Site directed chromosomal marking of a fluorescent pseudomonad isolated from the phytosphere of sugar beet; stability and potential for marker gene transfer.

A plasmid-free, non-pathogenic, ribosomal RNA group 1 fluorescent pseudomonad, Pseudomonas fluorescens SBW25, was selected from the microflora of sugar beet (Beta vulgaris) and modified to contain constitutively expressed marker genes. By site directed homologous recombination a KX cassette [kanamycin resistance (kanτ) and catechol 2,3 dioxygenase (xylE)] and a ZY cassette [lactose utilization (lacZY, β-galactosidase, lactose permease)] were introduced at least 1 Mbp apart on the 6.6 Mbp bacterial chromosome. Separate sites were selected to provide sensitive detection methods and allow assessments of marker gene stability of the genetically modified micro-organism (GMM), SBW25EeZY6KX, when it colonized the leaves and roots of sugar beet plants following seed inoculation.     

不同生境中桂花和夹竹桃叶际细菌的群落结构

植物叶际微生物多样性是目前植物-微生物关系研究的热点之一,但影响叶际微生物群落结构的主要因素目前还存在很大争议.本研究以生长在3处生境的桂花和夹竹桃为对象,基于高通量测序技术,分析2种植物叶际细菌的群落结构,探讨影响植物叶际细菌群落结构的主要因素.结果表明:来自3处生境的2种植物叶际细菌多样性无显著差异,构成叶际细菌群落的优势门主要包括放线菌门、拟杆菌门、衣原体门、蓝细菌门、厚壁菌门和变形菌门,优势属主要包括甲基杆菌属、鞘氨醇单胞菌属、薄层杆属、Polaromonas和无毛螺旋体属.植物种类、生境及二者的交互作用均能显著影响叶际细菌群落结构,其中生境的影响最大.     

Hormonema schizolunatum, a new species of dothideaceous black yeasts from phyllosphere

Two black yeast isolates from plants from the Canary Islands (Spain) are described and illustrated. Absence of Woronin bodies at simple septal pores, local coralloid terminal hyphal cells, indeterminate thallus maturation, the presence of budding cells and local conversion to meristematic growth all indicate a relationship to the Dothideaceae (Dothideales, Ascomycota). Morphological properties were consistent with the genus Hormonema Lagerberg & Melin, as defined by presence of percurrent conidiogenous loci alongside undifferentiated hyphae, and results of PCR-ribotyping supported this classification. The isolates were judged to belong to a hitherto undescribed species, characterized in particular by curved conidia soon developing transverse septa. The physiological profile of this species is also described.     

Rhizome phyllosphere oxygenation in Phragmites and other species in relation to redox potential, convective gas flow, submergence and aeration pathways

Underground rhizomes of emergent aquatic macrophytes are important for perennation, vegetative spread, competition and anchorage. In four species we examined the potential for the development of oxidized phyllospheres around rhizome apical buds, similar to the protective oxygenated rhizospheres around roots. Redox potentials and polarographic measurements of radial oxygen loss were recorded using platinum cathodes around the apical buds. The aeration pathway from atmosphere to phyllosphere was investigated anatomically and by applied pressurized gas flow. Redox potentials increased by +400, +45, +200 and +340 mV around rhizome apices of Phragmites australis, Oryza rhizomatis, Carex rostrata and Glyceria maxima, respectively. Radial oxygen loss from rhizome apices of Phragmites was increased by convective gas flow through the rhizome and by shoot de-submergence, and decreased by resistances applied within the aeration pathway and by shoot submergence. We conclude that oxygen passes via internal gas-space connections between aerial shoot, rhizome and underground buds and into the phyllosphere regions via scale-leaf stomata and surfaces on the buds. We suggest that oxidized phyllospheres may protect rhizome apices against phytotoxins in waterlogged soils, just as oxidized rhizospheres protect roots.     

Phyllosphere Bacteria Improve Animal Contribution to Plant Nutrition

Many plant species have evolved special adaptations for acquiring nitrogen in nutrient‐poor soils. In Brazilian savannas, the bromeliad Bromelia balansae (Bromeliaceae) is inhabited by mutualistic spiders (Psecas chapoda, Salticidae), which provide nutrients to the plant through their debris (feces, prey carcasses). In this study, we tested if bacteria present on the B. balansae phyllosphere improves plant nutrition and growth by mineralizing complex organic N compounds from spider debris that accumulate on the phyllosphere into simple compounds that may be absorbed easily by leaves. We conducted a greenhouse experiment by manipulating bacteria abundance on the bromeliad phyllosphere using antibiotics. Using isotopic mixed model equations, we demonstrated that debris from spiders contributed 10.7 ± 1.9 percent (mean ± standard error) of the N in bromeliads that had their bacterial abundance reduced. In contrast, spider feces contributed 27.1 ± 4.4 percent of bromeliad N in the presence of the entire bacterial assemblage. These bromeliads accumulated 57 percent more soluble protein and grew 13 percent more than bromeliads that were grown under reduced bacterial density. These results highlight the importance of mineralizing bacteria on phyllosphere as a mechanism of N uptake by bromeliads.     

Seasonal dynamic of the numbers of epiphytic yeasts

The numbers of epiphytic yeasts on the leaves and flowers of 25 plant species throughout their vegetation period was determined. The numbers of yeasts on the leaves were found to change regularly throughout the year. The average dynamics for all of the plant species investigated included an increase in yeast numbers during spring and summer with the maximum in late autumn and early winter. The character of the yeasts’ dynamics depends on the ecological characteristics of the plants and the duration of the ontogenesis of their leaves and flowers. Three types of dynamics of epiphytic yeasts were revealed: year-round with an increase in autumn-winter, year-round without visible changes, and seasonal with a terminal increase for annual plants.     

Culturable and metagenomic approaches of wheat bran and wheat straw phyllosphere’s highlight new lignocellulolytic microorganisms

The phyllosphere, defined as the aerial parts of plants, is one of the most prevalent microbial habitats on earth. The microorganisms present on the phyllosphere can have several interactions with the plant. The phyllosphere represents then a unique niche where microorganisms have evolved through time in that stressful environment and may have acquired the ability to degrade lignocellulosic plant cell walls in order to survive to oligotrophic conditions. The dynamic lignocellulolytic potential of two phyllospheric microbial consortia (wheat straw and wheat bran) has been studied. The microbial diversity rapidly changed between the native phyllospheres and the final degrading microbial consortia after 48 h of culture. Indeed, the initial microbial consortia was dominated by the Ralstonia (35·8%) and Micrococcus (75·2%) genera for the wheat bran and wheat straw whereas they were dominated by Candidatus phytoplasma (59%) and Acinetobacter (31·8%) in the final degrading microbial consortia respectively. Culturable experiments leading to the isolation of several new lignocellulolytic isolates (belonging to Moraxella and Atlantibacter genera) and metagenomic reconstruction of the microbial consortia highlighted the existence of an unpredicted microbial diversity involved in lignocellulose fractionation but also the existence of new pathways in known genera (presence of CE2 for Acinetobacter, several AAs for Pseudomonas and several GHs for Bacillus in different metagenomes-assembled genomes). The phyllosphere from agricultural co-products represents then a new niche as a lignocellulolytic degrading ecosystem.