Effect of pre-cooling on the shelf-life and quality of formosa papaya.

Papaya is a climacteric fruit, rapidly ripening after harvesting due to ethylene production and increased respiratory rate. This swift ripening results in softening of fruit tissues, shortening the fruit shelf life. Pre-cooling serves as an alternative to minimize fruit ripening and post-harvest losses by reducing metabolism. This study aimed to evaluate the effect of pre-cooling on the quality and conservation of Formosa 'Tainung I' papaya. Papayas at maturation stage II were obtained from a commercial orchard with conventional production. The experimental design was a completely randomized 4×6 split-plot scheme, with pre-cooling treatments (Control, without pre-cooling treatment; pre-cooling at 15 °C in a cold chamber; pre-cooling at 7 °C in a cold chamber; and forced-air cooling at 7 °C) in the plot, and days of storage (0, 7, 14, 21, 28, and 35 days) in the subplot. Pre-cooling effectively delayed the ripening and senescence of Formosa papaya, reducing the loss of green color and firmness. Regardless of the treatment used, chilling injury and incidence of fungi from the genus Fusarium and Alternaria limited the shelf life of Formosa 'Tainung I' papaya up to 21 days of storage. Additionally, the appearance of hardened regions in the pulp compromised the sensory quality of the fruits, necessitating further investigation into the causes of this disorder.

Formulation of the biological control yeast Meyerozyma caribbica by electrospraying process: effect on postharvest control of anthracnose in mango (Mangifera indica L.) and papaya (Carica papaya L.).

BACKGROUND: Microorganism for biological control of fruit diseases is an eco-friendly alternative to the use of chemical fungicides. RESULTS: This is the first study evaluating the electrospraying process to encapsulate the biocontrol yeast Meyerozyma caribbica. The effect of encapsulating material [Wey protein concentrate (WPC), Fibersol® and Trehalose], its concentration and storage temperature on the cell viability of M. caribbica, and in vitro and in vivo control of Colletotrichum gloeosporioides was evaluated. The processing with commercial resistant maltodextrin (Fibersol®) 30% (w/v) as encapsulating material showed the highest initial cell viability (95.97 ± 1.01%). The storage at 4 ± 1 °C showed lower losses of viability compared to 25 ± 1 °C. Finally, the encapsulated yeast with Fibersol 30% w/v showed inhibitory activity against anthracnose in the in vitro and in vivo tests, similar to yeast fresh cells. CONCLUSION: Electrospraying was a highly efficient process due to the high cell viability, and consequently, a low quantity of capsules is required for the postharvest treatment of fruits. Additionally, the yeast retained its antagonistic power during storage. © 2021 Society of Chemical Industry.

Growth behavior of low populations of Listeria monocytogenes on fresh-cut mango, melon and papaya under different storage temperatures.

The growth behavior of Listeria monocytogenes low population (1-4 cells/sample) on fresh-cut mango, melon, papaya and fruit mix stored at 4, 8, 12 and 16 °C was evaluated over 10 days. Mango showed the lowest counts for L. monocytogenes during 10 days regardless of storage temperature (<1.7 log cfu.g-1). Melon supported high bacterial growth over 10 days, reaching 5 log cfu.g-1 at 16 °C. Both the fruit and storage temperature influenced the Listeria low population growth potential (δ). Cumulative frequency distribution of L. monocytogenes showed that after 10 days, 100% of fresh-cut fruits and fruit mix stored at 4 °C remained ≤2 log cfu.g-1, while at 12 and 16 °C 100% of melon, papaya and fruit mix samples exceeded this limit. At 8 °C, 100% of mango and fruit mix samples remained below this limit after 10 days, whereas 100% of melon and papaya reached it after 7 days. Results indicate 4 °C as the ideal to store safely fresh-cut mango, melon, papaya and fruit mix for 10 days. Besides, 8 °C can also be an option, but not for melon and papaya. Findings highlight the ability of L. monocytogenes to survive and grow in fresh-cut fruits even at a very low initial population levels.

Mutualistic interaction of native Serratia marcescens UENF-22GI with Trichoderma longibrachiatum UENF-F476 boosting seedling growth of tomato and papaya.

A plethora of bacteria-fungal interactions occur on the extended fungal hyphae network in soil. The mycosphere of saprophytic fungi can serve as a bacterial niche boosting their survival, dispersion, and activity. Such ecological concepts can be converted to bioproducts for sustainable agriculture. Accordingly, we tested the hypothesis that the well-characterised beneficial bacterium Serratia marcescens UENF-22GI can enhance plant growth-promoting properties when combined with Trichoderma longibrachiatum UENF-F476. The cultural and cell interactions demonstrated S. marcescens and T. longibrachiatum mutual compatibility. Bacteria cells were able to attach, forming aggregates to biofilms and migrating through the fungal hyphae network. Long-distance bacterial migration through growing hyphae was confirmed using a two-compartment Petri dishes assay. Fungal inoculation increased the bacteria survival rates into the vermicompost substrate over the experimental time. Also, in vitro indolic compound, phosphorus, and zinc solubilisation bacteria activities increased in the presence of the fungus. In line with the ecophysiological bacteria fitness, the bacterium-fungal combination boosted tomato and papaya plantlet growth when applied into the plant substrate under nursery conditions. Mutualistic interaction between mycosphere-colonizing bacterium S. marcescens UENF-22GI and the saprotrophic fungi T. longibrachiatum UENF-F467 increased the ecological fitness of the bacteria alongside with beneficial potential for plant growth. A proper combination and delivery of mutual compatible beneficial bacteria-fungal represent an open avenue for microbial-based products for the biological enrichment of plant substrates in agricultural systems.

Biological prevention and control of pitaya fruit canker disease using endophytic fungi isolated from papaya.

Pitaya fruit canker is an important disease in pitaya production. Facilitating resistance through the application of biological control principles is a promising alternative to traditional control strategies. This study evaluated the induced resistance of Penicillium rolfsii, numbered Y17 isolated from papaya leaves in pitaya fruit, and evaluated the activity of the defense enzymes, total antioxidant capacity (T-AOC), and malondialdehyde (MDA) content of the treated fruit. The results demonstrate that treatment with Y17 effectively induced resistance of pitaya fruit to canker disease caused by Neoscytalidium dimidiatum, with an inhibition rate of 70.87%. In addition, Y17 notably improved the activities of peroxidase, catalase, and polyphenol oxidase as well as the T-AOC of the treated samples. Y17 treatment reduced the MDA content in these fruits. Taken together, our results suggest that Y17 treatment could trigger pitaya fruit defense responses and effectively induce resistance to fruit canker disease.

Crop performance and soil fertility improvement using organic fertilizer produced from valorization of Carica papaya fruit peel.

In recent times, research attention is focusing on harnessing agricultural wastes for the production of value-added products. In this study, the valorization of Carica papaya (Pawpaw) fruit peels was evaluated for the production of quality organic fertilizer via anaerobic digestion (AD) while the effects of the fertilizer on maize crop were also assessed. Pawpaw peel was first pretreated by thermo-alkaline methods before AD and analyses were carried out using standard methods. The resulting digestate was rich in nutrients and was dewatered to form solid organic fertilizer rich in microbes and soil nutrients. When applied to maize plants, organic fertilizer showed a better effect on plant traits than NPK 15-15-15 fertilizer and without fertilizer application. These were more pronounced at mid to high organic fertilizer applications (30-to-60-kg nitrogen/hectare (kg N/ha)) rate. Comparison between the values obtained from the field experiments reveals that the organic fertilizer showed better performance in all parameters such as the number of leaves, leaf area, plant height, stem girth, total shoot, and root biomass, and length of the root. However, the chemical fertilizer outperformed all the organic fertilizer applied rates in the average highest size of the corn ear by 1.4%. After harvesting, nutrient elements were found to have bioaccumulated in plant organs (leaves, stem, and root) with the highest values being 29.7 mg/L for nitrogen in the leaf and this value was reported from the experiment with 50 kg N/ha. For phosphorus and potassium, the highest concentrations of 7.05 and 8.4 mg/L were recorded in the plant' stem of the experiment with 50 kg N/ha. All the treated soils recorded an increase in values of all nutrient elements over the control with the highest values recorded in the experiment with 60 kg N/ha. In soil with 60 kg N/ha, the nitrogen, phosphorus, and potassium increased by 28, 40, and 22% respectively over the chemical fertilizer applied experiment while different levels of increases were also recorded for all other macro and microelements in all the experiments. Thus, agricultural practices by using anaerobic digestates as organic fertilizers is a sustainable method to overcome the dependence on inorganic fertilizers high rate.

Salmonella inactivation and sponge/microfiber mediated cross-contamination during papaya wash with chlorine or peracetic acid as sanitizer.

Imported papayas from Mexico have been implicated in multiple salmonellosis outbreaks in the United States in recent years. While postharvest washing is a critical process to remove latex, dirt, and microbes, it also has the potential of causing cross-contamination by foodborne pathogens, with sponge or other fibrous rubbing tools often questioned as potential harboring or transmitting risk. In this study, Salmonella inactivation and cross-contamination via sponges and microfiber wash mitts during simulated papaya washing and cleaning were investigated. Seven washing treatments (wash without sanitizer; wash at free chlorine 25, 50, and 100 mg/L, and at peracetic acid 20, 40, and 80 mg/L), along with unwashed control, were evaluated, using Salmonella strains with unique antibiotic markers differentially inoculated on papaya rind (serovars Typhimurium, Heidelberg, and Derby) and on wash sponge or microfiber (serovars Typhimurium, Newport, and Braenderup). Salmonella survival and transfer on papaya and on sponge/microfiber, and in wash water were detected using selective plating or enrichment. The washing and cleaning process reduced Salmonella on inoculated papayas by 1.69-2.66 and 0.69-1.74 log for sponge and microfiber cleaning, respectively, with the reduction poorly correlated to sanitizer concentration. Salmonella on inoculated sponge or microfiber was under detection limit (1.00 log CFU/cm2) by plate count, but remained recoverable by selective enrichment. Transference of Salmonella from inoculated papaya to sponge/microfiber, and vice versa, could be detected sporadically by selective enrichment. Sponge/microfiber mediated Salmonella cross-contamination from inoculated to uninoculated papayas was frequently detectable by selective enrichment, but rendered undetectable by wetting sponge/microfiber in sanitizing wash water (FC 25-100 mg/L or PAA 20-80 mg/L) between washing different papaya fruits. Therefore, maintaining adequate sanitizer levels and frequently wetting sponge/microfiber in sanitizing wash water can effectively mitigate risks of Salmonella cross-contamination associated with postharvest washing, especially with regard to the use of sponge or microfiber wash mitts.

Thermal favorability for the Oidium caricae and Asperisporium caricae in areas of edaphoclimatic aptitude for the Carica papaya.

Objective this study aimed to determine the thermal favorability for the oidium (Oidium caricae) and early blight (Asperisporium caricae) in areas of edaphoclimatic aptitude for the papaya (Carica papaya) in the Espírito Santo state, Brazil. The edaphoclimatic zoning was based on the overlapping of maps that characterize the average annual air temperature obtained by multiple linear regression, annual water deficiency calculated by the Thornthwaite and Matter method (1955) and favorable soil classes to the development of papaya. The results indicated that as regards the edaphoclimatic zoning for the papaya crop it was observed that 71.70% of the area is classified as apt for its development. In relation to agrometeorological favorability for the occurrence of fungal diseases, there was a favorability of 7.64% for the development without restrictions of the oidium and a predominance of 64,56% of favorability with thermal restriction. For the early blight fungus, it was observed that the zones of favorability without restriction correspond to 11.66% of the area apt for the papaya cultivation and that 55.13% of the area has favorability with restriction of humidity. The edaphoclimatic zoning for the papaya crop showed compatibility with the most productive areas of this crop in the state of Espírito Santo. The zoning of thermal favorability for the occurrence of papaya fungal diseases proved to be a valuable tool for studies of plant diseases, allowing the establishment of plans for the allocation of resistant varieties, in order to minimize the risks of loss of crop productivity due to the disease. This methodology presents potential to be used in other areas, cultures and phytopathological diseases.

Active coatings based on hydroxypropyl methylcellulose and silver nanoparticles to extend the papaya (Carica papaya L.) shelf life.

This study aimed to understand the effect of silver nanoparticles (AgNPs) on physiochemical properties of hydroxypropyl methylcellulose (HPMC) film-forming solutions (FFS) and nanocomposite films (NCF), as well as the efficacy of these materials to control the development of anthracnose caused by Colletotrichum gloeosporioides in papaya (Carica papaya L.). FFS were characterized by pH, particle size distribution, and rheology. In addition, thickness, morphology, water contact angle, barrier, chemical, crystallinity, thermal, and mechanical properties from NCF were investigated. The minimum inhibitory concentration of AgNPs against C. gloeosporioides was determined by in vitro test. FFS with 0.25 wt% of AgNPs were used as coatings in papayas inoculated with C. gloeosporioides. Finally, the physicochemical parameters were investigated during their storing up to 7 days at 10 °C, followed by 7 days at 20 °C. The presence of AgNPs impacted the thickness, morphology, moisture content, chemical bonds, crystalline structure, and thermal properties of films. Coatings with 0.25 wt% of AgNPs reduced the incidence and severity of C. gloeosporioides and avoided the weight loss of papayas during storing. The ripening of papaya occurred naturally, showing that the coating only delayed this process. Thus, HPMC-AgNPs coating can be an alternative to extend the papaya shelf life.

Coatings with chitosan and phenolic-rich extract from acerola (Malpighia emarginata D.C.) or jabuticaba (Plinia jaboticaba (Vell.) Berg) processing by-product to control rot caused by Lasiodiplodia spp. in papaya (Carica papaya L.) fruit.

This study evaluated if coatings with chitosan (Chi) and phenolic-rich extract from acerola (Malpighia emarginata D.C., PEA) or jabuticaba (Plinia jaboticaba (Vell.) Berg, PEJ) processing by-products are effective to control the development of rot caused by Lasiodiplodia pseudotheobromae, L. viticola, L. euphorbicola, L. theobromae and L. hormozganensis in papaya (Carica papaya L.) fruit. Effects of formulated coatings on some physicochemical parameters indicative of postharvest quality of papaya were investigated. Twenty-six different phenolics were found in PEA and PEJ, including flavonoids, stilbenes, tannins and phenolic acids. Chi (1-5 mg/mL), PEA and PEJ (25-100 mg/mL) separately caused mycelial growth inhibition on all isolates. Combinations of Chi (3 and 4 mg/mL) and PEA (50 and 75 mg/mL) or PEJ (75 and 100 mg/mL) had additive interactions. Coatings with Chi (4 mg/mL) and PEA (50 or 75 mg/mL) or PEA (75 or 100 mg/mL) inhibited rot development in papaya fruit infected with Lasiodiplodia isolates during 8 days of room temperature storage. Coatings with 4 mg/mL Chi and 75 mg/mL PEA or 100 mg/mL PEJ were the most effective to control rot development. These coatings did not affect negatively physicochemical parameters indicative of postharvest quality of papaya fruit during storage. Coatings with combined Chi and PEA or PEJ could be novel strategies to control postharvest rot caused by Lasiodiplodia in papaya fruit.

Colletotrichum gloesporioides inhibition using chitosan-Ruta graveolens L essential oil coatings: Studies in vitro and in situ on Carica papaya fruit.

In this study we assessed the efficacy of chitosan (CHI) (2%) emulsion added with Ruta graveolens L. essential oil (REO) at different concentrations (0.5%, 1.0% and 1.5%) to control C. gloesporioides grows both "in situ" and "in vitro" in papaya Maradol (Carica papaya L.). In vitro studies showed a decrease on fungal growth (mycelia diameter) with the increase of REO concentration, while 0.5% of REO induce a reduction of 56.42%, REO at 1.0% and 1.5% induced a reduction of 97%. Microscopic analysis showed irreversible deleterious morphological and ultrastructural alterations as well as changes in conidia morphology, and conidia germination inhibition up to 90%. Among the most abundant REO constituents, 2-Nonanol showed strong antifungal activity followed by 2-Undecanone, Benzyl acetate, 2-Nonanone, 2-Tridecanone and 2-Dodecanone. Studies "in situ" on papaya fruit during 12 days at 20 °C, showed a reduction of the C. gloesporioides lesion expansion by 50% using CHI-REO 0.5% emulsions and by 100% with treatments of CHI-REO 1.0 and 1.5%, in addition the emulsions were efficacious to reduce the fruit surface microbiota. On the other hand, physicochemical analysis of the papaya fruits demonstrated that CHI-REO emulsions treatment delayed papaya ripening without affecting the organoleptic characteristics. All these results demonstrated for the first time the application of coatings CHI-REO as a postharvest treatment for the control of anthracnose on papaya fruit.

Endomicrobial Community Profiles of Two Different Mealybugs: Paracoccus marginatus and Ferrisia virgata.

Mealybugs (Hemiptera: Coccomorpha: Pseudococcidae) harbour diverse microbial symbionts that play essential roles in host physiology, ecology, and evolution. In this study we aimed to reveal microbial communities associated with two different mealybugs, papaya mealybug (Paracoccus marginatus) and two-tailed mealybug (Ferrisia virgata) collected from the same host plant. Comparative analysis of microbial communities associated with these mealybugs revealed differences that appear to stem from phylogenetic associations and different nutritional requirements. This first report on both bacterial and fungal communities associated with these mealybugs provides a preliminary insight on factors affecting the endomicrobial communities. .

Oxidative foliar photo-necrosis produced by the bacteria Pseudomonas cedrina

BACKGROUND: Although bioactive metabolites capable of causing oxidative photo-necrosis in plant tissues have been identified in fungi, little is known about this type of mechanism in bacteria. These metabolites act as photosensitizers that generate reactive oxygen species (ROS) capable of causing damage to cells. In addition, these metabolites can pass into an energetically excited state when they receive some luminous stimulus, a condition in which they interact with other molecules present in the environment, such as molecular oxygen (O2), also known as triplet oxygen (3 O2), generating ROS. RESULTS: The suspension of the bacterial culture of Pseudomonas cedrina was shown to produce foliar necrosis in papaya leaves (Carica papaya L.) only in the presence of sunlight, which is evidence of photosensitizing mechanisms that generate singlet oxygen (1 O2). From the chemical study of extracts obtained from this bacteria, 3-(4-(2-carboxipropyl) phenyl) but-2-enoic acid (1) was isolated. This compound, in the presence of light and triplet oxygen (3 O2), was able to oxidize ergosterol to its peroxide, since it acted as a photosensitizer producing 1 O2, with which it was corroborated that a photosensitization reaction occurs, mechanism by which this bacterium could prove to cause oxidative foliar photo-necrosis. CONCLUSIONS: P. cedrina was able to induce oxidative foliar photo-necrosis because of its potential ability to produce photosensitizing metabolites that generate singlet oxygen in the plants it colonizes. Based on the above, it can be proposed that some bacteria can cause oxidative foliar photo-necrosis as an important mechanism in the pathogenesis of host species.

Genomic diversity of Salmonella enterica isolated from papaya samples collected during multiple outbreaks in 2017.

In 2017, the US Food and Drug Administration investigated the sources of multiple outbreaks of salmonellosis. Epidemiologic and traceback investigations identified Maradol papayas as the suspect vehicles. During the investigations, the genomes of 55 Salmonella enterica that were isolated from papaya samples were sequenced. Serovar assignments and phylogenetic analysis placed the 55 isolates into ten distinct groups, each representing a different serovar. Within-serovar SNP differences are generally between 0 and 20 SNPs, while the median between-serovar distance is 51 812 SNPs. We observed two groups with SNP distances between 21 and 100 SNPs. These relatively large within-serovar SNP distances may indicate that the isolates represent either diverse populations or multiple, genetically distinct subpopulations. Further inspection of these cases with traceback evidence allowed us to identify an 11th population. We observed that high levels of genomic diversity from individual firms is possible, with one firm yielding five of the ten serovars. Also, high levels of diversity are possible within small geographic regions, as five of the serovars were isolated from papayas that originated from farms located in Armería and Tecomán, Colima. In addition, we identified AMR genes that are present in three of the serovars studied here (aph(3')-lb, aph(6)-ld, tet(C), fosA7, and qnrB19) and we detected the presence of the plasmid IncHI2A among S. Urbana isolates.

A secreted protein of 15 kDa plays an important role in Phytophthora palmivora development and pathogenicity.

Phytophthora palmivora is a destructive oomycete plant pathogen with a wide host range. So far, little is known about the factors governing its infection structure development and pathogenicity. From the culture filtrate of a P. palmivora strain isolated from papaya, we identified a secreted glycoprotein of 15 kDa, designated as Ppal15kDa, using liquid chromatography tandem mass spectrometry. Two gene variants, Ppal15kDaA and Ppal15kDaB were amplified from a P. palmivora papaya isolate. Transient expression of both variants in Nicotiana benthamiana by agroinfiltration enhanced P. palmivora infection. Six Ppal15kDa mutants with diverse mutations were generated via CRISPR/Cas9-mediated gene editing. All mutants were compromised in infectivity on N. benthamiana and papaya. Two mutants with all Ppal15kDa copies mutated almost completely lost pathogenicity. The pathogenicity of the other four containing at least one wild-type copy of Ppal15kDa was compromised at varying levels. The mutants were also affected in development as they produced smaller sporangia, shorter germ tubes, and fewer appressoria. The affected levels in development corresponded to the levels of reduction in pathogenicity, suggesting that Ppal15kDa plays an important role in normal development of P. palmivora infection structures. Consistent with its role in infection structure development and pathogenicity, Ppal15kDa was found to be highly induced during appressorium formation. In addition, Ppal15kDa homologs are broadly present in Phytophthora spp., but none were characterized. Altogether, this study identified a novel component involved in development and pathogenicity of P. palmivora and possibly other Phytophthora spp. known to contain a Ppal15kDa homolog.

Application of coatings formed by chitosan and Mentha essential oils to control anthracnose caused by Colletotrichum gloesporioides and C. brevisporum in papaya (Carica papaya L.) fruit.

This study investigated the efficacy of coatings formed by chitosan (Chi) and Mentha piperita L. (MPEO) or M. × villosa Huds (MVEO) essential oil to control the development of antrachnnose in papaya fruit caused by Colletotrichum gloeosporioides and C. brevisporum. Chi (2.5-10 mg/mL), MPEO and MVEO (0.15-1.25 µL/mL) alone effectively inhibited the growth of C. gloeosporioides and C. brevisporum isolates in laboratory media. Combinations of Chi (5 and 7.5 mg/mL) and MPEO or MVEO (0.15-1.25 mL/mL) inhibited the growth of Colletotrichum isolates and mostly presented additive or synergistic interactions. Development of anthracnose lesions caused by C. gloeosporioides and C. brevisporum isolates was reduced by coatings formed by Chi (5 mg/mL) and MPEO or MVEO (0.3-1.25 µL/mL) combinations during storage (10 days, 25 ±â€¯0.5 °C). Decreases in anthracnose lesion development in papaya coated with Chi (5 mg/mL) and MPEO or MVEO (0.6 and 1.25 µL/mL) were similar or higher than those caused by a comercial fungicides formulation. The application of coatings formed by combinations of selected Chi and MPEO or MVEO concentrations could be considered an alternative strategy to control papaya anthracnose caused by C. gloeosporioides and C. brevisporum.

Evaluation of microbial communities in peels of Brazilian tropical fruits by amplicon sequence analysis.

Elucidation of the distinctive microbial taxonomic profiles of tropical fruit peels is the indispensable component of investigations aimed at the detection of microorganisms responsible for the post-harvest loss. The objective of the present work was to dissect the bacterial and fungal community of five tropical fruit peels (banana, guava, mango, papaya, and passion fruit) in wild (non-cultivated) and conventionally produced samples from Brazil. To that end, 16S rRNA-encoding gene and ITS rDNA amplicon analysis of the five tropical fruit peels were performed to discriminate the bacterial and fungal communities, respectively. The result showed that bacterial communities of the five types of fruit peels were by far more diversified than that of fungal communities, independent of the type of production system involved. Among the investigated fruits, non-cultivated papaya peels hosted the most diversified bacterial community while the least bacterial community diversity was found in the conventionally produced papaya fruit peels. The gene amplicon analysis clearly discriminated the bacterial community into their respective classes, while fungal communities were better classified in their phyla, yet with clearer component discrimination of fungal community based on the type of cultivation system practiced. Conventionally produced banana and non-cultivated passion fruit peels were characteristically dominated by fungal and bacterial groups, respectively. Overall, in conventionally produced fruit peels, bacterial community was mainly composed of Proteobacteria, Actinobacteria, and Bacilli. The result provided a broad microbial diversity profile that could be used as an important input for seeking alternative fruit spoilage control and post-harvest treatments.

Diverse cellular colonizing endophytic bacteria in field shoots and in vitro cultured papaya with physiological and functional implications.

The study was envisaged to assess the extent of normally uncultivable endophytic bacteria in field papaya plants and in vitro established cultures adopting cultivation vs molecular analysis and microscopy. Surface-sterilized axillary shoot-buds of papaya 'Arka Surya' revealed high bacterial diversity as per 16S rRNA metagene amplicon sequencing (6 phyla, 10 classes, 21 families) with an abundance of Pseudomonas (Gammaproteobacteria), which also formed a common contaminant for in vitro cultured field explants. Molecular analysis of seedling shoot-tip-derived healthy proliferating cultures of three genotypes ('Arka Surya', 'Arka Prabhath', 'Red Lady') with regular monthly subculturing also displayed high bacterial diversity (11-16 phyla, >25 classes, >50 families, >200 genera) about 12-18 months after initial establishment. 'Arka Surya' and 'Red Lady' cultures bore predominantly Actinobacteria (75-78%) while 'Arka Prabhath' showed largely Alphaproteobacteria corroborating the slowly activated Methylobacterium sp. Bright-field direct microscopy on tissue sections and tissue homogenate and epi-fluorescence microscopy employing bacterial DNA probe SYTO-9 revealed abundant intracellular bacteria embracing the next-generation sequencing elucidated high taxonomic diversity. Phylogenetic investigation of communities by reconstruction of unobserved states- PICRUSt- functional annotation suggested significant operational roles for the bacterial-biome. Metabolism, environmental information processing, and genetic information processing constituted major Kyoto Encyclopedia of Genes and Genomes KEGG attributes. Papaya stocks occasionally displayed bacterial growth on culture medium arising from the activation of originally uncultivable organisms to cultivation. The organisms included Bacillus (35%), Methylobacterium (15%), Pseudomonas (10%) and seven other genera (40%). This study reveals a hidden world of diverse and abundant conventionally uncultivable cellular-colonizing endophytic bacteria in field shoots and micropropagating papaya stocks with high genotypic similarity and silent participation in various plant processes/pathways.

Genetic variation of Colletotrichum magnum isolated from Carica papaya as revealed by DNA fingerprinting.

Mexico is one of the five largest producers of papaya worldwide, but losses caused by pathogens, mainly fungus, at the pre- and post-harvest stages are often more than 50% of the crop. Papaya anthracnose, caused by three different species of the Colletotrichum genus in Mexico, occupies a preponderant place in this problem. Although two of these species, C. gloeosporiodes and C. truncatum, have been characterized morphologically and genotypically, this has not occurred with C. magnum, the third species involved, about which there is very little information. Because of this, it is vital to know its genetic characterization, much more so considering that the studies carried out on the other two species reveal a wide genetic diversity, differences in pathogenicity and in the response to fungicides of the different strains characterized. In this work, Colletotrichum spp. isolates were collected at different papaya orchards in the south-southeast of Mexico. C. magnum isolates identified by species-specific primers were characterized by morphological and molecular approaches. Differences in colony characteristics resulted in five morphological groups. AP-PCR, DAMD and ISSR markers were found to be very efficient for revealing the interspecific variability of this species. The high genetic variability found in the accessions of C. magnum was linked to the geographical area where they were collected. Isolates from Chiapas State were the most variable, showing point mutations in the ITS1-ITS2 region. These results will enable a better phytosanitary management of anthracnose in papaya in this region of Mexico.

Anti-quorum sensing and antibiofilm potential of Alternaria alternata, a foliar endophyte of Carica papaya, evidenced by QS assays and in-silico analysis.

In the present study, secondary metabolites from an endophytic fungus, Alternaria alternata, colonizing Carica papaya, demonstrated antiquorum sensing properties against Pseudomonas aeruginosa. This study reports the antagonistic effects of fungal crude extract of A. alternata against the various quorum sensing (QS) associated virulent factors such as percentage decrease in production of pyocyanin, alginate, chitinase and rhamnolipid; significant decrease in proteases activity such as LasA protease activity, staphylolytic activity, Las B elastase; and a marked decrease in biofilm formation and associated factors such as exopolysaccharide (EPS) production and cell surface hydrophobicity (CSH). Further, motility pattern i.e., swimming and swarming was also found to be inhibited. This down regulation of QS and associated factors are further supported by in-silico analysis of interaction between QS receptor LasR and bioactive molecules viz., sulfurous acid, 2-propyl tridecyl ester and 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester present in fungal crude extract, found based on GCMS analysis, sketches the modulating ability of QS expression. This is the first report on an endophytic fungus of C. papaya having a role in QS inhibition against P. aeruginosa and lays a platform to explore further the endophytes for potent therapeutic agents in QS.