Adventitious root primordia formation and development in the stem of Ananas comosus var. bracteatus slip.

There are about 4-6 slips on a fruit, and they are good materials for effective regeneration of Ananas comosus var. bracteatus. Adventitious root (AR) induction is essential for the propagation of Ananas comosus var. bracteatus slips. Growth regulator treatment, and culture medium are imperative factors that affect slip growth and rooting. In order to screen the optimal methods for slips rooting and reveal the anatomic procedure of slip rooting, this study induced slip rooting by different treatment of growth regulator, culture medium, observed the slip stem structure, AR origination and formation procedure through paraffin sections. The results showed that, slip cuttings treated with 100 mg/L of Aminobenzotriazole (ABT) for 6 hrs, cultured in river sand: coconut chaff: garden soil 2:2:1 medium is the optimal method for rooting. The proper supplementary of ABT can enhance the soluble sugar content, soluble protein content, polyphenol oxidase (PPO) activity and peroxidase (POD) enzyme activity, which resulted in the improvement of rooting. The slip stem structure is quite different from other monocots, which consists of epidermis, cortex, and stele with vascular tissues distributed in the cortex and stele. The AR primordia originates from the parenchyma cells located on the borderline between the cortex and stele. The vascular tissues in the AR develop and are connected with vascular tissue of the stem before the AR grew out the stem. The number of primary xylem poles in AR is about 30.

Genome-wide analysis of AP2/ERF transcription factors in pineapple reveals functional divergence during flowering induction mediated by ethylene and floral organ development.

The APETALA2/ethylene-responsive factor (AP2/ERF) has important roles in regulating developmental processes and hormone signaling transduction in plants. Pineapple demonstrates a special sensitivity to ethylene, and AP2/ERFs may contribute to this distinct sensitivity of pineapples to ethylene. However, little information is available on the AP2/ERF of pineapple. In this study, 97 AP2/ERF family members were identified from the pineapple genome. The AcAP2/ERF superfamily could be further divided into five subfamilies, and different subfamily existed functional divergence in multifarious biological processes. ERF and RAV subfamily genes might play important roles in the process of ethylene response of pineapple; ERF and DREB subfamily genes had particular functions in the floral organ development. This study is the first to provide detailed information on the features of AP2/ERFs in pineapple, provide new insights into the potential functional roles of the AP2/ERF superfamily members, and will facilitate a better understanding of the molecular mechanism of flower in pineapple.

Strategies for vegetative propagation and viral cleaning of a miniature ornamental pineapple hybrid

This study assessed and compared different methods for vegetative propagation of a miniature ornamental pineapple hybrid (ORN-MUT), seeking to determine the best method for production of plantlets, as well as for removal of the PMWaV viral complex from plants cultured in vitro, for production of healthy parent plants. Pineapple wilt is a disease that can cause large economic and is caused by a viral complex called Pineapple mealybug wilt-associated virus (PMWaV). For this, four propagation methods were evaluated (conventional, stem sectioning, micropropagation and etiolation of nodal segments). The time necessary for each method and the number of plants formed were assessed. Stem tips (0.5 mm) were cultured and indexed for three PMWaV types. Conventional propagation produced 17 plantlets per plant in 566 days, stem sectioning produced 2.3 plantlets per stem in 591 days, while the conventional micropropagation technique produced 1,284 plants after four subcultures in 778 days. Stems etiolated for 60 days showed peak production in the second subculture, with 1,224 plants. This method required 883 days to obtain plants with ideal size for transplantation to the field. In turn, stems etiolated for 120 days produced 935 plants at the end of four subcultures, with peak output in the third subculture, in which the plants could be cultivated in the field after 943 days. Conventional micropropagation and etiolation for 60 days were the best methods for production of plantlets of the ORN-MUT hybrid. The results of this work showed that the cultivation of shoot tips is an efficient strategy to remove the PMWaV complex and obtain healthy mother plants and can be a useful tool for other varieties of pineapple.

A role for MIR828 in pineapple fruit development.

Chen et al. ( Nature Genet. 51: 1549-1558; Oct. 2019) sequenced Ananas comosus var. bracteatus accession CB5, cultivated for its bright pink-to-red colored fruit, and yellow-fleshed A. comosus accession F153, reporting an improved F153 reference assembly while annotating MICRORNA (MIRNA) loci and gene family expressions relevant to lignin and anthocyanin biosynthesis. An independent article (Xiong et al.Sci. Rep. 8: 1947; 2018) reported var. bracteatus MIRNAs but not MIR828, a negative regulator of anthocyanin and polyphenolics biosynthesis by targeting MYB transcription factors associated with UV light- and sugar-signaling in dicots. MIR828 has been reported in gymnosperms, Amborella (sister to flowering plants), and basal monocot orders Liliales, Asparagales, Zingiberales, Arecales, but not in the Poales, a sister order comprising grasses and ~3,000 species of bromeliads including pineapple. Here I show MIR828 exists in pineapple and directs post-transcriptional gene silencing of mRNAs encoding MYB family members with inferred function to regulate the conspicuous red fruit trait in var. bracteatus. MIR828 plesiomorphy (an ancient basal trait) may shed light on monocot apomorphic fruit development, postulated for 21 monocot families with fleshy fruits as due to homoplasy/convergence driven by tropical climate and/or enticements to vertebrate endozoic seed dispersers.

Genome-Wide Analysis of the YABBY Transcription Factor Family in Pineapple and Functional Identification of AcYABBY4 Involvement in Salt Stress.

The plant-specific transcription factor gene family, YABBY, belongs to the subfamily of zinc finger protein superfamily and plays an essential regulatory role in lateral organ development. In this study, nine YABBY genes were identified in the pineapple genome. Seven of them were located on seven different chromosomes and the remaining two were located on scaffold 1235. Through protein structure prediction and protein multiple sequence alignment, we found that AcYABBY3, AcYABBY5 and AcYABBY7 lack a C2 structure in their N-terminal C2C2 zinc finger protein structure. Analysis of the cis-acting element indicated that all the seven pineapple YABBY genes contain multiple MYB and MYC elements. Further, the expression patterns analysis using the RNA-seq data of different pineapple tissues indicated that different AcYABBYs are preferentially expressed in various tissues. RT-qPCR showed that the expression of AcYABBY2, AcYABBY3, AcYABBY6 and AcYABBY7 were highly sensitive to abiotic stresses. Subcellular localization in pineapple protoplasts, tobacco leaves and Arabidopsis roots showed that all the seven pineapple YABBY proteins were nucleus localized. Overexpression of AcYABBY4 in Arabidopsis resulted in short root under NaCl treatment, indicating a negative regulatory role of AcYABBY4 in plant resistance to salt stress. This study provides valuable information for the classification of pineapple AcYABBY genes and established a basis for further research on the functions of AcYABBY proteins in plant development and environmental stress response.

Aceitação sensorial do abacaxi “FRF 632”, resistente à fusariose, em diferentes estádios de maturação / Sensory acceptance of fusariosis resistant pineapple “FRF 632” at different maturation stages

O objetivo desse estudo foi avaliar a influência do ponto de colheita na qualidade sensorial e físico-química de abacaxi “FRF632”. Os frutos foram avaliados por 59 julgadores à cor, aroma, sabor, textura/firmeza, aceitação global e intenção de compra. Avaliou-se também a acidez titulável, o teor de sólidos solúveis e a relação sólidos solúveis/acidez titulável. Os consumidores gostaram dos frutos, com percentual de aprovação superior a 75% para todos os atributos. A doçura e a acidez dos frutos, nos estádios de maturação “amarelo” e “colorido”, foram consideradas ideias por um maior número de consumidores. Conclui-se que os consumidores preferiram consumir frutos colhidos nos estádios de maturação “colorido” e “amarelo”, pois os consideraram com melhor cor, sabor e aroma, doçura e acidez ideais, além de maior intenção de compra.

Evaluation of the micropropagation potential of curauá pineapple hybrids for fiber production / Avaliação do potencial de micropropagação de híbridos do abacaxi curauá para a produção de fibras

Plant fiber is a renewable and biodegradable material that can be used effectively to reinforce various composites. Pineapple hybrids selected for their fiber quality are in the phase of agronomic validation in Brazil by the Embrapa Cassava and Fruits research unit. The selection of a hybrid for large-scale fiber production depends on obtaining a large number of seedlings. This study evaluated the morphogenetic response and propagation potential of eight hybrids of Ananas comosus var. erectifolius, for the purpose of producing high-quality seedlings on a large scale. Stem and crown buds were reduced and placed in MS nutritive medium supplemented with BAP at 0.5 mg L-1, NAA at 0.01 mg L-1 and Phytagel® at 2.5 g L-1. After 45 days, the number of oxidized, contaminated and surviving buds was determined. Swollen buds and plantlets were transferred to a multiplication medium containing MS sucrose, salts and vitamins. The propagation potential was evaluated based on the geometric growth rate among sub-cultures. The FIB-NEG hybrid presented the best results for the establishment phase (40.28%). The best propagative potential was obtained from crown buds with the highest values for FIB-EST (3.93), FIB-MIN (3.91) and FIB-BOY (3.91) hybrids.

A fibra vegetal é uma fonte renovável, biodegradável e de excelente desempenho como reforço em compósitos variados. Híbridos selecionados pela qualidade de suas fibras estão em fase de validação agronômica na Embrapa Mandioca e Fruticultura e sua adoção para produção de fibra em larga escala depende de um elevado número de mudas. Este trabalho teve como objetivo avaliar a resposta morfogenética e o potencial propagativo de oito híbridos de Ananas comosus var. erectifolius, com a finalidade de produzir mudas de qualidade em larga escala. Gemas do caule e coroa foram reduzidas, introduzidas em meio nutritivo MS suplementado com BAP a 0,5 mg L-1, ANA a 0,01 mg L-1 e Phytagel® a 2,5 g L-1. Aos 45 dias foram avaliados o número de gemas oxidadas, contaminadas e sobreviventes. Gemas intumescidas e plantas formadas foram transferidas para o meio de multiplicação contendo sacarose, sais e vitaminas MS. Avaliou-se o potencial propagativo a partir de uma taxa de crescimento geométrico entre subcultivos. O híbrido FIB-NEG (40.28%) apresentou os melhores resultados em porcentagem para a fase de estabelecimento. O melhor potencial propagativo foi obtido a partir de gemas de coroa, com os valores mais elevados registrados para os híbridos FIB-EST (3.93), FIB-MIN (3.91) e FIB-BOY (3.91).

Genotypic and environmental effects on the level of ascorbic acid, phenolic compounds and related gene expression during pineapple fruit development and ripening.

Pineapple (Ananas comosus (L.) Merr.) is a non-climacteric tropical fruit whose ripening could be accompanied by oxidative processes and the concurrent activation of enzymatic and non-enzymatic reactive oxygen species (ROS) scavenging systems. To better understand the variability of these processes among climatic environments or genotypes in pineapple, the temporal expression dynamics for genes encoding oxidative and antioxidative stress enzymes were analyzed by real-time RT-PCR during fruit development and ripening, among three cultivars: Queen Victoria, Flhoran 41 and MD-2 hybrid, and in two climatic areas. Pineapple development and ripening involved changes in the levels of transcripts encoding for polyphenol oxidase and transcripts involved in the first steps of the phenylpropanoid pathway and in the balance of ROS, especially those encoding for ascorbate peroxydase and metallothioneins, regardless of the cultivar. Our results confirm the same dynamic in gene expression from the two environmental crop areas, however climatic conditions influenced the level of the expression of the major transcripts studied that were linked to these oxidative and antioxidant metabolisms. MT3a and MT3b transcripts were not influenced by genetic factor. The genetic effect was not significant on the various transcripts linked to the first steps of the phenylpropanoid pathway and to phenol oxidation, except 4CL ones. In ripe pineapple, highly significant relationships were found between the contents in antioxidant metabolites, i.e., ascorbic acid and total phenolic compounds, and the transcript levels of genes involved in the enzymatic ROS-scavenging system and in the biosynthesis or regeneration of ROS-scavenging compounds, like phenylpropanoids, ascorbic acid, metallothioneins.

Chemical profile of pineapple cv. Vitória in different maturation stages using electrospray ionization mass spectrometry.

BACKGROUND: Pineapple is the fruit of Ananas comosus var. comosus plant, being cultivated in tropical areas and has high energy content and nutritional value. Herein, 30 samples of pineapple cv. Vitória were analyzed as a function of the maturation stage (0-5) and their physico-chemical parameters monitored. In addition, negative-ion mode electrospray ionization mass spectrometry [ESI(-)FT-ICR MS] was used to identify and semi-quantify primary and secondary metabolites present in the crude and phenolic extracts of pineapple, respectively. RESULTS: Physico-chemical tests show an increase in the total soluble solids (TSS) values and in the TSS/total titratable acidity ratio as a function of the maturity stage, where a maximum value was observed in stage 3 (¾ of the fruit is yellow, which corresponds to the color of the fruit peel). ESI(-)FT-ICR MS analysis for crude extracts showed the presence mainly of sugars as primary metabolites present in deprotonated molecule form ([M - H]- and [2 M - H]- ions) whereas, for phenolic fractions, 11 compounds were detected, being the most abundant in the third stage of maturation. This behavior was confirmed by quantitative analysis of total polyphenols. CONCLUSION: ESI-FT-ICR MS was efficient in identifying primary (carbohydrates and organic acids) and secondary metabolites (13 phenolic compounds) presents in the crude and phenolic extract of the samples, respectively. © 2017 Society of Chemical Industry.

Integrated DNA methylome and transcriptome analysis reveals the ethylene-induced flowering pathway genes in pineapple.

Ethylene has long been used to promote flowering in pineapple production. Ethylene-induced flowering is dose dependent, with a critical threshold level of ethylene response factors needed to trigger flowering. The mechanism of ethylene-induced flowering is still unclear. Here, we integrated isoform sequencing (iso-seq), Illumina short-reads sequencing and whole-genome bisulfite sequencing (WGBS) to explore the early changes of transcriptomic and DNA methylation in pineapple following high-concentration ethylene (HE) and low-concentration ethylene (LE) treatment. Iso-seq produced 122,338 transcripts, including 26,893 alternative splicing isoforms, 8,090 novel transcripts and 12,536 candidate long non-coding RNAs. The WGBS results suggested a decrease in CG methylation and increase in CHH methylation following HE treatment. The LE and HE treatments induced drastic changes in transcriptome and DNA methylome, with LE inducing the initial response to flower induction and HE inducing the subsequent response. The dose-dependent induction of FLOWERING LOCUS T-like genes (FTLs) may have contributed to dose-dependent flowering induction in pineapple by ethylene. Alterations in DNA methylation, lncRNAs and multiple genes may be involved in the regulation of FTLs. Our data provided a landscape of the transcriptome and DNA methylome and revealed a candidate network that regulates flowering time in pineapple, which may promote further studies.

Mechanism of internal browning of pineapple: The role of gibberellins catabolism gene (AcGA2ox) and GAs.

Internal browning (IB), a physiological disorder (PD) that causes severe losses in harvested pineapple, can be induced by exogenous gibberellins (GAs). Over the years, studies have focused on roles of Gibberellin 2-oxidase (GA2oxs), the major GAs catabolic enzyme in plants, in the regulation of changes in morphology or biomass. However, whether GA2oxs could regulate PD has not been reported. Here, a full-length AcGA2ox cDNA was isolated from pineapple, with the putative protein sharing 23.59% to 72.92% identity with GA2oxs from five other plants. Pineapples stored at 5 °C stayed intact, while those stored at 20 °C showed severe IB. Storage at 5 °C enhanced AcGA2ox expression and decreased levels of a GAs (GA4) 'compared with storage at 20 °C. However, at 20 °C, exogenous application of abscisic acid (ABA) significantly suppressed IB. ABA simultaneously upregulated AcGA2ox and reduced GA4. Ectopic expression of AcGA2ox in Arabidopsis resulted in reduced GA4, lower seed germination, and shorter hypocotyls and roots, all of which were restored by exogenous GA4/7. Moreover, in pineapple, GA4/7 upregulated polyphenol oxidase, while storage at 5 °C and ABA downregulated it. These results strongly suggest the involvement of AcGA2ox in regulation of GAs levels and a role of AcGA2ox in regulating IB.

Postharvest internal browning of pineapple fruit originates at the phloem.

A typical symptom of postharvest chilling injury (PCI) in pineapple fruit (Ananas comosus (L.) Merr.) is internal browning (IB) near the fruit core. Since vascular bundles (VBs) are localized to this region, it was hypothesized that the VBs might be the site of IB. To test this, the anatomy and histochemistry of VBs during chilling stress in four pineapple cultivars with different levels of sensitivity to PCI were examined. Fruit were stored at 10°C for up to three weeks to stimulate translucency symptoms (TS; the initiation of IB). After three weeks of chilling exposure, the cultivars 'MD2' showed 0%, 'Pattavia' and 'Savee' showed 10-16%, and 'Trad Sri Thong' showed 100% TS and IB symptom. Scanning electron microscopy and in situ histochemical staining techniques that detect enzymes and substrates commonly associated with IB initiation were used in parallel. The TS of pineapple fruit coincided with the collapse of the phloem tissue. The VBs in the tissue where IB was initiated (i.e., the flesh adjacent to the core or F/C) had the highest activity of polyphenol oxidase, hydrogen peroxide, and phenolic compounds. The IB-resistant 'MD2' genotype had fewer VBs, but a greater proportion of sclerenchyma fibers (P<0.05) than did the susceptible 'Trad Sri Thong'. Based on these data, the first report of pineapple IB occurrence in the phloem was proposed.

Ripening-dependent metabolic changes in the volatiles of pineapple (Ananas comosus (L.) Merr.) fruit: II. Multivariate statistical profiling of pineapple aroma compounds based on comprehensive two-dimensional gas chromatography-mass spectrometry.

Ripening-dependent changes of pineapple volatiles were studied in a nontargeted profiling analysis. Volatiles were isolated via headspace solid phase microextraction and analyzed by comprehensive 2D gas chromatography and mass spectrometry (HS-SPME-GC×GC-qMS). Profile patterns presented in the contour plots were evaluated applying image processing techniques and subsequent multivariate statistical data analysis. Statistical methods comprised unsupervised hierarchical cluster analysis (HCA) and principal component analysis (PCA) to classify the samples. Supervised partial least squares discriminant analysis (PLS-DA) and partial least squares (PLS) regression were applied to discriminate different ripening stages and describe the development of volatiles during postharvest storage, respectively. Hereby, substantial chemical markers allowing for class separation were revealed. The workflow permitted the rapid distinction between premature green-ripe pineapples and postharvest-ripened sea-freighted fruits. Volatile profiles of fully ripe air-freighted pineapples were similar to those of green-ripe fruits postharvest ripened for 6 days after simulated sea freight export, after PCA with only two principal components. However, PCA considering also the third principal component allowed differentiation between air-freighted fruits and the four progressing postharvest maturity stages of sea-freighted pineapples.

Ripening-dependent metabolic changes in the volatiles of pineapple (Ananas comosus (L.) Merr.) fruit: I. Characterization of pineapple aroma compounds by comprehensive two-dimensional gas chromatography-mass spectrometry.

Qualitative ripening-dependent changes of pineapple volatiles were studied via headspace solid-phase microextraction and analyzed by comprehensive two-dimensional gas chromatography quadrupole mass spectrometry (HS-SPME-GC×GC-qMS). Early green-ripe stage, post-harvest ripened, and green-ripe fruits at the end of their commercial shelf-life were compared to air-freighted pineapples harvested at full maturity. In total, more than 290 volatiles could be identified by mass spectrometry and their linear retention indices. The majority of compounds comprise esters (methyl and ethyl esters of saturated and unsaturated fatty acids, acetates), terpenes, alcohols, aldehydes, 2-ketones, free fatty acids, and miscellaneous γ- and δ-lactones. The structured separation space obtained by GC×GC allowed revealing various homologous series of compound classes as well as clustering of sesquiterpenes. Post-harvest ripening increased the diversity of the volatile profile compared to both early green-ripe maturity stages and on-plant ripened fruits.

Authentication of pineapple (Ananas comosus [L.] Merr.) fruit maturity stages by quantitative analysis of γ- and δ-lactones using headspace solid-phase microextraction and chirospecific gas chromatography-selected ion monitoring mass spectrometry (HS-SPME-GC-SIM-MS).

Headspace solid phase microextraction and chirospecific gas chromatography-mass spectrometry in selected ion monitoring mode (HS-SPME-GC-SIM-MS) allowed quantitative determination of δ-lactones (δ-C8, δ-C10) and γ-lactones (γ-C6, γ-C8, γ-C10). A stable isotope dilution assay (SIDA) with d7-γ-decalactone as internal standard was used for quantitative analysis of pineapple lactones that was performed at three progressing post-harvest stages of fully ripe air-freighted and green-ripe sea-freighted fruits, covering the relevant shelf-life of the fruits. Fresh pineapples harvested at full maturity were characterised by γ-C6 of high enantiomeric purity remaining stable during the whole post-harvest period. In contrast, the enantiomeric purity of γ-C6 significantly decreased during post-harvest storage of sea-freighted pineapples. The biogenetical background and the potential of chirospecific analysis of lactones for authentication and quality evaluation of fresh pineapple fruits are discussed.

Influence of harvest maturity and fruit logistics on pineapple (Ananas comosus [L.] Merr.) volatiles assessed by headspace solid phase microextraction and gas chromatography-mass spectrometry (HS-SPME-GC/MS).

Profiling of volatiles from pineapple fruits was performed at four ripening stages using headspace solid-phase microextraction and gas chromatography-mass spectrometry (HS-SPME-GC/MS). In total, 142 volatiles were detected, of which 132 were identified. Multivariate data analysis was carried out to assess the effect of post-harvest storage on volatiles composition of green-ripe sea-freighted pineapple in comparison to air-freighted fruits harvested at full maturity. The latter fruits were characterised by volatiles described as potent odorants in pineapples, such as δ-octalactone, γ-lactones, 1-(E,Z)-3,5-undecatriene and 1,3,5,8-undecatetraene, as well as various methyl esters. In contrast, post-harvest storage of green-ripe sea-freighted fruits resulted in an increased formation of ethyl esters, acetates, acetoxy esters and alcohols, thus allowing the authentication of sea- and air-freighted pineapples, respectively. Particularly, compounds presumably derived from methyl-branched amino acid catabolism were identified in the fruits at later post-harvest stages. In addition, physicochemical traits were determined to characterise the fruit maturity stages.

Development and evaluation of a new method for sampling and monitoring the symphylid population in pineapple.

BACKGROUND: Symphylids (Hanseniella sp.) are polyphagous soilborne parasites. Today, symphylid populations on pineapple are monitored by observing root symptoms and the presence of symphylids at the bottom of basal leaves. The authors developed a reliable method with a bait and trap device to monitor symphylid populations in pineapple or fallow crops. The spatial distribution of the symphylid populations was evaluated using the variance/mean ratios and spatial analyses based on Moran's and Geary's indices. The method has been tested to monitor symphylid populations at different developmental stages of pineapple. RESULTS: Adding potato baits to the soil samples increased the trapping efficiency of symphylids when compared with 'soil only' and 'bait only' methods. The handling of the samples is also facilitated by the new device. Results showed that the vertical distribution of symphylids may be uniform deeply inside the soil profile under pineapple, up to 50 cm. Results showed that symphylid populations are highly aggregated, showing a spot area about 4-6 m wide for their development. CONCLUSION: The new method allows better and easier evaluation of symphylid populations. It may be very useful in the evaluation of new IPM methods to control symphylids under pineapple.

Structural interaction between GFP-labeled diazotrophic endophytic bacterium Herbaspirillum seropedicae RAM10 and pineapple plantlets 'Vitória'

The events involved in the structural interaction between the diazotrophic endophytic bacterium Herbaspirillum seropedicae, strain RAM10, labeled with green fluorescent protein, and pineapple plantlets 'Vitória' were evaluated by means of bright-field and fluorescence microscopy, combined with scanning electron microscopy for 28 days after inoculation. After 6 hours of inoculation, H. seropedicae was already adhered to the roots, colonizing mainly root hair surface and bases, followed by epidermal cell wall junctions. Bacteria adherence in the initial periods occurred mainly in the form of solitary cells and small aggregates with pleomorphic cells. Bacteria infection of root tissue occurred through the cavities caused by the disruption of epidermal cells during the emergence of lateral roots and the endophytic establishment by the colonization of intercellular spaces of the cortical parenchyma. Moreover, within 1 day after inoculation the bacteria were colonizing the shoots. In this region, the preferred sites of epiphytic colonization were epidermal cell wall junctions, peltate scutiform trichomes and non-glandular trichomes. Subsequently, the bacteria occupied the outer periclinal walls of epidermal cells and stomata. The penetration into the shoot occurred passively through stoma aperture followed by the endophytic establishment on the substomatal chambers and spread to the intercellular spaces of spongy chlorenchyma. After 21 days of inoculation, bacterial biofilm were seen at the root hair base and on epidermal cell wall surface of root and leaf, also confirming the epiphytic nature of H. seropedicae.

Growth of Byssochlamys nivea in pineapple juice under the effect of water activity and ascospore age

The study of thermal resistant mould, including Byssochlamys nivea, is of extreme importance since it has been associated with fruit and fruit products. The aim of this work is to analyze the influence of water activity (a w) and ascospore age (I) on the growth of Byssochlamys nivea in pineapple juice. Mold growth was carried out under different conditions of water activity (a w) (0.99, 0.96, 0.95, 0.93, 0.90) and ascospore age (I) (30, 51, 60, 69, 90 days). Growth parameters as length of adaptation phase (λ), maximum specific growth rate (µmax) and maximum diameter reached by the colony (λ) were obtained through the fit of the Modified Gompertz model to experimental data (measuring radial colony diameter). Statistica 6.0 was used for statistical analyses (significance level α = 0.05). The results obtained clearly showed that water activity is statistically significant and that it influences all growth parameters, while ascospore age does not have any statistically significant influence on growth parameters. Also, these data showed that by increasing a w from 0.90 to 0.99, the λ value substantially decreased, while µmax and λ values rose. The data contributed for the understanding of the behavior of B. nivea in pineapple juice. Therefore, it provided mathematical models that can well predict growth parameters, also helping on microbiological control and products' shelf life determination.

Características germinativas de sementes de Ananas ananassoides (Baker) L. B. Sm. (Bromeliaceae) / Germination characteristics of seeds of Ananas ananassoides (Baker) L. B. Sm. (Bromeliaceae)

Ananas ananassoides é uma planta perene com hábito de crescimento herbáceo e que tem enorme potencial paisagístico porque apresenta flores e brácteas com coloração exuberante e folhas com margens espinescentes que estão distribuídas em roseta. O objetivo deste experimento foi o de avaliar as características germinativas e o efeito de tratamentos pré-germinativos nas sementes da espécie. Primeiro, em um delineamento inteiramentecasualizado, foram investigados seis tratamentos em quatro repetições de 25 sementes. Os tratamentos constaram da imersão em água destilada à temperatura ambiente (25 ± 1,5°C) durante 24, 48 e 72h, imersão em água quente a 90°C durante 2 min. e imersão em ácido sulfúrico p.a. durante 2 min. e com posterior lavagem em água corrente por 24h que foram comparados com um controle. Depois de quatro meses de armazenamento, sementes intactas (controle) foram comparadas com sementes imersas em água a 90°C por 2 min. Assim, a maior porcentagem de germinação (92%) foi obtida com sementes recém-colhidas e imersas em água a 90oC durante 2 min., mas o armazenamento por quatro meses melhorou a porcentagem de germinação (G ≥ 96,0%). Os tempos médios de germinação não diferiram entre as sementes recém-colhidas (de 21,0 a 26,8 dias) e as armazenadas (entre 18,4 e 19,5 dias). A germinação das sementes recém-colhidas e armazenadas foi lenta e distribuída ao longo do tempo experimental.

Ananas ananassoides is a perennial plant with ornamental potential because of flowers and bracts with exuberant colors and herbaceous development where the leaves have spiny margins that are distributed in rosettes. The objective of this experiment with six completely randomized treatments and four replicates of 25 seeds was to evaluate the effects of seed soaking into distilled water at environmental temperature (25 ± 1.5°C) for 24, 48, and 72h; seed soaking into water at 90°C for 2 minutes; seed soaking into sulfuric acid p.a. for 2 minutes followed by tap water washing for 24h, and the control. Thereafter, 4-month-old seeds were immersed into water at 90°C for 2 minutes and compared to the intact seeds (control). Initially, the highest percentage of germination (G=92%) was evaluated in newly collected seeds that were immersed in water at 90°C for 2 minutes. No increases in the percentage of seed germination were detected by seed soakinginto water or sulfuric acid in comparison to the seeds that were immersed in water at 90°C for 2 minutes. However, the percentage of germination was higher than 96% when the seeds were stored for four months. No difference in the mean time to germination was detected between newly collected (between 21.0 and 26.8 days) and stored seeds (between 18.4 and 19.5 days). The germination of newly collected and stored seeds was slow and distributed throughout the experimental time