Whole-plant Response Research Articles

Investigation of resistance mechanisms to flucarbazone-sodium in wild oat (Avena fatua L.) from China

BackgroundWild oat (Avena fatua L.) is a self-pollinating, allohexaploid species in the family Gramineae (grasses), which is a malignant weed that mainly harms crops such as wheat. In recent years, a decline in the control efficiency of flucarbazone-sodium against wild oat has occurred in some regions of China.ResultsWe identified an ALS-resistant A. fatua population (R population). Whole-plant response assays revealed that the R population exhibited a moderate level of resistance (5.9-fold) to flucarbazone-sodium. Pre-treatment with malathion significantly reduced flucarbazone-sodium resistance in the R population. The known mutation sites and ALS gene relative expression that confer resistance to ALS inhibitor herbicides were not found in R population. Following flucarbazone-sodium treatment, the expression of eight genes related to metabolic enzymes was investigated using quantitative real-time PCR (qRT-PCR). CYP92A6 and the Aldo/keto reductase family were highly expressed in the R population after the application of flucarbazone-sodium.ConclusionsThe mechanism of flucarbazone-sodium resistance in A. fatua is mediated by NTSR, nor TSR. Two genes, CYP92A6 and the Aldo/keto reductase family, were discovered to be possibly related in the metabolism of NTSR in the A. fatua population, justifying more functional studies. The results will serve as a data resource for further studies on the molecular mechanisms of A. fatua to flucarbazone-sodium.

Empirical evidence and theoretical understanding of ecosystem carbon and nitrogen cycle interactions.

Interactions between carbon (C) and nitrogen (N) cycles in terrestrial ecosystems are simulated in advanced vegetation models, yet methodologies vary widely, leading to divergent simulations of past land C balance trends. This underscores the need to reassess our understanding of ecosystem processes, given recent theoretical advancements and empirical data. We review current knowledge, emphasising evidence from experiments and trait data compilations for vegetation responses to CO2 and N input, alongside theoretical and ecological principles for modelling. N fertilisation increases leaf N content but inconsistently enhances leaf-level photosynthetic capacity. Whole-plant responses include increased leaf area and biomass, with reduced root allocation and increased aboveground biomass. Elevated atmospheric CO2 also boosts leaf area and biomass but intensifies belowground allocation, depleting soil N and likely reducing N losses. Global leaf traits data confirm these findings, indicating that soil N availability influences leaf N content more than photosynthetic capacity. A demonstration model based on the functional balance hypothesis accurately predicts responses to N and CO2 fertilisation on tissue allocation, growth and biomass, offering a path to reduce uncertainty in global C cycle projections.

Integrated above and below-ground responses of the gypsum specialist Helianthemum squamatum (L.). to drought

Gypsum endemics (i.e. gypsophiles) have adapted to live in gypsum-rich soils where nutrient unbalance and drought can be extreme. Despite their relevance as plants adapted to extreme conditions, a complete analysis of the physiological responses of gypsophiles to drought is still lacking. Helianthemum squamatum (L.) Dum. Cours. is a conspicuous Iberian gypsophile that has been reported to use gypsum crystallization water during the driest period, but the mechanisms behind this process are unknown. To characterize gypsophile responses to drought and unravel the mechanisms underlying gypsum crystalline water use, H. squamatum plants were grown in pots with natural gypsum soil and gypsum soil with deuterium-labelled crystalline water. After three years, a drought experiment was carried out and whole-plant responses were investigated. Unexpectedly, the labelling treatment affected soil physicochemical characteristics and reduced microbial biomass and organic matter content, decreasing plant aerial biomass. H. squamatum plants did not use gypsum crystallization water during simulated drought neither in the labelled soil, nor in the natural one. Drought reduced plant transpiration, stomatal conductance, water use, photosynthetic rate and the foliar concentration of most elements except P and N, which were higher in the drought stressed plants. We detected increased root exudation of choline, an osmoprotector, by drought stressed plants. The drought treatment also affected the structure of microbial communities but did not reduce the relative abundance of functional microbial groups, highly adapted to the natural drought pulses. Our results highlight an integrated water-saving strategy of H. squamatum plants in the short-term, where responses at the leaf level are combined with belowground processes, like altered root exudation. Our findings also underline the remarkable resistance to drought of microbial communities present in gypsum soils.

Xylem structure and hydraulic function in roots and stems of chaparral shrub species from high and low elevation in the Sierra Nevada, California.

Xylem structure and hydraulics were compared between individuals at lower and upper elevation distribution limits for five chaparral shrub species along a steep transect in the southern Sierra Nevada, California, USA. Higher-elevation plants experienced frequent winter freeze-thaw events and increased precipitation. We hypothesized that environmental differences would lead to xylem trait differences between high and low elevations, but predictions were complicated because both water stress (low elevation) and freeze-thaw events (high elevation) may select for similar traits, such as narrow vessel diameter. We found significant changes in the ratio of stem xylem area to leaf area (Huber value) between elevations, with more xylem area required to support leaves at low elevations. Co-occurring species significantly differed in their xylem traits, suggesting diverse strategies to cope with the highly seasonal environment of this Mediterranean-type climate region. Roots were more hydraulically efficient and more vulnerable to embolism relative to stems, potentially due to roots being buffered from freeze-thaw stress, which allows them to maintain wider diameter vessels. Knowledge of the structure and function of both roots and stems is likely important in understanding whole-plant response to environmental gradients.

Linking whole-plant responses to cell physiology in glycophytes exposed to NaCl stress

Linking whole-plant responses to cell physiology in glycophytes exposed to NaCl stress

First Report of the Molecular Mechanism of Resistance to Tribenuron-Methyl in Silene conoidea L.

Silene conoidea L. is an annual troublesome broadleaf weed in winter wheat fields in China. In recent years, field applications of tribenuron-methyl have been ineffective in controlling S. conoidea in Hebei Province, China. The aim of this study was to determine the molecular basis of tribenuron-methyl resistance in S. conoidea. Whole-plant response assays revealed that the resistant population (R) exhibited a higher level of resistance (382.3-fold) to tribenuron-methyl. The R population also showed high cross-resistance to other acetolactate synthase (ALS) inhibitors, including imazethapyr, bispyribac-sodium and florasulam. However, the R population could be controlled by the field-recommended rates of bentazone, MCPA, fluroxypyr, carfentrazone-ethyl and bromoxynil. In vitro ALS activity assays indicated that the tribenuron-methyl I50 value for the R population was 18.5 times higher than those for the susceptible population (S). ALS gene sequencing revealed an amino acid mutation, Trp-574-Leu, in the R population. Pretreatment with the P450 inhibitor malathion indicated that the R population might have cytochrome P450-mediated metabolic resistance. These results suggest that the Trp-574-Leu mutation and P450-mediated enhanced metabolism coexist in S. conoidea to generate tribenuron-methyl resistance. This is the first time that target-site and non-target-site resistance to tribenuron-methyl has been reported in S. conoidea.

Target-site basis for resistance to flucarbazone- sodium in Japanese brome (Bromus japonicus Houtt.) in China

Japanese brome (Bromus japonicus Houtt.) is a troublesome annual weed and widely distributed in winter wheat (Triticum aestivum L.) fields in the North China Plain. A B. japonicus population (TJ06) suspected of resistance to acetolactate synthase (ALS) inhibitors was found in Tianjin, China. In this study, the TJ06 population with an Asp-376-Glu mutation in ALS gene was identified. TJ06 population developed 66.7-fold resistance to flucarbazone-sodium and exhibited obvious cross-resistance to other two ALS-inhibiting herbicides. The 50% plant growth reduction (GR50) to herbicides of pyroxsulam and mesosulfuron-methyl were 28.72 and 39.44 g ai ha-1, respectively. In in vitro ALS activity assays, the concentration of flucarbazone-sodium required to inhibit 50% ALS activity (I50) for TJ06 was 11.3-fold greater than that for a susceptible population (TJ01), which was highly correlated with that of whole-plant response experiments and indicated that the Asp-376-Glu mutation leading to resistance reduced sensitivity of the ALS enzyme to flucarbazone- sodium. Besides, one derived cleaved amplified polymorphic sequence (dCAPS) marker was designed to quickly detect Asp376 mutation in ALS gene of B. japonicus.

First person – Mitchell Booth

ABSTRACTFirst Person is a series of interviews with the first authors of a selection of papers published in Biology Open, helping researchers promote themselves alongside their papers. Mitchell Booth is first author on ‘ Tissue-specific transcriptome profiles identify functional differences key to understanding whole-plant response to life in variable salinity’, published in BiO. Mitchell is a PhD student in the lab of Elizabeth Sinclair at the University of Western Australia Oceans Institute, using omics to understand the mechanisms of life in a changing world.

Tissue-specific transcriptome profiles identify functional differences key to understanding whole plant response to life in variable salinity.

ABSTRACTPlants endure environmental stressors via adaptation and phenotypic plasticity. Studying these mechanisms in seagrasses is extremely relevant as they are important primary producers and functionally significant carbon sinks. These mechanisms are not well understood at the tissue level in seagrasses. Using RNA-seq, we generated transcriptome sequences from tissue of leaf, basal leaf meristem and root organs of Posidonia australis, establishing baseline in situ transcriptomic profiles for tissues across a salinity gradient. Samples were collected from four P. australis meadows growing in Shark Bay, Western Australia. Analysis of gene expression showed significant differences between tissue types, with more variation among leaves than meristem or roots. Gene ontology enrichment analysis showed the differences were largely due to the role of photosynthesis, plant growth and nutrient absorption in leaf, meristem and root organs, respectively. Differential gene expression of leaf and meristem showed upregulation of salinity regulation processes in higher salinity meadows. Our study highlights the importance of considering leaf meristem tissue when evaluating whole-plant responses to environmental change.This article has an associated First Person interview with the first author of the paper.

Occurrence of Bensulfuron-Methyl Resistance and Target-Site Resistance Mechanisms in Ammannia auriculata Biotypes from Paddy Fields.

Ammanniaauriculata is a troublesome broadleaf weed, widely distributed in the paddy fields of southern China. In this study, 10 biotypes of A. auriculata were sampled from Yangzhou City, China, where the paddy fields were seriously infested with A. auriculata, and their resistance levels to acetolactate synthase (ALS) inhibitor bensulfuron-methyl were determined. The whole-plant response assays showed that nine A. auriculata biotypes were highly resistant (from 16.4- to 183.1-fold) to bensulfuron-methyl in comparison with a susceptible YZ-S biotype, and only one YZ-6 biotype was susceptible. ALS gene sequencing revealed that three ALS gene copies existed in A. auriculata, and four different amino acid substitutions (Pro197-Leu, -Ala, -Ser, and -His) at site 197 in the AaALS1 or 2 genes were found in eight resistant biotypes. In addition, no amino acid mutations in three ALS genes were found in the YZ-3 biotype. These results suggested that target-site mutations or non-target-site resistance mechanisms were involved in tested resistant A. auriculata biotypes. Finally, a cleaved amplified polymorphic sequence (CAPS) marker was identified to rapidly detect the Pro197 mutations in A. auriculata.

Plant metabolism and defence strategies in the flowering stage: Time-dependent responses of leaves and flowers under attack.

Plants developing into the flowering stage undergo major physiological changes. Because flowers are reproductive tissues and resource sinks, strategies to defend them may differ from those for leaves. Thus, this study investigates the defences of flowering plants by assessing processes that sustain resistance (constitutive and induced) and tolerance to attack. We exposed the annual plant Brassica nigra to three distinct floral attackers (caterpillar, aphidand bacterial pathogen)and measured whole-plant responses at 4, 8and 12 days after the attack. We simultaneously analysed profiles of primary and secondary metabolites in leaves and inflorescencesand measured dry biomass of roots, leavesand inflorescences as proxies of resource allocation and regrowth. Regardless of treatments, inflorescences contained 1.2 to 4 times higher concentrations of primary metabolites than leaves, and up to 7 times higher concentrations of glucosinolates, which highlights the plant's high investment of resources into inflorescences. No induction of glucosinolates was detected in inflorescences, but the attack transiently affected the total concentration of soluble sugars in both leaves and inflorescences. We conclude that B.nigra evolved high constitutive rather than inducible resistance to protect their flowers; plants additionally compensated for damage by attackers viathe regrowth of reproductive parts. This strategy may be typical of annual plants.

Multiple resistance of silky windgrass to acetolactate synthase- and acetyl-CoA synthase–inhibiting herbicides

AbstractField and pot experiments were conducted in Greece to study the occurrence of resistance in silky windgrass to acetolactate synthase (ALS)- and acetyl-CoA synthase (ACCase)-inhibiting herbicides. Twenty-four populations of silky windgrass were examined in whole-plant response experiments. High levels of field-evolved resistance to chlorsulfuron (0% to 28% control in terms of fresh weight reduction) with the recommended field rates were confirmed in most silky windgrass populations. However, other ALS inhibitors, such as pyroxsulam and a premix of mesosulfuron-methyl and iodosulfuron, provided adequate control (76% to 100% in terms of fresh weight reduction) of most populations, except eight silky windgrass populations that were found to be cross-resistant to all ALS-inhibiting herbicides tested (i.e., chlorsulfuron, commercial mixture of mesosulfuron-methyl plus iodosulfuron, and pyroxsulam). Conversely, most silky windgrass populations were controlled effectively (90% to 100% in terms of fresh weight reduction) with the recommended field rates of ACCase inhibitors cycloxydim, clethodim, and pinoxaden, but five populations were also found to be resistant to clodinafop-propargyl (10% to 68% control in terms of fresh weight reduction). The ALS gene sequencing of the eight silky windgrass populations, with cross-resistance to ALS inhibitors, revealed a point mutation at the Pro-197 position, causing amino acid substitution by Ser or Thr in the ALS enzyme. Overall, chlorsulfuron and clodinafop-propargyl were selecting agents of field-evolved multiple resistance to ALS- and ACCase-inhibiting herbicides in five silky windgrass populations. As the available postemergence-applied chemistries/modes of action registered for grass weed control in cereals are rather limited, adopting integrated management practices and implementing proactive and reactive measures to delay the evolution of resistant populations is essential.

Target-site cross-resistance to ALS inhibitors in johnsongrass originating from Greek cornfields

AbstractFive johnsongrass populations collected from corn grown in northern Greece were studied to elucidate the levels and mechanisms of resistance to acetolactate synthase (ALS)- and acetyl-CoA carboxylase (ACCase)-inhibiting herbicides. Whole-plant response assays indicated that two populations were highly cross-resistant to all ALS inhibitors tested (foramsulfuron, nicosulfuron, rimsulfuron, and imazamox) but were effectively controlled by the recommended rate of the ACCase-inhibiting herbicides propaquizafop and clethodim. TheALSgene sequence revealed a point mutation that resulted in the substitution of Trp574 by Leu in the ALS enzyme, suggesting that the resistance mechanism is target-site mediated. These findings highlight a serious threat against the sustainable use of the ALS-inhibiting herbicides in controlling johnsongrass and other grass weeds in cornfields, suggesting rotational use of herbicides with different modes of action, along with the use of nonchemical methods, for viable Johnsongrass management.

Volatile-mediated plant-plant interactions: volatile organic compounds as modulators of receiver plant defence, growth, and reproduction.

It is firmly established that plants respond to biotic and abiotic stimuli by emitting volatile organic compounds (VOCs). These VOCs provide information on the physiological status of the emitter plant and are available for detection by the whole community. In the context of plant-plant interactions, research has focused mostly on the defence-related responses of receiver plants. However, responses may span hormone signalling and both primary and secondary metabolism, and ultimately affect plant fitness. Here we present a synthesis of plant-plant interactions, focusing on the effects of VOC exposure on receiver plants. An overview of the important chemical cues, the uptake and conversion of VOCs, and the adsorption of VOCs to plant surfaces is presented. This is followed by a review of the substantial VOC-induced changes to receiver plants affecting both primary and secondary metabolism and influencing plant growth and reproduction. Further research should consider whole-plant responses for the effective evaluation of the mechanisms and fitness consequences of exposure of the receiver plant to VOCs.

Whole-plant compensatory responses of isoprene emission from hybrid poplar seedlings exposed to elevated ozone

It is still unclear whether the responses of isoprene (ISO) emission to elevated O3 vary with biological organization level (i.e. leaf and whole-plant). To study such responses and the possible reasons explaining their variation, we investigated the effect of O3 (CF: charcoal-filtered ambient air; E-O3: non-filtered ambient air enriched with O3) on ISO emission rate (ISOrate), net photosynthetic rate (Pn), leaf nitrogen and carbon contents, and leaf growth traits in poplar seedlings (Populus deltoides cv. 55/56 × P. deltoides cv. Imperial) during one growing season. Opposite effects of E-O3 on Pn were found between upper leaves (positive effect) and lower leaves (negative effect). Compared to CF, E-O3 significantly decreased leaf mass per area, number of leaves, and leaf biomass, but increased leaf nitrogen content and individual leaf size. In the framework of such compensatory responses, poplar seedlings further increased ISOrate in upper leaves and decreased ISOrate in lower leaves, thus preventing significant decrease in the overall whole-plant ISOrate by E-O3. The measured whole-plant ISOrate also showed that the simplistic estimation approaches based on the linear regression between chlorophyll content indicated by soil plant analysis development meter (SPAD value) and leaf-level ISOrate could not accurately reflect the true response of whole plant to elevated O3. For more accurate predictions, the potential ISO compensatory response to increasing O3 concentration should be incorporated into the climate biogeochemical models related to ISO emission.

Artificial top-light is more efficient for tomato production than inter-light

Studies of whole-plant responses of tomato to light environments are limited and cannot be extrapolated from observations of seedlings or short-term crops in growth chambers. Effects of artificial light sources like high pressure sodium (HPS) and light emitting diodes (LED) are mainly studied as supplement to sunlight in greenhouses. Since natural sunlight is almost neglectable in Norway during wintertime, we could study effects of different types of artificial light on crop growth and production in tomato. The goal of this experiment was to quantify the effects of artificial HPS top-light, installed at the top of the canopy, and LED inter-light, installed between plant rows, on fresh and dry matter production and fruit quality of greenhouse tomatoes under controlled and documented conditions. Our aim was to optimize yield under different light conditions, while avoiding an unfavourable source-sink balance. Tomato plants were grown under HPS top light with an installed capacity of 161, 242 and 272 W m−2 combined with LED inter-light with an installed capacity of 0, 60 or 120 W m−2. We used stem diameter as a trait to regulate air temperature in different light treatments in order to retain plant vigour. Results show that both HPS top light and LED inter-light increased tomato yield. However, the positive effect of supplemental LED inter-light decreased at higher amounts of HPS top light. Under the conditions in this experiment, with neglectable incoming solar radiation, an installed amount of 242 Watt m-2 HPS top light and a daily light integral (DLI) of 30 mol m-2 day-1 resulted in best light use efficiency (in gram fresh tomato per mol). Addition of LED inter-light to HPS top light reduced light use efficiency. Results show that winter production using artificial light in Norway is more energy efficient compared to production under sunlight in southern countries. Results can be used for modelling purposes.

Transcriptional regulation of ZIP genes is independent of local zinc status in Brachypodium shoots upon zinc deficiency and resupply.

The biological processes underlying zinc homeostasis are targets for genetic improvement of crops to counter human malnutrition. Detailed phenotyping, ionomic, RNA-Seq analyses and flux measurements with 67 Zn isotope revealed whole-plant molecular events underlying zinc homeostasis upon varying zinc supply and during zinc resupply to starved Brachypodium distachyon (Brachypodium) plants. Although both zinc deficiency and excess hindered Brachypodium growth, accumulation of biomass and micronutrients into roots and shoots differed depending on zinc supply. The zinc resupply dynamics involved 1,893 zinc-responsive genes. Multiple zinc-regulated transporter and iron-regulated transporter (IRT)-like protein (ZIP) transporter genes and dozens of other genes were rapidly and transiently down-regulated in early stages of zinc resupply, suggesting a transient zinc shock, sensed locally in roots. Notably, genes with identical regulation were observed in shoots without zinc accumulation, pointing to root-to-shoot signals mediating whole-plant responses to zinc resupply. Molecular events uncovered in the grass model Brachypodium are useful for the improvement of staple monocots.

Cannabis Yield, Potency, and Leaf Photosynthesis Respond Differently to Increasing Light Levels in an Indoor Environment.

Since the recent legalization of medical and recreational use of cannabis (Cannabis sativa) in many regions worldwide, there has been high demand for research to improve yield and quality. With the paucity of scientific literature on the topic, this study investigated the relationships between light intensity (LI) and photosynthesis, inflorescence yield, and inflorescence quality of cannabis grown in an indoor environment. After growing vegetatively for 2 weeks under a canopy-level photosynthetic photon flux density (PPFD) of ≈425 μmol·m−2·s−1 and an 18-h light/6-h dark photoperiod, plants were grown for 12 weeks in a 12-h light/12-h dark “flowering” photoperiod under canopy-level PPFDs ranging from 120 to 1,800 μmol·m−2·s−1 provided by light emitting diodes. Leaf light response curves varied both with localized (i.e., leaf-level) PPFD and temporally, throughout the flowering cycle. Therefore, it was concluded that the leaf light response is not a reliable predictor of whole-plant responses to LI, particularly crop yield. This may be especially evident given that dry inflorescence yield increased linearly with increasing canopy-level PPFD up to 1,800 μmol·m−2·s−1, while leaf-level photosynthesis saturated well-below 1,800 μmol·m−2·s−1. The density of the apical inflorescence and harvest index also increased linearly with increasing LI, resulting in higher-quality marketable tissues and less superfluous tissue to dispose of. There were no LI treatment effects on cannabinoid potency, while there were minor LI treatment effects on terpene potency. Commercial cannabis growers can use these light response models to determine the optimum LI for their production environment to achieve the best economic return; balancing input costs with the commercial value of their cannabis products.

A Multi-Species Analysis Defines Anaplerotic Enzymes and Amides as Metabolic Markers for Ammonium Nutrition.

Nitrate and ammonium are the main nitrogen sources in agricultural soils. In the last decade, ammonium (NH4+), a double-sided metabolite, has attracted considerable attention by researchers. Its ubiquitous presence in plant metabolism and its metabolic energy economy for being assimilated contrast with its toxicity when present in high amounts in the external medium. Plant species can adopt different strategies to maintain NH4+ homeostasis, as the maximization of its compartmentalization and assimilation in organic compounds, primarily as amino acids and proteins. In the present study, we report an integrative metabolic response to ammonium nutrition of seven plant species, belonging to four different families: Gramineae (ryegrass, wheat, Brachypodium distachyon), Leguminosae (clover), Solanaceae (tomato), and Brassicaceae (oilseed rape, Arabidopsis thaliana). We use principal component analysis (PCA) and correlations among metabolic and biochemical data from 40 experimental conditions to understand the whole-plant response. The nature of main amino acids is analyzed among species, under the hypothesis that those Asn-accumulating species will show a better response to ammonium nutrition. Given the provision of carbon (C) skeletons is crucial for promotion of the nitrogen assimilation, the role of different anaplerotic enzymes is discussed in relation to ammonium nutrition at a whole-plant level. Among these enzymes, isocitrate dehydrogenase (ICDH) shows to be a good candidate to increase nitrogen assimilation in plants. Overall, metabolic adaptation of different carbon anaplerotic activities is linked with the preference to synthesize Asn or Gln in their organs. Lastly, glutamate dehydrogenase (GDH) reveals as an important enzyme to surpass C limitation during ammonium assimilation in roots, with a disparate collaboration of glutamine synthetase (GS).

New field wind manipulation methodology reveals adaptive responses of steppe plants to increased and reduced wind speed

BackgroundWind strongly impacts plant growth, leaf traits, biomass allocation, and stem mechanical properties. However, whether there are common whole-plant wind responses among different plant species is still unclear. We tested this null hypothesis by exposing four eudicot steppe species to three different wind treatments in a field experiment: reduced wind velocity using windbreaks, ambient wind velocity, and enhanced wind velocity through a novel methodology using wind-funneling baffles.ResultsAcross the four species, wind generally decreased plant height, projected crown area, and stepwise bifurcation ratio, and increased root length and stem base diameter. In contrast, the response patterns of shoot traits, especially mechanical properties, to wind velocity were idiosyncratic among species. There was no significant difference in total biomass among different treatments; this might be because the negative effects on heat dissipation and photosynthesis of low wind speed during hot periods, could counteract positive effects during favorable cooler periods.ConclusionsThere are common wind response patterns in plant-size-related traits across different steppe species, while the response patterns in shoot traits vary among species. This indicates the species-specific ways by which plants balance growth and mechanical support facing wind stress. Our new field wind manipulation methodology was effective in altering wind speed with the intended magnitude. Especially, our field wind-funneling baffle system showed a great potential for use in future field wind velocity enhancement. Further experiments are needed to reveal how negative and positive effects play out on whole-plant performance in response to different wind regimes, which is important as ongoing global climatic changes involve big changes in wind regimes.