Plant Sexual Reproduction Research Articles

Effects of Biological Factors and Temperature Elevation on Stigmatic Receptivity and Pollen Tube Growth in Diploid and Hexaploid Plum Genotypes [Prunus salicina and Prunus insititia

ABSTRACTPremise of the study. Biological factors and environmental stresses are predominant elements affecting the progamic phase of sexual plant reproduction. Thus, new knowledge is needed to select and develop cultivars adapted to these changes. This work was conducted to evaluate the pollen growth dynamics and the stigmatic receptivity at normal and elevated temperatures, in 12 Tunisian plum genotypes selected based on six different biological factors. Methods. The pistils were hand‐pollinated using their own pollen, and then the dynamics of pollen tube growth and stigma receptivity of Tunisian plum genotypes were surveyed under ambient temperature and under 35 °C. Key results. Results showed that the differences in flower size, geographical origin and drought tolerance did neither affect pollen growth dynamics nor stigma receptivity. However, the polyploidization enlarged the stigmatic receptivity but does not affect the pollen dynamics. Flowering time and wild plantation fasten the pollen growth and enlarge the stigmatic receptivity. We assume that, in particular case, the flowering plants seem to opt for an enlarged stigmatic receptivity and a rapid pollen growth as reproductive strategies to ‘hold on for dear life’. Experiments conducted at 35 °C showed that the temperature elevation accelerates pollen growth dynamics, while it shortens the stigmatic receptivity period. In particular, two accessions: the wild plum ‘Sauvage’ and the late flowering ‘Tasstour Hamra Tardive’ showed faster pollen growth and enlarged stigmatic receptivity, compared to other accessions. Conclusion. The main relevant conclusion of this work resides in the identification of the best adapted Tunisian genotypes that could be selected as pollen sources to extend the pollination period. Especially, accessions presenting a tolerance to high‐temperature deserve serious attention.

Transcriptome profiling of foxtail millet (Setaria italica) pollen and anther

Pollen development and germination play a crucial role in the sexual reproduction of plants. This study analysis of transcriptional dynamics of foxtail millet pollen with other tissues and organs (ovule, glume, seedling and root) through RNA-sequencing revealed that a total of 940 genes were up-regulated in foxtail millet pollen. Based on this, we analyzed the genes involved in pollen tube growth of receptor kinases and small peptides, calcium signaling, small G proteins, vesicle transport, cytoskeleton, cell wall correlation, and transcription factors that are up-regulated in pollen. At the same time, we compared the gene expression of foxtail millet pollen and mature anthers, and found that a large number of transcription factors were specific expressed in mature anthers. In addition, we verified the accuracy of the transcriptome data using RT-qPCR. Finally, employed the antisense Oligonucleotide (as-ODN) system found that inhibiting SiPME67 expression would cause abnormal growth of pollen tube subapical. In summary, we preliminarily analyzed the genes that were up-regulated in foxtail millet pollen, which provided a reference for understanding the male sterility mechanism of foxtail millet in the future and theoretical basis for creating new male sterility lines.

Phosphatidic Acid Signaling in Modulating Plant Reproduction and Architecture.

Phosphatidic acid (PA) is an important class of signaling lipids involved in various biological processes in plants. Functional characterization of the mutants of PA's metabolizing enzymes coupled with lipidomics and protein-lipid interaction analyses have revealed that PA signaling is involved in plant response to biotic and abiotic stress. Moreover, PA and its metabolizing enzymes have been found to affect various reproductive steps, including gametogenesis, pollen tube growth, self-incompatibility, haploid embryo formation, embryogenesis, and seed development. They also play a significant role in shaping plant reproductive and root architecture. Recent results have also started to shed light on the multifaceted modes of PA's action, but much remains to be elucidated. Here, we will review the recent findings of PA and its metabolizing enzymes to highlight their effects on plant sexual reproduction and architecture and discuss the possible mechanisms underlying PA's actions in these processes and future directions.

Cell fate determination during sexual plant reproduction.

The flowering plant life cycle is completed by an alternation of diploid and haploid generations. The diploid sporophytes produce initial cells that undergo meiosis and produce spores. From haploid spores, male or female gametophytes, which produce gametes, develop. The union of gametes at fertilization restores diploidy in the zygote that initiates a new cycle of diploid sporophyte development. During this complex process, cell fate determination occurs at each of the critical stages and necessarily underpins successful plant reproduction. Here, we summarize available knowledge on the regulatory mechanism of cell fate determination at these critical stages of sexual reproduction, including sporogenesis, gametogenesis, and early embryogenesis, with particular emphasis on regulatory pathways of both male and female gametes before fertilization, and both apical and basal cell lineages of a proembryo after fertilization. Investigations reveal that cell fate determination involves multiple regulatory factors, such as positional information, differential distribution of cell fate determinants, cell-to-cell communication, and cell type-specific transcription factors. These factors temporally and spatially act for different cell type differentiation to ensure successful sexual reproduction. Thesenew insights into regulatory mechanisms underlying sexual cell fate determination not only updates our knowledge on sexual plant reproduction, but also provides new ideas and tools for crop breeding.

Indirect effects of warming via phenology on reproductive success of alpine plants

Abstract Climate warming has induced pronounced shifts in the phenology of alpine plants worldwide, yet the impact of these changes on plant reproduction remains unclear, although phenology plays a vital role in reproduction. Based on a 7‐year field warming and altered precipitation experiment initiated in 2017, we measured three reproductive phenological events and seven reproductive traits of six dominant species, belonging to two flowering functional groups, to assess the effects of climate change on reproductive phenology and the reproductive consequence of changing the phenology in an alpine meadow on the eastern Tibetan Plateau from 2021 to 2023. Warming significantly advanced the start of flowering in early‐spring flowering (ESF) and mid‐summer flowering (MSF) plants, while the fruiting period of ESF plants remained stable. Furthermore, warming led to a decrease in the number of seeds and productivity of alpine plants, while seed size remained stable. Phenological changes regulate the reproductive success of alpine plants. Specifically, the start of flowering for ESF plants and the start of fruiting for MSF plants regulated the number of seeds and productivity of the two flowering functional groups, respectively. The increase in phenological niche overlap and intensification of interspecific competition, caused by different phenological responses of the flowering functional groups to warming, also are key factors contributing to the decline in seed production of alpine plants. Synthesis. These results demonstrate that the reproductive success of alpine plants would be decreased by earlier reproduction under climate warming, and alpine plants tend to maintain stable seed size in response to climate change. Our study emphasizes the important regulatory and indicative role of reproductive phenology in the sexual reproduction of alpine plants under future climate change.

Warming reduces mid-summer flowering plant reproductive success through advancing fruiting phenology in an alpine meadow.

Changes in reproductive phenology induced by warming are happening across the globe, with significant implications for plant sexual reproduction. However, the changes in plant reproductive output (number of flowers and fruits) and success (successful fruits/total flowers) in response to climate change have not been well characterized. Here, we conducted a warming and altered precipitation experiment in an alpine meadow on the eastern Tibetan Plateau to investigate the effects of climate change on the reproductive phenology and success of six common species belonging to two flowering functional groups. We found that warming advanced the start of flowering and the start of fruiting in mid-summerflowering plants. Warming reduced the reproductive output of early-spring flowering plants but did not change their reproductive success. The effects of warming and altered precipitation on the reproductive output and success of mid-summer flowering plants were year-dependent, and the fruiting phenology regulated the response of the mid-summer flowering plant's reproductive success to climate change. These findings highlight the critical role of fruiting phenology in the reproductive success of alpine plants and imply that alpine plants may reduce their fitness by producing fewer flowers and fruits under climate warming, especially for later flowering plants.

Invasion stage and competition intensity co-drive reproductive strategies of native and invasive saltmarsh plants: Evidence from field data

Biological invasion poses a significant threat to biodiversity conservation and also results in substantial economic loss including the excessive cost of management to control it. Still, its impact on plant sexual reproduction strategies remains underexplored in natural settings. We conducted a field experiment on native Phragmites australis and invasive Spartina alterniflora in Bohai Bay and assessed plant size (aboveground biomass and height) and sexual reproduction (ear biomass, reproductive allocation, etc.) in conjunction with water and soil properties. The results showed that during the early stage of invasion, the two species declined in size and sexual reproduction, with S. alterniflora showing a lesser decline than P. australis. However, in the late stage of invasion, S. alterniflora maintained its plant size by reducing its investment in sexual reproduction. Moreover, significant reproductive allometries were demonstrated by S. alterniflora under different competition intensities. P. australis displayed heightened sensitivity to water properties and soil non-resource conditions, while S. alterniflora adapted its inherent traits and environmental tolerance. S. alterniflora allocated more resources to thriving as an individual, while P. australis prioritized reproduction by increasing seed production. Overall, this study revealed the reproductive strategies that invasive and native species employ in response to competition and environmental factors, thereby offering crucial insights for conservation and management efforts.

Quantitative proteomic analysis reveals hub proteins for high temperature-induced male sterility in bread wheat (Triticum aestivum L.).

High-temperature (HT) stress can induce male sterility in wheat; however, the underlying mechanisms remain poorly understood. This study examined proteomic alterations across three developmental stages between normal and HT-induced male-sterile (HT-ms) anthers in wheat. Utilizing tandem mass tags-based proteomics, we identified 2532 differentially abundant proteins (DAPs): 27 in the tetrad stage, 157 in the binuclear stage, and 2348 in the trinuclear stage. Analyses through Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways indicated significant enrichment of these DAPs in seven pathways, namely phenylpropanoid biosynthesis, flavonoid biosynthesis, sphingolipid metabolism, MAPK signaling pathway, starch and sucrose metabolism, response to heat, and response to reactive oxygen species (ROS). Our results indicated the downregulation of DAPs associated with phenylpropanoid biosynthesis and starch and sucrose metabolism, which aligns with anther indehiscence and the lack of starch in HT-ms anthers. By contrast, DAPs in the ROS pathway were upregulated, which aligns with excessive ROS accumulation in HT-ms anthers. Additionally, we conducted protein-protein interaction analysis for the DAPs of these pathways, identifying 15 hub DAPs. The abundance of these hub proteins was confirmed through qRT-PCR, assessing mRNA expression levels of the corresponding transcripts. Collectively, these results offer insights into the molecular mechanisms underlying HT-induced male sterility in wheat at the proteomic level, providing a valuable resource for further research in plant sexual reproduction.

Alpine grassland community productivity and diversity differences influence significantly plant sexual reproduction strategies.

Whether and how community structure variation affects plant sexual reproduction is crucial for understanding species' local adaptation and plant community assembly, but remains unrevealed. In Qinghai-Tibetan grassland communities that differed in aboveground biomass (AGB) and species diversity, we found significant influence of AGB on both species' reproductive biomass allocation (RBA) and flowering and fruiting time, but of species diversity only on species' reproductive time. In high-AGB or high-diversity communities, smaller and earlier flowering species generally advanced their reproductive phenology and increased their reproductive allocation for maximizing their reproductive success, whereas larger and later flowering species delayed their reproductive phenology and decreased their reproductive allocation for maximizing their vegetative growth and resource competition. This change in reproductive allocation with the variation in community structures was more pronounced in nonclonal as compared to clonal plant species. Thus, we evidence an important influence of community structure on plant sexual reproduction strategies, and the pattern of the influence depends largely on species biological attributes.

RNA HELICASE 32 is essential for female gametophyte development in Arabidopsis

The normal progression of mitotic cycles and synchronized development within female reproductive organs are pivotal for sexual reproduction in plants. Nevertheless, our understanding of the genetic regulation governing mitotic cycles during the haploid phase of higher plants remains limited. In this study, we characterized RNA HELICASE 32 (RH32), which plays an essential role in female gametogenesis in Arabidopsis. The rh32 heterozygous mutant was semi-sterile, whereas the homozygous mutant was nonviable. The rh32 mutant allele could be transmitted through the male gametophyte, but not the female gametophyte. Phenotypic analysis revealed impaired mitotic progression, synchronization, and cell specification in rh32 female gametophytes, causing the arrest of embryo sacs. In the delayed pollination test, none of the retarded embryo sacs developed into functional female gametophytes, and the vast majority of rh32 female gametophytes were defective in the formation of the large central vacuole. RH32 is strongly expressed in the embryo sac. Knock-down of RH32 resulted in the accumulation of unprocessed 18 S pre-rRNA, implying that RH32 is involved in ribosome synthesis. Based on these findings, we propose that RH32 plays a role in ribosome synthesis, which is critical for multiple processes in female gametophyte development.

When Size Matters: New Insights on How Seed Size Can Contribute to the Early Stages of Plant Development.

The seed habit is the most complex and successful method of sexual reproduction in vascular plants. It represents a remarkable moment in the evolution of plants that afterward spread on land. In particular, seed size had a pivotal role in evolutionary success and agronomic traits, especially in the field of crop domestication. Given that crop seeds constitute one of the primary products for consumption, it follows that seed size represents a fundamental determinant of crop yield. This adaptative feature is strictly controlled by genetic traits from both maternal and zygotic tissues, although seed development and growth are also affected by environmental cues. Despite being a highly exploited topic for both basic and applied research, there are still many issues to be elucidated for developmental biology as well as for agronomic science. This review addresses a number of open questions related to cues that influence seed growth and size and how they influence seed germination. Moreover, new insights on the genetic-molecular control of this adaptive trait are presented.

Knockout of stigmatic ascorbate peroxidase 1 (APX1) delays pollen rehydration and germination by mediating ROS homeostasis in Brassica napus L.

The successful interaction between pollen and stigma is a critical process for plant sexual reproduction, involving a series of intricate molecular and physiological events. After self-compatible pollination, a significant reduction in reactive oxygen species (ROS) production has been observed in stigmas, which is essential for pollen grain rehydration and subsequent pollen tube growth. Several scavenging enzymes tightly regulate ROS homeostasis. However, the potential role of these ROS-scavenging enzymes in the pollen-stigma interaction in Brassica napus remains unclear. Here, we showed that the activity of ascorbate peroxidase (APX), an enzyme that plays a crucial role in the detoxification of hydrogen peroxide (H2O2), was modulated depending on the compatibility of pollination in B. napus. We then identified stigma-expressed APX1s and generated pentuple mutants of APX1s using CRISPR/Cas9 technology. After compatible pollination, the BnaAPX1 pentuple mutants accumulated higher levels of H2O2 in the stigma, while the overexpression of BnaA09.APX1 resulted in lower levels of H2O2. Furthermore, the knockout of BnaAPX1 delayed the compatible response-mediated pollen rehydration and germination, which was consistent with the effects of a specific APX inhibitor, ρ-Aminophenol, on compatible pollination. In contrast, the overexpression of BnaA09.APX1 accelerated pollen rehydration and germination after both compatible and incompatible pollinations. However, delaying and promoting pollen rehydration and germination did not affect the seed set after compatible and incompatible pollination in APX1 pentuple mutants and overexpression lines, respectively. Our results demonstrate the fundamental role of BnaAPX1 in pollen rehydration and germination by regulating ROS homeostasis during the pollen-stigma interaction in B. napus.

Genome-Wide Identification and Expression Profiles of IMB Genes Reveal Their Potential Roles in the Gametophytic Sexual Reproduction Process of Camellia sinensis

It is of great significance to explore the molecular mechanism of gametophytic sexual reproduction in the genetic improvement and breeding of tea plants [Camellia sinensis (L.) O. Kuntze]. Imported beta family members (IMBs) are a class of widely distributed nucleoplasmic transport receptor proteins in eukaryotes, affecting plant development and reproduction, and participating in flowering time and sexual reproduction. Still missing, though, is a thorough examination of IMB members in tea plants. Here, seven members of the IMB gene family were screened by a genome-wide investigation in tea plants. These members were scattered unevenly throughout five chromosomes. All of them contained the conserved KAP95 and HEAT Repeat domains. Additionally, the promoter regions of CsIMBs harbored cis-acting elements associated with plant hormones, light, and abiotic stress responses. In order to further confirm the function of CsIMBs in the sexual reproduction of tea plants, the expression patterns of CsIMBs in different flower development stages and the ovary (before and after pollination) were analyzed. The expression results highlighted that CsIMBs were related to the fertility and fruiting of tea plants. Furthermore, five of the seven CsIMBs (CsIMB1a, CsIMB1b, CsIMB2, CsIMB3a, and CsIMB3b) were found to be localized in the nucleus revealed by subcellular localization analysis. These results offer a comprehensive characterization of IMB genes as well as insights into the potential roles of CsIMBs participating in the gametophytic sexual reproduction of C. sinensis.

Fire effects on pollination and plant reproduction: a quantitative review.

Fire may favour plant flowering by opening the vegetation and increasing abiotic resource availability. Increased floral display size can attract more pollinators and increase the absolute fruit and seed production immediately after the fire. However, anthropogenic increases in fire frequency may alter these responses. We aim to assess the effects of fire on pollination and reproductive success of plants at the global scale. We performed a systematic literature review and meta-analyses to examine overall fire effects as well as different fire parameters on pollination and on plant reproduction. We also explored to what extent the responses vary among pollinators, pollination vectors, plant regeneration strategies, compatibility systems, vegetation types and biomes. Most studies were conducted in fire-prone ecosystems. Overall, single fires increased pollination and plant reproduction but this effect was overridden by recurrent fires. Floral visitation rates of pollinators were enhanced immediately following a wildfire, and especially in bee-pollinated plants. Fire increased the absolute production of fruits or seeds but not the fruit or seed set. The reproductive benefits were mostly observed in wind-pollinated (graminoids), herbaceous and resprouter species. Finally, fire effects on pollination were positively correlated with fire effects on plant reproductive success. Fire has a central role in pollination and plant sexual reproduction in fire-prone ecosystems. The increase in the absolute production of fruits and seeds suggests that fire benefits on plant reproduction are likely driven by increased abiotic resources and the consequent floral display size. However, reproduction efficiency, as measured by fruit or seed set, does not increase with fire. In contrast, when assessed on the same plant simultaneously, fire effects on pollination are translated into reproduction. Increased fire frequency due to anthropogenic changes can alter the nature of the response to fire.

Two Growing-Season Warming Partly Promoted Growth but Decreased Reproduction and Ornamental Value of Impatiens oxyanthera.

Climate warming profoundly affects the vegetative growth, flowering phenology and sexual reproduction of plants; therefore, it affects the ornamental value of wild flowers. Despite this, the extent and mechanism of the impact remain unclear. Here, we conducted a warming experiment for two growing seasons (increases of 1.89 °C in 2017 and 2.37 °C in 2018) with infrared heaters to examine the effects of warming on the ornamental value of the wild flower Impatiens oxyanthera, endemic to China, in Mount Emei. We evaluated the comprehensive ornamental value based on plant morphology and flowering characteristics using the analytic hierarchy process (AHP) and disentangled the impact of the two traits on ornamental value using principal component analysis (PCA) and the partial least squares structural equation model (PLS-SEM) under ambient and warming treatments. We hypothesized that warming would reduce the ornamental value of I. oxyanthera in terms of plant morphology and flowering traits. Our results showed that warming significantly decreased plant height and crown width and increased branch number and single-leaf area. Warming also decreased vexillum length, corolla tube length, nectar spur length and pedicel length. In addition, warming shortened flowering duration per plant and reduced flower number, while there was no significant effect on flower longevity and flower color at full-bloom stage between the control and warming treatment. Therefore, the comprehensive ornamental value under warming was lower than that under the control. Pedicel length, flower color, flower longevity and flowering duration per plant were the main factors affecting the comprehensive ornamental value. The PLS-SEM showed that warming had an indirect negative effect on ornamental value via direct negative effects on flowering traits. Collectively, these results indicate that, although promoting vegetative growth, short-term warming significantly decreased the ornamental value of I. oxyanthera due to warming-caused smaller flowers and shorter flowering duration.

Functional Characterization of CsSWEET5a, a Cucumber Hexose Transporter That Mediates the Hexose Supply for Pollen Development and Rescues Male Fertility in Arabidopsis.

Pollen cells require large amounts of sugars from the anther to support their development, which is critical for plant sexual reproduction and crop yield. Sugars Will Eventually be Exported Transporters (SWEETs) have been shown to play an important role in the apoplasmic unloading of sugars from anther tissues into symplasmically isolated developing pollen cells and thereby affect the sugar supply for pollen development. However, among the 17 CsSWEET genes identified in the cucumber (Cucumis sativus L.) genome, the CsSWEET gene involved in this process has not been identified. Here, a member of the SWEET gene family, CsSWEET5a, was identified and characterized. The quantitative real-time PCR and β-glucuronidase expression analysis revealed that CsSWEET5a is highly expressed in the anthers and pollen cells of male cucumber flowers from the microsporocyte stage (stage 9) to the mature pollen stage (stage 12). Its subcellular localization indicated that the CsSWEET5a protein is localized to the plasma membrane. The heterologous expression assays in yeast demonstrated that CsSWEET5a encodes a hexose transporter that can complement both glucose and fructose transport deficiencies. CsSWEET5a can significantly rescue the pollen viability and fertility of atsweet8 mutant Arabidopsis plants. The possible role of CsSWEET5a in supplying hexose to developing pollen cells via the apoplast is also discussed.

Discussing the basic ecological relations of sand dune vegetation process

Dune is often considered as a degraded ecosystem. Natural vegetation restoration and stable artificial vegetation construction are the basic means restoring dune ecosystem. Based on long-term study of dune ecosystem, by taking into consideration both the philosophical principles of unity of opposites and dynamic change, and related ecological theories, we put forward some ecological relations that should be paid attention to in the study of vegetation assembly from the perspective of the uniqueness of dune ecosystem. We discussed the necessity of coupling relationships of scale-pattern-process and the transformation of synergy-tradeoff relationships, interpreted the importance of distinguishing sand dune stabilized and shifting phases, disturbance and stress, wind erosion and sand burial in the study of vegetation process. We further explored the applied value of niche law or neutral law in the study of dune vegetation process. Finally, we discussed the issues that should be paid attention to in the study of dune vegetation process from the aspects of adaptability to aeolian activities and drought tolerance, physiological and reproductive process, sexual and asexual reproduction of plants. This study would provide theoretical supports for vegetation restoration and stable vegetation construction of dune ecosystem.

Hormone and transcriptomic analysis revealed that ABA and BR are key factors in the formation of inter‐subgeneric hybridization barrier in water lily

AbstractUnderstanding the process of signal communication between pollen and stigma is of significant importance for plant sexual reproduction. In the case of inter‐subgeneric hybridization in water lily, a pre‐fertilization hybridization barrier exists, the regulatory mechanism of which remains unclear. In this study, we conducted hormone and transcriptome analyses of unpollinated stigmas (Mock), self‐pollinated stigmas (SP), cross‐pollinated stigmas within the same subgenus (CP), and inter‐subgenus cross‐pollination stigmas (ISCP) in water lily to elucidate the formation mechanism of the inter‐subgeneric hybridization barrier. Our results indicated that the lack of abscisic acid (ABA) and brassinolide (BR) in ISCP stigmas are key factors contributing to the formation of the inter‐subgeneric hybridization barrier in water lily. Exogenous application of ABA and BR can help overcome the barrier between inter‐subgeneric water lily crosses. Through transcriptome analysis, we identified nine candidate genes involved in regulating the inter‐subgeneric hybridization barrier in water lily. In addition, we further demonstrated the importance of the NCED2‐mediated ABA synthesis pathway in the pollination process through AS‐ODN technology. Our study confirms that ABA and BR are critical for breaking the inter‐subgeneric hybridization barrier. The identification of the nine candidate genes provides important clues for further research on the hybridization recognition mechanism in water lily.

Unequal carbon and nitrogen translocation between ramets affects sexual reproductive performance of the clonal grass Leymus chinensis under nitrogen addition

Sexual reproduction is crucial for population continuity in clonal plants. The effect of nutrient translocation between ramets on sexual reproduction of clonal plants under nitrogen addition remains unclear. In this study, we focused on clonal fragments of Leymus chinensis reproductive ramets with different number of vegetative ramets connected to tillering nodes. A series of pot experiments was conducted under nitrogen addition, including 13C and 15N bidirectional labelling of vegetative ramets and reproductive ramets at the milk-ripe stage, determination of the 13C and 15N amount translocated, and assessment of the quantitative characteristics, nitrogen and carbon concentrations of reproductive ramets and vegetative ramets. Nitrogen addition promoted the translocation of 13C while inhibiting 15N between vegetative ramets and reproductive ramets. With an increase in the number of connected vegetative ramets, the 13C translocated by reproductive ramets and the 15N translocated by reproductive and vegetative ramets gradually increased. The translocation of 13C and 15N between vegetative and reproductive ramets was bidirectional and unequal. The translocated amount of 13C and 15N from reproductive ramets to vegetative ramets was always higher than that from vegetative ramets to reproductive ramets. Nitrogen addition did not prominently affect the sexual reproductive performance of L. chinensis, whereas the number of connected vegetative ramets both positively and negatively affected sexual reproductive performance. Ramet biomass is an important driver of nutrient acquisition by L. chinensis ramets. We demonstrate for the first time that unequal nutrient translocation between ramets affects sexual reproductive performance in L. chinensis. The findings contribute to an enhanced understanding of the reproductive strategies of clonal plant populations in future environments.

The Role of Reactive Oxygen Species in the In Vitro Germination and Growth of the Petunia (Petunia hybrida E. Vilm.) Male Gametophyte

The in vitro growth of the pollen tube (PT), an object of comprehensive and intensive research, is a model for studying the mechanisms of sexual reproduction in higher plants. We have studied the potential role of reactive oxygen species (ROS) in the in vitro germination and growth maintenance of the petunia (Petunia hybrida E. Vilm.) male gametophyte. The exogenous treatment with H2O2 influences the PT germination and polar growth in vitro. The addition of H2O2 to culture medium increases both the percentage of pollen grain germination and the PT length in the case of long cultivation, but inhibits both processes during the first hour of cultivation. This suggests that endogenous ROS play a decisive role in the early stages of pollen germination, with the sensitivity to endogenous ROS emerging later over the course of their growth. The addition of diphenylene iodonium chloride (DPI), a NADPH oxidase inhibitor, considerably decreases both the germination and the growth of the petunia male gametophyte at low concentrations (0.1 μM), and completely arrests the growth at high concentrations (1 μM). ROS are necessary for polar growth of the petunia male gametophyte; they are secreted in the early stages of pollen grain activation and are further localized to the initiation of the PT, mainly in the PT apical part, during polar growth, as confirmed with the help of intravital fluorescence microscopy.