 and RrCaM2 were clustered as 2 different subgroups. Besides, RrCaM1 and RrCaM2 shared one orthologous duplication pair with Fragaria vesca, respectively. RrCaM1 and Pyrus bretschneideri had 2 orthologous duplication pairs. CaMs genetic relationship among R. roxburghii, P. bretschneideri and Fragaria vesca was close. The results of RNA-Seq and qPCR indicated that the expression of RrCaM2 gene decreased significantly with the fruit development of R. roxburghii, while RrCaM1 was the opposite. Cis-acting element prediction and qPCR analysis showed that the promoter region of RrCaM1 contained 8 response elements such as hormone and light, while RrCaM2 lacked cis-acting elements responsive to gibberellin (GA₃), low temperature and drought, which also showed completely different response patterns to salicylic acid (SA), abscisic acid (ABA), and GA₃ treatments.The vitamin C (ascorbate, AsA) content and RrCaMs gene expression of R. roxburghii fruit were significantly decreased by chlorpromazine (CPZ) treatment (P<0.05). RrCaMs had a significant positive correlation with the expression of the L-galactose phosphatase (GPP) gene in AsA biosynthesis pathway and the dehydroascorbate reductase (DHAR) gene in recycling pathway. It indicates that it may be involved in the regulation of AsA accumulation in R. roxburghii fruit. This study found that the RrCaMs gene family played potentially important roles in fruit development, AsA accumulation and hormone response of R. roxburghii, and provides reference for the functional study of this gene family.

Uncaria rhynchophylla is one of commonly used staple Chinese medicinal materials in China, mainly used for headache, Alzheimer's disease and hypertension. Its active ingredients are mainly terpenoid indole alkaloids (TIAs). Strictosidine synthase (STR) is a key enzyme in the biosynthesis pathway of terpenoid indole alkaloids. So far, it has not been reported in U. rhynchophylla. In order to search for STR genes involved in the TIAs biosynthesis of U. Rhynchophylla. LC-MS method was used to measure the content of rhyncholphylline and isorhyncophylline in root, leaf, stem hook and capsule of U. rhynchophylla. Total RNA from different parts of U. rhynchophylla were compared by 4 extraction methods. UrSTR gene, which may be involved in the biosynthesis of rhyncholphylline and isorhyncophylline, was cloned and bioinformatics analysis was carried out. The expression patterns of UrSTR in different parts of U. rhynchophylla were detected by qPCR. The results were as follows: (1) The contents of rhyncholphylline and isorhyncophylline were the highest in capsules and the lowest in roots through LC-MS method. (2) The improved CTAB method was the best extraction method for different parts of U. rhynchophylla. (3) The total length of UrSTR was 1 122 bp, encoding a peptide chain of 374 amino acids with a transmembrane helical structure. The theoretical molecular weight of UrSTR was 41.446 kD, and the theoretical isoelectric point was 7.549. The evolution of UrSTR gene was clustered with Coffea arabica and C. canephora, showing a close genetic relationship. (4) The results of qPCR showed that UrSTR gene was mainly expressed in capsule and the lowest expression was found in root. The analysis showed that the expression of UrSTR gene was consistent with the trend of the content of rhyncholphylline/isorhyncophylline, indicating that UrSTR gene may be one of the candidate genes for U. rhynchophylla involved in the biosynthesis of TIAs. This study provides reference for the functional verification of UrSTR gene and the analysis of U.rhynchophylla alkaloid synthesis pathway.

It is critical to the actual output of animal husbandry that the sex control of animals be properly managed. A frequent commodity in the manufacturing of sex-controlled semen is used to regulate the sex ratio of their progeny. The commercial use of sex-controlled semen is mostly concentrated in dairy cows (Bos taurus), and sex-controlled frozen semen plays an important role in the propagation of improved breeds of dairy cows. Both the mechanical damage and the reduction of sperm motility after sorting are inevitable, so immunological methods to sort X/Y sperm is an ideal method. Semen from 10 healthy male boars was collected to study the gene expression of proteins on the sex chromosomes of porcine (Sus scrofa) sperm and its biological function, as well as to search for possible antigens for sex control in porcine sperm. Flow cytometry was used to remove seminal plasma and separate the sperm into X/Y sperm. The protein expression in the isolated porcine sperm was analyzed qualitatively and quantitatively by Label-free proteomics technology, and the expressed genes distributed on the sex chromosome were extracted and analyzed their biological function. Simultaneously, the protein expressed on the sex chromosome was predicted the transmembrane region by TMHMM 2.0. Subcellular localization was predicted by WoLF PSORT, signal peptide was predicted by SignalP 5.0; and some selected antigens were validated. A total of 1 925 proteins and 13 558 peptides were obtained by proteinomic sequencing. According to the results of this study, there were 2 976 proteins-coding genes, of which 67 were expressed by sex-specific genes, with 65 X and 2 Y chromosomes genes. The GO and KEGG analyses of the genes expressed on allosome revealed that they were mostly engaged in the energy generation pathway associated with sperm motility. 45 proteins were expected to contain the transmembrane structure, 47 proteins were located on the plasma membrane or extracellular region, and 28 proteins could generate signal peptides (secretory signal peptides/signal peptides, Sec/SPI). The proteins expressed in deoxyribonuclease like 1 (DNASE1L1), ATPase H⁺ transporting accessory protein 1 (ATP6AP1), tetraspanin 6 (TSPAN6), NADH: ubiquinone oxidoreductase subunit A1 (NDUFA1), phosphoribosyl pyrophosphate synthetase 2 (PRPS2), NADPH oxidase heavy chain subunit (GP91-PHOX), B cell receptor associated protein 31 (BCA31), sperm acrosome associated 5 (SPACA5), TIMP metallopeptidase inhibitor 1 (TIMP1), peroxiredoxin 4 (PRDX4) and apolipoprotein O like (APOOL) were selected as candidate antigens. The proteins ATP6AP1, NDUFA1, PRPS2, GP91-PHOX and TIMP1 were identified by Western blot. This study provides a reference for studying the genes expressed on the neutral chromosomes of porcine sperm and the strategies of sex control of porcine sperm.

β2 integrins αD (ITGAD) is a subunit of heterodimeric surface receptor, and plays a critical role in the regulation of lipid metabolism and blood glucose balance. This work aimed to detect the candidate molecular markers for marker-assisted selection (MAS) in goats (Capra hircus).In this study, the Insertion/Deletion (InDel) polymorphism of ITGAD gene in 3 goat breeds (355 in total) was identified through DNA pooling technology and high-resolution melting (HRM) analysis . The results showed that a novel InDel locus (NC_030832.1: g.15243_15244 ins C) was detected. Furthermore, in 3 goat breeds, the polymorphic information content (PIC) of the InDel locus was 0.289~0.369. The InDel locus was consistent with Hardy-Weinberg in 3 experimental populations. Association analysis showed that this InDel locus was significantly associated with the huckle bone width of Fuqing goat and the body height, body length, and chest circumference of Jianyang Daer goat (P<0.05). It was also extremely significantly correlated with chest depth, chest width, cannon circumference, and cannon circumference index of Jianyang Daer goat (P<0.01). Moreover, the II genotype conferred positive effects on growth traits compared with the DD genotype. The present study found the significant association of ITGAD InDel locus with the growth traits in goats, which could be adopted as the molecular genetic marker for growth traits of goats.

The copper metabolism Murr1 domain (COMMD) protein family is an important biologic part of the nuclear factor-κB (NF-κB) pathway, sodium transport, copper regulation, and adaptive regulation in hypoxia. Commd9 cDNA in high-alkaline-tolerant fish, Leuciscus waleckii, was cloned firstly and given the name LwCommd9 (GenBank No. MT013199) by rapid-amplification of cDNA ends (RACE) to find out what Commd9 does in alkaline environment. The results showed that the full-length cDNA sequence of the Commd9 gene was 938 bp in L. waleckii, which had 5 bp of 5' non-coding region, 339 bp of 3' non-coding region, and 594 bp of the ORF fragment could code for 197 amino acids that had a predicted molecular weight of about 21.79 kD and a theoretical isoelectric point of 5.63. Phylogenetic analysis revealed that the amino acid sequence of LwCommd9 showed high similarity with carp, particularly with Anabarilius grahami up to 93.37%. qPCR showed that LwCommd9 were expressed in virous organs, such as significant increase in liver and kidney (P<0.05). In addition, LwCommd9 expression in liver was significantly down regulated (P<0.01) in alkaline water species (alkali-water, AW), but that was highly up regulated (P<0.01) in freshwater species (FW) and showed in kidney with the same trend. The above results indicate that the Commd9 gene in L. waleckii might play a critial role in the process of high-alkaline adaptation, and the in-depth mechanisms related to this pathway can be explored deeply.

Quasipaa spinosa, a characteristic economic amphibian in China, have both gastronomic and medicinal values. As a sexual dimorphic feature of frogs, the spines of Q. spinosa skin derivatives are of great significance to improve the success rate of holding and also an important classification and identification feature of Tribe Paini. The molecular mechanism of its generation and changes will help to clarify the phylogenetic relationship of the amphibian Tribe Paini. In order to explore the molecular mechanism of the seasonal changes in spines of the Q. spinosa, this study performed transcriptome sequencing on 3 sets of samples from the breast skin of the male Q. spinosa during the breeding period and post-breeding period and the abdomen skin of the male Q. spinosa during the breeding period. A total of 100 312 Unigenes were obtained. After annotation, the result showed that the expression of multiple keratinizing protein genes significantly increased in the breast skin of male Q. spinosa during the breeding period. Genes related to apoptosis, cell proliferation and sex hormone synthesis were found in the differentially expressed genes of chest spiny skin in breeding period and post-breeding period. KEGG analysis showed that multiple differentially expressed genes were found to be enriched in the intercellular lipid metabolism of the skin stratum corneum. It was speculated that the increased keratin gene expression during the breeding period was one of the direct causes of the formation of spines. The reduction of lipid synthesis in the stratum corneum of the skin and the slowdown of cell proliferation might cause the spines to become smaller. In addition, the expression of genes related to sex hormone synthesis decreased during the breedding period to post-breeding period, indicating that the seasonal changes in spines of the Q. spinosa might be regulated by hormone changes. The 15 differentially expressed genes were verified by qPCR, the results showed that the transcriptome results were reliable. This study elucidates the molecular mechanism of spine formation and seasonal changes in Q. spinosa, the findings provide the basis for taxonomic classification of Q. spinosa, and a reference for related candidate genes and metabolic pathways for the seasonal variation and sexual dimorphism of skin derivatives of other vertebrates.

Cry toxin is the main insecticidal protein produced by Bacillus thuringiensis (Bt) strain, Cry4Ba and Cry11Aa of which are highly toxic to the larvae of Aedes aegypti. Cry toxin infects the insect midgut epithelial cells by binding to the specific membrane receptors. The midgut immune factors can recognize and interact with Cry toxin, and further drive the immune system to resist Bt infection. Therefore, in order to further explore the molecular interaction mechanism of Cry4Ba and Cry11Aa toxins to Aedes aegypti, the GST-Cry4Ba and GST-Cry11Aa fusion proteins were prepared for the GST-pull down assay, and the Cry4Ba and Cry11Aa protoxins were prepared for the Co-immunoprecipitation assay. The 133 Cry4Ba-interacting proteins and 237 Cry11Aa-interacting proteins were identified from the midgut brush border membrane vesicles (BBMVs) by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and database searching. Bioinformatics analysis revealed that at least 16 membrane-binding or transmembrane proteins, including 5 aminopeptidases N, 1 alkaline phosphatase, 1 cadherin-like, 2 α-amylases, 1 class B scavenger receptor, 2 voltage-dependent anion-selective channel proteins and 4 actins. In addition, 7 Cry-binding proteins related to immune or maintenance of cell homeostasis were screened, including 1 peptidoglycan recognition protein (short), 1 apolipophorin, 1 hemomucin, 1 serpin and 3 heat shock proteins. These results provided the necessary reference for functional verification of Bt receptors and the immune response mechanism against Bt in Aedes aegypti.

RNA methylation is the most important form of RNA modification, and N6-methyladenosine (m⁶A) is the most prevalent post-transcriptional modification of eukaryotic mRNA, which plays an important role in the regulation of growth and development of organisms. RNA methyltransferase acts as the major regulators of RNA methylation with few documented functional dissections focused on phytopathogenic fungi of plants. In the present study, structure and function of RNA methyltransferase in plant pathogenic fungi were explored via bioinformatics methods. The results showed that a total of 159 RNA methyltransferase genes were identified in 106 phytopathogenic fungi. The analysis of physicochemical properties showed that the average length of amino acids (aa) encoded by 159 phytopathogenic fungi RNA methyltransferases was about 470 aa (ranging from 179 to 1206 aa), the average molecular weight was about 52.64 kD (variation range 20.04~134.35 kD), and the average isoelectric point was about 7.26 (variation range 4.90~10.28). The evolutionary relationship of phytopathogenic fungi RNA methyltransferases was relatively conservative. The RNA methyltransferases which in the same phylum were generally clustered in the same clade, and the conservation motifs in same clade had highly similarity. Furthermore, Real-time qPCR method was used to analyze the expression pattern of gene encoding RNA methyltransferases in Setosphaeria turcica (StMETTL1) at different developmental stages. It was found that the expression level of StMETTL1 was significantly increased in infected nails stage (P<0.05), suggesting that StMETTL1 may play an important role in the process of S. turcica infection of the host. This study provides data support for further understanding the role of m⁶A in plant pathogenic fungi.

The tolC gene is widespread in gram-negative bacteria, and TolC family protein (TolC) expressed by tolC is an outer membrane channel protein, which mediates the extracellular transport of various compounds and plays an important role in the biological functions of bacteria. The identification of the characteristic of tolC gene of Gallibacterium anatis and its protein would play crucial roles in efforts to prevent and control this disease. In order to analyze the structural characteristics of gene and amino acid and potential as an antigen of tolC of G. anatis, in this study, the sequences and phylogenetic analysis of tolC gene of 28 G. anatis isolates were carried out. Taking G. anatis PDS-RZ-1-SLG as the research object, the physicochemical properties and structures of TolC were predicted by bioinformatics methods. In addition, the tolC gene sequences were cloned and expressed, and its antigenic properties were analyzed. The results showed that the tolC gene, with 1 371 bp in length, was ubiquitous in Chinese isolates of G. anatis and its nucleotide sequences were highly conserved (similarity: 92.1%~100%), which had a close relationship with G. genomosp 1 and 2. Moreover, the tolC gene of G. anatis encoded a protein of 456 amino acids, with a calculated molecular mass of 51.1 kD, and the TolC protein was a stable, hydrophilic, α-type and secretory membrane protein without transmembrane domains, but with a signal peptide sequence and 10 main B cell epitopes. SDS-PAGE and Western blot analysis showed the recombinant protein was about 45 kD, and the recombinant protein could react specifically with positive serum of G. anatis and the polyclonal anti-TolC antiserum raised in rabbits had a specific binding with G. anatis, which verified the reactogenicity and immunogenicity of the TolC protein. This study provides basic information for further research on the biological functions of the TolC, and establishes a theoretical basis for development of new vaccines against G. anatis.

Avian is a valuable animal model, and plays a significant part in agriculture and biomedicine. In recent years, newly developed programmable gene editing tools such as TALEN and CRISPR/Cas9 have been successfully adopted in avian species. These gene modification technologies are expected to be applied to practical uses, such as enhancing economic traits of poultry, investigating virus-receptor interaction, developing vector vaccines, improving poultry welfare, and producing pharmaceutical proteins and antibodies in eggs, etc. In this review, we present a concise description of the existing approaches of genome editing in bird species, and the application of CRISPR/Cas9 in advancing poultry breeding and production, as well as in poultry research. In addition, we discuss the potential use of gene-edited poultry model in various industries in the future. This review will lead to improve economic traits of poultry and control avian viral diseases by means of gene editing techniques.

Telosma mosaic virus (TelMV), East Asian passiflora virus (EAPV), and Passion fruit severe mottle-associated virus (PFSMaV) belonging to the genus Potyvirus have seriously damaged the passionflower (Passiflora edulis) industry in China. So far, there are no effective agents for preventing and curing plant viruses. Thus, Developments of viral detection techniques are crucial for establishing scientific prevention and control system of plant viral diseases. In this study, field-collected passionflower leaves, co-infected with TelMV, EAPV and PFSMaV were used for virus particle purification. Purified and mixed virions of these 3 potyviruses were used as the immunogen to immunize 4 BALB/c mice (Mus musculus). Four hybridoma strains (11A12, 6D8, 6D12 and 17E4) secreting specific and broad-spectrum monoclonal antibodies against above 3 infecting passionflower plant potyviruses were obtained by the hybridoma technique including cell fusion, hybridoma cell screening, antibody detection, cell cloning and ascites preparation. Western blot assay showed that the 4 monoclonal antibodies could simultaneously recognize capsid proteins of TelMV, EAPV and PFSMaV. Further, the produced monoclonal antibodies was used as the detection antibodies to develop two serological assays, i.e. dot enzyme-linked immunosorbent assay (Dot-ELISA) and Tissue print-ELISA for the broad-spectrum, specific and sensitive detection of these 3 potyviruses. The two established serological methods could simply, quickly, specifically, sensitively, broadly and efficiently detect TelMV, EAPV and PFSMaV in passionflower samples. Meanwhile, there were negative reactions when they detected the diseased leaves infected with Plumpox virus (PPV), Potato virus Y (PVY), or Potato virus A (PVA), Turnip mosaic virus (TuMV), Passiflora latent virus (PLV), Cucumber mosaic virus (CMV) or uninfected passionflower leaves. Sensitivity analysis assays showed that the virus could be detected in infected plant crude extracts diluted up to 1∶20 480 (W/V, g/mL), i.e. in 0.097 6 ng virus-infected plant tissues of the absolute detection amount by the established Dot-ELISA serological assay. The detection results of field samples showed that the two developed serological assays could accurately, sensitively and broadly monitor these 3 potyviruses in passionflower samples. The coincidence rate of serological and RT-PCR detection results of field samples was 100%. This study provides technical support for the detection and diagnosis of these 3 potyviruses, production of virus-free passionflower seeds and seedlings, and the scientific prevention and control of these viral diseases.