The crotonylated and succinylated proteins of jujube involved in phytoplasma-stress responses.

BACKGROUND: Protein posttranslational modifications (PTMs) are fast and early responses to environmental changes, including pathogen infection. Jujube witches' broom (JWB) is a phytoplasma disease causing great economic loss in jujube production. After phytoplasma infection, the transcriptional, translational, and metabolic levels in jujube were activated, enabling it to survive during phytoplasma invasion. However, no study has yet reported on PTMs in jujube. Lysine crotonylation (Kcr) and lysine succinylation (Ksu) have been popular studies in recent years and their function in plant phytoplasma-stress responses remains unclear. RESULTS: Here, 1656 crotonylated and 282 succinylated jujube proteins were first identified under phytoplasma-stress, of which 198 were simultaneously crotonylated and succinylated. Comparative analysis revealed that 656 proteins, 137 crotonylated and 43 succinylated proteins in jujube were regulated by phytoplasma infection, suggesting that Kcr was more universal than Ksu. Kcr differentially expressed proteins (DEPs) were related to ribosomes, photosynthetic and carbon metabolism, while Ksu DEPs were mainly involved in carbon metabolism, the TCA cycle and secondary metabolite biosynthesis. The crosstalk network among proteome, crotonylome and succinylome showed that DEPs related to ribosomal, peroxidases and glutathione redox were enriched. Among them, ZjPOD51 and ZjPHGPX2 significantly increased at the protein and Kcr level under phytoplasma-stress. Notably, 7 Kcr sites were identified in ZjPHGPX2, a unique antioxidant enzyme. After inhibitor nicotinamide (NAM) treatment, GPX enzyme activity in jujube seedlings was reduced. Further, site-directed mutagenesis of key Kcr modification sites K130 and/or K135 in ZjPHGPX2 significantly reduced its activity. CONCLUSIONS: This study firstly provided large-scale datasets of Kcr and Ksu in phytoplasma-infected jujube and revealed that Kcr modification in ZjPHGPX2 positively regulates its activity.

The 'Candidatus Phytoplasma ziziphi' effectors SJP1/2 negatively control leaf size by stabilizing the transcription factor ZjTCP2 in jujube.

Phytoplasmas manipulate host plant development to benefit insect vector colonization and their own invasion. However, the virulence factors and mechanisms underlying small-leaf formation caused by jujube witches' broom (JWB) phytoplasmas remain largely unknown. Here, effectors SJP1 and SJP2 from JWB phytoplasmas were identified to induce small-leaf formation in jujube (Ziziphus jujuba). In vivo interaction and expression assays showed that SJP1 and SJP2 interacted with and stabilized the transcription factor ZjTCP2. Overexpression of SJP1 and SJP2 in jujube induced ZjTCP2 accumulation. In addition, the abundance of miRNA319f_1 was significantly reduced in leaves of SJP1 and SJP2 transgenic jujube plants and showed the opposite pattern to the expression of its target, ZjTCP2, which was consistent with the pattern in diseased leaves. Overexpression of ZjTCP2 in Arabidopsis promoted ectopic leaves arising from the adaxial side of cotyledons and reduced leaf size. Constitutive expression of the miRNA319f_1 precursor in the 35S::ZjTCP2 background reduced the abundance of ZjTCP2 mRNA and reversed the cotyledon and leaf defects in Arabidopsis. Therefore, these observations suggest that effectors SJP1 and SJP2 induced small-leaf formation, at least partly, by interacting with and activating ZjTCP2 expression both at the transcriptional and the protein level, providing new insights into small-leaf formation caused by phytoplasmas in woody plants.

Potential Plant-To-Plant Transmission: Shared Endophytic Bacterial Community Between Ziziphus lotus and Its Parasite Cuscuta epithymum.

Microbiota associated with host-parasite relationships offer an opportunity to explore interactions among plants, parasites, and microbes, thereby contributing to the overall complexity of community structures. The dynamics of ecological interactions between parasitic plants and their hosts in arid environments remain largely understudied, especially in Africa. This study aimed to examine the bacterial communities of Cuscuta epithymum L. (clover dodder), an epiphytic parasitic plant, and its host, Ziziphus lotus L. (jujuba), in an arid environment. Our goal was to uncover the ecological complexities of microbial communities within the framework of plant-plant interactions. We conducted a comprehensive analysis of the bacterial composition and diversity within populations of the C. epithymum parasite, the infected- and non-infected jujuba host, and their interface at the shoots of the host. This involved amplicon sequencing, targeting the V5-V6 regions of the 16S rRNA gene. A total of 5680 amplicon sequence variants (ASVs) were identified, with Pseudomonadota, Bacillota, and Actinobacteriota being prevalent phyla. Among the bacterial communities, three genera were dominant: Cutibacterium, Staphylococcus, and Acinetobacter. Interestingly, analyses of alpha-diversity (p = 0.3 for Shannon index and p = 0.5 for Simplon index) and beta-diversity (PERMANOVA, with p-values of 0.6 and 0.3) revealed no significant differences between Cuscuta-infected and non-infected jujube shrubs, suggesting a shared shoot endophytic bacteriome. This finding advances our comprehension of microbial communities linked to plant-parasite interactions in the arid environments of Africa. Further research on various hosts is required to confirm plant-to-plant bacterial transmission through Cuscuta infection. Additionally, studies on functional diversity, cytology, ecophysiology and the mechanisms by which bacterial communities transferred between host and parasite are necessary.

Molecular characterization of a novel partitivirus isolated from the phytopathogenic fungus Aplosporella javeedii.

Aplosporella javeedii is a pathogenic fungus that causes canker and dieback of jujube in China. In this study, we report a new mycovirus, Aplosporella javeedii partitivirus 1 (AjPV1), isolated from A. javeedii strain NX55-3. The AjPV1 genome contains two double-stranded RNA elements (dsRNA1 and dsRNA2). The size of dsRNA1 is 2,360 bp, and it encodes a putative RNA-dependent RNA polymerase (RdRp), while dsRNA2 is 2,301 bp in length and encodes a putative capsid protein (CP). The sequences of RdRp and CP have significant similarity to those of members of the family Partitiviridae. Sequence alignment and phylogenetic analysis showed that AjPV1 is a new member of the family Partitiviridae that is related to members of the genus Betapartitivirus. To our knowledge, AjPV1 is the first mycovirus reported from A. javeedii.

Isolation and characterization of endophytic bacteria for controlling root rot disease of Chinese jujube.

AIM: Fusarium oxysporum is the primary pathogen causing root rot disease that severely affects cultivation of jujube fruit in the Xinjiang province of China. The aim of this study was to identify endophytic bacteria in healthy jujube organs that could effectively suppress F. oxysporum growth. METHODS AND RESULTS: Different plant organs (leaves, twigs and roots) were collected from healthy Chinese jujube cultivated in southern Xinjiang province of China. The endophytic bacterium Brevibacterium halotolerans JZ7 was selected for its strong antagonistic activity and growth-promoting characteristics. Gas-chromatography mass-spectrometry analysis showed that acetoin, 2,3-butanediol and fenretinide were the three dominant volatile organic compounds produced by strain JZ7. Fenretinide strongly suppressed spore germination of F. oxysporum in vitro. Pot experiments showed that strain JZ7 colonized both the roots and rhizosphere soil of Chinese jujube and significantly reduced F. oxysporum level in jujube rhizosphere soil. CONCLUSION: We demonstrated that B. halotolerans JZ7 can be developed into a biological control agent to combat root rot disease of Chinese jujube in the Xinjiang province of China. SIGNIFICANCE AND IMPACT OF THE STUDY: The suggested strategy for biological control of jujube root rot disease is fully in accordance with the current principles of sustainability.

Core RNA Interference Genes Involved in miRNA and Ta-siRNA Biogenesis in Hops and Their Expression Analysis after Challenging with Verticillium nonalfalfae.

RNA interference is an evolutionary conserved mechanism by which organisms regulate the expression of genes in a sequence-specific manner to modulate defense responses against various abiotic or biotic stresses. Hops are grown for their use in brewing and, in recent years, for the pharmaceutical industry. Hop production is threatened by many phytopathogens, of which Verticillium, the causal agent of Verticillium wilt, is a major contributor to yield losses. In the present study, we performed identification, characterization, phylogenetic, and expression analyses of three Argonaute, two Dicer-like, and two RNA-dependent RNA polymerase genes in the susceptible hop cultivar Celeia and the resistant cultivar Wye Target after infection with Verticillium nonalfalfae. Phylogeny results showed clustering of hop RNAi proteins with their orthologues from the closely related species Cannabis sativa, Morus notabilis and Ziziphus jujuba which form a common cluster with species of the Rosaceae family. Expression analysis revealed downregulation of argonaute 2 in both cultivars on the third day post-inoculation, which may result in reduced AGO2-siRNA-mediated posttranscriptional gene silencing. Both cultivars may also repress ta-siRNA biogenesis at different dpi, as we observed downregulation of argonaute 7 in the susceptible cultivar on day 1 and downregulation of RDR6 in the resistant cultivar on day 3 after inoculation.

Genome-wide identification of MAPKKK genes and their responses to phytoplasma infection in Chinese jujube (Ziziphus jujuba Mill.).

BACKGROUND: Mitogen-activated protein kinase (MAPK) cascades play vital roles in signal transduction in response to a wide range of biotic and abiotic stresses. In a previous study, we identified ten ZjMAPKs and five ZjMAPKKs in the Chinese jujube genome. We found that some members of ZjMAPKs and ZjMAPKKs may play key roles in the plant's response to phytoplasma infection. However, how these ZjMAPKKs are modulated by ZjMAPKKKs during the response process has not been elucidated. Little information is available regarding MAPKKKs in Chinese jujube. RESULTS: A total of 56 ZjMAPKKKs were identified in the jujube genome. All of these kinases contain the key S-TKc (serine/threonine protein kinase) domain, which is distributed among all 12 chromosomes. Phylogenetic analyses show that these ZjMAPKKKs can be classified into two subfamilies. Specifically, 41 ZjMAPKKKs belong to the Raf subfamily, and 15 belong to the MEKK subfamily. In addition, the ZjMAPKKKs in each subfamily share the same conserved motifs and gene structures. Only one pair of ZjMAPKKKs (15/16, on chromosome 5) was found to be tandemly duplicated. Using qPCR, the expression profiles of these MAPKKKs were investigated in response to infection with phytoplasma. In the three main infected tissues (witches' broom leaves, phyllody leaves, and apparently normal leaves), ZjMAPKKK26 and - 45 were significantly upregulated, and ZjMAPKKK3, - 43 and - 50 were significantly downregulated. ZjMAPKKK4, - 10, - 25 and - 44 were significantly and highly induced in sterile cultivated tissues infected by phytoplasma, while ZjMAPKKK6, - 7, - 17, - 18, - 30, - 34, - 35, - 37, - 40, - 41, - 43, - 46, - 52 and - 53 were significantly downregulated. CONCLUSIONS: For the first time, we present an identification and classification analysis of ZjMAPKKKs. Some ZjMAPKKK genes may play key roles in the response to phytoplasma infection. This study provides an initial understanding of the mechanisms through which ZjMAPKKKs are involved in the response of Chinese jujube to phytoplasma infection.

Genome-wide analysis of the WRKY gene family and their positive responses to phytoplasma invasion in Chinese jujube.

BACKGROUND: The WRKY gene family is one of the most important families in higher plants. As transcription factors, they actively respond to biotic and abiotic stress and are also involved in plant development. Chinese jujube (Ziziphus jujuba Mill.) is the largest type of dried fruit tree in China in terms of production, but its production is largely limited by phytoplasma infection, and the information about the role of WRKY genes under phytoplasma stress was still limited. RESULTS: We identified 54 ZjWRKYs in the jujube genome and classified them into three subgroups according to conserved WRKY domains and zinc-finger structure. 41 ZjWRKYs were distributed on 11 of 12 pseudo chromosomes in Chinese jujube. The majority of ZjWRKYs were highly expressed in the seven examined tissues, indicating that they play multiple roles in these vegetative and reproductive organs. Transcriptome data showed that most of the characterised ZjWRKYs were highly expressed at later stages of fruit development. RT-qPCR demonstrated that the expression of 23 ZjWRKYs changed following phytoplasma infection, suggesting that they are involved in signalling pathways that respond to phytoplasma stress. Then, STRING analysis and yeast two-hybrid screening proved that some ZjWRKY proteins were interacting with ZjMAPKK proteins, which were also involved in phytoplasma invasion. Moreover, their differential expressions were further confirmed in resistant and susceptible jujube varieties under phytoplasma stress. These results suggest that ZjWRKYs play significant roles in phytoplasma tolerance and should be crucial candidate genes for jujube-phytoplasma interaction. CONCLUSIONS: 54 ZjWRKYs in Chinese jujube were identified and classified into three subgroups. 41 ZjWRKYs were unevenly distributed along the chromosomes. The majority of ZjWRKYs were highly expressed in various tissues. Most of the ZjWRKYs were positive responses to phytoplasma invasion, and that provided candidate genes for the future studies of jujube-phytoplasma interaction.

Genome-wide analysis of the bHLH gene family in Chinese jujube (Ziziphus jujuba Mill.) and wild jujube.

BACKGROUND: The bHLH (basic helix-loop-helix) transcription factor is one of the largest families of transcription factors in plants, containing a large number of members with diverse functions. Chinese jujube (Ziziphus jujuba Mill.) is the species with the highest economic value in the family Rhamnaceae. However, the characteristics of the bHLH family in the jujube genome are still unclear. Hence, ZjbHLHs were first searched at a genome-wide level, their expression levels under various conditions were investigated systematically, and their protein-protein interaction networks were predicted. RESULTS: We identified 92 ZjbHLHs in the jujube genome, and these genes were classified into 16 classes according to bHLH domains. Ten ZjbHLHs with atypical bHLH domains were found. Seventy ZjbHLHs were mapped to but not evenly distributed on 12 pseudo- chromosomes. The domain sequences among ZjbHLHs were highly conserved, and their conserved residues were also identified. The tissue-specific expression of 37 ZjbHLH genes in jujube and wild jujube showed diverse patterns, revealing that these genes likely perform multiple functions. Many ZjbHLH genes were screened and found to be involved in flower and fruit development, especially in earlier developmental stages. A few genes responsive to phytoplasma invasion were also verified. Based on protein-protein interaction prediction and homology comparison, protein-protein interaction networks composed of 92 ZjbHLHs were also established. CONCLUSIONS: This study provides a comprehensive bioinformatics analysis of 92 identified ZjbHLH genes. We explored their expression patterns in various tissues, the flowering process, and fruit ripening and under phytoplasma stress. The protein-protein interaction networks of ZjbHLHs provide valuable clues toward further studies of their biological functions.

Tracing Rhizophagus irregularis isolate IR27 in Ziziphus mauritiana roots under field conditions.

Arbuscular mycorrhizal fungi (AMF) play a major role as biofertilizer for sustainable agriculture. Nevertheless, it is still poorly documented whether inoculated AMF can successfully establish in field soils as exotic AMF and improve plant growth and productivity. Further, the fate of an exogenous inoculum is still poorly understood. Here, we pre-inoculated two cultivars (Tasset and Gola) of the fruit tree Ziziphus mauritiana (jujube) with the exotic AM fungus Rhizophagus irregularis isolate IR27 before transplantation in the field. In two experiments, tracking and quantification of R. irregularis IR27 were assessed in a 13-month-old jujube and an 18-month-old jujube in two fields located in Senegal. Our results showed that the inoculant R. irregularis IR27 was quantitatively traced and discriminated from native R. irregularis isolates in roots by using a qPCR assay targeting a fragment of the RNA polymerase II gene (RPB1), and that the inoculum represented only fractions ranging from 11 to 15% of the Rhizophagus genus in the two plantations 13 and 18 months after transplantation, respectively. This study validates the use of the RPB1 gene as marker for a relative quantification of a mycorrhizal inoculant fungus isolate in the field.

[Investigation of aflatoxins,mycobiota,and toxigenic fungi during post-harvest handling of Ziziphi Spinosae Semen].

Ziziphi Spinosae Semen is one of the Chinese herbal medicine being susceptible to aflatoxins contamination. To investigate the sources of aflatoxins contamination and toxigenic fungi species on Ziziphi Spinosae Semen,32 samples were collected from multiple steps during the post-harvest processing in this study. Aflatoxins in these samples were determined by immunoaffinity column and HPLC coupled with post-column photochemical derivatization. The dilution-plate method was applied to the fungi isolation. The isolated fungi strains were identified by morphological characterization and molecular approaches. The results showed that aflatoxins were detected in 28 samples from every step during the processing of Ziziphi Spinosae Semen. Three samples were detected with aflatoxin B_1 and 2 samples with both aflatoxin B_1 and total aflatoxin exceeding the limit of Chinese Pharmacopoeia. Especially the samples from the washing step,with the highest detected amounts of AFB_1 and AFs were reached 94. 79,121. 43 µg·kg~(-1),respectively. All 32 samples were contaminated by fungi. The fungal counts on the newly harvested samples were 2. 20 × 10~2 CFU·g~(-1). Moreover,it increased as tphreocessing progresses,and achieved 1. 16×10~6 CFU·g~(-1) after washing. A total of 321 isolates were identified to 17 genera. Aspergillus flavus was the main source of aflatoxins during the processing and storage of Ziziphi Spinosae Semen. One isolate of A. flavus was confirmed producing AFB_1 and AFB_2. The fungal count was significantly increased by composting,and Aspergillus was the predominant genus after shell breaking. The contamination level of aflatoxins was increased by composting and washing.

Comparative genome analysis of jujube witches'-broom Phytoplasma, an obligate pathogen that causes jujube witches'-broom disease.

BACKGROUND: JWB phytoplasma is a kind of insect-transmitted and uncultivable bacterial plant pathogen causeing a destructive Jujube disease. To date, no genome information about JWB phytoplasma has been published, which hindered its characterization at genomic level. To understand its pathogenicity and ecology, the genome of a JWB phytoplasma isolate jwb-nky was sequenced and compared with other phytoplasmas enabled us to explore the mechanisms of genomic rearrangement. RESULTS: The complete genome sequence of JWB phytoplasma (jwb-nky) was determined, which consisting of one circular chromosome of 750,803 bp with a GC content of 23.3%. 694 protein-encoding genes, 2 operons for rRNA genes and 31 tRNA genes as well as 4 potential mobile units (PMUs) containing clusters of DNA repeats were identified. Based on PHIbaes analysis, a large number of genes were genome-specific and approximately 13% of JWB phytoplasma genes were predicted to be associated with virulence. Although transporters for maltose, dipeptides/oligopeptides, spermidine/putrescine, cobalt, Mn/Zn and methionine were identified, KEGG pathway analysis revealed the reduced metabolic capabilities of JWB phytoplasma. Comparative genome analyses between JWB phytoplasma and other phytoplasmas shows the occurrence of large-scale gene rearrangements. The low synteny with other phytoplasmas indicated that the expansion of multiple gene families/duplication probably occurred separately after differentiation. CONCLUSIONS: In this study, the complete genome sequence of a JWB phytoplasma isolate jwb-nky that causing JWB disease was reported for the first time and a number of species-specific genes were identified in the genome. The study enhanced our understandings about genomic basis and the pathogenicity mechanism of this pathogen, which will aid in the development of improved strategies for efficient management of JWB diseases.

Changing Host Photosynthetic, Carbohydrate, and Energy Metabolisms Play Important Roles in Phytoplasma Infection.

Phytoplasmas parasitize plant phloem tissue and cause many economically important plant diseases. Jujube witches'-broom disease is a destructive phytoplasma disease of Chinese jujube (Ziziphus jujuba). To elucidate the influence of phytoplasma on host photosynthetic, carbohydrate and energy metabolisms, four types of jujube tissues showing disease symptoms with different severity were investigated at the structural, physiological, and molecular levels. Quantitative real-time PCR and high-performance liquid chromatography results showed that the down-regulation of genes related to photosynthesis and the lower contents of chlorophyll in diseased leaves. This clearly inhibited the light-harvesting and photosystem II activity of photosynthesis; however, overexpression of genes related to starch, sucrose and glucose synthesis led to higher contents of these carbohydrates. Meanwhile, transmission electron microscopy images revealed that dense amounts of phytoplasmas accumulated in the sieve elements of diseased petiole phloem, and the structure of the grana and stroma lamellae of chloroplasts in the diseased leaves was destroyed. Phytoplasma infection inhibited photosynthesis and led to abnormal carbohydrate accumulation in the diseased leaves. Furthermore, comparative metabolite analysis indicated that phytoplasma infection also stimulated amino acids and energy metabolisms of the diseased leaves. Continually inhibiting the photosynthetic process and stimulating carbohydrate and energy metabolisms of diseased trees may exhaust their nutrients. Our results highlight the importance of changing host metabolisms during the pathogenic process.

[Kinetic models for determination of yeast in fresh jujube using near infrared spectroscopy].

The objectives of this study were: (1) to optimize a near-infrared (NIR) spectroscopy model for fresh jujube stored at room temperature to predict the quality change (yeast growth), (2) to establish a kinetic model of yeast growth for fresh jujubes at room temperature according to NIR spectroscopy data and storage time, and (3) to predict the shelf life of fresh jujube at room temperature. The Lizao samples of fresh jujubes were adopted as the research object in the study. The NIR spectral data were achieved before yeast infection level measured. In order to optimize the NIR model, the pretreatment techniques such as Savitzky-Golay smoothing (S-G smoothing), multiplicative scatter correction (MSC), first derivative (1-Der) and second derivative (2-Der) were compared with the raw spectra by using a statistical software package (Unscrambler 9.8), and the regression coefficient (RC) method was used to choose the characteristic wavenumber. Multiple linear regression (MLR) was applied as NIR modeling method. According to the predicted yeast infection level using NIR model, the chemical kinetic models of spectral data and storage time at room temperature with zero-order and first-order reaction were established by using a statistical software package (SPSS 18). The shelf life could be predicted based on the chemical kinetic model. The results showed that the characteristic wave numbers of 10 300, 8 330, 6 900, 5 666, 5 150 and 4 060 cm(-1) in the whole near-infrared range with MSC technique could be chosen to model the quality deterioration of fresh jujube at room temperature. The NIR model that produced the best prediction had the form of B = 320.027 - 233.920(chi1) - 206.663(chi2) - 61.584(chi3) - 14.847(chi4) - 2.680(chi5) - 9.131(chi6), where B is yeast value (lg/cfu x g(-1)), chi1-chi6 are absorbance value of characteristic wavenumber. The correlation coefficient of calibration (R(c)) was 0.950, the root mean square error of calibration (RMSEC) was 2. 560, the correlation coefficient of prediction (R(p)) was 0.863, and the root mean square error of prediction (RMSEP) was 2.447. The zero-order reaction kinetic model performed better than the first-order model. The zero-order reaction kinetic model of yeast growth with storage time was predicted by B(t) = 171.395-124.445(chi1) - 109.945(chi2) - 32.763(chi3) - 7.899(chi4) - 1.426(chi5) - 4.857(chi6) + 0.045t with a correlation coefficient of 0.996. Based on the linear correlation between the NIR measurement and storage time, the shelf life of fresh jujube at room temperature was predicted to be 8 days for the yeast infection level less than 10 cfu x g(-1). The study showed that the NIR when combed with kinetic models could be used as a non-destructive, rapid method to detect the yeast growth in fresh jujube, and to predict the shelf life and ensure the quality and safety of fresh jujube.

The effect and mechanism of Debaryomyces nepalensis and six strains of winter jujube epiphytic bacteria in building a synthetic community to control post-harvest black spot disease of winter jujube.

The study aimed to develop a synthetic microbial community capable of managing postharvest black spot disease in winter jujube. The research revealed that treatment with Debaryomyces nepalensis altered the surface microbial community, reducing the presence of harmful fungi such as Alternaria, Penicillium, Fusarium, and Botrytis, while boosting beneficial bacteria like Pantoea, Bacillus, Staphylococcus, and Pseudomonas, leading to a decreased decay rate in date fruits. A synthetic community was crafted, integrating D. nepalensis with seven other bacterial strains selected for their abundance, compatibility, culturability, and interactions. This community was refined through homo-pore damage experiments and safety assessments to a final formulation consisting of D. nepalensis and six other bacteria: Bacillus subtilis, Bacillus velezensis, Staphylococcus arlettae, Staphylococcus gallinarum, Pseudomonas sp., and Pseudomonas psychrotolerans. Fruit inoculation tests demonstrated that this synthetic community (6 + 1) significantly lowered the incidence and size of black spot lesions compared to single-strain treatments. By the 10th day of storage, the incidence was 69.23 % lower than the control and 52.94 % lower than the group treated solely with D. nepalensis. Mechanistic studies of the synthetic community's antibacterial effects showed that it can produce volatile compounds, proteases, and ß-1,3-glucanase to inhibit pathogen growth. Additionally, the community forms a biofilm to compete for nutrients and induce jujube resistance to disease.

Impact of ultra-high-pressure treatment on microbial community composition and flavor quality of jujube juice: Insights from high-throughput sequencing technology, intelligent bionic sensory system, and metabolomics approach.

Ultra-high-pressure (UHP1) technology for cold pasteurization is a viable alternative to traditional heat sterilization for preserving food nutrients and flavor compounds during fruit juice processing. In this study, cutting-edge techniques, including high-throughput sequencing technology, intelligent bionic sensory systems, and metabolomics, were used to examine the impact of UHP treatment on microbial community composition, odor, and taste quality of jujube juice. The UHP treatment demonstrated its effect by inducing a reddish-yellow color in the jujube juice, thereby enhancing its brightness, overall color, and stability. The most significant enhancement was observed at 330 MPa. The microorganisms responsible for spoilage and deterioration of jujube juice during storage were categorized into three clusters: bacterial clusters at 0-330 MPa, 360-450 MPa, and 480-630 Mpa. The results showed no distinct distribution patterns for fungi based on the pressure strength. The dominant bacterial genera were Lactobacillus, Nocardia, Achromobacter, Enterobacter, Pseudomonas, Mesorhizobium, and Rhodococcus, whereas the dominant fungal genera were yeast and mold. Notably, Lactobacillus, Achromobacter, Enterobacter, and Pseudomonas were responsible for the significant differences between the 360 MPa to 450 MPa and 480 MPa to 630 MPa clusters in terms of bacterial spoilage, whereas Torulaspora, Lodderomyces, Wickerhamomyces, and Fusarium were the primary fungal spoilage genera. UHP treatment exerted no significant impact on the taste of jujube juice but influenced its sourness. Treatment at 330 MPa had the most pronounced effect on the presence of aromatic compounds and other odorants, which were substantially increased. Further analysis revealed the prevalence of organic acids, such as malic acid, succinic acid, and tartaric acid, in jujube juice and demonstrated a consistent relationship between changes in organic acids and sourness. In addition, nine distinct odorants with VIP values greater than 1 were identified in the jujube juice. Among these, methyl acetate and methyl caproate exhibited substantial increases following the UHP treatment at 330 MPa.

Paenibacillus beijingensis sp. nov., a novel nitrogen-fixing species isolated from jujube garden soil.

A novel Gram-positive, rod-shaped, motile, spore-forming, nitrogen-fixing bacterium, designated strain 7188(T), was isolated from jujube rhizosphere soil in Beijing, China. The strain grew at 4-40 °C and pH 6-12, with an optimum of 30 °C and pH 7.0, respectively. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain 7188(T) is a member of the genus Paenibacillus. Levels of 16S rRNA gene sequence similarities between strain 7188(T) and the type strains of all recognized members of the genus Paenibacillus were below 96 %. The major cellular fatty acids were anteiso-C(15:0), anteiso-C(17:0) and C(16:0). The predominant menaquinone was MK-7. The DNA G+C content of strain 7188(T) was 60.3 mol%. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and unknown aminophospholipids. The diamino acid in the cell wall peptidoglycan is meso-diaminopimelic acid. On the basis of these results, strain 7188(T) is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus beijingensis sp. nov. is proposed. The type strain is 7188(T) (=ACCC 03082(T) = DSM 24997(T)).

Debaryomyces nepalensis reduces fungal decay by affecting the postharvest microbiome during jujube storage.

Microbial antagonists are effective and environmentally friendly in controlling postharvest diseases of fruit. The present study investigated the influence of D. nepalensis on epiphytic microbiome and postharvest decay of jujube. Results showed that D. nepalensis notably reduced fungal decay, maintained the fruit firmness and delayed discoloration. The epiphytic microbiome revealed that D. nepalensis changed the fungal communities, but few influence on bacterial communities were observed. D. nepalensis, as the dominant population in the treatment group, decreased the abundance of pathogenic fungi of Alternaria, Penicillium, Fusarium and Botrytis, while increased the beneficial bacteria of Pantoea. The canonical correspondence analysis revealed that Debaryomyces was negatively correlated with the decay rate, whereas Penicillium, Acremonium, Rhodosporidiobolus and Hansfordia were positively correlated. In conclusion, D. nepalensis altered the successional process of fungal and bacterial communities to reduce the decay rate of jujube during storage.

Ozone reduces the fruit decay of postharvest winter jujube by altering the microbial community structure on fruit surface.

Microbial community structure on fruit surface plays an important role in fruit decay during postharvest storage, although the underlying mechanism has not been fully elucidated. Winter jujube (Ziziphus jujuba Miller cv. Dongzao) is a unique fruit resource with high edible and commercial value in China, while postharvest decay has always been a severe problem leading to short shelf life and poor quality of fruit. Ozone treatment is regarded as one of the most effective means to control decay and extend shelf life because of its cost-effective and eco-friendly properties. In the present study, three concentrations of ozone (2.5, 5 and 10 µL L-1) were found to reduce significantly postharvest decay of winter jujube on days 10 and 15, which were produced from Huanghua City, Hebei, China. High-throughput sequencing revealed significant changes in the bacterial and fungal communities in response to the application of ozone treatment, while Didymella, Rhizopus, Alternaria, Phialemoniopsis and Mycosphaerella were found to be the most abundant in fungi, and Methylobacterium, Pseudomonas, Pantoea, Sphingomonas and Gluconobacter being the most abundant in bacteria. Results of linear discriminant analysis (LDA) effect size (LEfSe) indicated that ozone treatments considerably reduced the abundance of Rhizopus and Gluconobacter on the surface of winter jujube fruit. Furthermore, Pearson correlation analysis showed that Rhizopus was positively correlated with Gluconobacter (r = 0.97) while negatively correlated with Didymella (r = -0.96). By predicting the metabolic function, ozone may inhibit metabolic pathways including nucleoside and nucleotide biosynthesis, amino acid biosynthesis, fatty acid and lipid degradation, respiration, and electron transfer, thereby reducing the incidence of fruit decay and maintaining the firmness of winter jujube fruit.

Epsilon-poly-l-lysine (ε-PL) exhibits multifaceted antifungal mechanisms of action that control postharvest Alternaria rot.

ε-Poly-l-lysine (ε-PL) is a natural antimicrobial poly-cationic peptide widely applied as a natural preservative in the food industry, whereas its application in preventing postharvest loss of fruit was largely absent. This study investigated the antifungal activity of ε-PL and determined the possible mechanisms involved. The in vivo results indicated that 500 mg L-1 exogenous ε-PL treatment significantly inhibited black spot rot in apple, jujube, and tomato. The lesion diameter inhibition rate was range from 20.11% to 29.09% by 500 mg L-1 ε-PL treatment. ε-PL exerts antifungal activity against A. alternata in vitro, the half-inhibition concentration is 160.1 mg L-1. ε-PL induced morphology and ultrastructure change on the pathogen, which resulted in the inhibition of A. alternata. This was accomplished by disturbing pathogen membrane integrity and functionality. The fluorometric assay confirmed that ε-PL induced endogenous reactive oxygen species formation and accumulation in A. alternata and the elicited severe lipid peroxidation that caused membrane lesions. Further, ε-PL treatment enhanced the expression of genes involved in antioxidant metabolism and pathogenesis-related responses in apple fruit. These findings illustrated that ε-PL exhibits multifaceted antifungal activity by the direct effect on the pathogen as well as induce host defense responses. ε-PL may be conducive as a promising alternative for Alternaria rot management.