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Many previous studies conclude that pre-zygotic barriers such as mechanical isolation account for most reproductive isolation between pairs of taxa. However, the inheritance and persistence of barriers such as these after the first generation of hybridization is rarely quantified, even though it is a vital consideration in understanding gene flow potential. There is an asymmetrical pre-zygotic mechanical barrier to hybridization between Eucalyptus nitens and Eucalyptus globulus, which completely prevents small-flowered E. nitens pollen from mating with large E. globulus flowers, while the reverse cross is possible. We aimed to determine the relative importance of pre- and post-zygotic barriers in preventing gene flow following secondary contact between E. nitens and E. globulus, including the inheritance of barriers in advanced-generation hybrids. Experimental crossing was used to produce outcrossed E. nitens, E. globulus and their F1, F2, BCg and BCn hybrids. The strength and inheritance of a suite of pre- and post-zygotic barriers were assessed, including 20-year survival, growth and reproductive capacity. The mechanical barrier to hybridization was lost or greatly reduced in the F1 hybrid. In contrast, intrinsic post-zygotic barriers were strong and persistent. Line-cross analysis indicated that the outbreeding depression in the hybrids was best explained by epistatic loss. The removal of strong mechanical barriers between E. nitens and E. globulus allows F1 hybrids to act as a bridge for bi-directional gene flow between these species. However, strong and persistent post-zygotic barriers exist, meaning that wherever F1 hybridization does occur, intrinsic post-zygotic barriers will be responsible for most reproductive isolation in this system. This potential transient nature of mechanical barriers to zygote formation due to additive inheritance in hybrids appears under-appreciated, and highlights the often important role that intrinsic post-mating barriers play in maintaining species boundaries at zones of secondary contact.

The larger distribution area of asexuals compared with their sexual relatives in geographical parthenogenesis (GP) scenarios has been widely attributed to the advantages of uniparental reproduction and polyploidy. However, potential disadvantages of sexuals due to their breeding system have received little attention so far. Here, we study the breeding system of five narrowly distributed sexual lineages of Ranunculus notabilis s.l. (R. auricomus complex) and its effects on outcrossing, inbreeding, female fitness, and heterozygosity. We performed selfing and intra‐ and interlineage crossings by bagging 481 flowers (59 garden individuals) followed by germination experiments. We compared seed set and germination rates, and related them to genetic distance and genome‐wide heterozygosity (thousands of RADseq loci). Selfings (2.5%) unveiled a significantly lower seed set compared with intra‐ (69.0%) and interlineage crossings (69.5%). Seed set of intra‐ (65%) compared to interpopulation crossings (78%) was significantly lower. In contrast, all treatments showed comparable germination rates (32%–43%). Generalized linear regressions between seed set and genetic distance revealed positive relationships in general and between lineages, and a negative one within lineages. Seed set was the main decisive factor for female fitness. Germination rates were not related to genetic distance at any level, but were positively associated with heterozygosity in interlineage crossings. Experiments confirmed full crossability and predominant outcrossing among sexual R. notabilis s.l. lineages. However, up to 5% (outliers 15%–31%) of seeds were formed by selfing, probably due to semi‐self‐compatibility in a multi‐locus gametophytic SI system. Less seed set in intrapopulation crossings, and higher seed set and germination rates from crossings of genetically more distant and heterozygous lineages (interlineage) indicate negative inbreeding and positive outbreeding effects. In GP scenarios, sexual species with small and/or isolated populations can suffer from decreased female fitness due to their breeding system. This factor, among others, probably limits range expansion of sexuals. Potential disadvantages of sexual plant species due to their breeding system have received little attention in geographical parthenogenesis (GP) scenarios so far. We compared seed set and germination rates of five sexual lineages of Ranunculus notabilis s.l (R. auricomus complex) and related them to genetic distance and genome‐wide heterozygosity. Results suggest that inbreeding depression particularly affects intrapopulation (intralineage) crossings, and that positive outbreeding effects particularly influence interlineage crossings. In GP scenarios, sexual species with small and/or isolated populations can suffer from decreased female fitness due to their breeding system.

Reproductive barriers to gene flow play a key role in speciation. However, as it is not always feasible to study them directly, most studies rely on genetic divergence to infer species delimitation. In order to correlate genetic distances to reproductive incompatibilities, compact groups of closely related species are needed. In this work, we explored a species complex of Baikal amphipods (Crustacea: Amphipoda: Gammaroidea), Eulimnogammarus verrucosus. Three biological species (W, S, and E), geographically isolated in Baikal, had been found to have interspecific differences exceeding the patristic distance threshold of 0.16, and a postzygotic incompatibility had been confirmed for the closest pair, W and S. Here, we expanded our knowledge on geographical distribution of the species, discovering that secondary contact between the W and S species already occurs in natural conditions near the source of the Angara River. Our experiments have shown that the three species within the E. verrucosus species complex are separated by both prezygotic and postzygotic barriers. While neither of these barriers is absolute, their combination can ensure reproductive isolation upon secondary contact of the species. The experimental system we have developed in this and previous works can provide support for testing species delimitation hypotheses based on sequencing data and further extend these results to related species for which such experiments are unfeasible.

In this study, we define a chi-square random field on a multi-dimensional lattice points index set with a direct product covariance structure and consider the distribution of the maximum of this random field. We provide two approximate formulas for the upper tail probability of the distribution based on nonlinear renewal theory and an integral-geometric approach called the volume-of-tube method. This study is motivated by the detection problem of the interactive loci pairs which play an important role in forming biological species. The joint distribution of scan statistics for detecting the pairs is regarded as the chi-square random field above, and hence the multiplicity-adjusted p -value can be calculated using the proposed approximate formulas. By using these formulas, we examine the data of Mizuta, Harushima and Kurata (Proc Nat Acad Sci USA 107(47):20417–20422, 2010 ) who reported a new interactive loci pair of rice inter-subspecies.

Experimental hybridization of X0 and XY chromosome races of the brachypterous grasshopper P. sapporensis did not reveal pre-zygotic reproductive isolation. However, a partial zygotic barrier was found between the X0-standard race from Shimokawa and XY-standard chromosome race from Akan. Approximately 40% of embryos from females crossed with males from other chromosome races developed parthenogenetically, the remaining embryos were normal heterozygotes. Adult F1 males and females from crosses of this type had properly developed testes and ovaries. Non-sister associations and other irregularities in meiosis were not observed in male meiosis. Crossing experiments demonstrated that hybrids between X0 and XY races occur to some extent. The absence of a hybrid zone between the X0 and XY chromosome races may be the result of selection against heterozygotes. Crosses between the XY-Tanno and X0-standard (Teine) subraces resulted in F1 and F2 generations in spite of the many chromosome differences between them such as a X-A translocation and fixed pericentric inversions in four pairs of autosomes. These results do not support the hypothesis that chromosomal differences play a key role in restricting gene flow between the X0 and XY races of P. sapporensis.

In view of the key role of undergraduate experimental teaching reform in cultivating high-quality talents with both innovative spirit and practical ability, this paper deeply discusses multi-dimensional reform strategies. Specifically, the teaching mode of \"double teachers for every student\" is innovatively introduced, and scientific research projects are deeply integrated into undergraduate experimental teaching, aiming at realizing the modern development of teaching content and the diversified expansion of teaching methods. By designing and applying the undergraduate experimental teaching platform for intelligent limb rehabilitation training based on the concept of \"medical-engineering interdisciplinary crossing\", it not only builds a bridge for students to contact cutting-edge scientific research and strengthen practical skills, but also provides valuable ideas and practical models for the innovation of undergraduate experimental teaching. In the future, with the continuous optimization and upgrading of platform functions, it is expected to provide students with a richer and richer learning experience and comprehensively promote students' overall quality.

1. Millions of dollars are spent on wildlife crossing structures intended to reduce the barrier effects of roads on wildlife. However, we know little about the degree to which these structures facilitate dispersal and gene flow. 2. Our study incorporates two elements that are rarely used in the evaluation of wildlife crossing structures: an experimental design including a before and after comparison, and the use of genetic techniques to demonstrate effects on gene flow at both population and individual levels. We evaluated the effect of wildlife crossing structures (canopy bridges and glider poles) on a gliding mammal, the squirrel glider (Petaurus norfolcensis). We genotyped 399 individuals at eight microsatellite markers to analyse population structure, first-generation migrants and parentage relationships. 3. We found that the freeway was not a complete genetic barrier, with a strong effect evident at only one site. We hypothesise that the presence of corridors alongside the freeway and throughout the surrounding landscape facilitated circuitous detours for squirrel gliders. 4. Installing a crossing structure at the location with a strong barrier effect restored gene flow within just 5 years of mitigation. 5. Synthesis and applications. Our study highlights the importance of using genetic techniques not just to evaluate the success of road-crossing structures for wildlife, but also to guide their placement within the landscape. Managers wishing to reduce the effects of linear infrastructure on squirrel gliders and other arboreal mammals should aim to preserve and enhance vegetation along roadsides and within centre medians, as well as mitigate large gaps by implementing wildlife crossing structures.

Participatory plant breeding (PPB) has been practiced for several reasons, including sociological, humanitarian, and egalitarian. This paper aims to demonstrate that PPB should be practiced simply because it increases plant breeding efficiency, which is defined as (i) the ratio between the number of varieties adopted and the number of crosses made, (ii) the response to selection, and (iii) the benefit/cost ratio, not only as often done by public breeding programs, by the number of varieties released by public breeding programs. After reviewing the reasons for the lack of adoption of several of the varieties released, and the theoretical basis justifying the use of correlated response to measure selection gains, the issue of the benefit/cost ratio is discussed within the context of adoption rates. The assumption is that without adoption, no benefit will occur. Ways to increase the three measures of breeding efficiency are discussed in detail, and the conclusion is made that the three measures can be increased by combining decentralized selection with farmers' participation in a PPB program. The essential features of a PPB program are reviewed, including the experimental designs and statistical analysis used to increase the precision of on-farm trials. The findings show that PPB increases breeding efficiency, both in terms of response to selection measured over the time from the initial cross to adoption and in terms of the benefit/cost ratio as a consequence of a higher adoption rate. In a PPB program, adoption starts during the process of selection and precedes variety release. In a conventional plant breeding (CPB) program, the sequence is reversed. Other differences between a PPB and a CPB program include the increase of agrobiodiversity, which in a PPB program is higher because of the rapid spatiotemporal turnover of varieties, and because the seed of new varieties is readily available to farmers, thus contributing to food security. Because of its decentralized nature, PPB can accommodate organic farms and become easily adopted by both national and international public breeding programs as one way of adapting crops to climate changes.

The goal of many plant scientists' research is to explain natural phenotypic variation in terms of simple changes in DNA sequence. Traditionally, linkage mapping has been the most commonly employed method to reach this goal: experimental crosses are made to generate a family with known relatedness, and attempts are made to identify cosegregation of genetic markers and phenotypes within this family. In vertebrate systems, association mapping (also known as linkage disequilibrium mapping) is increasingly being adopted as the mapping method of choice. Association mapping involves searching for genotype-phenotype correlations in unrelated individuals and often is more rapid and cost-effective than traditional linkage mapping. We emphasize here that linkage and association mapping are complementary approaches and are more similar than is often assumed. Unlike in vertebrates, where controlled crosses can be expensive or impossible (e.g., in humans), the plant scientific community can exploit the advantages of both controlled crosses and association mapping to increase statistical power and mapping resolution. While the time and money required for the collection of genotype data were critical considerations in the past, the increasing availability of inexpensive DNA sequencing and genotyping methods should prompt researchers to shift their attention to experimental design. This review provides thoughts on finding the optimal experimental mix of association mapping using unrelated individuals and controlled crosses to identify the genes underlying phenotypic variation.

The last 20 years have seen a dramatic increase in efforts to mitigate the negative effects of roads and traffic on wildlife, including fencing to prevent wildlife-vehicle collisions and wildlife crossing structures to facilitate landscape connectivity. While not necessarily explicitly articulated, the fundamental drivers behind road mitigation are human safety, animal welfare, and/or wildlife conservation. Concomitant with the increased effort to mitigate has been a focus on evaluating road mitigation. So far, research has mainly focussed on assessing the use of wildlife crossing structures, demonstrating that a broad range of species use them. However, this research has done little to address the question of the effectiveness of crossing structures, because use of a wildlife crossing structure does not necessarily equate to its effectiveness. The paucity of studies directly examining the effectiveness of crossing structures is exacerbated by the fact that such studies are often poorly designed, which limits the level of inference that can be made. Without well performed evaluations of the effectiveness of road mitigation measures, we may endanger the viability of wildlife populations and inefficiently use financial resources by installing structures that are not as effective as we think they are. In this paper we outline the essential elements of a good experimental design for such assessments and prioritize the parameters to be measured. The framework we propose will facilitate collaboration between road agencies and scientists to undertake research programs that fully evaluate effectiveness of road mitigation measures. We discuss the added value of road mitigation evaluations for policy makers and transportation agencies and provide recommendations on how to incorporate such evaluations in road planning practices.