Reptilian transferrins: evolution of disulphide bridges and conservation of iron-binding center

Transferrins, found in invertebrates and vertebrates, form a physiologically important family of proteins playing a major role in iron acquisition and transport, defense against microbial pathogens, growth and differentiation. These proteins are bilobal in structure and each lobe is composed of two domains divided by a cleft harboring an iron atom. Vertebrate transferrins comprise of serotransferrins, lactoferrins and ovotransferrins. In mammals serotransferrins transport iron in physiological fluids and deliver it to cells, while lactoferrins scavenge iron, limiting its availability to invading microbes. In oviparous vertebrates there is only one transferrin gene, expressed either in the liver to be delivered to physiological fluids as serotransferrin, or in the oviduct with a final localization in egg white as ovotransferrin. Being products of one gene sero- and ovotransferrin are identical at the amino-acid sequence level but with different, cell specific glycosylation patterns. Our knowledge of the mechanisms of transferrin iron binding and release is based on sequence and structural data obtained for human serotransferrin and hen and duck ovotransferrins. No sequence information about other ovotransferrins was available until our recent publication of turkey, ostrich, and red-eared turtle (TtrF) ovotransferrin mRNA sequences [Ciuraszkiewicz, J., Olczak, M., Watorek, W., 2006. Isolation, cloning and sequencing of transferrins from red-eared turtle, African ostrich and turkey. Comp. Biochem. Physiol. 143 B, 301-310]. In the present paper, ten new reptilian mRNA transferrin sequences obtained from the Nile crocodile (NtrF), bearded dragon (BtrF), Cuban brown anole (AtrF), veiled and Mediterranean chameleons (VtrF and KtrF), sand lizard (StrF), leopard gecko (LtrF), Burmese python (PtrF), African house snake (HtrF), and grass snake (GtrF) are presented and analyzed. Nile crocodile and red-eared turtle transferrins have a disulphide bridge pattern identical to known bird homologues. A partially different disulphide bridge pattern was found in the Squamata (snakes and lizards). The possibility of a unique interdomain disulphide bridge was predicted for LtrF. Differences were found in iron-binding centers from those of previously known transferrins. Substitutions were found in the iron-chelating residues of StrF and TtrF and in the synergistic anion-binding residues of NtrF. In snakes, the transferrin (PtrF, HtrF and GtrF) N-lobe "dilysine trigger" occurring in all other known transferrins was not found, which indicates a different mechanism of iron release.     

Generation of transgenic Lolium temulentum plants by Agrobacterium tumefaciens-mediated transformation

Lolium temulentum L. (Darnel ryegrass) has been proposed to be used as a model species for functional genomics studies in forage and turf grasses, because it is a self-fertile, diploid species with a short life cycle and is closely related to other grasses. Embryogenic calluses were induced from mature embryos of a double haploid line developed through anther culture. The calluses were broken up into small pieces and used for Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105 harboring pCAMBIA1301 and pCAMBIA1305.2 vectors were used to infect embryogenic callus pieces. Hygromycin was used as a selection agent in stable transformation experiments. Hygromycin resistant calluses were obtained after 4–6 weeks of selection and transgenic plants were produced in 10–13 weeks after Agrobacterium-mediated transformation. Fertile plants were readily obtained after transferring the transgenics to the greenhouse. Transgenic nature of the regenerated plants was demonstrated by Polymerase chain reaction (PCR), Southern hybridization analysis, and GUS staining. Progeny analysis showed Mendelian inheritance of the transgenes. The transformation system provides a valuable tool for functionality tests of candidate genes in forage and turf grasses.     

Impacts of clearing,fragmentation and disturbance on the bird fauna of Eucalypt savanna woodlands in central Queensland,Australia

Abstract This study reports on the responses of bird assemblages to woodland clearance, fragmentation and habitat disturbance in central Queensland Australia, a region exposed to very high rates of vegetation clearance over the last two to three decades. Many previous studies of clearing impacts have considered situations where there is a very sharp management contrast between uncleared lands and cleared areas: in this situation, the contrast is more muted, because both cleared lands and uncleared savanna woodlands are exposed to cattle grazing, invasion by the exotic grass Cenchrus ciliaris and similar fire management. Bird species richness (at the scale of a 1‐ha quadrat) was least in cleared areas (8.1 species), then regrowth areas (14.6 species), then uncleared woodlands (19.9 species). Richness at this scale was unrelated to woodland fragment size, connectivity or habitat condition; but declined significantly with increasing abundance of miners (interspecifically aggressive colonial honeyeaters). At whole of patch scale, richness increased with fragment size and decreased with abundance of miners. This study demonstrates complex responses of individual bird species to a regional management cocktail of disturbance elements. Of 71 individual bird species modelled for woodland fragment sites, the quadrat‐level abundance of 40 species was significantly related to at least one variable representing environmental position (across a rainfall gradient), fragment condition, fragment size and/or connectivity. This study suggests that priorities for conservation management include: cessation of broad‐scale clearing; increased protection for regrowth (particularly where this may bolster connectivity and/or size of woodland fragments); control of miners; maintenance of fallen woody debris in woodlands; increase in fire frequency; and reduction in the incidence of grazing and exotic pasture grass.     

Genetic relationships among Eurasian Puccinellia distans genotypes

Puccinellia distans (Jacq.) Parl. is a common grass species found throughout the world. It can grow in arid and saline environments as well as under toxic boron concentrations. In this work we performed sequence related amplified polymorphism (SRAP) marker analysis on 20 wild P. distans genotypes to understand the genetic relationships among different genotypes and subspecies. We tested 119 SRAP primer pairs and found that 43 were polymorphic. The molecular data were then analyzed to determine the genetic relationships and population structure of the genotypes. We were able to trace the origin of genotypes that were carried to distant locations or gathered for research purposes. We also found that geographical distance between genotypes was not an important determinant of genetic relationships as even distantly located Puccinellia genotypes were closely related. As P. distans is known to be tolerant to salinity stress and toxic mineral concentrations, the findings of this work can be used as a starting point for selection of genotypes that should be tested under such conditions.     

Evolutionary divergence of β–expansin structure and function in grasses parallels emergence of distinctive primary cell wall traits

Expansins are wall‐loosening proteins that promote the extension of primary cell walls without the hydrolysis of major structural components. Previously, proteins from the EXPA (α–expansin) family were found to loosen eudicot cell walls but to be less effective on grass cell walls, whereas the reverse pattern was found for EXPB (β–expansin) proteins obtained from grass pollen. To understand the evolutionary and structural bases for the selectivity of EXPB action, we assessed the extension (creep) response of cell walls from diverse monocot families to EXPA and EXPB treatments. Cell walls from Cyperaceae and Juncaceae (families closely related to grasses) displayed a typical grass response (‘β–response’). Walls from more distant monocots, including some species that share with grasses high levels of arabinoxylan, responded preferentially to α–expansins (‘α–response’), behaving in this regard like eudicots. An expansin with selective activity for grass cell walls was detected in Cyperaceae pollen, coinciding with the expression of genes from the divergent EXPB–I branch that includes grass pollen β–expansins. The evolutionary origin of this branch was located within Poales on the basis of phylogenetic analyses and its association with the ‘sigma’ whole‐genome duplication. Accelerated evolution in this branch has remodeled the protein surface in contact with the substrate, potentially for binding highly substituted arabinoxylan. We propose that the evolution of the divergent EXPB–I group made a fundamental change in the target and mechanism of wall loosening in the grass lineage possible, involving a new structural role for xylans and the expansins that target them.     

禾草内生真菌对其它微生物的影响研究进展

禾草内生真菌在宿主植物的茎叶等地上组织中普遍存在,不仅能够提高禾草对生物与非生物逆境的抗性,而且能够对周围环境中的不同微生物类群产生影响。主要总结了禾草Neotyphodium/Epichlo内生真菌对病原真菌、丛枝菌根真菌和土壤微生物的影响及其作用机理。发现禾草内生真菌普遍存在对病原真菌的抑制作用,而对丛枝菌根真菌存在不对称的竞争作用,且因种类而异。禾草内生真菌对土壤微生物群落的作用则会随着土壤类型和时间等外界因素发生变化。禾草内生真菌对不同类群微生物的影响机制主要包括:通过生态位竞争、抑菌物质分泌、诱导抗病性等对病原真菌造成影响;通过根系化学物质释放、营养元素调节、侵染条件差异等对丛枝菌根真菌造成影响;通过根际沉积物和凋落物等对土壤微生物群落造成影响。禾草内生真菌产生的生物碱能提高宿主植物对包括昆虫在内草食动物采食的抗性,影响病原菌的侵入、定殖和扩展;根组织分泌物中包含次生代谢产物能够抑制菌根真菌、土传病原真菌及其它土壤微生物的侵染与群落组成;也可能通过次生代谢物影响禾草的其它抗性。因此,禾草内生真菌在植物-微生物系统中的作用应该给予更多的关注和深入研究。     

不同放牧制度下呼伦湖流域草原植被冠层截留

冠层截留是降雨过程中的水量分配和流域水平衡的一个重要组成部分,通过水浸泡法和降雨模拟实验研究呼伦湖流域草原3种放牧制度下(休牧、轮牧、自由放牧(超载放牧))植被冠层截留量的变化规律,并利用遥感解译植被归一化指数(NDVI),确定3种放牧制度下草原面积,估算呼伦湖流域草原降雨截留量。研究表明:在休牧、轮牧、自由放牧3种制度下,水浸泡法测定的截留量分别是0.468、0.320、0.271 mm。降雨模拟实验法测得的结果分别是0.957、0.613、0.431 mm。休牧、轮牧、自由放牧草场叶面积指数、盖度、容重、生物量等指标差异显著(P0.05),且单株植被高度、鲜重对截留量影响显著呈线性正相关关系。呼伦湖流域草原一次降雨量为大于等于30 mm全流域降雨,其植被截留量为6.462×106m3。     

Microsite determinants of variability in seedling and cutting establishment in tropical forest restoration plantations

Plantations are frequently established on abandoned pasture lands to speed forest recovery. This strategy requires matching a tree species mix with the prevailing microenvironmental conditions. In four degraded pastures of the Mexican Lacandon rainforest, we planted 2,400 trees of 6 species (Guazuma ulmifolia, Inga vera, Ochroma pyramidale, Trichospermum mexicanum, Bursera simaruba, and Spondias mombin) to (1) test survival, initial growth, and establishment costs; (2) evaluate whether vegetative cuttings outperform direct seeding or transplants of nursery‐raised seedlings; (3) determine tree response to herbaceous dominance and soil compaction; and (4) scrutinize the results' consistency across sites and sampling scales of tree–microenvironment interactions (individual tree vs. averaged plot responses). After 2 years, overall survival and growth rates were high for 2 of 3 nursery‐raised species. Contrary to expectations, all seedlings outperformed the cuttings while direct seeding resulted in a cost‐effective option of intermediate efficacy. The impact of soil resistance to root penetration on tree biomass accumulation was species dependent while bulk density was not relevant. Soil‐covering, herbaceous vegetation accelerated growth in 3 of 4 tested species during the dry season. At this initial stage of forest restoration in abandoned pastures, Guazuma and Trichospermum were the most restoration‐effective species. Costs can be reduced by using direct‐seeding Inga and avoiding weeding during the dry season. Finally, our results demonstrate how species selection trials can be misleading due to site variations in tree response and to sampling scales that fail to account for small‐scale environmental heterogeneity. We recommend ways to improve the design of restoration trials.     

The type of competition modulates the ecophysiological response of grassland species to elevated CO2 and drought

The effects of elevated CO₂ and drought on ecophysiological parameters in grassland species have been examined, but few studies have investigated the effect of competition on those parameters under climate change conditions. The objective of this study was to determine the effect of elevated CO₂ and drought on the response of plant water relations, gas exchange, chlorophyll a fluorescence and aboveground biomass in four grassland species, as well as to assess whether the type of competition modulates that response. Elevated CO₂ in well‐watered conditions increased aboveground biomass by augmenting CO₂ assimilation. Drought reduced biomass by reducing CO₂ assimilation rate via stomatal limitation and, when drought was more severe, also non‐stomatal limitation. When plants were grown under the combined conditions of elevated CO₂ and drought, drought limitation observed under ambient CO₂ was reduced, permitting higher CO₂ assimilation and consequently reducing the observed decrease in aboveground biomass. The response to climate change was species‐specific and dependent on the type of competition. Thus, the response to elevated CO₂ in well‐watered grasses was higher in monoculture than in mixture, while it was higher in mixture compared to monoculture for forbs. On the other hand, forbs were more affected than grasses by drought in monoculture, while in mixture the negative effect of drought was higher in grasses than in forbs, due to a lower capacity to acquire water and mineral nutrients. These differences in species‐level growth responses to CO₂ and drought may lead to changes in the composition and biodiversity of the grassland plant community in future climate conditions.     

Phytoavailability of Heavy Metals and Metalloids in Soils Irrigated with Wastewater,Akaki, Ethiopia: A Greenhouse Study

Irrigation with untreated wastewater from several industrial, commercial, and domestic discharges for decades caused accumulation of various heavy metals and metalloids in soils along the Akaki River in Ethiopia. Assessment of environmental threats and the potential phytoremediation of the soils require understanding of the toxic elements’ uptake and distribution in plant parts. Hence, a greenhouse study was performed to examine the phytoavailability and distribution of Cr, Ni, Co, Cu, Zn, Cd, Pb, Hg, Se, V, and As in forage grasses: Oat (Avena sativa), Rhodes grass (Chloris gayana), Setaria (Setaria sphacelata), and the legumes Alfalfa (Medicago sativa) and Desmodium (Desmodium unicinatum). The average contents of Cr, Ni, Co, Cu, Zn, Pb, Hg, Se, and V in the plants were generally higher than the background levels for forage grasses/legumes, and some of these elements were in the phytotoxic range. Root bioconcentration factor (BCF = root to soil concentration ratio) > 1 was observed for Cu (Oat, Rhodes, Desmodium, and Setaria: Fluvisol), Zn (Setaria: Fluvisol), Cd (Rhodes: Fluvisol; Setaria from both soils) and Hg (Oat and Alfalfa: Fluvisol). Alfalfa and Desmodium displayed translocation factor > 1 (TF = shoot to root concentration ratio) for most heavy metals. Most heavy metals/metalloids may pose a health threat to humans and stock via introduction to the food chain. The plant factors (species and plant part), soil factors (soil type, soil fractions, pH, and CEC), and their interactions significantly (p < 0.05) influenced plant heavy metal and metalloid levels. However, the role of plant part and species emerged as the most important on heavy metal uptake, translocation, sequestration, and ultimately transfer to the food chain. Accordingly, the uptake and distribution of heavy metals/metalloids in the plants reflect the potential environmental and health hazards attributable to the use of fodder grasses, legumes, and cultivation of vegetables in soils with polymetallic and metalloid contamination.