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A polyphasic approach was applied to characterize taxonomically a novel endophytic bacterial strain, designated as EP178 T , which was previously isolated from Passiflora incarnata leaves and characterized as plant-growth promoter. The strain EP178 T 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 (C 18:1 ω7c), summed feature 3 (C 16:1 ω6c /C 16:1 ω7c), and C 16:0 , and the predominant ubiquinone was Q-9. The phylogenetic and nucleotide-similarity analysis with 16S rRNA gene sequences showed that strain EP178 T 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 EP178 T and the closest type strain, P. oryzihabitans DSM 6835 T , 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 EP178 T represents a new species of the genus Pseudomonas, for which the name Pseudomonas flavocrustae sp. nov. was proposed. The type strain is EP178 T (= CBMAI 2609 T  = ICMP 24844 T  = MUM 23.01 T )., (© 2024. The Author(s).)

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., (© 2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

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., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

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., (© 2021 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.)

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.

Plants and endophytic microorganisms have coevolved unique relationships over many generations. Plants show a specific physiological status in each developmental stage, which may determine the occurrence and dominance of specific endophytic populations with a predetermined ecological role. This study aimed to compare and determine the structure and composition of cultivable and uncultivable bacterial endophytic communities in vegetative and reproductive stages (RS) of Passiflora incarnata. To that end, the endophytic communities were assessed by plating and Illumina-based 16S rRNA gene amplicon sequencing. Two hundred and four cultivable bacterial strains were successfully isolated. From the plant's RS, the isolated strains were identified mainly as belonging to the genera Sphingomonas, Curtobacterium, and Methylobacterium, whereas Bacillus was the dominant genus isolated from the vegetative stage (VS). From a total of 133,399 sequences obtained from Illumina-based sequencing, a subset of 25,092 was classified in operational taxonomy units (OTUs). Four hundred and sixteen OTUs were obtained from the VS and 66 from the RS. In the VS, the most abundant families were Pseudoalteromonadaceae and Alicyclobacillaceae, while in the RS, Enterobacteriaceae and Bacillaceae were the most abundant families. The exclusive abundance of specific bacterial populations for each developmental stage suggests that plants may modulate bacterial endophytic community structure in response to different physiological statuses occurring at the different plant developmental stages., (© 2019 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.)

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.

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.

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.

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.

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.

Previous works indicated a considerable variation in the pathogenicity, virulence, and host range of Oidium neolycopersici isolates causing tomato powdery mildew epidemics in many parts of the world. In this study, rDNA internal transcribed spacer (ITS) sequences, and amplified fragment length polymorphism (AFLP) patterns were analyzed in 17 O. neolycopersici samples collected in Europe, North America, and Japan, including those which overcame some of the tomato major resistance genes. The ITS sequences were identical in all 10 samples tested and were also identical to ITS sequences of eight previously studied O. neolycopersici specimens. The AFLP analysis revealed a high genetic diversity in O. neolycopersici and indicated that all 17 samples represented different genotypes. This might suggest the existence of either a yet unrevealed sexual reproduction or other genetic mechanisms that maintain a high genetic variability in O. neolycopersici. No clear correlation was found between the virulence and the AFLP patterns of the O. neolycopersici isolates studied. The relationship between O. neolycopersici and powdery mildew anamorphs infecting Aquilegia vulgaris, Chelidonium majus, Passiflora caerulea, and Sedum alboroseum was also investigated. These anamorphs are morphologically indistinguishable from and phylogenetically closely related to O. neolycopersici. The cross-inoculation tests and the analyses of ITS sequences and AFLP patterns jointly indicated that the powdery mildew anamorphs collected from the above mentioned plant species all represent distinct, but closely related species according to the phylogenetic species recognition. All these species were pathogenic only to their original host plant species, except O. neolycopersici which infected S. alboroseum, tobacco, petunia, and Arabidopsis thaliana, in addition to tomato, in cross-inoculation tests. This is the first genome-wide study that investigates the relationships among powdery mildews that are closely related based on ITS sequences and morphology. The results indicate that morphologically indistinguishable powdery mildews that differed in only one to five single nucleotide positions in their ITS region are to be considered as different taxa with distinct host ranges.

The introduction of anti-apoptotic genes into plants leads to resistance to environmental stress and broad-spectrum disease resistance. The anti-apoptotic gene (p35) from a baculovirus was introduced into the genome of passion fruit plants by biobalistics. Eleven regenerated plants showed the presence of the p35 gene by PCR and/or dot blot hybridization. Transcriptional analysis of regenerated plants showed the presence of specific p35 transcripts in 9 of them. Regenerated plants containing the p35 gene were inoculated with the cowpea aphid-borne mosaic virus (CABMV), the bacterium Xanthomonas axonopodis pv passiflorae, and the herbicide, glufosinate, (Syngenta). None of the plants showed resistance to CABMV. Regenerated plants (p35+) showed less than half of local lesions showed by non-transgenic plants when inoculated with X. axonopodis and some p35+ plants showed increased tolerance to the glufosinate herbicide when compared to non-transgenic plants.

The cultivated passion fruit (Passiflora edulis f. flavicarpa) is a cross-pollinated species native to South America. In the current study, a segregating F1 population derived from a single cross between the clones IAPAR-06 and IAPAR-123 was used to construct AFLP-based linkage maps and to map resistance genes to bacterial spot caused by Xanthomonas axonopodis pv. passiflorae. Linkage analysis was performed by the 2-way pseudo-testcross mapping method using markers that segregated in a 1:1 ratio. The IAPAR-06 linkage map was constructed using 115 markers, 112 of which were allocated to 9 linkage groups (LG) covering 790.2 cM. The map of IAPAR-123 was constructed using 140 markers, 138 of which were allocated to 9 LG covering 488.9 cM. In both maps, clusters of markers were detected, indicating that the AFLP markers were not distributed at random. Bacterial resistance was assessed by measuring the diseased leaf area after wound-inoculating the leaves of F1 plants. Quantitative resistance loci (QRLs) mapping was carried out by composite interval mapping and 1 QRL was detected, which explained 15.8% of the total phenotypic variation. The possibility of considering these data for marker-assisted selection in passion fruit breeding programs is discussed.

A single cross between two clones of passion fruit (Passiflora edulis Sims. f. flavicarpa Deg., 2n = 18) was selected for genetic mapping. The mapping population was composed of 90 F1 plants derived from a cross between 'IAPAR 123' (female parent) and 'IAPAR 06' (male parent). A total of 380 RAPD primers were analyzed according to two-way pseudo-testcross mapping design. The linkage analysis was performed using Mapmaker version 3.0 with LOD 4.0 and a maximum recombination fraction (theta) of 0.30. Map distances were estimated using the Kosambi mapping function. Linkage maps were constructed with 269 loci (2.38 markers/primer), of which 255 segregated 1:1, corresponding to a heterozygous state in one parent and null in the other. The linkage map for 'IAPAR123' consisted of 135 markers. A total of nine linkage groups were assembled covering 727.7 cM, with an average distance of 11.20 cM between framework loci. The sizes of the linkage groups ranged from 56 to 144.6 cM. The linkage map for 'IAPAR 06' consisted of 96 markers, covering 783.5 cM. The average distance between framework loci was 12.2 cM. The length of the nine linkage groups ranged from 20.6 to 144.2 cM. On average, both maps provided 61% genome coverage. Twenty-four loci (8.9%) remained unlinked. Among their many applications, these maps are a starting point for the identification of quantitative trait loci for resistance to the main bacterial disease affecting passion fruit orchards in Brazil, caused by Xanthomonas campestris pv. passiflorae, because parental genotypes exhibit diverse responses to bacterial inoculation.

The effects of mycorrhizal inoculation and increasing soil P levels on the expression of total proteins and peroxidase activity on passion fruit roots were evaluated. The experimental design was entirely at random, with four treatments of inoculation (a--control; b--Gigaspora albida; c--Scutellospora heterogama; d--mixture of G. albida, G. margarita, S. heterogama, and Glomus clarum) x three levels of soil P (4, 11, and 30 mg/dm3 of soil), each with three replicates. Plants were harvested 70 days after inoculation, when root colonization, shoot P level, protein content, and enzymatic activity of peroxidase (PAGE--7%) on root extract were evaluated. Regarding protein, there was no significant difference among the treatments, except between those roots receiving mixed inoculum and 11 mg P/dm3 of soil. Effect of P on protein concentration, when compared with the inoculation effect was observed. For peroxidase, there was an eletrophoretic band common to all treatments (rf: 0.43) and another that was absent only in noncolonized plants, grown in soil with lower P (rf: 0.46). Mycorrhizal specific bands were not present but a small decrease of intensity of bands in noncolonized plants was observed. Conversely, the control roots presented a single band (rf: 0.33) not observed in the other extracts, that may demonstrate an inhibitory effect of AMF on some host activities. The data showed the influence of P level in soil on the protein expression of roots, suggesting the influence of this nutrient on root genetic expression as well as on the mechanisms of symbiotic control/recognition.