Hybridization Barriers Research Articles

“In Vitro” Ovule Culture to Improve Genetic Variability in Hydrangea macrophylla

In flowering plants, such as Hydrangea macrophylla, the main breeding objective is to increase genetic variability in ornamental traits. This study investigates in vitro techniques, through ovule culture, to overcome the hybridization barriers and increase the efficiency of crossing in Hydrangea macrophylla in which breeding has been hampered by a fairly long breeding cycle and lack of information about its genetic resources. Two different types of media were compared, Gamborg B5 and Murashige and Skoog basal salts, to verify the germination rate of immature ovules in different intraspecific crosses. The germination rate and viability of the seedlings were influenced by the parental genotypes in the different combinations of crossing, highlighting, in some cases, the poor compatibility between some of them. The crossing combination “Parental A × Parental B”, showed the highest germinated ovules percentage (78.3%). The media used seem to less affect the ovule germination while mainly influencing the development and growth of the young seedlings and in particular the number of leaves, the branching attitude, and root length, with the Gamborg medium determining up to a 30% increase, compared to MS medium. In addition, we tested the effectiveness of using SSR markers to assess the parentage of the putative hybrids even though only three out of twelve SSR markers showed allelism. Although the number of SSR markers was low, they were allowed to profile the parentage according to Mendelian laws.

A recessive LRR-RLK gene causes hybrid breakdown in cotton.

An LRR-RLK gene causing interspecific hybrid breakdown between Gossypium. anomalum and G. hirsutum was identified by deploying a map-based cloning strategy. The self-destructing symptoms of hybrid incompatibility in most cases are attributed to autoimmunity. The cloning of genes responsible for hybrid incompatibility in cotton is helpful to clarify the mechanisms underlying hybrid incompatibility and can break the barriers in distant hybridization. In this study, a temperature-dependent lethality was identified in CSSL11-9 (chromosome segment substitution line) with Gossypium anomalum chromosome segment on chromosome A11. Transcriptome analysis showed the differentially expressed genes related to autoimmune responses were highly enriched, suggesting that expression of CSSL11-9 plant lethal gene activated autoimmunity in the absence of any pathogen or external stimulus, inducing programmed cell death (PCD) and causing a lethal phenotype. The lethal phenotype was controlled by a pair of recessive genes and then fine mapped between JAAS3191-JAAS3050 interval, which covered 63.87 kb in G. hirsutum genome and 98.66 kb in G. anomalum. We demonstrated that an LRR-RLK gene designated as hybrid breakdown 1 (GoanoHBD1) was the causal gene underlying this locus for interspecific hybrid incompatibility between G. anomalum and G. hirsutum. Silencing this LRR-RLK gene could restore CSSL11-9 plants from a lethal to a normal phenotype. Our findings provide new insights into reproductive isolation and may benefit cotton breeding.

Genetic and environmental manipulation of Arabidopsis hybridization barriers uncovers antagonistic functions in endosperm cellularization.

Speciation involves reproductive isolation, which can occur by hybridization barriers acting in the endosperm of the developing seed. The nuclear endosperm is a nutrient sink, accumulating sugars from surrounding tissues, and undergoes coordinated cellularization, switching to serve as a nutrient source for the developing embryo. Tight regulation of cellularization is therefore vital for seed and embryonic development. Here we show that hybrid seeds from crosses between Arabidopsis thaliana as maternal contributor and A. arenosa or A. lyrata as pollen donors result in an endosperm based post-zygotic hybridization barrier that gives rise to a reduced seed germination rate. Hybrid seeds display opposite endosperm cellularization phenotypes, with late cellularization in crosses with A. arenosa and early cellularization in crosses with A. lyrata. Stage specific endosperm reporters display temporally ectopic expression in developing hybrid endosperm, in accordance with the early and late cellularization phenotypes, confirming a disturbance of the source-sink endosperm phase change. We demonstrate that the hybrid barrier is under the influence of abiotic factors, and show that a temperature gradient leads to diametrically opposed cellularization phenotype responses in hybrid endosperm with A. arenosa or A. lyrata as pollen donors. Furthermore, different A. thaliana accession genotypes also enhance or diminish seed viability in the two hybrid cross-types, emphasizing that both genetic and environmental cues control the hybridization barrier. We have identified an A. thaliana MADS-BOX type I family single locus that is required for diametrically opposed cellularization phenotype responses in hybrid endosperm. Loss of AGAMOUS-LIKE 35 significantly affects the germination rate of hybrid seeds in opposite directions when transmitted through the A. thaliana endosperm, and is suggested to be a locus that promotes cellularization as part of an endosperm based mechanism involved in post-zygotic hybrid barriers. The role of temperature in hybrid speciation and the identification of distinct loci in control of hybrid failure have great potential to aid the introduction of advantageous traits in breeding research and to support models to predict hybrid admixture in a changing global climate.

Structural evidence for MADS-box type I family expansion seen in new assemblies of Arabidopsis arenosa and A. lyrata.

Arabidopsis thaliana diverged from A. arenosa and A. lyrata at least 6 million years ago. The three species differ by genome-wide polymorphisms and morphological traits. The species are to a high degree reproductively isolated, but hybridization barriers are incomplete. A special type of hybridization barrier is based on the triploid endosperm of the seed, where embryo lethality is caused by endosperm failure to support the developing embryo. The MADS-box type I family of transcription factors is specifically expressed in the endosperm and has been proposed to play a role in endosperm-based hybridization barriers. The gene family is well known for its high evolutionary duplication rate, as well as being regulated by genomic imprinting. Here we address MADS-box type I gene family evolution and the role of type I genes in the context of hybridization. Using two de-novo assembled and annotated chromosome-level genomes of A. arenosa and A. lyrata ssp. petraea we analyzed the MADS-box type I gene family in Arabidopsis to predict orthologs, copy number, and structural genomic variation related to the type I loci. Our findings were compared to gene expression profiles sampled before and after the transition to endosperm cellularization in order to investigate the involvement of MADS-box type I loci in endosperm-based hybridization barriers. We observed substantial differences in type-I expression in the endosperm of A. arenosa and A. lyrata ssp. petraea, suggesting a genetic cause for the endosperm-based hybridization barrier between A. arenosa and A.lyrata ssp. petraea.

ECS1 and ECS2 suppress polyspermy and the formation of haploid plants by promoting double fertilization.

The current pace of crop plant optimization is insufficient to meet future demands and there is an urgent need for novel breeding strategies. It was previously shown that plants tolerate the generation of triparental polyspermy-derived plants and that polyspermy can bypass hybridization barriers. Polyspermy thus has the potential to harness previously incompatible climate-adapted wild varieties for plant breeding. However, factors that influence polyspermy frequencies were not previously known. The endopeptidases ECS1 and ECS2 have been reported to prevent the attraction of supernumerary pollen tubes by cleaving the pollen tube attractant LURE1. Here, we show that these genes have an earlier function that is manifested by incomplete double fertilization in plants defective for both genes. In addition to supernumerary pollen tube attraction, ecs1 ecs2 mutants exhibit a delay in synergid disintegration, are susceptible to heterofertilization, and segregate haploid plants that lack a paternal genome contribution. Our results thus uncover ECS1 and ECS2 as the first female factors triggering the induction of maternal haploids. Capitalizing on a high-throughput polyspermy assay, we in addition show that the double mutant exhibits an increase in polyspermy frequencies. As both haploid induction and polyspermy are valuable breeding aims, our results open new avenues for accelerated generation of climate-adapted cultivars.

Genetic variation in a tepary bean (Phaseolus acutifolius A. Gray) diversity panel reveals loci associated with biotic stress resistance.

Tepary bean (Phaseolus acutifolius A. Gray), indigenous to the arid climates of northern Mexico and the Southwest United States, diverged from common bean (Phaseolus vulgaris L.), approximately 2 million years ago and exhibits a wide range of resistance to biotic stressors. The tepary genome is highly syntenic to the common bean genome providing a foundation for discovery and breeding of agronomic traits between these two crop species. Although a limited number of adaptive traits from tepary bean have been introgressed into common bean, hybridization barriers between these two species required the development of bridging lines to alleviate this barrier. Thus, to fully utilize the extant tepary bean germplasm as both a crop and as a donor of adaptive traits, we developed a diversity panel of 422 cultivated, weedy, and wild tepary bean accessions which were then genotyped and phenotyped to enable population genetic analyses and genome-wide association studies for their response to a range of biotic stressors. Population structure analyses of the panel revealed eight subpopulations and the differentiation of botanical varieties within P. acutifolius. Genome-wide association studies revealed loci and candidate genes underlying biotic stress resistance including quantitative trait loci for resistance to weevils, common bacterial blight, Fusarium wilt, and bean common mosaic necrosis virus that can be harnessed not only for tepary bean but also common bean improvement.

Seedlessness Trait and Genome Editing-A Review.

Parthenocarpy and stenospermocarpy are the two mechanisms underlying the seedless fruit set program. Seedless fruit occurs naturally and can be produced using hormone application, crossbreeding, or ploidy breeding. However, the two types of breeding are time-consuming and sometimes ineffective due to interspecies hybridization barriers or the absence of appropriate parental genotypes to use in the breeding process. The genetic engineering approach provides a better prospect, which can be explored based on an understanding of the genetic causes underlying the seedlessness trait. For instance, CRISPR/Cas is a comprehensive and precise technology. The prerequisite for using the strategy to induce seedlessness is identifying the crucial master gene or transcription factor liable for seed formation/development. In this review, we primarily explored the seedlessness mechanisms and identified the potential candidate genes underlying seed development. We also discussed the CRISPR/Cas-mediated genome editing approaches and their improvements.

Functional, communicative, and hybrid barriers to accessing mental health care in LGBTQ+ communities

ABSTRACT Understanding how LGBTQ+ populations experience common mental health care barriers reveals uncommon ways these barriers interact, layer, and compound to increase health disparities. This mixed methods study organised 250 open ended responses into 30 codes situated along a functional to communicative spectrum. The codes revealed sub-categories and relationships between barrier groupings which highlight ways mental health care obstacles for sexual and gender minorities span categories and are operationalised in unique ways that compound constraints. Organizing LGBTQ+ mental health care access obstacles along a continuum that spans functional barriers (i.e. time, money, transportation) to communicative ones (i.e. stigma, trust) revealed a hybrid category (i.e. providers, bureaucracy) where functional and communicative barriers overlap, mesh and operate simultaneously. Lack of access to trained mental health care providers who offer affirming, appropriate care sits at the centre of this web of constraints and as such is influential in a wide array of mental health care issues. Prioritising mental health care workforce training in LGBTQ+ specific competencies has the potential to create a ‘ripple effect’ that mitigates interconnected mental health care barriers throughout this spectrum.

The Hybridization Barrier between Herbaceous Medicago sativa and Woody M. arborea Is Weakened by Reproductive Abnormalities in M. sativa Seed Parents.

Historically, crosses between Medicago sativa (alfalfa) and M. arborea with alfalfa as the seed parent failed, as did crosses using M. arborea as the seed parent. Thus, a reproductive barrier kept the two species isolated until early in this century. The breakthrough came when alfalfa seed parents were identified in Wisconsin USA and Queensland AU that produced partial hybrids (hereafter hybrids). The hybrids were obtained by making large numbers of crosses on selected alfalfa parents. This was the first level of weakening the crossing barrier as reported in Plants in 2013. Further weakening of the barrier is reported herein whereby more hybrids were obtained with fewer crosses. This was accomplished by pedigree selection for new alfalfa seed parents and by using a product of the first hybrids called Alborea. New alfalfa seed parents were crossed with M. arborea, and Alborea parents were backcrossed to M. arborea. Hybrid plants were produced with fewer crosses in both cases. These hybrids, like the first hybrids, have mostly alfalfa traits but also have traits from M. arborea. It was theorized early on that the alfalfa component could be explained by 2n eggs in the alfalfa parents that were fertilized by normal n gametes from M. arborea. Evidence that the Wisconsin alfalfa and Alborea seed parents did in fact produce 2n eggs was reported in Plants in 2022. Moreover, they produced 2n eggs at approximately the same frequency that they produced hybrids. As reported herein, Alborea parents produced the highest frequency of hybrids and thus had the weakest barrier. Importantly, they also have the highest frequency of 2n eggs. It was determined that alfalfa and Alborea parents that produce 2n eggs and hybrids, also produce 2n pollen. In effect, an experiment was undertaken in reverse showing that 2n pollen could be used to screen for plants that produce hybrids. In the thousands of crosses made over the years, fertilization of normal n eggs in alfalfa parents always failed. Normal meiosis appears to be the main barrier to producing interspecific hybrids in our case. Fertilization of abnormal 2n eggs ensures sufficient alfalfa genetic material to continue embryogenesis. Evidently, the meiotic abnormality of 2n eggs is the major factor that weakens the crossing barrier.

The effect of grafting on pollen fertility and seed production of diploid interspecific hybrid and tetraploid eggplant genotypes

Solanum melongena is susceptible to diverse diseases and parasites, in particular the wilts (bacterial, Fusarium and Verticillium), nematodes, and some insects. Solanum torvum is robust to Verticillium and some bacterial wilts and root-knot nematode. It was reported that interspecific hybrid plants originated out of the hybridization of these species were sterile. In our previous studies that have been carried out to overcome this interspecific hybridization barrier, only from Faselis F1 that was used as the female parent, many interspecific hybrid seeds were obtained and 9.07% of the seeds grew into plants in vitro. The percentage of pollen viability and germination of tetraploid interspecific hybrid (amphidiploid) genotypes that have been produced by in vitro colchicine treatment, were 6.8 and 3.4 fold of its source diploid genotype, respectively. However, the increment was not high enough to overcome the interspecific hybridization barrier between S. torvum and S. melongena. In this study, the changes in pollen fertility of diploid interspecific hybrids and tetraploid plants, which have grafted on Pala or Faselis F1 , were investigated. Pollen viability and germination percentage in diploid genotypes were not affected by grafting, although they increased significantly (51.54% and 119.73%, respectively) in tetraploid genotypes by grafting. It was determined that the fertile diploid rootstock used for the tetraploid genotypes that produced by chromosome doubling from cultured eggplants could increase the number of seeds per fruit. It was concluded that some fertile diploid varieties can be used as rootstocks in order to increase pollen viability, germination rate and seed yield of tetraploid genotypes with low pollen yield obtained from cultivated eggplant by chromosome doubling.

Comprehensive Assessment of Intergeneric Cross-compatibility of Six Fruit Tree Species in the Tribe Maleae (Rosaceae) Based on <i>In Vivo</i> Pollen Tube Growth and Field Pollination

The development of intergeneric hybrids for horticultural crops has been attempted to introduce new quality and resistance traits and to enlarge the gene pool. Interspecific and intergeneric hybridization are often hindered by incompatibility reactions occurring at various stages of hybridization, from early pollination to initial growth, and the reproductive stages of the progeny. In this study, we investigated intergeneric and interspecific cross-compatibility among six species in the tribe Maleae (Rosaceae), namely, Pyrus communis (European pear), P. pyrifolia (Japanese pear), Malus × domestica (apple), Eriobotrya japonica (loquat), Cydonia oblonga (quince), and Pseudocydonia sinensis (Chinese quince). In vivo pollen tube growth tests showed the presence of a postmating, prezygotic barrier in many cross-combinations, in which cross-compatibility was regulated by both genetic distance and crossing direction. Strong hybridization barriers were observed in pollen tube growth and fruit setting when intergeneric hybridization was performed with E. japonica, a species phylogenetically distant from the others studied. Different compatibility reactions in reciprocal crosses were observed in some intergeneric hybridizations; C. oblonga as a pollen donor was incompatible with P. sinensis, whereas the reciprocal cross was compatible, resulting in the development of hybrid seedlings. Furthermore, the pollen tube growth rate differed among Pyrus species when pollinated on the apple pistils, suggesting divergence of cross-compatibility response in a specific linage. Factors affecting intergeneric hybridization are discussed with reference to the genetic distance between species and morphological characteristics such as pistil length. Our comprehensive assessment of intergeneric cross-compatibility will help provide a way to overcome crossing barriers and develop new hybrid crops in the tribe Maleae.

Ethohydraulic Experiments Investigating Retention Rates of an Electrified Bar Rack

Bar racks at water intakes of hydropower plants serve mainly to protect the turbines from floating debris. Additionally, they can be utilized to protect downstream migrating fish in order to prevent a potentially harmful turbine passage. The Bar Rack FishProtector consists of a common bar rack equipped with electrodes mounted on the upstream side of the bars. The application of a low voltage current at the electrodes creates an electric field in the water which is actively avoided by fish. Thus, a hybrid barrier consisting of a mechanical barrier and a behavioral barrier is formed. An unscaled model of a Bar Rack FishProtector (bar spacing sb = 50 mm, bar thickness tb = 20 mm) was used in field experiments to investigate the retention rate in an experimental setup with only one possible migration route (downstream, rack passage) and an average flow velocity of 0.43 m/s. Ethohydraulic experiments were performed with three indicator species barbel (Barbus barbus), bream (Abramis brama) and roach (Rutilus rutilus) and additionally perch (Perca fluviatilis) in selected trials. The twelve trials included four reference trials without electric field present (Nday = 2, Nnight = 2) and eight trials with electric field (Nday = 6, Nnight = 2). The results show that the experimental retention rate could be increased significantly by the application of an electrical field during the night and during the day with an even more pronounced effect during the night. The differences between the functionality of the system during the day and at night as well as other influencing parameters are discussed. No significant influence of the applied voltage on the electrodes or significant influence of fish size could be identified.

Renewable hybrid roadside barrier: Optimization of timber thickness

Researchers have recently focused on new and original roadside barriers that prioritize aesthetic, and environmental concerns by employing natural materials. In this study, the safety performance (Acceleration Severity Index (ASI), Theorical Head Impact Velocity (THIV)), structural performance (Working Width (W), Exit Angle (α)), and failure analysis (visual deformation) of a newly developed Renewable Hybrid Barrier (RHB) system at different timber thicknesses were tried to be determined by pendulum crash test and Finite Element (FE) models. The FE models were calibrated and validated based on pendulum crash test results, and then the most suitable timber thickness in terms of safety and structural performance was determined via FE analyses. The results revealed that as the timber thickness decreased, the safety parameters, such as ASI and THIV, decreased, thus the barrier safety increased. However, it was observed that the deflection and deformations in the barrier increased as the timber thickness decreased. In this sense, the safest and the most structurally durable barrier was determined through conducting virtual optimization tests. Studies on diversification of the usage areas of natural/renewable materials should be increased in the future.

5-Methyl-cytosine stabilizes DNA but hinders DNA hybridization revealed by magnetic tweezers and simulations.

5-Methyl-cytosine (5mC) is one of the most important DNA modifications and plays versatile biological roles. It is well known that 5mC stabilizes DNA duplexes. However, it remains unclear how 5mC affects the kinetics of DNA melting and hybridization. Here, we studied the kinetics of unzipping and rezipping using a 502-bp DNA hairpin by single-molecule magnetic tweezers. Under constant loading rates, 5mC increases the unzipping force but counterintuitively decreases the rezipping force at various salt and temperature conditions. Under constant forces, the non-methylated DNA hops between metastable states during unzipping and rezipping, which implies low energy barriers. Surprisingly, the 5mC DNA can't rezip after fully unzipping unless much lower forces are applied, where it rezips stochastically in a one-step manner, which implies 5mC kinetically hinders DNA hybridization and high energy barriers in DNA hybridization. All-atom molecular dynamics simulations reveal that the 5mC kinetically hinders DNA hybridization due to steric effects rather than electrostatic effects caused by the additional methyl groups of cytosines. Considering the possible high speed of DNA unzipping and zipping during replication and transcription, our findings provide new insights into the biological roles of 5mC.

An Effective Green Porous Structural Adhesive for Thermal Insulating, Flame-Retardant, and Smoke-Suppressant Expandable Polystyrene Foam

To develop an efficient way to overcome the contradiction among flame retardancy, smoke suppression, and thermal insulation in expanded polystyrene (EPS) foams, which are widely used insulation materials in buildings, a novel “green” porous bio-based flame-retardant starch (FRS) coating was designed from starch modified with phytic acid (PA) that simultaneously acts as both a flame retardant and an adhesive. This porous FRS coating has open pores, which in combination with the closed cells formed by EPS beads, create a hierarchically porous structure in FRS–EPS that results in superior thermal insulation with a lower thermal conductivity of 27.0 mW·(m·K) − 1 . The resultant FRS–EPS foam showed extremely low heat-release rates and smoke-production release, indicating excellent fire retardancy and smoke suppression. The specific optical density was as low as 121, which was 80.6% lower than that of neat EPS, at 624. The FRS–EPS also exhibited self-extinguishing behavior in vertical burning tests and had a high limiting oxygen index (LOI) value of 35.5%. More interestingly, after being burnt with an alcohol lamp for 30 min, the top side temperature of the FRS–EPS remained at only 140 °C with ignition, thereby exhibiting excellent fire resistance. Mechanism analysis confirmed the intumescent action of FRS, which forms a compact phosphorus-rich hybrid barrier, and the phosphorus-containing compounds that formed in the gas phase contributed to the excellent flame retardancy and smoke suppression of FRS–EPS. This novel porous biomass-based FRS system provides a promising strategy for fabricating polymer foams with excellent flame retardancy, smoke suppression, and thermal insulation.

Improvement and Reduction of Self-Heating Effect in AlGaN/GaN HEMT Devices

GaN is one of the third-generation broadband semiconductor materials developed rapidly in recent years, and Algan/GanHEMT has a broad application prospect in the fields of high temperature, high power, high frequency and radiation resistance, etc. In recent years, gallium nitride based high electron mobility transistors have been widely used in emerging industries, such as 5G technology, new energy vehicles, unmanned aircraft and other fields, due to their high power and high voltage resistance. However, due to the high power density of HEMT devices, the self-heating effect will lead to a significant increase in the junction temperature of the device, which will seriously affect the performance, reliability, and lifetime of the device. In this paper, the temperature characteristics of GaN high electron mobility transistors (GaN HEMTs) are studied, and the effect of self-heating effect on GaN HEMT devices is analyzed. In order to reduce the device temperature and improve the reliability of the device, a new device structure is proposed in this paper. The new structure replaces the conventional silicon and Si3N4 with highly thermally conductive diamond and SiC as the substrate and passivation layer of the device, which facilitates the heat dissipation from the substrate and passivation layer. Also, the new structure employs a hybrid barrier layer and field plate. Simulation results show that the new structure has about 30% lower temperature peak, 47% higher output current, 28% higher transconductivity, and up to 18.22% higher current collapse rate compared to the conventional structure.

Diverged subpopulations in tropical Urochloa (Brachiaria) forage species indicate a role for facultative apomixis and varying ploidy in their population structure and evolution.

Urochloa (syn. Brachiaria) is a genus of tropical grasses sown as forage feedstock, particularly in marginal soils. Here we aimed to clarify the genetic diversity and population structure in Urochloa species to understand better how population evolution relates to ploidy level and occurrence of apomictic reproduction. We explored the genetic diversity of 111 accessions from the five Urochloa species used to develop commercial cultivars. These accessions were conserved from wild materials collected at their centre of origin in Africa, and they tentatively represent the complete Urochloa gene pool used in breeding programmes. We used RNA-sequencing to generate 1.1 million single nucleotide polymorphism loci. We employed genetic admixture, principal component and phylogenetic analyses to define subpopulations. We observed three highly differentiated subpopulations in U. brizantha, which were unrelated to ploidy: one intermixed with U. decumbens, and two diverged from the former and the other species in the complex. We also observed two subpopulations in U. humidicola, unrelated to ploidy; one subpopulation had fewer accessions but included the only characterized sexual accession in the species. Our results also supported a division of U. decumbens between diploids and polyploids, and no subpopulations within U. ruziziensis and U. maxima. Polyploid U. decumbens are more closely related to polyploid U. brizantha than to diploid U. decumbens, which supports the divergence of both polyploid groups from a common tetraploid ancestor and provides evidence for the hybridization barrier of ploidy. The three differentiated subpopulations of apomictic polyploid U. brizantha accessions constitute diverged ecotypes, which can probably be utilized in hybrid breeding. Subpopulations were not observed in non-apomictic U. ruziziensis. Sexual Urochloa polyploids were not found (U. brizantha, U. decumbens) or were limited to small subpopulations (U. humidicola). The subpopulation structure observed in the Urochloa sexual-apomictic multiploidy complexes supports geographical parthenogenesis, where the polyploid genotypes exploit the evolutionary advantage of apomixis, i.e. uniparental reproduction and clonality, to occupy extensive geographical areas.

Characteristics of Interspecific Hybridization and Inbred Progeny of Pumpkin (Cucurbita moschata Duch.) and Winter Squash (Cucurbita maxima Duch.)

Hybrid incompatibility and F1 hybrid dysgenesis in the interspecific hybridization between pumpkin (Cucurbita moschata Duch.) and winter squash (Cucurbita maxima Duch.) have been previously noted. For this reason, it is difficult to obtain F2 generations due to F1 sterility. However, back-crossing and add-crossing can be utilized to overcome these incompatibility barriers in interspecific hybridization. To date, few studies have focused on parental selection, the back-crossing process, and offspring characteristics related to interspecific hybridization. Hence, we explored the F1 seed acquisition rate, plant characteristics, and F2 generation fertility after interspecific back-crossing and add-crossing hybridization of C. moschata and C. maxima. Moreover, as a female parent, X-1 (C. moschata) yielded F1 seeds when crossed with winter squash (C. maxima) 10-04-3, 10-37, or 10-05-2. BC1F1 seeds and plants could be obtained with winter squash (C. maxima) as the recurrent parent. Some healthy F2 seeds and plants were obtained from the interspecific hybrids, including ZJ-13, ZJ-8, and ZJ-11. Further fruit nutrition quality analysis showed that the starch, polysaccharide, cellulose, and dry material contents of ZJ-7 and ZJ-8 were significantly higher than those of the parent pumpkin and winter squash lines. The bioflavonoid content of ZJ-8 was lower than that of its parents, and its soluble protein was at the median value. Meanwhile, the bioflavonoid content of ZJ-1 was lower than that of its parents, but its soluble protein was significantly higher. These results suggest that back-crossing and multi-crossing can overcome the barriers to interspecific crossing of C. moschata × C. maxima. Some interspecific hybrid fruits had nutritional contents much higher than those of their parent lines. Additionally, X-1 (C. maxima) was found to act as a bridge species in interspecific hybridization. Collectively, in this study, the barriers to interspecific hybridization of C. moschata × C. maxima were addressed through crossing methods and choice of parents, and the obtained results are expected to provide novel support for interspecific hybrid breeding between C. moschata and C. maxima.

Morphological, ecological and geographic differences between diploids and tetraploids of Symphytum officinale (Boraginaceae) justify both cytotypes as separate species.

Polyploidization is generally considered to be an important evolutionary driver affecting the genetic diversity, that can alter the morphology, phenology, physiology or ecology of plants, which in turn may make the taxonomy of polyploids more difficult. One such example is the Symphytum officinale complex, a polyploid species group represented by three major cytotypes: tetraploids (2n = 48), less common, geographically restricted diploids (2n = 24) and hypotetraploids (2n = 40). In most European floras only one polymorphic species, S. officinale, is widely recognized, while the particular cytotypes are usually considered conspecific. Our study provided a thorough evaluation of the ploidy level diversity, morphological and ecological variation, with a special attempt to clarify the status of ‘white-flowered’ diploids. Using flow cytometry, we identified three cytotypes: widespread tetraploids (76.1 %); less frequent diploids (23.6 %) with scattered distribution across the range of tetraploids and confined only to several areas of Europe; and extremely rare triploids (0.3 %). Diploids and tetraploids showed diffuse parapatric pattern of distribution, with only four mixed-cytotype populations (2.7 %) found, but almost entirely without triploids, suggesting reproductive isolation between di- and tetraploids. Niche of diploids falls nearly completely within the niche of tetraploids that showed niche expansion. Tetraploids also showed a shift in niche optimum towards a less continental and colder climate, coupled with expansion to more disturbance-prone sites with higher nutrient availability. Diploids were clearly distinguishable morphologically from tetraploids. The morphological differentiation of studied cytotypes appears to be taxonomically significant, especially in combination with ecological differences and the apparent presence of hybridization barriers. Both cytotypes should be treated as separate species (i.e. S. bohemicum and S. officinale s. str.).

Implementation of patient-reported outcomes for symptom management in oncology practice through the SIMPRO research consortium: a protocol for a pragmatic type II hybrid effectiveness-implementation multi-center cluster-randomized stepped wedge trial

BackgroundMany cancer patients experience high symptom burden. Healthcare in the USA is reactive, not proactive, and doctor-patient communication is often suboptimal. As a result, symptomatic patients may suffer between clinic visits. In research settings, systematic assessment of electronic patient-reported outcomes (ePROs), coupled with clinical responses to severe symptoms, has eased this symptom burden, improved health-related quality of life, reduced acute care needs, and extended survival. Implementing ePRO-based symptom management programs in routine care is challenging. To study methods to overcome the implementation gap and improve symptom control for cancer patients, the National Cancer Institute created the Cancer-Moonshot funded Improving the Management of symPtoms during And following Cancer Treatment (IMPACT) Consortium.MethodsSymptom Management IMplementation of Patient Reported Outcomes in Oncology (SIMPRO) is one of three research centers that make up the IMPACT Consortium. SIMPRO, a multi-disciplinary team of investigators from six US health systems, seeks to develop, test, and integrate an electronic symptom management program (eSyM) for medical oncology and surgery patients into the Epic electronic health record (EHR) system and associated patient portal. eSyM supports real-time symptom tracking for patients, automated clinician alerts for severe symptoms, and specialized reports to facilitate population management. To rigorously evaluate its impact, eSyM is deployed through a pragmatic stepped wedge cluster-randomized trial. The primary study outcome is the occurrence of an emergency department treat-and-release event within 30 days of starting chemotherapy or being discharged following surgery. Secondary outcomes include hospitalization rates, chemotherapy use (time to initiation and duration of therapy), and patient quality of life and satisfaction. As a type II hybrid effectiveness-implementation study, facilitators and barriers to implementation are assessed throughout the project.DiscussionCreating and deploying eSyM requires collaboration between dozens of staff across diverse health systems, dedicated engagement of patient advocates, and robust support from Epic. This trial will evaluate eSyM in routine care settings across academic and community-based healthcare systems serving patients in rural and metropolitan locations. This trial’s pragmatic design will promote generalizable results about the uptake, acceptability, and impact of an EHR-integrated, ePRO-based symptom management program.Trial registrationClinicalTrials.gov NCT03850912. Registered on February 22, 2019. Last updated on November 9, 2021.