Retrotransposons and multilocus sequence analysis reveals diversity and genetic variability in endophytic fungi-associated with Serjania laruotteana Cambess.

The composition of endophytic communities is dynamic and demonstrates host specificity; besides, they have great intra- and interspecific genetic variability. In this work, we isolated leaf endophytic fungi from Serjania laruotteana, identify them using multilocus analysis, and evaluate the genetic variability using IRAP (inter-retrotransposon amplified polymorphism) and REMAP (retrotransposon-microssatellite amplified polymorphism). A total of 261 fungi were isolated and 58 were identified. Multilocus phylogenetic analysis using the partial sequences from the ITS1-5.8S-ITS2 regions, elongation factor 1-alpha, ß-tubulin, actin, glyceraldehyde-3-phosphate dehydrogenase, and calmodulin genes identify that most strains belonged to the Colletotrichum and Diaporthe genera, other isolated genera were Xylaria, Phyllosticta, Muyocopron, Fusarium, Nemania, Plectosphaerella, Corynespora, Bipolaris, and Curvularia. The IRAP and REMAP analyzes were performed with Colletotrichum and Diaporthe genera and showed 100% of polymorphism and high intra- and interspecific variability. This is the first report of the diversity of endophytic fungi from S. laruotteana. In addition, it demonstrated that the IRAP and REMAP can be used to distinguish morphologically similar lineages, revealing differences even strains of the same species.

The role of cell wall polysaccharides disassembly in Lasiodiplodia theobromae-induced disease occurrence and softening of fresh longan fruit.

Cell wall polysaccharides in fruits act a pivotal role in their resistance to fungal invasion. Lasiodiplodia theobromae (Pat.) Griff. & Maubl. is a primary pathogenic fungus causing the spoilage of fresh longan fruit. In this study, the influences of L. theobromae inoculation on the disassembly of cell wall polysaccharides in pericarp of fresh longans and its association with L. theobromae-induced disease and softening development were investigated. In contrast to the control, samples with L. theobromae infection showed more severe disease development, lower firmness, lower amounts of cell wall materials, covalent-soluble pectin, ionic-soluble pectin, cellulose and hemicellulose, whereas higher value of water-soluble pectin, higher activities of cell wall polysaccharide-disassembling enzymes (cellulase, ß-galactosidase, polygalacturonase and pectinesterase). These findings revealed that cell wall polysaccharides disassembly induced by enzymatic manipulation was an essential pathway for L. theobromae to infect harvested longans, and thus led to the disease occurrence and fruit softening.

Both artificial root exudates and natural Koelreuteria paniculata exudates modify bacterial community structure and enhance phenanthrene biodegradation in contaminated soils.

Polycyclic aromatic hydrocarbons (PAHs) are environmental pollutants that represent a serious threat to the health of humans and ecosystems. The effects of plant root and artificial root exudates (ARE) on the biodegradation of phenanthrene (PHE) and their impact on soil bacterial community structure was the focus of this work using four treatments for 180 days. Treatments included; control treatment (CK), low concentration of ARE (AREL), high concentration of ARE (AREH), and planting Koelreuteria paniculata saplings (KOE). The diversity and composition of soil bacterial community were analyzed using high-throughput sequencing. The results showed that KOE treatments had the most significant effect on the biodegradation of PHE compared to controls. ARE treatments had the similar effects on the biodegradation of PHE in soil with high efficiency in AREH than AREL. Both KOE and ARE treatments reduced diversity of bacterial community but increased the abundance of PAHs degrading bacterial populations within representative phyla, including Proteobacteria and Firmicutes. During the study, the total bacterial OTUs showed the number of unique genus types initially increased, then lowered in the later stages of the incubation process. Specific bacterial populations enriched by the treatments and supported by the exudates seemed to determine the biodegradation of PHE and not the overall bacterial diversity.

Extension of shelf life of semi-dry longan pulp with gaseous chlorine dioxide generating film.

A packaging system using gaseous chlorine dioxide generating film (CDGF) in a sealed container was developed to extend the shelf life of semi-dry longan pulp (moisture content 38.8 wt%; aw0.8). The antimicrobial properties, formation of chloroxyanion residues and effects of CDGF on the quality of semi-dry longan pulp were investigated. CDGF was triggered by the moisture vapor from semi-dry longan pulp in the sealed container and released gaseous ClO2 into the headspace of the container. The antifungal test showed that CDGF significantly inactivated artificially inoculated molds in semi-dry longan pulp and achieved reductions of over 3 log CFU/g after 28 days storage at room temperature (25 °C). CDGF reduced total aerobic bacterial populations by over 6.4 log CFU/g and maintained these population levels at around 2.0 log CFU/g throughout the 180-day storage period at room temperature. The residual concentrations of chloride, chlorate and perchlorate in longan pulp increased and then decreased during the 180-day storage. Residual chloride levels were maintained at 1.5 mg/g after Day 120 and residual chlorate and perchlorate levels were not detected after Day 120 and Day 180, respectively, in CDGF-treated samples. CDGF treatments reduced total polyphenol content but didn't have any significant impact on the levels of polysaccharides in samples. There were no significant differences between CDGF-treated and control samples in color changes during storage. The content of 5-hydroymethylfurfural (5-HMF) in both samples increased during storage, suggesting that the Maillard reaction occurred. This study demonstrated an effective approach to develop a new antimicrobial packaging system for semi-dry longan pulp.

Lasiodiplodia theobromae (Pat.) Griff. & Maubl. reduced energy status and ATPase activity and its relation to disease development and pericarp browning of harvested longan fruit.

This study aimed to investigate the effects of Lasiodiplodia theobromae (Pat.) Griff. & Maubl (L. theobromae) inoculation on the energy status and activity of adenosine triphosphatase (ATPase) during L. theobromae-induced disease development and pericarp browning of harvested 'Fuyan' longan (Dimocarpus longan Lour. cv. Fuyan) fruit. The results showed that, compared to the control longans, L. theobromae-inoculated longans displayed higher indices of fruit disease and pericarp browning, lower pericarp ATP and ADP contents, higher AMP content, lower level of energy charge, as well as lower activities of Ca2+-ATPase, Mg2+-ATPase and H+-ATPase in membranes of plasma, vacuole, and mitochondria. These results indicated that the infection of L. theobromae reduced energy status and ATPase activities, caused ions disorder, damaged the integrity and function of the cell and organelles including vacuole and mitochondria in pericarp of longan fruit, which contributed to L. theobromae-promoted disease development and pericarp browning of harvested longan fruit during storage.

The Changes in Metabolisms of Membrane Lipids and Phenolics Induced by Phomopsis longanae Chi Infection in Association with Pericarp Browning and Disease Occurrence of Postharvest Longan Fruit.

This study investigated the changes in metabolisms of membrane lipids and phenolics caused by Phomopsis longanae Chi infection in association with pericarp browning and fruit disease occurrence of postharvest longans. Compared with the uninoculated-longans, the longans inoculated by P. longanae exhibited higher cellular membrane permeability; higher PLD, lipase, and LOX activities; and higher levels of saturated fatty acids (SFAs) and phosphatidic acid but lower levels of phosphatidylinositol, phosphatidylcholine, and unsaturated fatty acids (USFAs). Additionally, the longans inoculated by P. longanae showed higher activities of POD and PPO but a lower amount of total phenolics. These findings suggested that infection of P. longanae enhanced activities of PLD-, lipase-, and LOX- stimulated degradations of membrane lipids and USFAs, which destroyed the integrity of the cell membrane structure, resulting in enzymatic browning by contact of phenolics with POD and PPO, and resulting in reduction of resistance to pathogen infection and accordingly accelerated disease occurrence of postharvest longan fruit.

Fermentation process optimization and chemical constituent analysis on longan (Dimocarpus longan Lour.) wine.

Based on single factor and orthogonal experiments, optimal fermentation conditions for longan wine were Saccharomyces cerevisiae strain of Lalvin KD, juice content of 70% and alcohol content of 10°. Sixteen amino acids were detected. Proline, alanine, glutamic acid and aspartic acid contents were relatively high. Sixty-three volatile aroma compounds were identified using solid-phase micro extraction and gas chromatography (SPME-GC). Ethyl lactate content was the highest, followed by octanoic acid ethyl ester, isoamyl alcohol and decanoic acid ethyl ester. Main functional components were polysaccharides. Longan wine polysaccharide (LWP) with molecular weight 10-30 kDa exhibited the highest hypoglycemic and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activities. 10-30 kDa polysaccharides mainly consisted of glucose, mannose, galactose, arabinose, galacturonic acid and glucuronic acid in molar ratio of 167.72:3.38:3.13:3.46:2.33:1. Infrared and nuclear magnetic resonance spectra confirmed that the sugar ring of 10-30 kDa polysaccharides was in the 〈beta〉-configuration.

Lasiodiplodia theobromae (Pat.) Griff. & Maubl.-induced disease development and pericarp browning of harvested longan fruit in association with membrane lipids metabolism.

Effects of Lasiodiplodia theobromae inoculation on disease development, pericarp browning and membrane lipids metabolism of harvested "Fuyan" longan fruit were studied. Compared with control fruit, L. theobromae-inoculated longans showed higher fruit disease index, pericarp browning index and cell membrane permeability, as well as higher activities of phospholipase D, lipase and lipoxygenase. Additionally, there were lower contents of membrane phospholipids but higher content of phosphatidic acid, and lower level of unsaturated fatty acids but higher level of saturated ones with lower ratio of unsaturated fatty acid to saturated fatty acid and lower index of unsaturated fatty acids in pericarp of L. theobromae-inoculated longans. These results suggested that L. theobromae-induced disease development and pericarp browning of harvested longans might be attributed to the damaged cellular membrane structural integrity, induced by the activated membrane lipids-degrading enzymes increasing the degradation of membrane phospholipids and unsaturated fatty acids in pericarp of harvested longan fruit.

The roles of ROS production-scavenging system in Lasiodiplodia theobromae (Pat.) Griff. & Maubl.-induced pericarp browning and disease development of harvested longan fruit.

Effects of Lasiodiplodia theobromae on reactive oxygen species (ROS) production-scavenging system during L. theobromae-induced pericarp browning and disease development of harvested "Fuyan" longans were investigated. Compared with control longans, L. theobromae-inoculated longans exhibited higher pericarp browning index and fruit disease index, higher pericarp O2- generation rate and MDA content. Moreover, L. theobromae infection also resulted in lower contents of pericarp AsA and GSH, lower levels of pericarp DPPH radical scavenging ability and reducing power. Additionally, L. theobromae infection decreased the activities of pericarp SOD, CAT and APX from day 2 to day 5. These findings suggested that L. theobromae-induced pericarp browning and disease development of harvested longans might be due to reduction of ROS scavenging ability and increase in ROS production, which might stimulate membrane lipid peroxidation, disrupt cellular membrane structure, and cause the loss of cellular compartmentalization and disease resistance, in turn, resulting in pericarp browning and disease development.

Effect of a saprophytic fungus on the growth and the lead uptake, translocation and immobilization in Dodonaea viscosa.

Phytoremediation is a feasible alternative to remediate soils polluted with toxic elements, which can be enhanced by manipulating plant-microbe interactions. Regarding this, free-living saprophytic fungi that interact beneficially with roots have been scarcely studied. Thus, the aim of this study was to assess the effect of a saprophytic fungus, Lewia sp., on the plant growth and the ability of Dodonaea viscosa to phytoaccumulate or phytostabilize soluble and insoluble sources of lead in a solid support. The growth of D. viscosa was influenced by both Pb and Lewia sp. While seedlings exposed to Pb showed a decrease in biomass production, in seedlings grown without Pb the biomass was stimulated by Lewia sp. The fungus strongly stimulated the weight-to-length ratio in roots. Regardless of the treatment, D. viscosa accumulated 4.4-6.5 times more Pb in roots than in shoots, conducting to low translocation factors (< 0.2). The presence of Lewia sp. significantly improved Pb accumulation, achieving high bioconcentration factors (> 22), which was attributed to an increased bioavailability and uptake of Pb due to the fungus. This study demonstrated that Lewia sp. could improve Pb-phytostabilization by D. viscosa in soils polluted with soluble and insoluble forms of Pb.

New Meliolaceae from the Brazilian Atlantic forest 1. Species on hosts in the families Asteraceae, Burseraceae, Euphorbiaceae, Fabaceae and Sapindaceae.

Five new species, two new varieties and three newly reported taxa belonging to the Meliolaceae were collected in fragments of Atlantic forest from Minas Gerais, Brazil, in association with native plants and are described and illustrated herein. The newly described species are Appendiculella eupatorii, Meliola cassiae-ferrugineae, M. mutisiae, M. peruiferae, M. vernaliae. The new varieties are M. garugae var. protii and M. paullinifolii var. rubiginosae. These taxa are reported in Brazil for the first time: Asteridiella cyclopoda, A. entebbeensis var. codiaei and Meliola pazschkeana var. macropoda. We studied other species belonging in Meliolaceae collected on hosts belonging to the Asteraceae, Burseraceae, Euphorbiaceae, Fabaceae (Caesalpinioideae and Papilionoideae) and Sapindaceae in Brazil.

Sawadaea koelreuteriae comb, nov., a powdery mildew of Koelreuteria paniculata.

A powdery mildew parasitizing Koelreuteria spp. was first described under the name Uncinula koelreuteriae Miyake and later transferred to the genus Typhulochaeta. Based on morphological and molecular data of several herbarium specimens collected from Korea, the generic placement of Typhulochaeta is discussed and T. koelreuteriae is combined in the genus Sawadaea. Redescription and epitypification of this species is provided hereby.

Role of mycorrhizal colonization in plant establishment on an alkaline gold mine tailing.

The potential role of arbuscular mycorrhizal fungi (AMF) in the revegetation of an alkaline gold mine tailing was studied in Barberton, South Africa. The tailing, characterized by a slow spontaneous plant succession, is colonized by the shrub Dodonaea viscosa and the grasses, Andropogon eucomus and Imperata cylindrica, all colonized by AMF. The effectiveness of mycorrhizal colonization in grasses was tested under laboratory conditions using fungal isolates of various origins. Both grasses were highly mycorrhiza dependent, and the presence of mycorrhizal colonization significantly increased their biomass and survival rates. The fungi originating from the gold tailing were better adapted to the special conditions of the tailing than the control isolate. Although the total colonization rate found for native fungi was lower than for fungi from non-polluted sites, they were more vital and more effective in promoting plant growth. The results obtained might serve as a practical approach to the phytostabilization of alkaline gold tailings.

Effect of Lactobacillus plantarum and chitosan in the reduction of browning of pericarp Rambutan (Nephelium lappaceum).

The effects of Lactobacillus plantarum alone or in combination with chitosan were evaluated on quality and color retention in rambutan fruits (Nephelium lappaceum) stored at 25 degrees C and 10 degrees C with 75+/-2.5% of relative humidity for 10 and 15 days, respectively. The development of the microorganisms was evidenced by viability analyses and lactic acid production. The application of L. plantarum significantly improved color retention (a* and L*), and reduced weight losses. The lactobacilli, alone or in combination with chitosan, preserved fruit quality characteristics such as firmness, total soluble solids and titratable acidity. The lactobacilli application on rambutan pericarp produced acidification of pericarp and avoided the browning; thereby desiccation was prevented due to biofilm formation.

Taxol determination from Pestalotiopsis pauciseta, a fungal endophyte of a medicinal plant.

Taxol is the most effective antitumor agent developed in the past three decades. It has been used for effective treatment of a variety of cancers. A taxol-producing endophytic fungus Pestalotiopsis pauciseta (strain CHP-11) was isolated from the leaves of Cardiospermum helicacabum and screened for taxol production. The fungus was identified based on the morphology of the fungal culture and the characteristics of the spores and screened for taxol production. The amount of taxol produced by this endophytic fungus was quantified by HPLC and it produced 113.3 microg/L, thus the fungus can serve as a potential material for fungus engineering to improve taxol production. This fungal taxol also had strong anticancer activity against some cancer cells viz., BT 220, H116, Int 407, HL 251 and HLK 210 tested by Apoptotic assay and it is indicated that with the increase of taxol concentration from 0.005-0.05 micromol/L, taxol induced increased cell death through apoptosis.

Growth and survival of seedlings of native plants in an impoverished and highly disturbed soil following inoculation with arbuscular mycorrhizal fungi.

We investigated whether arbuscular mycorrhizas influenced growth and survival of seedlings in an extremely impoverished and highly disturbed soil. Seedlings of four plants species native to the site were either inoculated with native sporocarpic arbuscular mycorrhizal (AM) fungi or fertilised prior to transplanting, and followed over 86 weeks at the site. One treatment was also irrigated with N-rich leachate from the site. In a laboratory experiment, seedlings were fertilised with excess P for 6 weeks, and location of the P store determined. Growth and survival of AM and fertilised seedlings were similar at the site. Inoculated mycorrhizal fungi and roots appeared to extend into the surrounding soil together. P concentration in leaves of all plants was extremely low. Irrigation with leachate increased growth of seedlings. In the laboratory experiment, significantly more P was stored in roots than shoots. We suggest that successful revegetation of extremely disturbed and impoverished sites requires selection of mycorrhizal fungi and plants to suit the edaphic conditions and methods of out-planting.