Identification and Description of a New Pathogen Causing Flower Dry Rot on Passiflora edulis in China.

Passiflora edulis Sims (passion fruit) is an economically important fruit crop. However, a new flower dry rot has occurred in orchards located in Zhanjiang, China, and has led to serious production loss. Its disease incidence is approximately 30 to 40%. A total of 221 isolates of Fusarium sp. were obtained from samples of three types of symptomatic flowers. Three representative single-spore isolates (PaB-1, PaB-2, and PaB-3) from each type were used for pathogenicity tests, multilocus phylogenetic analyses, and morphological descriptions. Pathogenicity tests of buds of 5-month-old P. edulis plants showed symptoms similar to those observed in nature, and Koch's postulates were achieved. By comparing 36 typical species from the FUSARIUM-ID database, multilocus phylogenetic analyses showed that the sequences of TEF1, RPB2, and ITS of these isolates belong to the Incarnatum clade of the F. incarnatum-equiseti species complex (FIESC-17-a) with an independent branch. Therefore, the pathogenic isolates were identified as F. pernambucanum (FIESC-17-a). Moreover, in this study, the conidial anastomosis tubes were first observed in the FIESC. This is the first report of flower dry rot on P. edulis caused by F. pernambucanum. Further studies should be performed to determine effective disease management strategies.

The yeast community of Conotelus sp. (Coleoptera: Nitidulidae) in Brazilian passionfruit flowers (Passiflora edulis) and description of Metschnikowia amazonensis sp. nov., a large-spored clade yeast.

Species of the nitidulid beetle Conotelus found in flowers of Convolvulaceae and other plants across the New World and in Hawaii consistently harbour a yeast community dominated by one or more large-spored Metschnikowia species. We investigated the yeasts found in beetles and flowers of cultivated passionfruit in Rondônia state, in the Amazon biome of Brazil, where a Conotelus species damages the flowers and hinders fruit production. A sample of 46 beetles and 49 flowers yielded 86 and 83 yeast isolates, respectively. Whereas the flower community was dominated by Kodamaea ohmeri and Kurtzmaniella quercitrusa, the major yeasts recovered from beetles were Wickerhamiella occidentalis, which is commonly isolated from this community, and a novel species of large-spored Metschnikowia in the arizonensis subclade, which we describe here as Metschnikowia amazonensis sp. nov. Phylogenetic analyses based on barcode sequences (ITS-D1/D2) and a multigene alignment of 11,917 positions (genes ura2, msh6, and pmt2) agreed to place the new species as a sister to Metschnikowia arizonensis, a rare species known only from one locality in Arizona. The two form sterile asci when mated, which is typical of related members of the clade. The α pheromone of the new species is unique but typical of the subclade. The type of M. amazonensis sp. nov. is UFMG-CM-Y6309T (ex-type CBS 16156T , mating type a), and the designated allotype (mating type α) is UFMG-CM-Y6307A (CBS 16155A ). MycoBank MB 833560.

Endophytic fungi from Passiflora incarnata: an antioxidant compound source.

Endophytes are considered one of the most important microbial resources for obtaining biomolecules of therapeutic use. Passiflora incarnata, widely employed by the pharmaceutical industry, shows therapeutic effects on anxiety, nervousness, constipation, dyspepsia and insomnia based on their antioxidant compounds. In this study, from 315 endophytic fungi isolated from P. incarnata leaves, 60 were selected to determinate presence of chemical constituents related with antioxidant activity, based on their production of soluble pigments. The promising fungi were evaluated specifically on their potential to produce phenolic compounds, flavonoids and for antioxidant activity. Five isolates significantly produced flavonoids and phenolic compounds in the ethyl acetate and n-Butanol extracts, also saponins and high antioxidant activity against the DPPH (2.2-diphenyl-1-picrylhydrazyl) free radical. A strain of Aspergillus nidulans var. dentatus (former Emericella dentata) was able to produce tannins as well; its butanolic extract was very similar than the BHT (butylated hydroxytoluene) (94.3% × 94.32%) and Rutin (95.8%) reference substances in the DPPH radical scavenging. Similarly, a Chaetomium strain exhibited 93.6% and 94.7% of antioxidant activity in their ethyl acetate and butanolic fractions, respectively. The chromatographic analysis of the ethyl acetate fraction from the Aspergillus strain revealed the production of orcinol (3.19%). Four-methoxymethylphenol (4.79%), sorbicillin (33.59%) and ergosterol (23.08%) was produced by Trichoderma longibrachiatum and isopropenyl-1,4-dimethyl-1,2,3,3a,4,5,6,7-octahydroazulene were found in two Fusarium oxysporum strains. The phytochemical screening showed that all analyzed fungi were able to produce a kind of secondary metabolite (phenols, flavonoids, tannins and/or saponins). The study shows a great underexplored potential for industrial application of P. incarnata endophytes.

Is there an impact of the dairy matrix on the survival of Lactobacillus casei Lc-1 during shelf life and simulated gastrointestinal conditions?

BACKGROUND: The selection of the food matrix to be used as a vehicle for a probiotic culture is important because its chemical composition and physicochemical characteristics can affect probiotic survival during the shelf-life of the product and under simulated gastrointestinal conditions (SGIC). The present study aimed to evaluate the influence of the dairy matrix (chocolate fermented milk beverage, chocolate flan or passion fruit flan) on the survival of Lactobacillus casei Lc-1 during refrigerated storage (4 °C for 21 days) and SGIC. RESULTS: Chocolate fermented milk beverage and chocolate and passion fruit flans could be considered as matrices for the incorporation of L. casei, providing suitable counts (6.38-7.84 log cfu g-1 ) during storage. The type of matrix had an impact on the inicial probiotic counts in the products and on the probiotic resistance to the SGIC. The chocolate fermented milk beverage presented higher initial probiotic counts (7.72 versus 6.65-7.28 log cfu g-1 ). The higher pH (5.3-6.8), solid matrix and increased fat content (65.0-72.9 g 100 g-1 ) contributed to the higher resistance to the SGIC of the chocolate and passion fruit flans, allowing recovery of viability during the enteric phase (increases of 1-1.5 log cycles). CONCLUSION: The type of dairy matrix has an impact on the inicial probiotic counts in the products and on the probiotic resistance to the SGIC. Chocolate and passion fruit flans proved to be more suitable options than chocolate fermented milk beverage for the incorporation of Lactobacillus casei Lc-1. © 2019 Society of Chemical Industry.

Use of mycorrhizal fungi releases the application of organic fertilizers to increase the production of leaf vitexin in yellow passion fruit.

BACKGROUND: Low-cost organic fertilizers, such as coconut powder and vermicompost, and arbuscular mycorrhizal fungi (AMF) may benefit the Passiflora edulis f. flavicarpa plant. However, it has not been established whether the joint application of these inputs may increase the production of vitexin and other molecules associated with the phytotherapeutic properties of this plant. Here, we tested the hypothesis that the application of AMF and organic fertilizers maximizes the production of bioactive compounds in leaves of P. edulis. RESULTS: The inoculation of Acaulospora longula into P. edulis grown in fertilization-free soil promoted an increase of 86% in the concentration of leaf vitexin, 10.29% in the concentration of total phenols, and 13.78% in the concentration of total tannins in relation to the AMF-free control, rendering soil fertilization superfluous. CONCLUSION: The application of A. longula increases the production of foliar biomolecules, such as vitexin, in yellow passion fruit plants. Thus, the addition of coconut powder and vermicompost to the substrate composition is not necessary, leading to the commercialized production of phytomass in the herbal medicines industry. © 2019 Society of Chemical Industry.

Orange and Passion Fruit Wastes Characterization, Substrate Hydrolysis and Cell Growth of Cupriavidus necator, as Proposal to Converting of Residues in High Value Added Product.

Brazil is the world's largest producer of orange and passion fruit, which are destined mainly for industrialization, generating grand volumes of wastes. The solid portion of these residues is a rich source of pectin - composed mainly of galacturonic acid and neutral sugars, which through the hydrolysis process can be used in biological conversion processes, as the production of polyhydroxyalkanoates (PHAs). This way, we characterized these wastes, followed by the extraction and hydrolysis of pectin for employ as a substrate for the cell growth of Cupriavidus necator. The results confirmed the large portion of pectin (almost 40 g.100g-1) and soluble sugars, present in these wastes. The hydrolyzed extract showed as a good source of carbon for the cell growth of C. necator with YX/S 0.56 and 0.44, µMax 0.27 and 0.21 for orange and passion fruit wastes respectively, similar to other carbon sources. This way, the extraction and hydrolysis of orange and passion fruit wastes for the cellular growth of C. necator, can be a good alternative to converting of residues in high value added product.

Production of biomolecules of interest to the anxiolytic herbal medicine industry in yellow passionfruit leaves (Passiflora edulis f. flavicarpa) promoted by mycorrhizal inoculation.

BACKGROUND: Our contemporary way of life has led us to consume high amounts of chemically-synthesized allopathic medicinal products and anxiolytics to which a viable alternative is the use of Passiflora-based herbal medicines with composition containing vitexin, a flavonoid with anxiolytic and antidepressant properties. Arbuscular mycorrhizal fungi (AMF) are known for enhancing the production of biomolecules, however, increase production of phytochemistry in Passiflora edulis f. flavicarpa has not been reported in the literature. Our aim was to select AMF to benefit the production of vitexin in leaves of P. edulis by inoculating seedlings in the region of roots with Acaulospora longula, Claroideoglomus etunicatum and Gigaspora albida. RESULTS: The inoculation increased the concentration of vitexin in 63.64% and the inoculation with A. longula also increased the content of flavonoids and total saponins in the leaves in relation to the control. CONCLUSION: The increase in the production of vitexin in the leaf in response to the inoculation with AMF, with emphasis to A. longula, interests the pharmaceutical industry and can generate profit to the production of yellow passionfruit-based anxiolytic herbal medicine. © 2019 Society of Chemical Industry.

Activity of Antarctic fungi extracts against phytopathogenic bacteria.

This study aims to obtain secondary metabolites extracts from filamentous fungi isolated from soil and marine sediments from Antarctic ecosystems and to assess its potential antibacterial activity on Xanthomonas euvesicatoria and Xanthomonas axonopodis pv. passiflorae (phytopathogenic bacteria causing diseases in pepper and tomato and passionfruit, respectively). Among the 66 crude intracellular and extracellular extracts obtained from fungi recovered from soil and 79 obtained from marine sediment samples, 25 showed the ability to prevent the growth of X. euvesicatoria in vitro and 28 showed the ability to prevent the growth of X. axonopodis pv. passiflorae in vitro. Intracellular and extracellular extracts from soil fungi inhibited around 97% of X. euvesicatoria and 98% of X. axonopodis pv. passiflorae at 2·1 mg ml-1 . The average inhibition rates against X. euvesicatoria and X. axonopodis pv. passiflorae for intracellular and extracellular extracts from marine sediments fungi were around 96 and 97%, respectively, at 3·0 mg ml-1 . Extracts containing secondary metabolites with antimicrobial activity against X. euvesicatoria and X. axonopodis pv. passiflorae were obtained, containing possible substitutes for the products currently used to control these phytopathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Micro-organisms from extreme ecosystems, such as the Antarctic ecosystem, need to survive in harsh conditions with low temperatures, low nutrients and high UV radiation. Micro-organisms adapt to these conditions evolving diverse biochemical and physiological adaptations essential for survival. All this makes these micro-organisms a rich source of novel natural products based on unique chemical scaffolds. Discovering novel bioactive compounds is essential because of the rise in antibiotic-resistant micro-organisms and the emergence of new infections. Fungi from Antarctic environments have been proven to produce bioactive secondary metabolites against various micro-organisms, but few studies have shown activity against Xanthomonas phytopathogens.

Grafting height does not affect Fusarium wilt control or horticultural performance of Passiflora gibertii N.E.Br. rootstock.

The influence of the grafting height (5, 10, 20 and 30 cm above the root collar) of P. edulis on P. gibertii was evaluated on the incidence of Fusarium wilt and horticultural performance. Plants of P. gibertii grafted on P. edulis and non-grafted plants of both species were also studied. In addition, histopathological studies were also performed on the roots of non-grafted P. edulis collected at three severity stages of Fusarium wilt. In greenhouse, the graft take was inversely related to the grafting height in general. In the field conditions, the plant growth of P. gibertii grafted on P. edulis was superior to its reciprocal grafting, even though the former combination was susceptible to Fop. Plants of P. edulis grafted on P. gibertii at all grafting heights did not present symptoms of Fop, and the number of fruit yield and quality were equivalent, but plant growth was decreased in relation to the non-grafted plants. Starch depletion in the root system of P. edulis was directly related to the severity of the Fusarium wilt. P. gibertii was confirmed as a Fusarium wilt resistant rootstock of P. edulis, with minimal influence of the grafting height for the control of the disease.

Physicochemical and Microbiological Quality of Dietetic Functional Mixed Cerrado Fruit Jam during Storage.

The objective of the research was to evaluate changes of dietetic functional mixed cerrado fruit jam (marolo, sweet passion fruit, and soursop) processed in a vacuum pot and stored for 180 days in BODs at 25°C and 35°C. The parameters evaluated were pH, soluble solids (SS), titratable acidity (TA), total sugars (TS), total carotenoids (TC), total phenolics (TP), vitamin C, antioxidant activity (DPPH), and microbiological analysis. There was a significant effect of storage time on pH, SS, TA, TC, TS, and TP. Vitamin C and DPPH showed an effect for the temperature x storage time interaction. Statistical models are not adjusted for pH and SS, presenting an average of 4.15 and 61%, respectively. Carotenoids decreased up to105 days; total sugars increased up to 105 days. The TP, vitamin C, and DPPH, at the temperatures evaluated, showed a decrease up to 105 days. Yeasts and filamentous fungi were not detected.

Arbuscular mycorrhizal fungi and vermicompost to maximize the production of foliar biomolecules in Passiflora alata Curtis seedlings.

BACKGROUND: Arbuscular mycorrhizal fungi (AMF) are soil organisms that associate with plant roots and promote plant growth, increasing the concentration of secondary metabolites, which are molecules of interest to the pharmaceutical industry. Passiflora alata is a Brazilian medicinal plant that is used as a raw material for anxiolytic phytotherapeutic agents. The anxiolytic properties are related to the presence of biomolecules in the plant material, principally flavonoids. The objective of this study was to evaluate the effect of inoculation with AMF and organic fertilization on the production of soluble carbohydrates and total foliar proteins, phenols and flavonoids in P. alata seedlings. RESULTS: There was an effect of the tested treatments on the analyzed variables. Seedlings inoculated and cultivated in soil to which 0.15 kg vermicompost kg(-1) had been added showed increased growth and production of primary and secondary metabolites compared with treatments with lower levels of manure, which did not differ from those cultivated in soil with 0.2 kg vermicompost kg(-1). CONCLUSION: The inoculation of P. alata seedlings with Gigaspora albida is an alternative to maximize the production of pharmacologically important foliar biomolecules, especially flavonoids, with benefits that vary in accordance with the fertility of the soil.

Pseudomonas flavocrustae sp. nov., an endophyte with plant growth promoting traits isolated from Passiflora incarnata.

A polyphasic approach was applied to characterize taxonomically a novel endophytic bacterial strain, designated as EP178T, which was previously isolated from Passiflora incarnata leaves and characterized as plant-growth promoter. The strain EP178T forms Gram stain-negative and rod-shaped cells, and circular and yellow-pigmented colonies. Its growth occurs at 10-37 °C, at pH 6.0-8.0, and tolerates up to 7% (w/v) NaCl. The major cellular fatty acids found were summed feature 8 (C18:1 ω7c), summed feature 3 (C16:1 ω6c /C16:1 ω7c), and C16:0, and the predominant ubiquinone was Q-9. The phylogenetic and nucleotide-similarity analysis with 16S rRNA gene sequences showed that strain EP178T belongs to Pseudomonas genus. The genomic-based G + C content was 65.5%. The average nucleotide identity and digital DNA-DNA hybridization values between strains EP178T and the closest type strain, P. oryzihabitans DSM 6835T, were 92.6% and 52.2%, respectively. Various genes associated with plant-growth promoting mechanisms were annotated from genome sequences. Based on the phenotypic, genomic, phylogeny and chemotaxonomic data, strain EP178T represents a new species of the genus Pseudomonas, for which the name Pseudomonas flavocrustae sp. nov. was proposed. The type strain is EP178T (= CBMAI 2609T = ICMP 24844T = MUM 23.01T).

'Candidatus Phytoplasma sudamericanum', a novel taxon, and strain PassWB-Br4, a new subgroup 16SrIII-V phytoplasma, from diseased passion fruit (Passiflora edulis f. flavicarpa Deg.).

Symptoms of abnormal proliferation of shoots resulting in formation of witches'-broom growths were observed on diseased plants of passion fruit (Passiflora edulis f. flavicarpa Deg.) in Brazil. RFLP analysis of 16S rRNA gene sequences amplified in PCRs containing template DNAs extracted from diseased plants collected in Bonito (Pernambuco) and Viçosa (Minas Gerais) Brazil, indicated that such symptoms were associated with infections by two mutually distinct phytoplasmas. One phytoplasma, PassWB-Br4 from Bonito, represents a new subgroup, 16SrIII-V, in the X-disease phytoplasma group ('Candidatus Phytoplasma pruni'-related strains). The second phytoplasma, PassWB-Br3 from Viçosa, represents a previously undescribed subgroup in group 16SrVI. Phylogenetic analyses of 16S rRNA gene sequences were consistent with the hypothesis that strain PassWB-Br3 is distinct from previously described 'Ca. Phytoplasma' species. Nucleotide sequence alignments revealed that strain PassWB-Br3 shared less than 97.5 % 16S rRNA gene sequence similarity with previously described 'Ca. Phytoplasma' species. The unique properties of its DNA, in addition to natural host and geographical occurrence, support the recognition of strain PassWB-Br3 as a representative of a novel taxon, 'Candidatus Phytoplasma sudamericanum'.

Arbuscular mycorrhizal fungi inoculation stimulates the production of foliar secondary metabolites in Passiflora setacea DC.

Passiflora setacea DC. growing is of interest to the herbal industries since in its leaves are produced secondary metabolites that confer antioxidant, anxiolytic, and antidepressant properties in Passiflora. Therefore, it is important to search for sustainable alternatives that aim to enhance the production of these compounds to add value to the phytomass, such as the inoculation with arbuscular mycorrhizal fungi (AMF) and the application of coconut coir dust, which has not been reported to P. setacea yet. The aim was to select the efficient combination of AMF and coconut coir dust to increase the compounds' production and optimize the antioxidant activity in P. setacea leaves. The P. setacea seedlings that were cultivated in substrates without coconut coir dust and colonized by Gigaspora albida N.C. Schenck & G.S. Sm. produced more total saponins (1,707.43%), total tannins (469.98%), and total phenols (85.81%), in comparison to the non-mycorrhizal plants, in addition to enhancing the glomalin-related soil proteins. On the other hand, in general, the use of coir dust as a substrate has not been shown to increase the production of these bioactive compounds. It is concluded that the production of P. setacea seedlings using G. albida is an alternative to offer phytomass to the herbal medicines industry based on passion fruit.

Characterization of cellular, biochemical and genomic features of the diazotrophic plant growth-promoting bacterium Azospirillum sp. UENF-412522, a novel member of the Azospirillum genus.

Given their remarkable beneficial effects on plant growth, several Azospirillum isolates currently integrate the formulations of various commercial inoculants. Our research group isolated a new strain, Azospirillum sp. UENF-412522, from passion fruit rhizoplane. This isolate uses carbon sources that are partially distinct from closely-related Azospirillum isolates. Scanning electron microscopy analysis and population counts demonstrate the ability of Azospirillum sp. UENF-412522 to colonize the surface of passion fruit roots. In vitro assays demonstrate the ability of Azospirillum sp. UENF-412522 to fix atmospheric nitrogen, to solubilize phosphate and to produce indole-acetic acid. Passion fruit plantlets inoculated with Azospirillum sp. UENF-41255 showed increased shoot and root fresh matter by 13,8% and 88,6% respectively, as well as root dry matter by 61,4%, further highlighting its biotechnological potential for agriculture. We sequenced the genome of Azospirillum sp. UENF-412522 to investigate the genetic basis of its plant-growth promotion properties. We identified the key nif genes for nitrogen fixation, the complete PQQ operon for phosphate solubilization, the acdS gene that alleviates ethylene effects on plant growth, and the napCAB operon, which produces nitrite under anoxic conditions. We also found several genes conferring resistance to common soil antibiotics, which are critical for Azospirillum sp. UENF-412522 survival in the rhizosphere. Finally, we also assessed the Azospirillum pangenome and highlighted key genes involved in plant growth promotion. A phylogenetic reconstruction of the genus was also conducted. Our results support Azospirillum sp. UENF-412522 as a good candidate for bioinoculant formulations focused on plant growth promotion in sustainable systems.

Genetic diversity and a PCR-based method for Xanthomonas axonopodis detection in passion fruit.

Xanthomonas axonopodis pv. passiflorae causes bacterial spot in passion fruit. It attacks the purple and yellow passion fruit as well as the sweet passion fruit. The diversity of 87 isolates of pv. passiflorae collected from across 22 fruit orchards in Brazil was evaluated using molecular profiles and statistical procedures, including an unweighted pair-group method with arithmetical averages-based dendrogram, analysis of molecular variance (AMOVA), and an assigning test that provides information on genetic structure at the population level. Isolates from another eight pathovars were included in the molecular analyses and all were shown to have a distinct repetitive sequence-based polymerase chain reaction profile. Amplified fragment length polymorphism technique revealed considerable diversity among isolates of pv. passiflorae, and AMOVA showed that most of the variance (49.4%) was due to differences between localities. Cluster analysis revealed that most genotypic clusters were homogeneous and that variance was associated primarily with geographic origin. The disease adversely affects fruit production and may kill infected plants. A method for rapid diagnosis of the pathogen, even before the disease symptoms become evident, has value for producers. Here, a set of primers (Xapas) was designed by exploiting a single-nucleotide polymorphism between the sequences of the intergenic 16S-23S rRNA spacer region of the pathovars. Xapas was shown to effectively detect all pv. passiflorae isolates and is recommended for disease diagnosis in passion fruit orchards.

The effect of the root-colonizing Piriformospora indica on passion fruit (Passiflora edulis) development: Initial defense shifts to fitness benefits and higher fruit quality.

Passion fruit (Passiflora edulis) has an important economic value as exotic ingredient in juice blends. We inoculated the passion fruit cultivar Passiflora edulis Sims f. edulis's roots with the beneficial root-colonizing fungus Piriformospora indica under greenhouse conditions. The experiments were performed at three different locations and times (between 2017 and 2019). After transient initial growth retardation associated with a mild salicylic-acid (SA)-dependent defense activation and reduced sucrose metabolism, plant performance and growth are promoted during later stages. The elevated SA level in the aerial parts stimulates the plant immune system and promotes pathogen resistance in the adult plants and the fruit peels. P. indica stimulates the fruit size and fruit quality, and the higher amounts of defense-related secondary metabolites in the peels restrict growth of herbivorous insect larvae fed with peel extracts. We conclude that application of P. indica to passion fruits stimulates the plants' immune system and improves the fruits' quality.

Kinetic study of selected microorganisms and quality attributes during cold storage of mango and passion fruit smoothie subjected to dimethyl dicarbonate.

The objectives of this research were to study the effect of DMDC (0-250 ppm) on quality and shelf life of mango and passion fruit smoothie during cold storage. The correlation between microbial population (total microorganisms, yeast and mold, E. coli and S. aureus) and DMDC concentration using zero-order kinetic and first-order kinetic was also determined. In addition, the effect of DMDC compared with pasteurization (90 °C, 100 s) on quality of mixed mango and passion fruit smoothie during the cold storage (4 °C) was studied. The results showed that microbial inactivation was best-described by first-order kinetic model due to a higher coefficient of determination (R2). In addition, DMDC did not affect the decreasing trend of total soluble solid, color difference (∆E*) and total phenolic compound as compared to control during the cold storage. DMDC also hindered the increasing trend in microbial population and prevented the loss of antioxidant activity (DPPH and FRAP assays) and total flavonoid content and decreased the PPO activity as compared with the control during the cold storage. In summary, DMDC showed the potential to maintain the quality and to extend the shelf life of mango and passion fruit smoothie during cold storage.

Diversity and characteristics of culturable endophytic bacteria from Passiflora edulis seeds.

Defense compounds generally inhibit microbial colonization of plants. In this study, we examined the presence of endophytes in Passiflora edulis seeds that accumulate resveratrol and piceatannol at extremely high levels as defense compounds. Interestingly, although no microbial colonies appeared on an agar growth medium from the cut or homogenized seeds, colonies were generated from cut seedlings derived from the seeds. A total of 19 bacterial strains were isolated, of which 15 were classified as Gram-positive. As we hypothesized that extremely high levels of piceatannol in the seeds would inhibit the growth of endophytes cultured directly from the seeds, we examined the antimicrobial activity of this compound against the isolated bacteria. Piceatannol exerted bacteriostatic rather than bactericidal effects on most of the bacteria tested. These results suggest that the bacteria remain static in the seeds due to the presence of piceatannol and are transmitted to the seedlings during the germination process, enabling colonies to be established from the seedlings on the agar medium. We also investigated the biocatalytic activity of the isolated bacteria toward resveratrol and piceatannol. One bacterium, Brevibacterium sp. PE28-2, converted resveratrol and piceatannol to their respective derivatives. This strain is the first endophyte shown to exhibit such activity.

Alicyclobacillus acidoterrestris in pasteurized exotic Brazilian fruit juices: isolation, genotypic characterization and heat resistance.

In this study, the population of Alicyclobacillus spp. was estimated in pasteurized exotic Brazilian fruit juices using the most probable number (MPN) technique followed by biochemical tests. Pasteurized passion fruit (n = 57) and pineapple (n = 50) juices were taken directly from Brazilian manufacturers. While Alicyclobacillus spp. was isolated from passion fruit juice, the microorganism was not found in any pineapple juice samples. A higher incidence of Alicyclobacillus was observed in samples taken in June and July (dry months in Brazil) in comparison to the other months (March, April, May and August), and the highest Alicyclobacillus counts were recovered from these samples(>23 MNP/100 mL). Sixteen (n = 16) Alicyclobacillus strains were typed using the randomly amplified polymorphic DNA method (RAPD-PCR). RAPD-PCR revealed great genetic similarity between the passion fruit juice strains and Alicyclobacillus acidoterrestris DSM 2498. The heat resistance of three isolates was determined, and the mean D(95°) (1.7 min) and z (7.6 °C) values in the passion fruit juice were not significantly different (p > 0.05) from those obtained for the DSM 2498 strain (D(95°) = 1.5 min and z = 7.1 °C). This is the first report on the isolation of A. acidoterrestris from exotic fruit juices such as passion fruit juice. It is worth pointing out the importance of applying good agricultural practices in the field and applying controls for the fruit selection and washing steps, as well as controlling the time/temperature conditions for pasteurization so as to reduce the incidence and chances of A. acidoterrestris spoilage in these juices.