Complete genome sequence analysis of Pestalotiopsis microspora, a fungal pathogen causing kiwifruit postharvest rots.

BACKGROUND: The postharvest rot of kiwifruit is one of the most devastating diseases affecting kiwifruit quality worldwide. However, the genomic basis and pathogenicity mechanisms of kiwifruit rot pathogens are lacking. Here we report the first whole genome sequence of Pestalotiopsis microspora, one of the main pathogens causing postharvest kiwifruit rot in China. The genome of strain KFRD-2 was sequenced, de novo assembled, and analyzed. RESULTS: The genome of KFRD-2 was estimated to be approximately 50.31 Mb in size, with an overall GC content of 50.25%. Among 14,711 predicted genes, 14,423 (98.04%) exhibited significant matches to genes in the NCBI nr database. A phylogenetic analysis of 26 known pathogenic fungi, including P. microspora KFRD-2, based on conserved orthologous genes, revealed that KFRD-2's closest evolutionary relationships were to Neopestalotiopsis spp. Among KFRD-2's coding genes, 870 putative CAZy genes spanned six classes of CAZys, which play roles in degrading plant cell walls. Out of the 25 other plant pathogenic fungi, P. microspora possessed a greater number of CAZy genes than 22 and was especially enriched in GH and AA genes. A total of 845 transcription factors and 86 secondary metabolism gene clusters were predicted, representing various types. Furthermore, 28 effectors and 109 virulence-enhanced factors were identified using the PHI (pathogen host-interacting) database. CONCLUSION: This complete genome sequence analysis of the kiwifruit postharvest rot pathogen P. microspora enriches our understanding its disease pathogenesis and virulence. This study establishes a theoretical foundation for future investigations into the pathogenic mechanisms of P. microspora and the development of enhanced strategies for the efficient management of kiwifruit postharvest rots.

Fluorescence hyperspectral imaging technology combined with chemometrics for kiwifruit quality attribute assessment and non-destructive judgment of maturity.

Dry matter content (DMC), firmness and soluble solid content (SSC) are important indicators for assessing the quality attributes and determining the maturity of kiwifruit. However, traditional measurement methods are time-consuming, labor-intensive, and destructive to the kiwifruit, leading to resource wastage. In order to solve this problem, this study has tracked the flowering, fruiting, maturing and collecting processes of Ya'an red-heart kiwifruit, and has proposed a non-destructive method for kiwifruit quality attribute assessment and maturity identification that combines fluorescence hyperspectral imaging (FHSI) technology and chemometrics. Specifically, first of all, three different spectral data preprocessing methods were adopted, and PLSR was used to evaluate the quality attributes (DMC, firmness, and SSC) of kiwifruit. Next, the differences in accuracy of different models in discriminating kiwifruit maturity were compared, and an ensemble learning model based on LightGBM and GBDT models was constructed. The results indicate that the ensemble learning model outperforms single machine learning models. In addition, the application effects of the 'Convolutional Neural Network'-'Multilayer Perceptron' (CNN-MLP) model under different optimization algorithms were compared. To improve the robustness of the model, an improved whale optimization algorithm (IWOA) was introduced by modifying the acceleration factor. Overall, the IWOA-CNN-MLP model performs the best in discriminating the maturity of kiwifruit, with Accuracytest of 0.916 and Loss of 0.23. In addition, compared with the basic model, the accuracy of the integrated learning model SG-MSC-SEL was improved by about 12%-20 %. The research findings will provide new perspectives for the evaluation of kiwifruit quality and maturity discrimination using FHSI and chemometric methods, thereby promoting further research and applications in this field.

Genome-wide identification of kiwifruit K+ channel Shaker family members and their response to low-K+ stress.

BACKGROUND: 'Hongyang' kiwifruit (Actinidia chinensis cv 'Hongyang') is a high-quality variety of A. chinensis with the advantages of high yield, early ripening, and high stress tolerance. Studies have confirmed that the Shaker K+ genes family is involved in plant uptake and distribution of potassium (K+). RESULTS: Twenty-eight Shaker genes were identified and analyzed from the 'Hongyang' kiwifruit (A. chinensis cv 'Hongyang') genome. Subcellular localization results showed that more than one-third of the AcShaker genes were on the cell membrane. Phylogenetic analysis indicated that the AcShaker genes were divided into six subfamilies (I-VI). Conservative model, gene structure, and structural domain analyses showed that AcShaker genes of the same subfamily have similar sequence features and structure. The promoter cis-elements of the AcShaker genes were classified into hormone-associated cis-elements and environmentally stress-associated cis-elements. The results of chromosomal localization and duplicated gene analysis demonstrated that AcShaker genes were distributed on 18 chromosomes, and segmental duplication was the prime mode of gene duplication in the AcShaker family. GO enrichment analysis manifested that the ion-conducting pathway of the AcShaker family plays a crucial role in regulating plant growth and development and adversity stress. Compared with the transcriptome data of the control group, all AcShaker genes were expressed under low-K+stress, and the expression differences of three genes (AcShaker15, AcShaker17, and AcShaker22) were highly significant. The qRT-PCR results showed a high correlation with the transcriptome data, which indicated that these three differentially expressed genes could regulate low-K+ stress and reduce K+ damage in kiwifruit plants, thus improving the resistance to low-K+ stress. Comparison between the salt stress and control transcriptomic data revealed that the expression of AcShaker11 and AcShaker18 genes was significantly different and lower under salt stress, indicating that both genes could be involved in salt stress resistance in kiwifruit. CONCLUSION: The results showed that 28 AcShaker genes were identified and all expressed under low K+ stress, among which AcShaker22 was differentially and significantly upregulated. The AcShaker22 gene can be used as a candidate gene to cultivate new varieties of kiwifruit resistant to low K+ and provide a reference for exploring more properties and functions of the AcShaker genes.

Genomes of diverse Actinidia species provide insights into cis-regulatory motifs and genes associated with critical traits.

BACKGROUND: Kiwifruit, belonging to the genus Actinidia, represents a unique fruit crop characterized by its modern cultivars being genetically diverse and exhibiting remarkable variations in morphological traits and adaptability to harsh environments. However, the genetic mechanisms underlying such morphological diversity remain largely elusive. RESULTS: We report the high-quality genomes of five Actinidia species, including Actinidia longicarpa, A. macrosperma, A. polygama, A. reticulata, and A. rufa. Through comparative genomics analyses, we identified three whole genome duplication events shared by the Actinidia genus and uncovered rapidly evolving gene families implicated in the development of characteristic kiwifruit traits, including vitamin C (VC) content and fruit hairiness. A range of structural variations were identified, potentially contributing to the phenotypic diversity in kiwifruit. Notably, phylogenomic analyses revealed 76 cis-regulatory elements within the Actinidia genus, predominantly associated with stress responses, metabolic processes, and development. Among these, five motifs did not exhibit similarity to known plant motifs, suggesting the presence of possible novel cis-regulatory elements in kiwifruit. Construction of a pan-genome encompassing the nine Actinidia species facilitated the identification of gene DTZ79_23g14810 specific to species exhibiting extraordinarily high VC content. Expression of DTZ79_23g14810 is significantly correlated with the dynamics of VC concentration, and its overexpression in the transgenic roots of kiwifruit plants resulted in increased VC content. CONCLUSIONS: Collectively, the genomes and pan-genome of diverse Actinidia species not only enhance our understanding of fruit development but also provide a valuable genomic resource for facilitating the genome-based breeding of kiwifruit.

Microbial community diversity analysis of kiwifruit pollen and identification of potential pathogens.

The kiwifruit industry typically uses commercial pollen for artificial pollination. However, during the collection of male flowers and pollen production, pollen can be easily contaminated by pathogenic bacteria that cause diseases such as canker and flower rot. Consequently, it is crucial to understand the structure of the pollen microbial community. This study employed Illumina high-throughput sequencing technology to analyze the fungal and bacterial composition in pollen samples from various regions in Shaanxi Province. Concurrently, potential pathogenic strains were isolated using traditional microbial isolation and cultivation techniques, and their molecular identification was performed through 16S rDNA sequence analysis. A tieback test was conducted on healthy branches to verify the pathogenicity of the strains. The results revealed a rich diversity of fungi and bacteria in kiwifruit pollen. At the phylum level, pollen fungi were mainly distributed in Ascomycota, and bacteria were mainly distributed in Proteobacteria and Firmicutes. The dominant fungal genera were Mycosphaerella, Aspergillus, and Cladosporium; the dominant bacterial genera were Weissella, Pantoea, Enterobacter, and Pseudomonas, respectively. Additionally, both Erwinia persicina and Pseudomonas fluorescens, isolated from pollen, exhibited high pathogenicity toward healthy kiwifruit branches. These findings contribute to a deeper understanding of the microbial diversity in commercial kiwifruit pollen used for mass pollination.

Honeybee (Apis mellifera L.) pollination enhances the yield and flavor quality of kiwifruit.

Pollination is essential for achieving high yields and enhancing the quality of kiwifruit cultivation, both of which significantly influence growers' interests and consumers' preferences. However, compared to studies on yield, there are fewer studies exploring the impact of pollination methods on the flavor of kiwifruit Actinidia chinensis Planchon. This study examined the effects of bee (Apis mellifera L.) pollination and artificial pollination on the yield and flavor of kiwifruit in the main producing areas of China. Compared with those pollinated artificially, bee-pollinated kiwifruit exhibited a greater fruit set rate, heavier fruit weight, and greater number of seeds. Notably, the number of seeds was positively correlated with fruit weight in bee-pollinated kiwifruit, whereas no such correlation was detected in artificially pollinated fruit. Bee pollination not only enhanced the yield but also improved the flavor of kiwifruit. Specifically, bee-pollinated kiwifruit contained higher levels of sucrose and lower concentrations of glucose and fructose, while the acid content was less affected by pollination methods. Furthermore, significant differences were observed in the volatile organic compound (VOC) levels in kiwifruit subjected to different pollination treatments, with bee-pollinated fruit exhibiting a superior flavor. Our findings provide new insights into the beneficial role of bee pollination in enhancing kiwifruit yield and quality, underscoring the crucial importance of bees in kiwifruit pollination.

Integrated transcriptome and targeted metabolome analyses provide insights into flavonoid biosynthesis in kiwifruit (Actinidia chinensis).

So far, a variety of metabolite components of kiwifruit have been elucidated. However, the identification and analysis of flavonoids in different tissues of kiwifruit are rarely carried out. In this study, we performed transcriptome and metabolome analyses of roots (Gkf_R), stems (Gkf_T), leaves (Gkf_L), and fruits (Gkf_F) to provide insights into the differential accumulation and regulation mechanisms of flavonoids in kiwifruit. Results showed that a total of 301 flavonoids were identified, in four tissues with different accumulation trends, and a large proportion of flavonoids had high accumulation in Gkf_L and Gkf_R. A total of 84 genes have been identified involved in the flavonoid biosynthesis pathway, and the expression levels of five LAR, two DFR, and one HCT were significantly correlated with the accumulation of 16 flavonoids and co-localized in the flavonoid biosynthesis pathway. In addition, a total of 2362 transcription factor genes were identified, mainly MYBs, bHLHs, ERFs, bZIPs and WRKYs, among which the expression level of bHLH74, RAP2.3L/4L/10L, MYB1R1, and WRKY33 were significantly correlated with 25, 56, 43, and 24 kinds of flavonoids. Our research will enrich the metabolomic data and provide useful information for the directed genetic improvement and application in the pharmaceutical industry of kiwifruit.

Changes in fruit quality properties and phytochemical substances of kiwifruit (Actinidia deliciosa) grown in different agro-ecological conditions during cold storage.

BACKGROUND: The changes in the physical structures of the products are the first things that consumers pay attention to. Therefore, it is essential and significant importance to take measures to improve the storage conditions of products and to minimize quality losses. The main objective of the study was to evaluate the effects of agro-ecological conditions on bioactive compounds and fruit quality of kiwifruit during cold storage. The 'Hayward' kiwifruit cultivar grown in Ordu, Giresun, Samsun, Rize, and Yalova provinces of Türkiye were kept at 0 ± 0.5 °C and relative humidity of 90 ± 5% for 150 d. RESULTS: The kiwifruit obtained from the provinces of Yalova, Ordu, and Giresun experienced the least weight loss during cold storage. Kiwifruit from Samsun and Yalova provinces had the lowest fruit firmness, while those from Giresun had the highest on 150th d. The changes were observed in the skin and flesh colors of the kiwifruit belonging to all cultivation areas. The amount of vitamin C increased throughout the study in all ecological conditions, but the Yalova province's kiwifruit was found to have the highest levels. Additionally, in all ecologies, kiwifruit showed an increase in antioxidant activity, total phenolics, and total flavonoids, all known to have beneficial effects on human health. The total antioxidant activity and total phenolics were highest in the kiwifruit of Yalova province, but the total flavonoids were found in the kiwifruit of Rize and Ordu provinces. CONCLUSION: The study's results revealed that kiwifruit's bioactive compounds and quality parameters may vary depending on the cultivation area. Additionally, it can be stated that Yalova province kiwifruit experiences the least amount of postharvest quality losses.

Sugar-conditioned honey bees can be biased towards a nectarless dioecious crop.

The targeted pollination strategy has shown positive results in directing honey bees to crop flowers offering nectar along with pollen as reward. Kiwifruit is a functionally dioecious species, which relies on bees to transport pollen from staminate to pistillate nectarless flowers. Following the targeted pollination procedures recently validated, we first developed a mimic odor (KM) based on kiwifruit floral volatiles for which bees showed the highest level of generalization to the natural floral scent, although the response towards pistillate flowers was higher than towards staminate flowers. Then, in the field, feeding colonies KM-scented sucrose solution resulted in higher amounts of kiwifruit pollen collected by honey bees compared to control colonies fed unscented sucrose solution. Our results support the hypothesis that olfactory conditioning bees biases their foraging preferences in a nectarless crop, given the higher visitation to target flowers despite having provided the mimic odor paired with a sugar reward.

Genome sequencing and CAZymes repertoire analysis of Diaporthe eres P3-1W causing postharvest fruit rot of 'Hongyang' kiwifruit in China.

Postharvest rot caused by various fungal pathogens is a damaging disease affecting kiwifruit production and quality, resulting in significant annual economic losses. This study focused on isolating the strain P3-1W, identified as Diaporthe eres, as the causal agent of 'Hongyang' postharvest rot disease in China. The investigation highlighted cell wall degrading enzymes (CWDEs) as crucial pathogenic factors. Specially, the enzymatic activities of cellulase, ß-galactosidase, polygalacturonase, and pectin methylesterases peaked significantly on the second day after infection of D. eres P3-1W. To gain a comprehensive understanding of these CWDEs, the genome of this strain was sequenced using PacBio and Illumina sequencing technologies. The analysis revealed that the genome of D. eres P3-1W spans 58,489,835 bp, with an N50 of 5,939,879 bp and a GC content of 50.7%. A total of 15,407 total protein-coding genes (PCGs) were predicted and functionally annotated. Notably, 857 carbohydrate-active enzymes (CAZymes) were identified in D. eres P3-1W, with 521 CWDEs consisting of 374 glycoside hydrolases (GHs), 108 carbohydrate esterase (CEs) and 91 polysaccharide lyases (PLs). Additionally, 221 auxiliary activities (AAs), 91 glycosyltransferases (GTs), and 108 carbohydrate binding modules (CBMs) were detected. These findings offer valuable insights into the CAZymes of D. eres P3-1W.

Effect of sequential fermentation with indigenous non-Saccharomyces cerevisiae combinations and Saccharomyces cerevisiae on the chemical composition and aroma compounds evolution of kiwifruit wine.

To unlock the potential of indigenous non-Saccharomyces cerevisiae and develop novel starters to enhance the aromatic complexity of kiwifruit wine, Zygosaccharomyces rouxii, Pichia kudriavzevii and Meyerozyma guilliermondii were pairwise combined and then used in sequential fermentation with Saccharomyces cerevisiae. The impact of different starter cultures on the chemical composition and flavor profile of the kiwifruit wines was comprehensively analyzed, and the aroma evolution during alcoholic fermentation was investigated by examining the changes in key volatiles and their loss rates. Compared with Saccharomyces cerevisiae, mixed starter cultures not only improve antioxidant capacity but also increase esters and alcohols yields, presenting intense floral and fruity aromas with high sensory acceptability. The results indicated that sequential inoculation of non-Saccharomyces cerevisiae combination and Saccharomyces cerevisiae promoted the development of volatiles while maintaining the stability of key aroma compounds in the winemaking environment and reducing the aroma loss rates during alcoholic fermentation.

Genome-Wide Identification of the DOF Gene Family in Kiwifruit (Actinidia chinensis) and Functional Validation of AcDOF22 in Response to Drought Stress.

DNA-binding one zinc finger (DOF) transcription factors are crucial plant-specific regulators involved in growth, development, signal transduction, and abiotic stress response generation. However, the genome-wide identification and characterization of AcDOF genes and their regulatory elements in kiwifruit (Actinidia chinensis) has not been thoroughly investigated. In this study, we screened the kiwifruit genome database and identified 42 AcDOF genes (AcDOF1 to AcDOF42). Phylogenetic analysis facilitated the categorization of these genes into five subfamilies (DOF-a, DOF-b, DOF-c, DOF-d, and DOF-e). We further analyzed the motifs, conserved domains, gene structures, and collinearity of the AcDOFgene family. Gene ontology (GO) enrichment analysis indicated significant enrichment in the "flower development" term and the "response to abiotic stress" category. Promoter prediction analysis revealed numerous cis-regulatory elements related to responses to light, hormones, and low-temperature and drought stress in AcDOF promoters. RNA-seq expression profiles demonstrated the tissue-specific expression of AcDOF genes. Quantitative real-time PCR results showed that six selected genes (AcDOF04, AcDOF09, AcDOF11, AcDOF13, AcDOF21, and AcDOF22) were differentially induced by abscisic acid (ABA), methyl jasmonate (MeJA), and cold, salt, and drought stresses, with AcDOF22 specifically expressed at high levels in drought-tolerant cultivars. Further experiments indicated that transient AcDOF22 overexpression in kiwifruit leaf disks reduced water loss and chlorophyll degradation. Additionally, AcDOF22 was localized to the nucleus and exhibited transcriptional activation, enhancing drought resistance by activating the downstream drought marker gene AcDREB2A. These findings lay the foundation for elucidating the molecular mechanisms of drought resistance in kiwifruit and offer new insights into drought-resistant breeding.

Physiological disorders in cold-stored 'Autumn Sense' hardy kiwifruit depend on the storage temperature and the modulation of targeted metabolites.

This study aimed to elucidate the effects of storage temperature on various fruit quality attributes, physiological disorders, and associated metabolites in the 0.5, 3, or 10 °C stored hardy kiwifruit. Peel pitting, which was highest in the 0.5 °C stored fruit, was identified as a chilling injury symptom of hardy kiwifruit. Proline and branched-chain amino acid contents showed higher values at 0.5 °C stored fruit as chilling responses. On the other hand, fruit shriveling and decay were highest in the 10 °C after 5 weeks of storage. The 10 °C storage induced fruit ripening during 3 weeks, but fruit shriveling and decay were severe after 5 weeks of storage. Therefore, storing the 'Autumn Sense' hardy kiwifruit at proper temperatures would be more beneficial, as it alters targeted metabolites and helps reduce the incidence of physiological disorders during cold storage.

Dynamic m6A mRNA methylation reveals the involvement of AcALKBH10 in ripening-related quality regulation in kiwifruit.

Kiwifruit ripening is a complex and highly coordinated process that occurs in conjunction with the formation of fruit edible quality. The significance of epigenetic changes, particularly the impact of N6-methyladenosine (m6A) RNA modification on fruit ripening and quality formation, has been largely overlooked. We monitored m6A levels and gene expression changes in kiwifruit at four different stages using LC-MS/MS, MeRIP, RNA-seq, and validated the function of AcALKBH10 through heterologous transgenic expression in tomato. Notable m6A modifications occurred predominantly at the stop codons and the 3' UTRs and exhibited a gradual reduction in m6A levels during the fruit ripening process. Moreover, these m6A modifications in the aforementioned sites demonstrated a discernible inverse relationship with the levels of mRNA abundance throughout the ripening process, suggesting a repression effect of m6A modification in the modulation of kiwifruit ripening. We further demonstrated that AcALKBH10 rather than AcECT9 predominantly regulates m6A levels in ripening-related genes, thereby exerting the regulatory control over the ripening process and the accumulation of soluble sugars and organic acids, ultimately influencing fruit ripening and quality formation. In conclusion, our findings illuminate the epi-regulatory mechanism involving m6A in kiwifruit ripening, offering a fresh perspective for cultivating high-quality kiwifruit with enhanced nutritional attributes.

Transcriptome analysis reveals ozone treatment maintains ascorbic acid content in fresh-cut kiwifruit by regulating phytohormone signalling pathways.

Ascorbic acid (AsA) is an indicator of the nutritional value of freshly cut kiwifruit during storage at 4℃, and its degradation can be inhibited after ozone treatment (1 mg/L, 10 min). The aim of this study was to elucidate the regulatory mechanism affecting AsA metabolism in fresh-cut kiwifruit after ozone treatment. In this study, ozone treatment not only prevented the decrease in AsA/dehydroascorbic acid and delayed the accumulation of total soluble solids/titratable acidity, but also altered phytohormone levels differently. Transcriptomic profiling combined with cis-acting element and correlation analysis were performed to reveal that abscisic acid and salicylic acid synergistically delay AsA degradation under ozone-treatment conditions. Actinidia03760, encoding ascorbate peroxidase, could be specifically recognized by the bZIP transcription factor and is considered a key candidate gene for further research. Collectively, ozone treatment is a promising method for preserving AsA content and improving the nutrition of fresh-cut kiwifruit.

Functional characterization of AcWRKY94 in response to Pseudomonas syringae pv. actinidiae in kiwifruit.

WRKY transcription factors are essential for coping with various biotic stresses. Pseudomonas syringae pv. actinidiae (Psa)-induced kiwifruit canker is a major problem restricting kiwifruit yield. Nevertheless, it's unclear how the kiwifruit WRKY genes respond to Psa. Through genome-wide identification, 112 WRKY members were found in 'Hongyang' genome in this work. Promoter analysis revealed that there were many cis-acting elements associated with stress responses in the AcWRKY gene's promoter region. According to transcriptomic analysis, 90 of the AcWRKY genes were differently expressed following Psa, salicylic acid (SA), or methyl jasmonate (MeJA) treatments. Almost all group III WRKYs were responsive to at least one of these treatments, with tissue-specific expression patterns. Quantitative RT-PCR study provided more evidence that Psa and SA treatments significantly induced the expression of the group III WRKY gene AcWRKY94, whereas MeJA treatment repressed it. AcWRKY94 was a transcriptionally active protein localized in the nucleus. Transient overexpression of AcWRKY94 in the leaves of 'Hongyang' enhanced the resistance of kiwifruit to Psa. Overexpression of AcWRKY94 in kiwifruit callus remarkably promoted the expression of PR and JAZ genes associated with SA and JA signals, respectively. These data imply that AcWRKY94 controls the signaling pathway dependent on SA and JA, thereby enhancing resistance to Psa. Taken together, this study establishes the basis for functional research on WRKY genes and provides important information for elucidating the resistance mechanism of kiwifruit canker disease.

Hydrogen sulfide enhances kiwifruit resistance to soft rot by regulating jasmonic acid signaling pathway.

As the third active gas signal molecule in plants, hydrogen sulfide (H2S) plays important roles in physiological metabolisms and biological process of fruits and vegetables during postharvest storage. In the present study, the effects of H2S on enhancing resistance against soft rot caused by Botryosphaeria dothidea and the involvement of jasmonic acid (JA) signaling pathway in kiwifruit during the storage were investigated. The results showed that 20 µL L-1 H2S fumigation restrained the disease incidence of B. dothidea-inoculated kiwifruit during storage, and delayed the decrease of firmness and the increase of soluble solids (SSC) content. H2S treatment increased the transcription levels of genes related to JA biosynthesis (AcLOX3, AcAOS, AcAOC2, and AcOPR) and signaling pathway (AcCOI1, AcJAZ5, AcMYC2, and AcERF1), as well as the JA accumulation. Meanwhile, H2S promoted the expression of defense-related genes (AcPPO, AcSOD, AcGLU, AcCHI, AcAPX, and AcCAT). Correlation analysis revealed that JA content was positively correlated with the expression levels of JA biosynthesis and defense-related genes. Overall, the results indicated that H2S could promote the increase of endogenous JA content and expression of defense-related genes by regulating the transcription levels of JA pathway-related genes, which contributed to the inhibition on the soft rot occurrence of kiwifruit.

Kiwifruit-Agaricus blazei intercropping effectively improved yield productivity, nutrient uptake, and rhizospheric bacterial community.

Intercropping systems have garnered attention as a sustainable agricultural approach for efficient land use, increased ecological diversity in farmland, and enhanced crop yields. This study examined the effect of intercropping on the kiwifruit rhizosphere to gain a deeper understanding of the relationships between cover plants and kiwifruit in this sustainable agricultural system. Soil physicochemical properties and bacterial communities were analyzed using the Kiwifruit-Agaricus blazei intercropping System. Moreover, a combined analysis of 16S rRNA gene sequencing and metabolomic sequencing was used to identify differential microbes and metabolites in the rhizosphere. Intercropping led to an increase in soil physicochemical and enzyme activity, as well as re-shaping the bacterial community and increasing microbial diversity. Proteobacteria, Bacteroidota, Myxococcota, and Patescibacteria were the most abundant and diverse phyla in the intercropping system. Expression analysis further revealed that the bacterial genera BIrii41, Acidibacter, and Altererythrobacter were significantly upregulated in the intercropping system. Moreover, 358 differential metabolites (DMs) were identified between the monocropping and intercropping cultivation patterns, with fatty acyls, carboxylic acids and derivatives, and organooxygen compounds being significantly upregulated in the intercropping system. The KEGG metabolic pathways further revealed considerable enrichment of DMs in ABC transporters, histidine metabolism, and pyrimidine metabolism. This study identified a significant correlation between 95 bacterial genera and 79 soil metabolites, and an interactive network was constructed to explore the relationships between these differential microbes and metabolites in the rhizosphere. This study demonstrated that Kiwifruit-Agaricus blazei intercropping can be an effective, labor-saving, economic, and sustainable practice for reshaping bacterial communities and promoting the accumulation and metabolism of beneficial microorganisms in the rhizosphere.

Analysis of spatiotemporal changes mechanism of cell wall biopolymers and monosaccharide components in kiwifruit during Botryosphaeria dothidea infection.

To further reveal the interaction mechanism between plants and pathogens, this study used confocal Raman microscopy spectroscopy (CRM) combined with chemometrics to visualize the biopolymers distribution of kiwifruit cell walls at different infection stages at the cellular micro level. Simultaneously, the changes in the content of various monosaccharides in fruit were studied at the molecular level using high-performance liquid chromatography (HPLC). There were significant differences in the composition of various nutrient components in the cell wall structure of kiwifruit at different infection times after infection by Botryosphaeria dothidea. PCA could cluster samples with infection time of 0-9 d into different infection stages, and SVM was used to predict the PCA classification results, the accuracy >96 %. Multivariate curve resolution-alternating least squares (MCR-ALS) helped to identify single substance spectra and concentration signals from mixed spectral signals. The pure substance chemical imaging maps of low methylated pectin (LMP), high methylated pectin (HMP), cellulose, hemicellulose, and lignin were obtained by analyzing the resolved concentration data. The imaging results showed that the lignin content in the kiwifruit cell wall increased significantly to resist pathogens infection after the infection of B. dothidea. With the development of infection, B. dothidea decomposed various substances in the host cell walls, allowing them to penetrate the interior of fruit cells. This caused significant changes in the form, structure, and distribution of various chemicals on the fruit cell walls in time and space. HPLC showed that glucose was the main carbon source and energy substance obtained by pathogens from kiwifruit during infection. The contents of galactose and arabinose, which maintained the structure and function of the fruit cell walls, decreased significantly and the cell wall structure was destroyed in the late stage of pathogens infection. This study provided a new perspective on the cellular structure changes caused by pathogenic infection of fruit and the defense response process of fruit and provided effective references for further research on the mechanisms of host-pathogen interactions in fruit infected by pathogens.

Visible light-responsive TiO2-based hybrid nanofiller reinforced multifunctional chitosan film for effective fruit preservation.

In this study, we developed a multifunctional chitosan film with visible light-responsive photocatalytic properties by incorporating a novel nanofiller-a nanohybrid particle of poly(tannic acid) (PTA) and TiO2 (TP-NPs). Firstly, the hybridization of TiO2 with PTA not only improved its dispersion but also obtained TP-NPs with smaller band gaps (from 3.11 eV to 1.55 eV) and higher separation efficiency of photogenerated e--h+ (about 1.5-fold enhancement), thereby producing more reactive oxygen species and enhancing the antibacterial efficacy (compared with TiO2, the antibacterial effect of TP-NPs on Staphylococcus aureus and Escherichia coli was heightened by about 2 times under visible light for 1 h). Secondly, TP-NPs were hydrogen bonded with chitosan, strengthening its mechanical and barrier properties, while imparting exceptional antibacterial efficacy. Moreover, the multifunctional properties enabled the active film to effectively delay the quality deterioration of grapes and kiwifruit. Hence, this study presented a multifunctional active packaging film tailored for fruit preservation.