Methanotrophs and methylobacteria are found in woody plant tissues within the winter period

Samples of tree seeds, buds, and needles collected within the winter period at ambient temperatures from –11 to –17°C were analyzed for the presence of methylotrophic microflora. Thin sections of blue spruce needles were found to contain bacteria morphologically close to pink-pigmented methylobacteria. The methylobacteria that were isolated in pure cultures from samples of linden seeds and buds and pine and blue spruce needles, as well as of lilac, maple, and apple buds, were classified into the genera Methylobacterium and Paracoccus based on the data of morphological studies, enzyme assay, and DNA-DNA hybridization analysis. The methanotrophs that were isolated in pure cultures from samples of linden buds and blue spruce needles were referred to the genus Methylocystis based on the data of morphological studies, enzyme assay, DNA-DNA hybridization, and the phylogenetic analysis of the particulate methane monooxygenase gene pmoA sequences. The inference is made that aerobic methylotrophic bacteria are permanently associated with plants. At the beginning of the vegetative period in spring, the phyllosphere of coniferous and deciduous trees is colonized by methylotrophic bacteria that have wintered inside plant tissues.Translated from Mikrobiologiya, Vol. 73, No. 6, 2004, pp. 817–824.Original Russian Text Copyright © 2004 by Doronina, Ivanova, Suzina, Trotsenko.     

植物的不同组织对氰化物降解的可行性研究

氰化物是目前世界范围内最常使用的提取黄金等贵重金属的沥滤剂,其对自然生态环境的污染和破坏以及对人畜的毒性作用是众所周知的,氰化物污染的治理也就成为了环境工作者关注的焦点课题。植物细胞的悬浮液通常用于研究污染物的降解及在植物体内的生化反应机制。本实验直接用三种杨柳科植物的不同组织(根须、嫩茎、老茎和叶片)来观察和测定植物细胞线粒体中的氰丙氨酸合成酶(β-cyanoalanine synthase)转化氰化物的可行性。实验是在一封闭的玻璃器皿(100mL)中进行的(100mL的氰化钾溶液中加入1.0g(鲜重)的植物组织,氰化钾溶液的浓度0.444～0.457 CN mg·L~(-1))。在为期28 h的时间内,水溶液中的氰化物22.13～67.04％被植物的不同组织去除。最快的氰化物去除率发现在美洲黑杨(Populus deltoides)的叶片实验组(1.9449 mg CN/kg(鲜重)·h),苏柳(Salix matssudana Koidz×Salix alba L)的叶片实验组次之(1.7259 mg CN/kg(鲜重)·h),最慢的氰化物去除率发现在苏柳的嫩茎实验组(0.4934 mg CN/kg(鲜重)·h)。实验结果表明,选用植物的组织同样可以观察和测定污染物在植物体内的转化,特别是植物的叶片表现的尤其敏感。     

Activities of de-N-glycosylation are ubiquitously found in tomato plant

Activities of two de-N-glycosylation enzymes, PNGase (peptide N ⁴(N-acetyl-glucosaminyl)asparagine amidase) and ENGase (endo N-acetyl-β-D-glucosaminidase), involved in the release of N-glycans from N-glycoproteins, were monitored in several organs of tomato plants (Lycopersicon esculentum, Mill., cv. Dombito) with a fluorescence-HPLC procedure using a resofurin-labelled N-glycopeptide substrate. PNGase and ENGase activities were detected in every organ assayed but with quantitative differences. The highest activities were found in the youngest parts of the plant, i.e. apical buds, flowers and leaf blades. PNGase activities were consistently higher than ENGase activities (three-fold in average). Both de-N-glycosylation activities were associated with high levels of proteins and protease activities. During fruit growth and ripening, these three parameters decreased notably. The ubiquitous detection of these enzyme activities in the different organs is probably associated with the previously characterized unconjugated N-glycans in tomato. The possible role of PNGase and ENGase degradation products (i.e. unconjugated N-glycans) are discussed in relation with their biological functions in plant development.     

A bioassay for insect deterrent compounds found in plant and animal tissues

A general field bioassay for detecting biologically active compounds in plants and insects has been developed and tested for efficacy and sensitivity. Methanolic extracts, in sucrose solution, of 20 plant and six caterpillar species were offered to the ponerine ant Paraponera clavata and the feeding preferences observed. The bioassay resulted in the detection of nine plant and three caterpillar species with ant-deterrent extracts, and 11 plant and three caterpillar species with neutral or attractant extracts. All of the plants showing ant-deterrent characteristics which had been chemically investigated in our laboratory, or for which chemical literature was available, contained secondary metabolites of known deterrence. Both naturally occurring and artificial differences in chemical concentrations could be detected using the bioassay. The method provides a means of screening plants and insects for compounds that are insect anti-feedants or that can modify insect behaviour.     

Variation in woody plant mortality and dieback from severe drought among soils, plant groups, and species within a northern Arizona ecotone

Vegetation change from drought-induced mortality can alter ecosystem community structure, biodiversity, and services. Although drought-induced mortality of woody plants has increased globally with recent warming, influences of soil type, tree and shrub groups, and species are poorly understood. Following the severe 2002 drought in northern Arizona, we surveyed woody plant mortality and canopy dieback of live trees and shrubs at the forest–woodland ecotone on soils derived from three soil parent materials (cinder, flow basalt, sedimentary) that differed in texture and rockiness. Our first of three major findings was that soil parent material had little effect on mortality of both trees and shrubs, yet canopy dieback of trees was influenced by parent material; dieback was highest on the cinder for pinyon pine (Pinus edulis) and one-seed juniper (Juniperus monosperma). Ponderosa pine (Pinus ponderosa) dieback was not sensitive to parent material. Second, shrubs had similar mortality, but greater canopy dieback, than trees. Third, pinyon and ponderosa pines had greater mortality than juniper, yet juniper had greater dieback, reflecting different hydraulic characteristics among these tree species. Our results show that impacts of severe drought on woody plants differed among tree species and tree and shrub groups, and such impacts were widespread over different soils in the southwestern U.S. Increasing frequency of severe drought with climate warming will likely cause similar mortality to trees and shrubs over major soil types at the forest–woodland ecotone in this region, but due to greater mortality of other tree species, tree cover will shift from a mixture of species to dominance by junipers and shrubs. Surviving junipers and shrubs will also likely have diminished leaf area due to canopy dieback.     

Ethylene and extraneous material formation in woody tissues

Chloroethyl phosphonic acid (CEPA) was administered to the cambial region of Eucalyptus astringens, E. calophylla, E. sieberi and E. sideroxylon. The kino which exuded has been associated with the liberation of ethylene from the CEPA in the sap stream. The kinos from E. astringens and E. calophylla were chemically identical with those obtained naturally from these species. The composition of the polyphenols formed in the sapwood of Rhus after administration of CEPA had a similar chromatographic pattern to that obtained from the heartwood. In both these situations, the induced polyphenols differ from those normally present in the tissues in that the former contain acetate engendered moieties. CEPA produced copious amounts of carbohydrate gum from an apricot branch, and eucalypt sapwood blocks ventilated with ethylene contained a greater proportion of tyloses than the controls. The evidence supports the view that ethylene is an active intermediate in the formation of extraneous components. Under natural conditions, ethylene could arise as a result of injury or physiological stress such as water shortage.     

百山祖常绿阔叶林灌草层物种组成和 分布的10年动态

森林群落的动态受多方面因素的影响。林下的草本植物和木本植物幼苗幼树对于森林的更新及动态具有重要作用。为了解百山祖常绿阔叶林群落下层植物物种组成和分布的动态变化, 我们分析了2003-2013年间百山祖5 ha森林动态样地灌草层物种(包括DBH < 1 cm的木本植物和所有草本植物)在物种重要值、α多样性以及物种分布等方面的变化。结果表明: (1) 10年间, 群落灌草层植物个体数减少了28.7%, 其中草质藤本植物个体数下降比例最大(76.9%), 而乔木的幼苗和幼树下降比例最小(14.4%); (2)整个灌草层的物种数明显下降, 同时乔木的幼苗和幼树及直立灌木的均匀度均下降, 而草本植物均匀度有所上升; (3)除短尾柯(Lithocarpus brevicaudatus)、叶萼山矾(Symplocos phyllocalyx)、显脉野木瓜(Stauntonia conspicua)和福建悬钩子(Rubus fujianensis)等植物重要值逆势上扬外, 直立灌木物种重要值10年间的变化值与其在2003年时的重要值呈显著正相关, 而其余各类生活型植物都显示, 初始重要值越高的物种在10年后下降幅度越大; (4)物种分布与生境类型的相关性变化复杂, 除与山谷生境正相关的物种数量增加外, 研究期间与生境存在显著相关性的物种数都有所减少; (5) 10年来仅陡坡处物种多样性指数没有显著下降。总体来说, 百山祖常绿阔叶林灌草层群落仍未达到顶极状态, 2003-2013年间正缓慢向顶极群落演替。群落内部的种间和种内竞争应是当前群落动态的主导因素。     

Resprouting as a life history strategy in woody plant communities

Resprouting is an efficient means by which woody plants regain biomass lost during disturbance, but there is a life history trade-off that occurs in all disturbance regimes between investment in the current generation through resprouting vs investment in future generations at the same or more distant sites. The relative allocation to resprouting vs seeding in woody plant communities is dictated by the nature of disturbance regimes. Resprouting is the predominant response to the least severe disturbance regimes, but is also a common response in disturbance regimes of high severity, those that destroy most or all above-ground biomass, and which occur at medium to high frequency. The response to disturbance either by resprouting or seeding is dictated by the site's productivity. We present a comprehensive model for relative allocation to resprouting vs seeding across a range of disturbance regimes. Competition between plants that mostly seed vs those that mostly resprout should accentuate differences in allocation along a gradient of disturbance frequency. However the patchy nature of disturbance in time and space, coupled with gene flow among populations undergoing different disturbance regimes, ensures that it is unlikely that either resprouting or seeding will be the sole response in most plant communities at most disturbance frequencies. Additional influences on resprouting in woody plant communities include changes in allocation during the lifespan of individual plants and phylogenetic constraints that are expressed as biogeographic patterns.     

Methanotrophs and Methanogens in Masonry

Methanotrophs were present in 48 of 225 stone samples which were removed from 19 historical buildings in Germany and Italy. The average cell number of methanotrophs was 20 CFU per g of stone, and their activities ranged between 11 and 42 pmol of CH₄ g of stone⁻¹ day⁻¹. Twelve strains of methane-oxidizing bacteria were isolated. They belonged to the type II methanotrophs of the genera Methylocystis, Methylosinus, and Methylobacterium. In masonry, growth substrates like methane or methanol are available in very low concentrations. To determine if methane could be produced by the stone at rates sufficient to support growth of methanotrophs, methane production by stone samples under nonoxic conditions was examined. Methane production of 0.07 to 215 nmol of CH₄ g of stone⁻¹ day⁻¹ was detected in 23 of 47 stone samples examined. This indicated the presence of the so-called “mini-methane”-producing bacteria and/or methanogenic archaea. Methanotrophs occurred in nearly all samples which showed methane production. This finding indicated that methanotrophs depend on biogenic methane production in or on stone surfaces of historical buildings.     

Effects of associative pseudomonads and methylobacteria on plant growth and resistance to phytopathogens and xenobiotics

The in vivo and in vitro interactions between tobacco (Nicotiana tabacum L.), tomato (Lycopersicon esculentum Mill.), cabbage (Brassica oleracea var. capitata L.), rape (B. napus L.), and the common ice plant (Mesembryanthemum crystallinum L.) and bacteria Pseudomonas aureofaciens, P. putida, and Methylovorus mays were studied. Stable associations of these microorganisms with plants are demonstrated. Colonized plants were characterized by accelerated growth, more efficient rooting, better adaptation to in vivo conditions, and enhanced resistance to bacterial and fungal phytopathogens (Erwinia carotovora, Sclerotinia sclerotiorum and Phytophthora infestans). Plants colonized by bacteria resistant to kanamycin and naphthalene can grow steadily on the medium containing these compounds. The results obtained indicate a promising usage of beneficial associative microorganisms for the development of technologies for plant protection against biotic and abiotic stressors.     

Measurement of light within thin plant tissues with fiber optic microprobes

Vogelmann, T. C., Knapp, A. K., McClean, T. M. and Smith, W. K. 1988. Measurement of light within thin plant tissues with fiber optic microprobes. - Physiol. Plant. 72: 623–630.
The measurement of light with fiber optic microprobes has been extended to thin (200–300 μm) plant tissue samples. To test the method, light measurements were made in thin aqueous films and paradermal sections from 10-day-old etiolated Cucurbita pepo L. cv. Fordhook cotyledons. The measurements obtained were highly reproducible. Paradermal sections of spongy mesophyll that were irradiated with collimated light scattered light more effectively than the palisade layer of intact cotyledons. These results demonstrate that different plant tissues have different light scattering characteristics. The successful extension of the fiber optic microprobe technique to thin systems makes it possible to examine the optical properties of different cell layers within leaves and other plant organs.     

Ecosystem responses to woody plant encroachment in a semiarid savanna rangeland

Woody plant encroachment alters the structure and function of rangeland ecosystems. The objective of this study was to explore the association between woody plant encroachment and various ecosystem properties (i.e. vascular plant species diversity, richness, evenness, soil organic matter, herbaceous biomass, leaf litter and bare ground cover) in a semiarid savanna rangeland, and also to test whether the relationships were influenced by woody species composition, elevation and site. We carried out a vegetation survey in four rangeland sites in the lower Omo region of southwestern Ethiopia, and regressed each one of the ecosystem properties, separately, against woody plant density, elevation and site using multiple linear regressions. We found that vascular plant species diversity, richness and evenness increased with woody plant density, most likely due to increased spatial heterogeneity and soil microclimate improvement. Bare ground cover increased significantly, whereas herbaceous biomass and soil organic matter did not respond to woody encroachment. In a subsequent investigation, we used a redundancy analysis to assess whether ecosystem properties were influenced by the identity of encroaching woody plant species. Species diversity and richness responded positively to Lannea triphylla, whereas leaf litter responded positively to Grewia tenax and G. villosa. Our findings suggest that woody plant encroachment in a semiarid rangeland does alter ecosystem properties. However, its impact is highly variable, influenced by a set of factors including the level of encroachment and identity of encroaching woody species.     

Soil C and N responses to woody plant expansion in a mesic grassland

Changes in land management and reductions in fire frequency have contributed to increased cover of woody species in grasslands worldwide. These shifts in plant community composition have the potential to alter ecosystem function, particularly through changes in soil processes and properties. In semi-arid grasslands, the invasion of shrubs and trees is often accompanied by increases in soil resources and more rapid N and C cycling. We assessed the effects of shrub encroachment in a mesic grassland in Kansas (USA) on soil CO₂ flux, extractable inorganic N, and N mineralization beneath shrub communities (Cornus drummondii) and surrounding undisturbed grassland sites. In this study, a shift in plant community composition from grassland to shrubland resulted in a 16% decrease in annual soil CO₂ flux(4.78 kg CO₂ m^–2 year^–1 for shrub dominated sites versus 5.84 kg CO₂ m^–2 year^–1 for grassland sites) with no differences in total soil C or N or inorganic N. There was considerable variability in N mineralization rates within sites, which resulted in no overall difference in cumulative N mineralized during this study (4.09 g N m^–2 for grassland sites and 3.03 g N m^–2 for shrub islands). These results indicate that shrub encroachment into mesic grasslands does not significantly alter N availability (at least initially), but does alter C cycling by decreasing soil CO₂ flux.     

Monoterpene engineering in a woody plant Eucalyptus camaldulensis using a limonene synthase cDNA

Metabolic engineering aimed at monoterpene production has become an intensive research topic in recent years, although most studies have been limited to herbal plants including model plants such as Arabidopsis. The genus Eucalyptus includes commercially important woody plants in terms of essential oil production and the pulp industry. This study attempted to modify the production of monoterpenes, which are major components of Eucalyptus essential oil, by introducing two expression constructs containing Perilla frutescens limonene synthase ( PFLS ) cDNA, whose gene products were designed to be localized in either the plastid or cytosol, into Eucalyptus camaldulensis . The expression of the plastid-type and cytosol-type PFLS cDNA in transgenic E. camaldulensis was confirmed by real-time polymerase chain reaction (PCR). Gas chromatography with a flame ionization detector analyses of leaf extracts revealed that the plastidic and cytosolic expression of PFLS yielded 2.6- and 4.5-times more limonene than that accumulated in wild-type E. camaldulensis , respectively, while the ectopic expression of PFLS had only a small effect on the emission of limonene from the leaves of E. camaldulensis. Surprisingly, the high level of PFLS in Eucalyptus was accompanied by a synergistic increase in the production of 1,8-cineole and α-pinene, two major components of Eucalyptus monoterpenes. This genetic engineering of monoterpenes demonstrated a new potential for molecular breeding in woody plants.     

Thidiazuron: a potent cytokinin for woody plant tissue culture

Thidizuron (TDZ) is among the most active cytokinin-like substances for woody plant tissue culture. It facilitates efficient micropropagation of many recalcitrant woody species. Low concentrations (<1 µM) can induce greater axillary proliferation than many other cytokinins; however, TDZ may inhibit shoot elongation. In some cases it is necessary to transfer shoots to an elongation medium containing a lower level of TDZ and/or a less active cytokinin. At concentrations higher than 1 µM, TDZ can stimulate the formation of callus, adventitious shoots or somatic embryos. Subsequent rooting of microshoots may be unaffected or slightly inhibited by prior exposure to TDZ. The main undesirable side effect of TDZ is that cultures of some species occasionally form fasciated shoots. The high cytokinin activity and positive response of woody species to TDZ have established it as among the most active cytokinins forin vitro manipulation of many woody species.