Browsing by Author "Wisniewski, Michael"
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- Assembly and dynamics of the apple carposphere microbiome during fruit development and storageZhimo, V. Yeka; Kumar, Ajay; Biasi, Antonio; Abdelfattah, Ahmed; Sharma, Vijay Kumar; Salim, Shoshana; Feygenberg, Oleg; Bartuv, Rotem; Freilich, Shiri; Whitehead, Susan R.; Wisniewski, Michael; Droby, Samir (Frontiers, 2022-08-09)Microbial communities associated with fruit can contribute to quality and pathogen resistance, but little is known about their assembly and dynamics during fruit development and storage. Three apple cultivars growing under the same environmental conditions were utilized to examine the apple carposphere microbiome composition and structure at different developmental stages and storage. There was a significant effect (Adonis, p <= 0.001) of fruit genotype and its developmental stages and storage times on the fruit surface microbial assemblage and a strong temporal microbial community succession was detected (Mantel test: R <= 0.5, p = 0.001) in both bacterial and fungal communities. A set of 15 bacterial and 35 fungal core successional taxa and members exhibiting differential abundances at different fruit stages were identified. For the first time, we show the existence of underlying universal dynamics in the assembly of fruit-associated microbiomes. We also provide evidence of strong microbial cross-domain associations and uncover potential microbe-microbe correlations in the apple carposphere. Together our findings shed light on how the fruit carposphere assemble and change over time, and provide new insights into fruit microbial ecology.
- Changes in the Fungal Community Assembly of Apple Fruit Following Postharvest Application of the Yeast Biocontrol Agent Metschnikowia fructicolaBiasi, Antonio; Zhimo, V. Yeka; Kumar, Ajay; Abdelfattah, Ahmed; Salim, Shoshana; Feygenberg, Oleg; Wisniewski, Michael; Droby, Samir (MDPI, 2021-10-04)Recently, increasing focus has been placed on exploring fruit microbiomes and their association with their hosts. Investigation of the fruit surface microbiome of apple has revealed variations in the composition and structure depending on management practices, phenological stages, and spatial distribution on the fruit itself. However, the fate of the fruit surface microbiome assembly and dynamics in apple following interventions such as the application of biocontrol agents remains unknown. The objective of the study was to explore the effect of a postharvest application of a yeast biocontrol agent, Metschnikowia fructicola, on the composition of the epiphytic fungal microbiota on apples during cold storage. Our results demonstrated that the applied biocontrol agent, M. fructicola, persisted in high abundance (>28% relative abundance) on the fruit surface throughout the storage period. The biocontrol application significantly decreased the richness and caused a significant shift in the overall composition and structure of the fungal microbiome relative to untreated or water-treated controls. The yeast application reduced the abundance of several apple fungal pathogens, namely, Alternaria, Aspergillus, Comoclatris, Stemphylium, Nigrospora, Penicillium, and Podosphaera, throughout the cold storage period.
- Contrasting effects of genotype and root size on the fungal and bacterial communities associated with apple rootstocksLiu, Jia; Abdelfattah, Ahmed; Wasserman, Birgit; Wisniewski, Michael; Droby, Samir; Fazio, Gennaro; Mazzola, Mark; Wu, Xuehong (Oxford University Press, 2022-01-05)The endophytic microbiome of plants is believed to have a significant impact on its physiology and disease resistance, however, the role of host genotype in determining the composition of the endophytic microbiome of apple root systems remains an open question that has important implications for defining breeding objectives. In the current study, the bacterial and fungal microbiota associated with four different apple rootstocks planted in April, 2018 in the same soil environment and harvested in May, 2019 were evaluated to determine the role of genotype on the composition of both the bacterial and fungal communities. Results demonstrated a clear impact of genotype and root size on microbial composition and diversity. The fungal community was more affected by plant genotype whereas the bacterial community was shaped by root size. Fungal and bacterial abundance was equal between different-sized roots however, significantly higher microbial counts were detected in rhizosphere samples compared to root endosphere samples. This study provides information that can be used to develop a comprehensive and readily applicable understanding of the impact of genotype and environmental factors on the establishment of plant microbiome, as well as its potential function and impact on host physiology.
- Effect of Oligogalacturonides on Seed Germination and Disease Resistance of Sugar Beet Seedling and RootZhao, Can; Wu, Chunyan; Li, Kuikui; Kennedy, John F.; Wisniewski, Michael; Gao, Lihong; Han, Chenggui; Liu, Jia; Yin, Heng; Wu, Xuehong (MDPI, 2022-07)Oligogalacturonides (OGs) are a bioactive carbohydrate derived from homogalacturonan. The OGs synthesized in this study significantly inhibited the mycelial growth of Rhizoctonia solani AG-4HGI in vitro, even at a low concentration (10 mg/L). The seed vigor test demonstrated that the application of 50 mg/L OGs to sugar beet seeds significantly increased average germination percentage, germination energy, germination index, and seedling vigor index. The same concentration of OGs also improved the seedling emergence percentage of sugar beet when seeds were sown in soil inoculated with D2 and D31 isolates, respectively. The lesion diameter on mature sugar beet roots caused by R. solani AG-4HGI isolates D2 and D31 also decreased by 40.60% and 39.86%, respectively, in sugar beets roots first treated with 50 mg/mL OGs in the wound site, relative to lesion size in untreated/pathogen inoculated wounds. Sugar beet roots treated with 50 mg/mL OGs prior to inoculation with the D2 isolate exhibited up-regulation of the defense-related genes glutathione peroxidase (GPX) and superoxide dismutase (SOD) by 2.4- and 1.6-fold, respectively, relative to control roots. Sugar beet roots treated with 50 mg/mL OGs prior to inoculation with D31 exhibited a 2.0- and 1.6-fold up-regulation of GPX and SOD, respectively, relative to the control. Our results indicate that OGs have the potential to be used for the protection of sugar beet against R. solani AG-4HGI.
- Effect of Washing, Waxing and Low-Temperature Storage on the Postharvest Microbiome of AppleAbdelfattah, Ahmed; Whitehead, Susan R.; Macarisin, Dumitru; Liu, Jia; Burchard, Erik; Freilich, Shiri; Dardick, Christopher; Droby, Samir; Wisniewski, Michael (MDPI, 2020-06-23)There is growing recognition of the role that the microbiome plays in the health and physiology of many plant species. However, considerably less research has been conducted on the postharvest microbiome of produce and the impact that postharvest processing may have on its composition. Here, amplicon sequencing was used to study the effect of washing, waxing, and low-temperature storage at 2 °C for six months on the bacterial and fungal communities of apple calyx-end, stem-end, and peel tissues. The results of the present work reveal that tissue-type is the main factor defining fungal and bacterial diversity and community composition on apple fruit. Both postharvest treatments and low temperature storage had a strong impact on the fungal and bacterial diversity and community composition of these tissue types. Distinct spatial and temporal changes in the composition and diversity of the microbiota were observed in response to various postharvest management practices. The greatest impact was attributed to sanitation practices with major differences among unwashed, washed and washed-waxed apples. The magnitude of the differences, however, was tissue-specific, with the greatest impact occurring on peel tissues. Temporally, the largest shift occurred during the first two months of low-temperature storage, although fungi were more affected by storage time than bacteria. In general, fungi and bacteria were impacted equally by sanitation practices, especially the epiphytic microflora of peel tissues. This research provides a foundation for understanding the impact of postharvest management practices on the microbiome of apple and its potential subsequent effects on postharvest disease management and food safety.
- Efficacy of the biocontrol agent Trichoderma hamatum against Lasiodiplodia theobromae on macadamiaLi, Xiaojiao; Leng, Jinsong; Yu, Longfeng; Bai, Haidong; Li, Xiaojun; Wisniewski, Michael; Liu, Jia; Sui, Yuan (Frontiers, 2022-08)Macadamia (Macadamia integrifolia) trees are an important source of revenue in rainforest ecosystems. Their nuts are rich in vitamins, minerals, fiber, antioxidants, and monounsaturated oils. The fungus Lasiodiplodia theobromae, however, is a major disease problem, causing kernel rot and other disease symptoms. In the present study, a dual confrontation assay was used to evaluate the inhibitory effect of an endophytic strain of Trichoderma hamatum C9 from macadamia root against L. theobromae. Volatiles and cell-free culture filtrate of T. hamatum were also used to assess their antifungal activity against L. theobromae. Results suggested that T. hamatum exhibited a significant inhibitory effect against L. theobromae in vitro. Further results of a biocontrol assay indicated that a spray treatment of T. hamatum conidial suspension significantly decreased the size of lesions caused by artificially inoculated L. theobromae on macadamia leaves, as well as the disease index in young trees inoculated with L. theobromae, relative to sterile water controls. Collectively, our findings indicate that T. hamatum C9 represents a potential biocontrol agent that can be used to manage L. theobromae on macadamia.
- Evidence for host-microbiome co-evolution in appleAbdelfattah, Ahmed; Tack, Ayco J. M.; Wasserman, Birgit; Liu, Jia; Berg, Gabriele; Norelli, John; Droby, Samir; Wisniewski, Michael (Wiley, 2022-06)Plants evolved in association with a diverse community of microorganisms. The effect of plant phylogeny and domestication on host-microbiome co-evolutionary dynamics are poorly understood. Here we examined the effect of domestication and plant lineage on the composition of the endophytic microbiome of 11 Malus species, representing three major groups: domesticated apple (M. domestica), wild apple progenitors, and wild Malus species. The endophytic community of M. domestica and its wild progenitors showed higher microbial diversity and abundance than wild Malus species. Heirloom and modern cultivars harbored a distinct community composition, though the difference was not significant. A community-wide Bayesian model revealed that the endophytic microbiome of domesticated apple is an admixture of its wild progenitors, with clear evidence for microbiome introgression, especially for the bacterial community. We observed a significant correlation between the evolutionary distance of Malus species and their microbiome. This study supports co-evolution between Malus species and their microbiome during domestication. This finding has major implications for future breeding programs and our understanding of the evolution of plants and their microbiomes.
- Fox Hunting in Wild Apples: Searching for Novel Genes in Malus SieversiiWisniewski, Michael; Artlip, Timothy; Liu, Jia; Ma, Jing; Burchard, Erik; Norelli, John; Dardick, Christopher (MDPI, 2020-12-14)Malus sieversii is considered the progenitor of modern apple (Malus pumila) cultivars and to represent a valuable source of genetic diversity. Despite the importance of M. sieversii as a source of disease resistance, stress tolerance, and novel fruit traits, little is known about gene function and diversity in M. sieversii. Notably, a publicly annotated genome sequence for this species is not available. In the current study, the FOX (Full-length cDNA OvereXpressing) gene hunting system was used to construct a library of transgenic lines of Arabidopsis in which each transgenic line overexpresses a full-length gene obtained from a cDNA library of the PI619283 accession of M. sieversii. The cDNA library was constructed from mRNA obtained from bark tissues collected in late fall–early winter, a time at which many abiotic stress-adaptative genes are expressed. Over 4000 apple FOX Arabidopsis lines have been established from the pool of transgenic seeds and cDNA inserts corresponding to various Gene Ontology (GO) categories have been identified. A total of 160 inserts appear to be novel, with no or limited homology to M. pumila, Arabidopsis, or poplar. Over 1300 lines have also been screened for freezing resistance. The constructed library of transgenic lines provides a valuable genetic resource for exploring gene function and diversity in Malus sieversii. Notably, no such library of t-DNA lines currently exists for any Malus species.
- Global analysis of the apple fruit microbiome: are all apples the same?Abdelfattah, Ahmed; Freilich, Shiri; Bartuv, Rotem; Zhimo, V. Yeka; Kumar, Ajay; Biasi, Antonio; Salim, Shoshana; Feygenberg, Oleg; Burchard, Erik; Dardick, Christopher; Liu, Jia; Khan, Awais; Ellouze, Walid; Ali, Shawkat; Spadaro, Davide; Torres, Rosario; Teixido, Neus; Ozkaya, Okan; Buehlmann, Andreas; Vero, Silvana; Mondino, Pedro; Berg, Gabriele; Wisniewski, Michael; Droby, Samir (2021-03-18)We present the first worldwide study on the apple (Malus x domestica) fruit microbiome that examines questions regarding the composition and the assembly of microbial communities on and in apple fruit. Results revealed that the composition and structure of the fungal and bacterial communities associated with apple fruit vary and are highly dependent on geographical location. The study also confirmed that the spatial variation in the fungal and bacterial composition of different fruit tissues exists at a global level. Fungal diversity varied significantly in fruit harvested in different geographical locations and suggests a potential link between location and the type and rate of postharvest diseases that develop in each country. The global core microbiome of apple fruit was represented by several beneficial microbial taxa and accounted for a large fraction of the fruit microbial community. The study provides foundational information about the apple fruit microbiome that can be utilized for the development of novel approaches for the management of fruit quality and safety, as well as for reducing losses due to the establishment and proliferation of postharvest pathogens. It also lays the groundwork for studying the complex microbial interactions that occur on apple fruit surfaces.
- Identification and Functional Analysis of NLP-Encoding Genes from the Postharvest Pathogen Penicillium expansumLevin, Elena; Raphael, Ginat; Ma, Jing; Ballester, Ana-Rosa; Feygenberg, Oleg; Norelli, John; Aly, Radi; Gonzalez-Candelas, Luis; Wisniewski, Michael; Droby, Samir (MDPI, 2019-06-15)Penicillium expansum is a major postharvest pathogen that infects different fruits, mainly through injuries inflicted during harvest or subsequent handling after harvest. Several effectors were suggested to mediate pathogenicity of P. expansum in fruit tissue. Among these effectors Nep1-like proteins (NLPs), produced by various microorganisms with different lifestyles, are known for their ability to induce necrosis in dicot plants and were shown to be involved in virulence of several plant-related pathogens. This study was aimed at the identification and functional characterization of two NLP genes found in the genome of P. expansum. The genes were designated Penlp1 and Penlp2 and were found to code type1 and type3 NLP respectively. Necrosis-inducing activity of the two proteins was demonstrated by transient expression in Nicotiana benthamiana leaves. While Penlp1 expression was induced during apple infection and in liquid culture, the highest level of Penlp2 expression was found in ungerminated spores. Deletion of Penlp1, but not Penlp2, resulted in reduced virulence on apples manifested by reduced rate of lesion development (disease severity).
- Infrared thermography of in situ natural freezing and mechanism of winter-thermonasty in Rhododendron maximumArora, Rajeev; Wisniewski, Michael; Tuong, Tan; Livingston, David (Wiley, 2023-03)Evergreen leaves of Rhododendron species inhabiting temperate/montane climates are typically exposed to both high radiation and freezing temperatures during winter when photosynthetic biochemistry is severely inhibited. Cold-induced "thermonasty, " that is, lamina rolling and petiole curling, can reduce the amount of leaf area exposed to solar radiation and has been associated with photoprotection in overwintering rhododendrons. The present study was conducted on natural, mature plantings of a cold-hardy and large-leaved thermonastic North American species (Rhododendron maximum) during winter freezes. Infrared thermography was used to determine initial sites of ice formation, patterns of ice propagation, and dynamics of the freezing process in leaves to understand the temporal and mechanistic relationship between freezing and thermonasty. Results indicated that ice formation in whole plants is initiated in the stem, predominantly in the upper portions, and propagates in both directions from the original site. Ice formation in leaves initially occurred in the vascular tissue of the midrib and then propagated into other portions of the vascular system/venation. Ice was never observed to initiate or propagate into palisade, spongy mesophyll, or epidermal tissues. These observations, together with the leaf- and petiole-histology, and a simulation of the rolling effect of dehydrated leaves using a cellulose-based, paper-bilayer system, suggest that thermonasty occurs due to anisotropic contraction of cell wall cellulose fibers of adaxial versus abaxial surface as the cells lose water to ice present in vascular tissues.
- Metagenomics Approaches for the Detection and Surveillance of Emerging and Recurrent Plant PathogensPiombo, Edoardo; Abdelfattah, Ahmed; Droby, Samir; Wisniewski, Michael; Spadaro, Davide; Schena, Leonardo (2021-01)Globalization has a dramatic effect on the trade and movement of seeds, fruits and vegetables, with a corresponding increase in economic losses caused by the introduction of transboundary plant pathogens. Current diagnostic techniques provide a useful and precise tool to enact surveillance protocols regarding specific organisms, but this approach is strictly targeted, while metabarcoding and shotgun metagenomics could be used to simultaneously detect all known pathogens and potentially new ones. This review aims to present the current status of high-throughput sequencing (HTS) diagnostics of fungal and bacterial plant pathogens, discuss the challenges that need to be addressed, and provide direction for the development of methods for the detection of a restricted number of related taxa (specific surveillance) or all of the microorganisms present in a sample (general surveillance). HTS techniques, particularly metabarcoding, could be useful for the surveillance of soilborne, seedborne and airborne pathogens, as well as for identifying new pathogens and determining the origin of outbreaks. Metabarcoding and shotgun metagenomics still suffer from low precision, but this issue can be limited by carefully choosing primers and bioinformatic algorithms. Advances in bioinformatics will greatly accelerate the use of metagenomics to address critical aspects related to the detection and surveillance of plant pathogens in plant material and foodstuffs.
- Microbial Biopesticides: Diversity, Scope, and Mechanisms Involved in Plant Disease ControlVero, Silvana; Garmendia, Gabriela; Allori, Enzo; Sanz, José María; Gonda, Mariana; Alconada, Teresa; Cavello, Ivana; Dib, Julián Rafael; Diaz, Mariana Andrea; Nally, Cristina; Pimenta, Raphael Sanzio; da Silva, Juliana Fonseca Moreira; Vargas, Marisol; Zaccari, Fernanda; Wisniewski, Michael (MDPI, 2023-03-19)Food losses, defined as a reduction in the quantity and quality of food during production and storage, impact food safety and security. Losses caused by plant pathogens are among the most significant. Chemical pesticides have been extensively used to prevent microbial diseases. Their toxicity and reduced efficacy, however, have encouraged investigators to develop alternatives. Alternatives based on microbial biopesticides tend to be safer and more environmentally benign than conventional pesticides. In recent years, formulations based on biopesticides have progressively increased in number and diversity and have attracted commercial interest. Understanding the mechanisms by which biopesticides control the disease is fundamental to achieving optimal disease control. Biocontrol mechanisms can be divided into two main categories: those related to the ability to inhibit pathogens or their virulence factors, and those that enhance host plant fitness and induce disease resistance. Here, the first type of strategy is reviewed, which is directly mediated by physical contact between biocontrol agents and pathogens or indirectly by exposure of a pathogen to antimicrobial or microbial-inhibiting compounds produced by the microbial antagonist. Mechanisms involving physical contact include mycophagy, destruction of pathogenic bacteria by bacteriophages or predation, and disease inhibition by topical applications of specific dsRNA. Indirect mechanisms that do not involve direct contact with a pathogen include the production of antimicrobial compounds, competition, and virulence factor suppression by quorum quenching. These topics are reviewed and discussed.