First Report of Rhizoctonia solani AG‐7 on Cotton in Egypt

Eighty‐two isolates of Rhizoctonia solani were recorded from roots of naturally‐infected seedlings of the Egyptian cotton (Gossypium barbadense L.). Anastomosis groups (AGs) of the isolates were determined by using 13 different AGs testers. Three (3.7%) of the isolates were identified as R. solani AG7, while the remaining isolates were belonging to the AG 2‐1, AG4 and AG5. The identification of the three isolates was based on the frequency of the C2 reaction with the AG7 tester isolate. No fusion was observed between AG7 and isolates representing the other 13 AGs. Colonies of AG7 isolates grown on potato dextrose agar (PDA), malt yeast agar (MYA) and melt peptone agar (MPA) were brown to dark brown with aerial mycelium and sclerotia. The isolates had pitted sclerotial clusters and brownish exudates after 21 days of culturing on PDA, but without clear zonation. Pathogenicity test under greenhouse conditions revealed that AG7 caused the common symptoms of damping–off, which included seed rot, lesions on the hypocotyls and root rot.     

Use of DAPI for Anastomosis Group Typing of Strains of the Fungus Rhizoctonia solani

Strains of Rhizoctonia solani, a common soil-borne, pathogenic fungus of plants, are assigned to one of 11 anastomosis groups (AGs) based on the occurrence of imperfect fusions (anastomoses) between hyphae of a non-typed strain and a tester strain of one of the 11 AG's. Imperfect fusion is characterized by the death of one or more cells in each of the hyphae involved in the fusion. Although hyphae from branches of the same strain of JR. solani may fuse with each other (self-fusion), cell death does not occur. Cell death is accompanied by nuclear degradation and granulation, or plasmolysis of the cytoplasm, which often is not visible using bright-field microscopy. When the DNA-binding fluorochrome DAPI (4', 6-diamidino-2-phenylindole) is used and the hyphal fusions viewed under fluorescence microscopy, no nuclei are observed in fused hyphal cells from two strains of the same AG of R. solani Because DAPI reacts only with living nuclei, lack of staining is presumptive evidence that the fused cells are dead as a result of imperfect fusion. The use of DAPI reduces the time required for making AG determinations compared to standard methods because it eliminates the need to assess cell wall dissolution and cytoplasmic fusion. Also, it is not necessary to trace the hyphae involved in the fusion to their respective origins to ensure that self-fusion has not occurred.     

Anastomosis Group Determination of Rhizoctonia solani Kühn (Telemorph: Thanatephorus cucumeris) Isolates from Tomatoes Grown in Aydin,Turkey and their Disease Reaction on Various Tomato Cultivars

In the Province of Aydin‐Turkey most frequently Fusarium spp. and secondly Rhizoctonia solani Kühn were isolated from the roots and crown of tomato plants. Based on affinities for hyphal fusion with test isolates, all R. solani isolates were identified as AG‐4. The tomato cultivars which were grown in soil infested with R. solani AG‐4 exhibited different reactions. From the point of symptom expression and the rate of seedling emergence Sunny 6066 F1 was found to be the most resistant cultivar, whereas Rio Grande, Rio Fuego, NDM 725, Interpeel and Konia were the most susceptible cultivars.     

Rhizoctonia solani injuries in oilseed crops in Finland and impacts of different crop management practices on disease incidence and severity

A disease survey in Finnish oilseed Brassica (OSR) fields in 2007–09 revealed the widespread occurrence and several fold increase of necrotic stem base lesions and severely injured blackened roots in comparison to a corresponding survey carried out in 1984–89. Rhizoctonia solani was the predominant fungi detected in the isolations and was followed by several species of Fusarium and Thielaviopsis basicola. In 60% of the samples all three species were detected together. Only the R. solani AG 2–1 strains isolated from OSR and other cruciferous hosts caused damping off or stem base symptoms on turnip rape in a greenhouse experiment. Therefore R. solani AG 2–1 was considered the main pathogen associated with the observed symptoms in OSR crops. Cultural practices changed significantly between the 1980s and 2000s. In the 2007–09 survey there was an increase in the cultivation of oilseed rape instead of turnip rape, increase in the use of no soil or reduced soil tillage and of chemical control of weeds, but a reduction in macronutrient fertilization, especially P and K, when compared to the 1980s survey. The risk for high incidence of stem base lesions and blackened roots was affected by different cultural practices. No tillage and maintaining sufficient soil pH and NPK fertilisation decreased the risk for both types of R. solani induced symptoms. Late sowing date increased the risk for high incidence of stem base lesions, while application of fungicides against Sclerotinia reduced it. The incidence of R. solani damages in many fields was very high in spite of relatively long crop rotations and therefore the average effect of crop rotation in the disease was insignificant. Current turnip rape cultivars are vulnerable to root rot while oilseed rape is vulnerable to stem base symptoms. The higher incidence of R. solani induced diseases could be associated with the decline in productivity of OSR crops in Finland. This study showed that cultural practices such as reduced or no soil tillage, adequate levels of pH and of NPK fertilization could reduce the severity of the symptoms in OSR fields.     

High levels of gene flow and heterozygote excess characterize Rhizoctonia solani AG-1 IA (Thanatephorus cucumeris) from Texas

To date, much of the genetics of the basidiomycete Thanatephorus cucumeris (anamorph = Rhizoctonia solani) remains unknown. Here, we present a population genetics study using codominant markers to augment laboratory analyses. Seven single-copy nuclear RFLP markers were used to examine 182 isolates of Rhizoctonia solani AG-1 IA collected from six commercial rice fields in Texas. Thirty-six multilocus RFLP genotypes were identified. Population subdivision analyses indicated a high degree of gene flow/migration between the six geographic populations. Tests for Hardy-Weinberg equilibrium (HWE) among the 36 multilocus RFLP genotypes revealed that four of the seven loci did not significantly differ from HWE. Subsequent analysis demonstrated that departures from HWE at the three remaining loci were due to an excess of heterozygotes. Data presented here suggest that R. solani AG-1 IA is actively outbreeding (heterothallic). Possible explanations for heterozygote excess, which was observed at all seven RFLP loci, are discussed.     

Long‐term nitrogen addition causes the evolution of less‐cooperative mutualists

Human activities have altered the global nitrogen (N) cycle, and as a result, elevated N inputs are causing profound ecological changes in diverse ecosystems. The evolutionary consequences of this global change have been largely ignored even though elevated N inputs are predicted to cause mutualism breakdown and the evolution of decreased cooperation between resource mutualists. Using a long‐term (22 years) N‐addition experiment, we find that elevated N inputs have altered the legume–rhizobium mutualism (where rhizobial bacteria trade N in exchange for photosynthates from legumes), causing the evolution of less‐mutualistic rhizobia. Plants inoculated with rhizobium strains isolated from N‐fertilized treatments produced 17–30% less biomass and had reduced chlorophyll content compared to plants inoculated with strains from unfertilized control plots. Because the legume–rhizobium mutualism is the major contributor of naturally fixed N to terrestrial ecosystems, the evolution of less‐cooperative rhizobia may have important environmental consequences.     

Long‐term nitrogen loading alleviates phosphorus limitation in terrestrial ecosystems

Increased human‐derived nitrogen (N) deposition to terrestrial ecosystems has resulted in widespread phosphorus (P) limitation of net primary productivity. However, it remains unclear if and how N‐induced P limitation varies over time. Soil extracellular phosphatases catalyze the hydrolysis of P from soil organic matter, an important adaptive mechanism for ecosystems to cope with N‐induced P limitation. Here we show, using a meta‐analysis of 140 studies and 668 observations worldwide, that N stimulation of soil phosphatase activity diminishes over time. Whereas short‐term N loading (≤5 years) significantly increased soil phosphatase activity by 28%, long‐term N loading had no significant effect. Nitrogen loading did not affect soil available P and total P content in either short‐ or long‐term studies. Together, these results suggest that N‐induced P limitation in ecosystems is alleviated in the long‐term through the initial stimulation of soil phosphatase activity, thereby securing P supply to support plant growth. Our results suggest that increases in terrestrial carbon uptake due to ongoing anthropogenic N loading may be greater than previously thought.     

Long‐term sperm storage in the brownbanded bamboo shark Chiloscyllium punctatum

This study investigated the birth of a brownbanded bamboo shark Chiloscyllium punctatum at the Steinhart Aquarium. Genetic analyses suggest this is the longest documented case of sperm storage for any species of shark (45 months).     

Long‐term culture optimization of human omentum fat‐derived mesenchymal stem cells

Recent scientific explorations in search of novel sources for autologous transplantation transpired an alternative source of MSCs (mesenchymal stem cells) derived from omentum fat. The scarcity of experimental evidences probing into the biosafety concerns of omentum fat‐derived MSC under prolonged culture conditions limits its applicability as an efficient tool in regenerative medicine. This study, thus, aims to optimize human omentum fat‐derived MSC in four different media [DMEM (Dulbecco's modified Eagle's medium) LG (low glucose), DMEM KO (knock out), α‐MEM (α‐minimal essential media) and DMEM F12] in the facets of phenotypic characterization, growth kinetics, differentiation and karyotyping under prolonged culture. The cells exhibited a similarity in expression profile for the majority of markers with evidential variations in certain markers. The relevance of omentum fat‐derived MSCs became evident from its triumphant differentiation potential and karyotypic stability substantiated even at later passage. The results obtained from growth curve and PDT (population doubling time) lead to optimization of appropriate media for omentum fat‐derived stem cell research, thereby bringing omentum fat into the forefront of regenerative medicine.     

Long‐term non‐invasive and continuous measurements of legume nodule activity

Symbiotic nitrogen fixation is a process of considerable economic, ecological and scientific interest. The central enzyme nitrogenase reduces H⁺ alongside N₂, and the evolving H₂ allows a continuous and non‐invasive in vivo measurement of nitrogenase activity. The objective of this study was to show that an elaborated set‐up providing such measurements for periods as long as several weeks will produce specific insight into the nodule activity's dependence on environmental conditions and genotype features. A system was developed that allows the air‐proof separation of a root/nodule and a shoot compartment. H₂ evolution in the root/nodule compartment can be monitored continuously. Nutrient solution composition, temperature, CO₂ concentration and humidity around the shoots can concomitantly be maintained and manipulated. Medicago truncatula plants showed vigorous growth in the system when relying on nitrogen fixation. The set‐up was able to provide specific insights into nitrogen fixation. For example, nodule activity depended on the temperature in their surroundings, but not on temperature or light around shoots. Increased temperature around the nodules was able to induce higher nodule activity in darkness versus light around shoots for a period of as long as 8 h. Conditions that affected the N demand of the shoots (ammonium application, Mg or P depletion, super numeric nodules) induced consistent and complex daily rhythms in nodule activity. It was shown that long‐term continuous measurements of nodule activity could be useful for revealing special features in mutants and could be of importance when synchronizing nodule harvests for complex analysis of their metabolic status.     

Predictors of Long‐term Weight Maintenance

Objective: The purpose of this study was to evaluate available variables of a long‐term weight maintenance study to investigate possible factors predisposing to weight regain after a period of weight loss. Research Methods and Procedures: The Maastricht Weight Maintenance Study is an ongoing longitudinal study of healthy men and women (29 men and 62 women; 18 to 65 years of age; BMI = 30.2 ± 3.1 kg/m²). A variety of parameters were measured before and after a very‐low‐energy diet and after a follow‐up of at least 2 years. Results: Mean weight loss was 7.9 ± 3.6 kg, and percent weight regain was 113.8 ± 98.1%. Percent BMI regain was negatively associated with an increase in dietary restraint (r = ?0.47, p < 0.05). Percent weight regain was negatively correlated with baseline resting metabolic rate (r = ?0.38, p = 0.01) and baseline fat mass (r = ?0.24, p = 0.05) and positively correlated with the magnitude of change in body weight (BW) expressed as maximum amplitude of BW (r = 0.21, p < 0.05). In addition, amplitude of BW was positively correlated with the frequency of dieting (r = 0.57, p < 0.01). Discussion: The best predictors for weight maintenance after weight loss were an increase in dietary restraint during weight loss, a high baseline resting metabolic rate, a relatively high baseline fat mass favoring a fat‐free mass–sparing effect during weight loss, a rather stable BW, and a low frequency of dieting. Therefore, BW maintenance after BW loss seems to be a multifactorial issue, including mechanisms that regulate an individuals’ energy expenditure, body composition, and eating behavior in such a way that energy homeostasis is maintained.     

Cryopreservation of seed of Western Australian native species

The ability of seed of native Western Australian species to be stored using cryopreservation methods was investigated by subjecting seed of 90 native species representing 84 genera and 33 families to storage in liquid nitrogen. Seed of 68 native Western Australian species were germinated after storage in liquid nitrogen for two weeks following treatments which involved direct plunging into liquid nitrogen or slow cooling at 0.4°C min^–1 in 15% dimethyl sulphoxide (DMSO) or slow cooling at 0.4°C min^–1 in 35% DMSO. The largest number of species (37) responded positively to direct plunging without pretreatments, with only 10 species responding to slow cooling in 15% DMSO. Thirty one species had enhanced germination and 10 species depressed germination after any of the liquid nitrogen treatments. There were no trends in a species ability to survive liquid nitrogen storage and freezing regime, moisture content, seed size or taxonomic relatedness. However, hard seeded species belonging to the families Caesalpinaceae and Papilionaceae showed a consistently high degree of tolerance to liquid nitrogen storage. Significant physical damage to seed and cotyledons only occurred in Templetonia retusa (Papilionaecae) and this was alleviated by nicking the seed coat. This study indicates that seed of a large proportion of native Western Australian species may be amenable to storage in liquid nitrogen and that at least 40% of the listed rare and endangered species of Western Australia could be maintained in this way.     

Long‐term impacts of agricultural practices and climatic variability on carbon storage in a permanent pasture

Intra‐ and interannual variability of precipitation can lead to major modifications of grassland production and carbon storage capacity. Greater understanding of how climatic variability affects net CO₂ exchange [i.e. net ecosystem exchange (NEE)] of grazed grasslands is important to adapt grassland management and reduce risks of carbon losses. Since 2002, we continuously measured NEE (i.e. eddy covariance technique) on an upland grassland site (7 ha), divided in two paddocks grazed by heifers (intensive: 1 LSU ha^?1 yr^?1, 213 kg N ha^?1 yr^?1 and extensive: 0.5 LSU ha^?1 yr^?1, no fertilization). For years with dry and warm growing seasons (i.e. 2003, 2005 and 2008), absolute annual NEE was higher in the intensive paddock compared with the extensive paddock. The opposite was observed during years of ample seasonal rainfall and soil moisture (i.e. 2004, 2006 and 2007). Contrasted management led to two distinct plant communities being different in leaf area index (LAI), soil bulk density and soil water holding capacity. Differences in annual NEEs could thus be assigned to interactions between in carbon and water fluxes during dry and wet growth periods. Dry growth periods led to a reduction in weekly gross primary productivity (GPP) in the extensively managed paddock, whereas the GPP was maintained in the intensive paddock. In turn, during wet growth periods, GPP was similar in both paddocks, whereas N amendment and frequent defoliation significantly increased ecosystem respiration in the intensive paddock, presumably through a higher heterotrophic respiration following on a better C substrate quality and availability (rhizodeposition and senescent fine roots). In the extensive paddock, where plant cover was denser (reducing soil temperature) and less decomposable, C losses through heterotrophic respiration were comparatively smaller under wet conditions. Our results demonstrate that grassland subjected to a moderately intensive management could be more resilient in terms of carbon storage during drought and heat waves, presumably because of a trade‐off between heterotrophic and autotrophic respiration.     

Long‐term datasets: From descriptive to predictive data using ecoinformatics

This Special Feature includes contributions on data‐processing of large ecological datasets under the heading ecoinformatics. Herewith the latter term is now al so established in the Journal of Vegetation Science. Ecoinfomatics is introduced as a rapid growing field within community ecology which is generating exciting new developments in ecology and in particular vegetation ecology. In our field, ecoinformatics deals with the understanding of patterns of species distributions at local and regional scales, and on the assemblages of species in relation to their properties, the local environment and their distribution in the region. Community ecology using ecoinformatics is related to bioinformatics, community ecology, biogeography and macroecology. We make clear how ecoinformatics in vegetation science and particularly the IAVS Working Group on Ecoinformatics has developed from the work of the old Working Group for Data Processing which was active during the 1970s and 1980s. Recent developments, including the creation of TURBOVEG and Syn Bio Sys in Europa and VEGBANK in the USA, form a direct link with these pioneer activities, both scientifically and personally. The contributions collected in this Special Feature present examples of seco‐infeveral types of the use of databases and the application of programmes and models. The main types are the study of long‐term vegetation dynamics in different cases of primary and secondary succession and the understanding of successional developments in terms of species traits. Among the future developments of great significance we mention the use of a variety of different large datasets for the study of the distribution and ecology and conservation of rare and threatened species.     

Long‐term changes in forest carbon under temperature and nitrogen amendments in a temperate northern hardwood forest

Currently, forests in the northeastern United States are net sinks of atmospheric carbon. Under future climate change scenarios, the combined effects of climate change and nitrogen deposition on soil decomposition, aboveground processes, and the forest carbon balance remain unclear. We applied carbon stock, flux, and isotope data from field studies at the Harvard forest, Massachusetts, to the ForCent model, which integrates above‐ and belowground processes. The model was able to represent decadal‐scale measurements in soil C stocks, mean residence times, fluxes, and responses to a warming and N addition experiment. The calibrated model then simulated the longer term impacts of warming and N deposition on the distribution of forest carbon stocks. For simulation to 2030, soil warming resulted in a loss of soil organic matter (SOM), decreased allocation to belowground biomass, and gain of aboveground carbon, primarily in large wood, with an overall small gain in total system carbon. Simulated nitrogen addition resulted in a small increase in belowground carbon pools, but a large increase in aboveground large wood pools, resulting in a substantial increase in total system carbon. Combined warming and nitrogen addition simulations showed a net gain in total system carbon, predominately in the aboveground carbon pools, but offset somewhat by losses in SOM. Hence, the impact of continuation of anthropogenic N deposition on the hardwood forests of the northeastern United States may exceed the impact of warming in terms of total ecosystem carbon stocks. However, it should be cautioned that these simulations do not include some climate‐related processes, different responses from changing tree species composition. Despite uncertainties, this effort is among the first to use decadal‐scale observations of soil carbon dynamics and results of multifactor manipulations to calibrate a model that can project integrated aboveground and belowground responses to nitrogen and climate changes for subsequent decades.     

Long‐term influence of sod cutting depth on the restoration of degraded wet heaths

Temperate heaths have an unfavorable conservation status in most European biogeographical regions. Increasing nitrogen levels promote competitive grass species such as Molinia caerulea, which is a main threat to heathland conservation in Europe. This article investigates the long‐term influence of sod cutting and the resulting changes in soil properties on the heath composition, integrity, and structure. In 15 nature reserves across the northern half of Belgium, we used (1) a large number of plots (203); (2) a broad range of sod cut depths (2–40 cm), and (3) a temporal dimension that describes how long the effects of sod cutting persist (census up to 19 years after sod cutting). Multivariate analyses were used in order to explore the influence of sod cut depth and time after sod cutting on the soil and vegetation properties. There was a positive relationship between sod cut depth and soil pH and water level, and a negative relationship with Al³⁺, NH₄⁺, and total organic matter (TOM). However, only a limited number of typical (target) species appeared after sod cutting, and then only weakly. Most of the time they remained a minor component of the restored vegetation. Moreover, M. caerulea reappeared and its cover significantly increased during the years following sod cutting. Although we were able to show that sod cut depth has a differential effect on soil properties and vegetation recovery, it also appeared that sod cutting does not restore wet heaths in the long term when applied in regions with high nitrogen deposition.     

Long‐term persistence of wildlife populations in a pastoral area

Facilitating coexistence between people and wildlife is a major conservation challenge in East Africa. Some conservation models aim to balance the needs of people and wildlife, but the effectiveness of these models is rarely assessed. Using a case‐study approach, we assessed the ecological performance of a pastoral area in northern Tanzania (Manyara Ranch) and established a long‐term wildlife population monitoring program (carried out intermittently from 2003 to 2008 and regularly from 2011 to 2019) embedded in a distance sampling framework. By comparing density estimates of the road transect‐based long‐term monitoring to estimates derived from systematically distributed transects, we found that the bias associated with nonrandom placement of transects was nonsignificant. Overall, cattle and sheep and goat reached the greatest densities and several wildlife species occurred at densities similar (zebra, wildebeest, waterbuck, Kirk's dik‐dik) or possibly even greater (giraffe, eland, lesser kudu, Grant's gazelle, Thomson's gazelle) than in adjacent national parks in the same ecosystem. Generalized linear mixed models suggested that most wildlife species (8 out of 14) reached greatest densities during the dry season, that wildlife population densities either remained constant or increased over the 17‐year period, and that herbivorous livestock species remained constant, while domestic dog population decreased over time. Cross‐species correlations did not provide evidence for interference competition between grazing or mixed livestock species and wildlife species but indicate possible negative relationships between domestic dog and warthog populations. Overall, wildlife and livestock populations in Manyara Ranch appear to coexist over the 17‐year span. Most likely, this is facilitated by existing connectivity to adjacent protected areas, effective anti‐poaching efforts, spatio‐temporal grazing restrictions, favorable environmental conditions of the ranch, and spatial heterogeneity of surface water and habitats. This long‐term case study illustrates the potential of rangelands to simultaneously support wildlife conservation and human livelihood goals if livestock grazing is restricted in space, time, and numbers.     

Long‐term carbon storage through retention of dissolved aromatic acids by reactive particles in soil

Soils retain large quantities of carbon, thereby slowing its return to the atmosphere. The mechanisms governing organic carbon sequestration in soil remain poorly understood, yet are integral to understanding soil‐climate feedbacks. We evaluated the biochemistry of dissolved and solid organic carbon in potential source and sink horizons across a chronosequence of volcanic soils in Hawai'i. The soils are derived from similar basaltic parent material on gently sloping volcanic shield surfaces, support the same vegetation assemblage, and yet exhibit strong shifts in soil mineralogy and soil carbon content as a function of volcanic substrate age. Solid‐state¹³carbon nuclear magnetic resonance spectra indicate that the most persistent mineral‐bound carbon is comprised of partially oxidized aromatic compounds with strong chemical resemblance to dissolved organic matter derived from plant litter. A molecular mixing model indicates that protein, lipid, carbohydrate, and char content decreased whereas oxidized lignin and carboxyl/carbonyl content increased with increasing short‐range order mineral content. When solutions rich in dissolved organic matter were passed through Bw‐horizon mineral cores, aromatic compounds were preferentially sorbed with the greatest retention occurring in horizons containing the greatest amount of short‐range ordered minerals. These minerals are reactive metastable nanocrystals that are most common in volcanic soils, but exist in smaller amounts in nearly all major soil classes. Our results indicate that long‐term carbon storage in short‐range ordered minerals occurs via chemical retention with dissolved aromatic acids derived from plant litter and carried along preferential flow‐paths to deeper B horizons.