UNDERSTANDING PROMISCUITY: WHEN IS SEEKING ADDITIONAL MATES BETTER THAN GUARDING AN ALREADY FOUND ONE?

Paternity protection and the acquisition of multiple mates select for different traits. The consensus from theoretical work is that mate‐guarding intensifies with an increasing male bias in the adult sex ratio (ASR). A male bias can thus lead to male monogamy if guarding takes up the entire male time budget. Given that either female‐ or male‐biased ASRs are possible, why is promiscuity clearly much more common than male monogamy? We address this question with two models, differing in whether males can assess temporal cues of female fertility. Our results confirm the importance of the ASR: guarding durations increase with decreasing female availability and increasing number of male competitors. However, several factors prevent the mating system from switching to male monogamy as soon as the ASR becomes male biased. Inefficient guarding, incomplete last male sperm precedence, any mechanism that allows sperm to fertilize eggs after the male's departure, and (in some cases) the unfeasibility of precopulatory guarding all help explain cases where promiscuity exists on its own or alongside temporally limited mate‐guarding. Shortening the window of fertilization shifts guarding time budgets from the postcopulatory to the precopulatory stage.     

‘Real time’ genetic manipulation: a new tool for ecological field studies

Field experiments with transgenic plants often reveal the functional significance of genetic traits that are important for the performance of the plants in their natural environments. Until now, only constitutive overexpression, ectopic expression and gene silencing methods have been used to analyze gene‐related phenotypes in natural habitats. These methods do not allow sufficient control over gene expression for the study of ecological interactions in real time, of genetic traits that play essential roles in development, or of dose‐dependent effects. We applied the sensitive dexamethasone (DEX)‐inducible pOp6/LhGR expression system to the ecological model plant Nicotiana attenuata and established a lanolin‐based DEX application method to facilitate ectopic gene expression and RNA interference‐mediated gene silencing in the field and under challenging conditions (e.g. high temperature, wind and UV radiation). Fully established field‐grown plants were used to silence phytoene desaturase and thereby cause photobleaching only in specific plant sectors, and to activate expression of the cytokinin (CK) biosynthesis gene isopentenyl transferase (ipt). We used ipt expression to analyze the role of CKs in both the glasshouse and the field to understand resistance to the native herbivore Tupiocoris notatus, which attacks plants at small spatial scales. By spatially restricting ipt expression and elevating CK levels in single leaves, damage by T. notatus increased, demonstrating the role of CKs in this plant–herbivore interaction at a small scale. As the arena of most ecological interactions is highly constrained in time and space, these tools will advance the genetic analysis of dynamic traits that matter for plant performance in nature.     

QTL‐seq: rapid mapping of quantitative trait loci in rice by whole genome resequencing of DNA from two bulked populations

The majority of agronomically important crop traits are quantitative, meaning that they are controlled by multiple genes each with a small effect (quantitative trait loci, QTLs). Mapping and isolation of QTLs is important for efficient crop breeding by marker‐assisted selection (MAS) and for a better understanding of the molecular mechanisms underlying the traits. However, since it requires the development and selection of DNA markers for linkage analysis, QTL analysis has been time‐consuming and labor‐intensive. Here we report the rapid identification of plant QTLs by whole‐genome resequencing of DNAs from two populations each composed of 20–50 individuals showing extreme opposite trait values for a given phenotype in a segregating progeny. We propose to name this approach QTL‐seq as applied to plant species. We applied QTL‐seq to rice recombinant inbred lines and F₂ populations and successfully identified QTLs for important agronomic traits, such as partial resistance to the fungal rice blast disease and seedling vigor. Simulation study showed that QTL‐seq is able to detect QTLs over wide ranges of experimental variables, and the method can be generally applied in population genomics studies to rapidly identify genomic regions that underwent artificial or natural selective sweeps.     

Site‐specific T–DNA integration in Arabidopsis thaliana mediated by the combined action of CRE recombinase and ϕC31 integrase

Random T–DNA integration into the plant host genome can be problematic for a variety of reasons, including potentially variable transgene expression as a result of different integration positions and multiple T–DNA copies, the risk of mutating the host genome and the difficulty of stacking well‐defined traits. Therefore, recombination systems have been proposed to integrate the T–DNA at a pre‐selected site in the host genome. Here, we demonstrate the capacity of the ?C31 integrase (INT) for efficient targeted T–DNA integration. Moreover, we show that the iterative site‐specific integration system (ISSI), which combines the activities of the CRE recombinase and INT, enables the targeting of genes to a pre‐selected site with the concomitant removal of the resident selectable marker. To begin, plants expressing both the CRE and INT recombinase and containing the target attP site were constructed. These plants were supertransformed with a T–DNA vector harboring the loxP site, the attB sites, a selectable marker and an expression cassette encoding a reporter protein. Three out of the 35 transformants obtained (9%) showed transgenerational site‐specific integration (SSI) of this T–DNA and removal of the resident selectable marker, as demonstrated by PCR, Southern blot and segregation analysis. In conclusion, our results show the applicability of the ISSI system for precise and targeted Agrobacterium‐mediated integration, allowing the serial integration of transgenic DNA sequences in plants.     

Phenoscope: an automated large‐scale phenotyping platform offering high spatial homogeneity

Increased phenotyping accuracy and throughput are necessary to improve our understanding of quantitative variation and to be able to deconstruct complex traits such as those involved in growth responses to the environment. Still, only a few facilities are known to handle individual plants of small stature for non‐destructive, real‐time phenotype acquisition from plants grown in precisely adjusted and variable experimental conditions. Here, we describe Phenoscope, a high‐throughput phenotyping platform that has the unique feature of continuously rotating 735 individual pots over a table. It automatically adjusts watering and is equipped with a zenithal imaging system to monitor rosette size and expansion rate during the vegetative stage, with automatic image analysis allowing manual correction. When applied to Arabidopsis thaliana, we show that rotating the pots strongly reduced micro‐environmental disparity: heterogeneity in evaporation was cut by a factor of 2.5 and the number of replicates needed to detect a specific mild genotypic effect was reduced by a factor of 3. In addition, by controlling a large proportion of the micro‐environmental variance, other tangible sources of variance become noticeable. Overall, Phenoscope makes it possible to perform large‐scale experiments that would not be possible or reproducible by hand. When applied to a typical quantitative trait loci (QTL) mapping experiment, we show that mapping power is more limited by genetic complexity than phenotyping accuracy. This will help to draw a more general picture as to how genetic diversity shapes phenotypic variation.     

Mate sampling and choosiness in the sand goby

To date, mate choice studies have mostly focused on establishing which mates are chosen or how the choices are performed. Here, we combined these two approaches by empirically testing how latency to mate is affected by various search costs, variation in mate quality and female quality in the sand goby (Pomatoschistus minutus). Our results show that females adjust their mating behaviour according to the costs and benefits of the choice situation. Specifically, they mated sooner when access to males was delayed and when the presence of other females presented a mate sampling cost. We also found a positive link between size variation among potential mating partners and spawning delay in some (but not all) experimental conditions. By contrast, we did not find the number of available males or the females'' own body size (‘quality’) to affect mating latency. Finally, female mating behaviour varied significantly between years. These findings are notable for demonstrating that (i) mate sampling time is particularly sensitive to costs and, to a lesser degree, to variation among mate candidates, (ii) females'' mating behaviour is sensitive to qualitative rather than to quantitative variation in their environment, and (iii) a snapshot view may describe mate sampling behaviour unreliably.     

The ideal free distribution as an evolutionarily stable strategy

We examine the evolutionary stability of strategies for dispersal in heterogeneous patchy environments or for switching between discrete states (e.g. defended and undefended) in the context of models for population dynamics or species interactions in either continuous or discrete time. There have been a number of theoretical studies that support the view that in spatially heterogeneous but temporally constant environments there will be selection against unconditional, i.e. random, dispersal, but there may be selection for certain types of dispersal that are conditional in the sense that dispersal rates depend on environmental factors. A particular type of dispersal strategy that has been shown to be evolutionarily stable in some settings is balanced dispersal, in which the equilibrium densities of organisms on each patch are the same whether there is dispersal or not. Balanced dispersal leads to a population distribution that is ideal free in the sense that at equilibrium all individuals have the same fitness and there is no net movement of individuals between patches or states. We find that under rather general assumptions about the underlying population dynamics or species interactions, only such ideal free strategies can be evolutionarily stable. Under somewhat more restrictive assumptions (but still in considerable generality), we show that ideal free strategies are indeed evolutionarily stable. Our main mathematical approach is invasibility analysis using methods from the theory of ordinary differential equations and nonnegative matrices. Our analysis unifies and extends previous results on the evolutionary stability of dispersal or state-switching strategies.     

Phenological responses to small-scale spatial variation in snowmelt timing reveal compensatory and conservative strategies in subarctic-alpine plants

Background: In tundra ecosystems, the adjustment of phenological events, such as bud burst, to snowmelt timing is crucial to the climatic adaptation of plants. Natural small-scale variations in microclimate potentially enable plant populations to persist in a changing climate.

Aims: To assess how plant phenology responds to natural differences in snowmelt timing.

Methods: We observed the timing of eight vegetative and reproductive phenophases in seven dwarf-shrub species in relation to differences in snowmelt timing on a small spatial scale in an alpine environment in subarctic Finland.

Results: Some species and phenophases showed accelerated development with later snowmelt, thus providing full or partial compensation for the shorter snow-free period. Full compensation resulted in synchronous occurrence of phenophases across the snowmelt gradient. In other species, there was no acceleration of development. The timing of phenophases varied between two consecutive years and two opposing mountain slope aspects.

Conclusions: The results have shown three distinct patterns in the timing of phenophases in relation to snowmelt and suggest alternative strategies for adaptation to snowmelt timing. These strategies potentially apply to other species and tundra ecosystems and provide a framework, enabling one to compare and generalise phenological responses to snowmelt timing under different future climate scenarios.     

Bioethanol potential from miscanthus with low ILUC risk in the province of Lublin,Poland

Increasing production of biofuels has led to concerns about indirect land‐use change (ILUC). So far, significant efforts have been made to assess potential ILUC effects. But limited attention has been paid to strategies for reducing the extent of ILUC and controlling the type of LUC. This case study assesses five key ILUC mitigation measures to quantify the low‐ILUC‐risk production potential of miscanthus‐based bioethanol in Lublin province (Poland) in 2020. In 2020, a total area of 196 to 818 thousand hectare of agricultural land could be made available for biomass production by realizing above‐baseline yield developments (95–413 thousand ha), increased food chain efficiencies (9–30 thousand ha) and biofuel feedstock production on underutilized lands (92–375 thousand ha). However, a maximum 203–269 thousand hectare is considered legally available (not protected) and biophysically suitable for miscanthus production. The resulting low‐ILUC‐risk bioethanol production potential ranges from 12 to 35 PJ per year. The potential from this region alone is higher than the national Polish target for second‐generation bioethanol consumption of 9 PJ in 2020. Although the sustainable implementation potential may be lower, the province of Lublin could play a key role in achieving this target. This study shows that the mitigation or prevention of ILUC from bioenergy is only possible when an integrated perspective is adopted on the agricultural and bioenergy sectors. Governance and policies on planning and implementing ILUC mitigation are considered vital for realizing a significant bioenergy potential with low ILUC risk. One important aspect in this regard is monitoring the risk of ILUC and the implementation of ILUC mitigation measures. Key parameters for monitoring are land use, land cover and crop yields.