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Grain amaranths (Amaranthus spp.) have been cultivated for thousands of years in Central and South America. Their grains are of high nutritional value, but the low yield needs to be increased by selection of superior genotypes from genetically diverse breeding populations. Amaranths are adapted to harsh conditions and can be cultivated on marginal lands although little is known about their physiology. The development of controlled growing conditions and efficient crossing methods is important for research on and improvement of this ancient crop. Grain amaranth was domesticated in the Americas and is highly self-fertilizing with a large inflorescence consisting of thousands of very small flowers. We evaluated three different crossing methods (open pollination, hot water emasculation and hand emasculation) for their efficiency in amaranth and validated them with genetic markers. We identified cultivation conditions that allow an easy control of flowering time by day length manipulation and achieved flowering times of 4 weeks and generation times of 2 months. All three different crossing methods successfully produced hybrid F1 offspring, but with different success rates. Open pollination had the lowest (10%) and hand emasculation the highest success rate (74%). Hot water emasculation showed an intermediate success rate (26%) with a maximum of 94% success. It is simple to perform and suitable for a more large-scale production of hybrids. We further evaluated 11 single nucleotide polymorphism (SNP) markers and found that they were sufficient to validate all crosses of the genotypes used in this study for intra- and interspecific hybridizations. Despite its very small flowers, crosses in amaranth can be carried out efficiently and evaluated with inexpensive SNP markers. Suitable growth conditions strongly reduce the generation time and allow the control of plant height, flowering time, and seed production. In combination, this enables the rapid production of segregating populations which makes amaranth an attractive model for basic plant research but also facilitates further the improvement of this ancient crop by plant breeding.

Small millets are nutri-rich, climate-resilient food and fodder crops. They include finger millet, proso millet, foxtail millet, little millet, kodo millet, browntop millet, and barnyard millet. They are self-pollinated crops and belong to the family Poaceae. Hence, to widen the genetic base, the creation of variation through artificial hybridization is a prerequisite. Floral morphology, size, and anthesis behavior cause major hindrances in recombination breeding through hybridization. Manual emasculation of florets is practically very difficult; therefore, the contact method of hybridization is widely followed. However, the success rate of obtaining true F 1 s is 2% to 3%. In finger millet, hot water treatment (52°C) for 3 to 5 min causes temporal male sterility. Chemicals such as maleic hydrazide, gibberellic acid, and ethrel at different concentrations aid in inducing male sterility in finger millet. Partial-sterile (PS) lines developed at the Project Coordinating Unit, Small Millets, Bengaluru are also in use. The percent seed set in crosses derived from PS lines ranged from 27.4 to 49.4, with an average of 40.10%. In proso millet, little millet, and browntop millet, apart from contact method, hot water treatment, hand emasculation, and the USSR method of hybridization are also followed. A newly developed modified crossing method known as the Small Millets University of Agricultural Sciences Bengaluru (SMUASB) method in proso and little millets has a success rate of 56% to 60% in obtaining true hybrids. Hand emasculation and pollination under the greenhouse and growth chamber in foxtail millet with a success rate of 75% seed set is suggested. In barnyard millet, hot water treatment (48°C to 52°C) for 5 min followed by the contact method is often practiced. Kodo millet being cleistogamous, mutation breeding is widely followed to create variation. Most commonly, hot water treatment is followed in finger millet and barnyard millet, SMUASB in proso, and little millet. Although no specific method is suitable for all small millets, it is essential to identify a trouble-free technique that produces maximum crossed seeds in all the small millets.

Analyses of gender-based violence during mass conflict have typically focused on violence committed against women. Violence perpetrated against men has only recently been examined as gender-based violence in its own right. Using narratives from 1,136 Darfuri refugees, we analyze patterns of gender-based violence perpetrated against men and boys during the genocide in Darfur. We examine how this violence emasculates men and boys through four mechanisms: homosexualization, feminization, genital harm, and sex-selective killing. In line with an interactionist approach, we demonstrate how genocidal violence is gendered and argue that perpetrators committing gender-based violence perform masculinity in accordance with hegemonic gender norms in Sudan. We also show how gender-based violence enacts, reinforces, and creates meaning on multiple levels in a matrix of mutually reinforcing processes that we term the gender-genocide nexus. By extending the gender–violence link to the context of mass atrocity, this study facilitates an understanding of the mechanisms through which gender inequalities can be reproduced and maintained in diverse situations and structures.

In animal-pollinated plants, direct and indirect selection for large and small flowers in predominantly outcrossing and selfing species, respectively, is a common consequence of pollen limitation (PL). However, many hermaphroditic species show a mixed-mating system known as delayed selfing, which provides reproductive assurance (RA) only when outcrossing is not realized. Although RA is expected to reduce pollinator-mediated selection towards larger flowers, the consequences of delayed selfing for selection on flower size in mixed-mating species remain overlooked. We investigated whether RA weakens selection on flower size in Tuberaria guttata, a mixed-mating annual herb. We related pollinator visitation rates to flower size and measured seed production in emasculated, hand cross-pollinated and intact (control) flowers in three natural populations. For each population, we estimated variation in PL and RA across individuals differing in flower size and phenotypic selection on this trait. Pollinator visitation increased and RA decreased with flower size in all populations. Increasing RA diminished but did not fully alleviate PL, because of early-acting inbreeding depression. In the least-visited and most pollen-limited population, RA increased seed production by >200 %, intensely counteracting the strong pollinator-mediated selection for larger corollas. In the most-visited population, however, RA increased seed production by an average of only 9 %. This population exhibited the largest fraction of individuals that showed a decrease in seed production due to selfing and the weakest pollinator-mediated selection on flower size. The results suggest that the balance between the extent of RA and outcrossing contributes to determine flower size in mixed-mating systems. Pollinator-mediated selection favours larger flowers by increasing outcrossed seeds, but the benefits of RA greatly lessen this effect, especially under severe conditions of pollen limitation. Our findings also indicate that a mixed-mating system can represent an 'evolutionary trap' under an adequate pollinator supply.

The plant hormone auxin is a vital component for plant reproduction as it regulates the development of both male and female reproductive organs, including ovules and gynoecia. Furthermore, auxin plays important roles in the development and growth of seeds and fruits. Auxin responses can be detected in ovules shortly after fertilization, and it has been suggested that this accumulation is a prerequisite for the developmental reprogramming of the ovules to seeds, and of the gynoecium to a fruit. However, the roles of auxin at the final stages of ovule development, and the sources of auxin leading to the observed responses in ovules after fertilization have remained elusive. Here we have characterized the auxin readout in Arabidopsis ovules, at the pre-anthesis, anthesis and in the immediate post-fertilization stages, using the auxin sensor. In addition we have mapped the expression of auxin biosynthesis and conjugation genes, as well as that of auxin transporting proteins, during the same developmental stages. These analyses reveal specific spatiotemporal patterns of the different auxin homeostasis regulators. Auxin biosynthesis genes and auxin transport proteins define a pre-patterning of vascular cell identity in the pre-anthesis funiculus. Furthermore, our data suggests that auxin efflux from the ovule is restricted in an anther-dependent manner, presumably to synchronize reproductive organ development and thereby optimizing the chances of successful fertilization. Finally, auxin biosynthesis together with reduced auxin conjugation and transport result in an enhanced auxin readout throughout the sporophytic tissues of the ovules soon after fertilization. Together, our results suggest a sophisticated set of regulatory cascades that allow successful fertilization and the subsequent transition of the female reproductive structures into seeds and fruits.

The rostellum, a small structure located above the stigma in the gynostemium of monandrous orchid flowers, separates the pollinia from the stigma, can prevent self-pollination, and plays an important role in pol- lination. It secretes a sticky exudate before and after anthesis, to which the pollinia are attached. Additionally,the viscidium, a sticky disk at the base of the stipe, also connects to the rostellum. This structure varies among species and holds taxonomic significance. For example, in Catasetum, the rostellum extends into antennae-like projections. After pollination, emasculation, or injury, the plant hormone ethylene is produced, triggering various post-pollination, post-emasculation, and post-injury processes, including changes in perianth color, senescence, and, ultimately, the death of floral segments that have completed their functions. Although the rostellum plays a significant role, it remains understudied, and many aspects of its function are still unknown. This review high- lights current knowledge and explores speculative hypotheses that may invite differing opinions and perspectives, which will likely be revised as future research deepens our understanding of its biological and ecological roles.

• Premise of the study: Hermaphroditism in plants can lead to gender conflicts, such as pollen discounting—the loss of siring opportunities following self-pollination, and ovule discounting—the loss of seed production opportunities when self-pollen tubes disable ovules through early inbreeding depression or late-acting self-incompatibility. If ovules are discounted by self-pollination, it can be predicted that emasculation (removal of self-pollen) should increase seed production, as long as pollinators are not deterred by emasculation.• Methods: Using the hermaphroditic Kniphofia linearifolia, which belongs to a lineage known to possess late-acting self-incompatibility, we performed hand-pollinations with either self- or cross-pollen and recorded pollen tube growth and seed production. We paired experimentally emasculated plants with non-emasculated controls and quantified pollinator visitation rates, fruit, and seed set in both groups.• Key Results: Ovules penetrated by tubes from self-pollen uniformly failed to develop into seeds, as expected from ovarian self-incompatibility (or strong early inbreeding depression). Experimental emasculation had no effect on the rate of visitation by birds, the primary pollinators of this species, but led to increased rejection by bees, which are secondary pollinators. Despite reducing visitation by some pollinators, emasculation led to increased seed production and reduced rates of seed abortion.• Conclusions: These results show that female performance in hermaphroditic flowers can be enhanced when flowers are emasculated, even if the overall number of pollinator visits is decreased. The most likely explanation for this effect of emasculation is that it reduces ovule discounting arising from pollinator-mediated self-pollination.

A vision-based autonomous system for emasculating okra enhances agriculture by enabling precise flower bud identification, overcoming the labor-intensive, error-prone challenges of traditional manual methods with improved accuracy and efficiency. This study presents a framework for an adaptive, automated bud identification method to assist the emasculation process, hybridized optical coherence tomography (OCT). Three YOLOv8 variants were evaluated for accuracy, detection speed, and frame rate to identify the most efficient model. To strengthen the findings, YOLO was hybridized with OCT, enabling non-invasive sub-surface verification and precise quantification of the emasculated depth of both sepal and petal layers of the flower bud. To establish a solid benchmark, gold standard color histograms and a digital imaging-based method under optimal lighting conditions with confidence scoring were also employed. The results demonstrated that the proposed method significantly outperformed these conventional frameworks, providing superior accuracy and layer differentiation during emasculation. Hence, the developed YOLOv8 hybridized OCT method for flower bud identification and emasculation offers a powerful tool to significantly improve both the precision and efficiency of crop breeding practices. This framework sets the stage for implementing scalable, artificial intelligence (AI)-driven strategies that can modernize and optimize traditional crop breeding workflows.

Fruit initiation in Arabidopsis (Arabidopsis thaliana) is generally repressed until fertilization occurs. However, mutations in AUXIN RESPONSE FACTOR8 (ARF8) uncouple fruit initiation from fertilization, resulting in the formation of seedless, parthenocarpic fruit. Here we induced parthenocarpy in wild-type Arabidopsis by introducing either the mutant genomic (g) Atarf8-4 sequence or gAtARF8:β-glucuronidase translational fusion constructs by plant transformation. Silencing of endogenous AtARF8 transcription was not observed, indicating that the introduced, aberrant ARF8 transcripts were compromising the function of endogenous ARF8 and/or associated factors involved in suppressing fruit initiation. To analyze the role of ARF8 in tomato (Solanum lycopersicum) we initially emasculated 23 tomato cultivars to test for background parthenocarpy. Surprisingly, all had a predisposition to initiate fertilization-independent fruit growth. Expression of gAtarf8-4 in transgenic tomato ('Monalbo') resulted in a significant increase in the number and size of parthenocarpic fruit. Isolation of tomato ARF8 cDNA indicated significant sequence conservation with AtARF8. SlARF8 may therefore control tomato fruit initiation in a similar manner as AtARF8 does in Arabidopsis. Two SlARF8 cDNAs differing in size by 5 bp were found, both arising from the same gene. The smaller cDNA is a splice variant and is also present in Arabidopsis. We propose that low endogenous levels of the splice variant products might interfere with efficient formation/function of a complex repressing fruit initiation, thereby providing an explanation for the observed ovary expansion in tomato and also Arabidopsis after emasculation. Increasing the levels of aberrant Atarf8-4 transcripts may further destabilize formation/function of the complex in a dosage-dependent manner enhancing tomato parthenocarpic fruit initiation frequency and size and mimicking the parthenocarpic dehiscent silique phenotype found in homozygous Atarf8-4 mutants. Collectively these data suggest that similar mechanisms involving auxin signaling exist to inhibit parthenocarpic fruit set in tomato and Arabidopsis.

Background and aims - The Orchidaceae family is vulnerable, because of the destruction of their habitat, as well as the extraction of individuals from natural populations. This is the case of the genus Rhynchostele Rchb. f.; among the actions considered important for appropriate conservation strategies for this genus is the generation of fundamental knowledge, such as on its reproductive biology. The objective of this work is to understand the mating system and reproductive success of Rhynchostele cervantesii, an endangered epiphytic orchid endemic to Mexico. Material and methods - Manual and open-pollination treatments were conducted during 2014 and 2015 in a cloud forest in Michoacan, Mexico. In each period, 30 to 40 randomly selected inflorescences were subjected to the following treatments: a) spontaneous-self-pollination, b) emasculation, c) self-pollination, d) cross-pollination, and e) open-pollination. The developed fruits were counted and harvested, the viability of the seeds was determined, through the observation and evaluation of embryos using microscopy. Key results - Significant differences were recorded between the treatments in both 2014 and 2015, with higher fruit production in cross-pollination than in self-pollination and natural-pollination. There were significant differences in seed viability, with higher values for seeds from open-pollination and cross- pollination and lower values for seeds from self-pollination. Conclusions - Rhynchostele cervantesii is a species that requires pollinators for sexual reproduction because there is no fruit production with spontaneous-self-pollination. Under pollen limitation, the fruit set of natural pollination was a lot lower than in cross-pollination although fruits were the same quality. R. cervantesii had a mixed mating system with a tendency to exogamy, presenting high values of female reproductive success compared to other tropical epiphytic orchid species reported in the literature.