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Nature Communications Jun 2024Flowering plants rely on the polarized growth of pollen tubes to deliver sperm cells (SCs) to the embryo sac for double fertilization. In pollen, the vegetative nucleus...
Flowering plants rely on the polarized growth of pollen tubes to deliver sperm cells (SCs) to the embryo sac for double fertilization. In pollen, the vegetative nucleus (VN) and two SCs form the male germ unit (MGU). However, the mechanism underlying directional transportation of MGU is not well understood. In this study, we provide the first full picture of the dynamic interplay among microtubules, actin filaments, and MGU during pollen germination and tube growth. Depolymerization of microtubules and inhibition of kinesin activity result in an increased velocity and magnified amplitude of VN's forward and backward movement. Pharmacological washout experiments further suggest that microtubules participate in coordinating the directional movement of MGU. In contrast, suppression of the actomyosin system leads to a reduced velocity of VN mobility but without a moving pattern change. Moreover, detailed observation shows that the direction and velocity of VN's movement are in close correlations with those of the actomyosin-driven cytoplasmic streaming surrounding VN. Therefore, we propose that while actomyosin-based cytoplasmic streaming influences on the oscillational movement of MGU, microtubules and kinesins avoid MGU drifting with the cytoplasmic streaming and act as the major regulator for fine-tuning the proper positioning and directional migration of MGU in pollen.
Topics: Microtubules; Actin Cytoskeleton; Kinesins; Pollen; Actomyosin; Pollen Tube; Cell Nucleus; Arabidopsis; Cytoplasmic Streaming; Germination
PubMed: 38937444
DOI: 10.1038/s41467-024-49323-9 -
Biology Jun 2024Pollinators are needed for the reproduction of , and only sphingid moths have mouthparts long enough to reach the nectar at the bottom of the species' long, twisted...
Pollinators are needed for the reproduction of , and only sphingid moths have mouthparts long enough to reach the nectar at the bottom of the species' long, twisted floral tube. Though plants produce many flowers over a period of several months, one observes very few fruits in nature. We asked: (1) Are plants self-compatible, or do they need pollen from another individual to set fruit and seed? (2) Are cross-pollinations between unrelated individuals more successful than crosses with relatives? (3) How does the relatedness of pollen and ovule parent plants affect fruit set, seed number, and seed quality? We investigated the breeding system of by collecting fruits from seven sites, growing plants and performing hand pollinations over a period of several years, collecting and measuring fruits and counting seeds. is self-incompatible, though some individuals produce fruit by self-pollination. Cross-pollinations between unrelated individuals set the most fruit (59%), and those that were self-pollinated set the least (9%). Fruit set from cross-pollinations between related individuals was intermediate (32%). Although the number of seeds per fruit did not differ significantly among pollination treatments, fruits from self-pollinations had substantially fewer viable seeds than outcrossed fruits, with fruits from sibling crosses being intermediate. There were higher levels of self-compatibility in the fragment populations compared with plants from intact habitats. Self-incompatibility may explain why fruit set is low in this plant species; future investigation into the breakdown of self-incompatibility in smaller populations is warranted.
PubMed: 38927303
DOI: 10.3390/biology13060423 -
Plant Physiology Jun 2024The intricate process of male gametophyte development in flowering plants is regulated by jasmonic acid (JA) signaling. JA signaling initiates with the activation of the...
The intricate process of male gametophyte development in flowering plants is regulated by jasmonic acid (JA) signaling. JA signaling initiates with the activation of the basic-helix-loop-helix (bHLH) transcription factor (TF), MYC2, leading to the expression of numerous JA-responsive genes during stamen development and pollen maturation. However, the regulation of JA signaling during different stages of male gametophyte development remains less understood. This study focuses on the characterization of the plant ARID-HMG DNA-BINDING PROTEIN 15 (AtHMGB15), and its role in pollen development in Arabidopsis (Arabidopsis thaliana). Phenotypic characterization of a T-DNA insertion line (athmgb15-4) revealed delayed bolting, shorter siliques, and reduced seed set in mutant plants compared to the wildtype. Additionally, AtHMGB15 deletion resulted in defective pollen morphology, delayed pollen germination, aberrant pollen tube growth, and a higher percentage of non-viable pollen grains. Molecular analysis indicated the down-regulation of JA biosynthesis and signaling genes in the athmgb15-4 mutant. Quantitative analysis demonstrated that jasmonic acid and its derivatives were approximately tenfold lower in athmgb15-4 flowers. Exogenous application of methyl jasmonate could restore pollen morphology and germination, suggesting that the low JA content in athmgb15-4 impaired JA signaling during pollen development. Furthermore, our study revealed that AtHMGB15 physically interacts with MYC2 to form a transcription activation complex. This complex promotes the transcription of key JA signaling genes, the R2R3-MYB TFs MYB21 and MYB24, during stamen and pollen development. Collectively, our findings highlight the role of AtHMGB15 as a positive regulator of the JA pathway, controlling the spatiotemporal expression of key regulators involved in Arabidopsis stamen and pollen development.
PubMed: 38922580
DOI: 10.1093/plphys/kiae355 -
Plant Physiology Jun 2024Pollen germination and pollen tube elongation require rapid phospholipid production and remodeling in membrane systems that involve both de novo synthesis and turnover....
Pollen germination and pollen tube elongation require rapid phospholipid production and remodeling in membrane systems that involve both de novo synthesis and turnover. Phosphatidic acid phosphohydrolase (PAH) and lysophosphatidylcholine acyltransferase (LPCAT) are two key enzymes in membrane lipid maintenance. PAH generates diacylglycerol (DAG), a necessary precursor for the de novo synthesis of phosphatidylcholine (PC), while LPCAT reacylates lysophosphatidylcholine (LPC) to PC and plays an essential role in the remodeling of membrane lipids. In this study, we investigated the synthetic defects of pah and lpcat mutations in sexual reproduction of Arabidopsis (Arabidopsis thaliana) and explored the prospect of pistil lipid provision to pollen tube growth. The combined deficiencies of lpcat and pah led to decreased pollen tube growth in the pistil and reduced male transmission. Interestingly, pistils of the lipid mutant dgat1 ameliorated the male transmission deficiencies of pah lpcat pollen. In contrast, pollination with a non-specific phospholipase C (NPC) mutant exacerbated the fertilization impairment of the pah lpcat pollen. Given the importance of DAG in lipid metabolism and its contrasting changes in the dgat1 and npc mutants, we further investigated whether DAG supplement in synthetic media could influence pollen performance. DAG was incorporated into phospholipids of germinating pollen and stimulated pollen tube growth. Our study provides evidence that pistil derived lipids contribute to membrane lipid synthesis in pollen tube growth, a hitherto unknown role in synergistic pollen-pistil interactions.
PubMed: 38917229
DOI: 10.1093/plphys/kiae276 -
Plant Communications Jun 2024Transient heatwaves occur more frequently with climate warming, yet their impacts on crop yield are severely underestimated and even overlooked. Heatwaves spanning mere... (Review)
Review
Transient heatwaves occur more frequently with climate warming, yet their impacts on crop yield are severely underestimated and even overlooked. Heatwaves spanning mere days or even hours during sensitive stages (e.g., microgametogenesis and flowering) can significantly reduce crop yield by disrupting plant reproduction. Recently, advancements in multi-omics and GWAS analysis have shed light on specific organs (e.g., pollen, lodicule, and style), key metabolic pathways (sugar and reactive oxygen species (ROS) metabolisms, Ca homeostasis), and essential genes for transient heatwaves responses during the most sensitive stages in many crops. Based on this, this review places particular emphasis on the crop's heat-sensitive stages, using pollen development, floret opening, pollination, and fertilization as the central narrative thread. Complementing by key parts such as lodicule and tapetum, the multifaceted effects of transient heatwaves and their molecular basis are systematically demonstrated. A number of heat-tolerant genes for these processes have been identified in major crops such as maize and rice. The mechanisms and key heat-tolerant genes shared over different stages potentially facilitate the improvement of heat-tolerant crops more precisely.
PubMed: 38915200
DOI: 10.1016/j.xplc.2024.101009 -
Plant Reproduction Jun 2024ARID-HMG DNA binding protein, AtHMGB15, regulates pollen development and pollen germination in Arabidopsis. Previous studies have shown that ARID-HMG DNA binding...
ARID-HMG DNA binding protein, AtHMGB15, regulates pollen development and pollen germination in Arabidopsis. Previous studies have shown that ARID-HMG DNA binding protein, AtHMGB15 regulate pollen development and pollen germination in Arabidopsis. Here, we performed transcriptome and cytological studies to understand the role of AtHMGB15 in regulating pollen wall morphology and the pollen tube germination rate. Our result showed abnormal vacuolization in the tapetal cells during anther maturation and prolonged PCD in AtHMGB15 loss-of-function mutant. The tapetum has the ability to perform both secretory and biosynthetic activities critical for pollen maturation and pollen viability. Interestingly, expression of PCD executer genes CEP1, MC9 and RNS3 were significant down-regulation of in athmgb15-4. The growth of pollen tubes is regulated by the actin cytoskeleton dynamics. To address the defect in pollen tube growth of athmgb15, we monitored the actin network in growing pollen tubes of wildtype and athmgb15-4 using Rhodamine-phalloidin fluorescence. Our results indicate a highly fragmented actin distribution in athmgb15-4 pollen tubes with a lesser number of long actin fibers and significantly low f-actin concentration at the apex. q-RTPCR further indicates significant downy-regulation of actin regulatory proteins VLN2 and PRF4. Collectively, our results suggest that AtHMGB15 being a nuclear architectural protein orchestrates high-order chromatin organization to promote the transcription of genes responsible for pollen development and pollen germination.
PubMed: 38904831
DOI: 10.1007/s00497-024-00505-x -
Plant Science : An International... Jun 2024CrRLK1L subfamily members are involved in diverse growth- and development-related processes in Arabidopsis. However, the functions of their counterparts in rice are...
CrRLK1L subfamily members are involved in diverse growth- and development-related processes in Arabidopsis. However, the functions of their counterparts in rice are unknown. Here, OsANX expression was detected in developing inflorescences, mature pollen grains, and growing pollen tubes, and it was localized to the plasma membrane in pollen grains and tobacco epidermal cells. Homozygous osanx progeny could not be segregated from the CRISPR/Cas9-edited mutants osanx-c1 and osanx-c2, and such progeny were segregated only occasionally from osanx-c3. Further, all three alleles showed osanx male but not female gamete transmission defects, in line with premature pollen tube rupture in osanx-c3. Additionally, osanx-c3 exhibited precocious flowering, excessively branched inflorescences, and an extremely low seed setting rate of 1.4 %, while osanx-c2 and osanx-c3 had no obvious defects in inflorescence development or the seed setting rate compared to wild-type Nipponbare (Nip). Consistent with this, the complemented line pPS1:OsANX-GFP/osanx-c2 (PSC), in which the lack of OsANX expression was inflorescence-specific, showed slightly earlier flowering and overly-branched panicles. Multiple inflorescence meristem transition-related and inflorescence architecture-related genes were expressed at higher levels in osanx-c3 than in Nip; thus, they may partially account for the aforementioned mutant phenotypes. Our findings broaden our understanding of the biological functions of OsANX in rice.
PubMed: 38901780
DOI: 10.1016/j.plantsci.2024.112162 -
Journal of Comparative Physiology. A,... Jun 2024As pollinators, bees are key to maintaining the biodiversity of angiosperm plants, and for agriculture they provide a billion-dollar ecosystem service. But they also...
As pollinators, bees are key to maintaining the biodiversity of angiosperm plants, and for agriculture they provide a billion-dollar ecosystem service. But they also compete for resources (primarily nectar and pollen), especially the highly social bees that live in perennial colonies. So, how do they organize their daily temporal activities? Here, we present a versatile, low-cost device for the continuous, automatic recording and data analysis of the locomotor activity in the colony-entrance tube of highly eusocial bees. Consisting of an in-house built block containing an infrared detector, the passage of bees in the colony entrance tunnel is registered and automatically recorded in an Arduino environment, together with concomitant recordings of temperature and relative humidity. With a focus on the highly diverse Neotropical stingless bees (Meliponini), we obtained 10-day consecutive recordings for two colonies each of the species Melipona quadrifasciata and Frieseomelitta varia, and also for the honey bee. The Lomb-Scargle periodogram analysis identified a predominant circadian rhythmicity for all three species, but also indications of ultradian rhythms. For M. quadrifasciata, which is comparable in size to the honey bee, we found evidence for a possibly anticipatory activity already before sunrise. As all three species also presented activity at night in the colony entrance tube, this also raises questions about sleep organization in social insects. The cost and versatility of the device and the open-source options for data analysis make this an attractive system for conducting studies on circadian rhythms in social bees under natural conditions, complementing studies on flower visits by these important pollinators.
PubMed: 38898188
DOI: 10.1007/s00359-024-01709-2 -
Plants (Basel, Switzerland) Jun 2024An initial cross of 'Johnblue' (Darrow's blueberry) × 'Red Sunset' (lingonberry) produced more than 30 true intersectional diploid hybrids as confirmed by molecular...
An initial cross of 'Johnblue' (Darrow's blueberry) × 'Red Sunset' (lingonberry) produced more than 30 true intersectional diploid hybrids as confirmed by molecular markers. The most vigorous of these hybrids was extensively evaluated. This hybrid, US 2535-A, was floriferous and morphologically intermediate to the respective parents. Examination of pollen suggested low male fertility. Numerous crosses using the hybrid as a female reflected similarly low fertility and potential crossing barriers. Stylar examination suggested blockage of pollen tube growth in self-pollinations and significantly retarded growth in backcross pollinations. Nonetheless, two confirmed hybrid offspring were produced using the F hybrid as a female in crosses with and , respectively. In a second set of crosses utilizing additional and genotypes, another 23 verified hybrids in seven parental combinations were produced. Hybrids such as the ones presented offer the potential for generating de novo interspecific fruit types in blueberry and/or broadening the adaptation of lingonberry.
PubMed: 38891380
DOI: 10.3390/plants13111572 -
Journal of Experimental Botany Jun 2024Major constituents of the plant cell walls are structural proteins that belong to the hydroxyproline-rich glycoprotein (HRGP) family. Leucine-rich repeat extensin (LRXs)...
Major constituents of the plant cell walls are structural proteins that belong to the hydroxyproline-rich glycoprotein (HRGP) family. Leucine-rich repeat extensin (LRXs) proteins contain a leucine-rich domain and a C-terminal domain with repetitive Ser-Pro(3-5) motifs that are potentially to be O-glycosylated. It has been demonstrated that pollen-specific LRX8-11 from Arabidopsis thaliana are necessary to maintain the integrity of the pollen tube cell wall during polarized growth. In HRGP including classical extensins (EXTs) and likely in LRXs, proline residues are converted to hydroxyproline by prolyl-4-hydroxylases (P4Hs), thus defining novel O-glycosylation sites. In this context, we aimed to determine whether hydroxylation and subsequent O-glycosylation of Arabidopsis pollen LRXs are necessary for their proper function and cell wall localization in pollen tubes. We hypothesized that pollen-expressed P4H4 and P4H6 catalyze the hydroxylation of the proline units present in Ser-Pro3-5 motifs of LRX8-LRX11. Here, we show the p4h4-1 p4h6-1 double mutant exhibits a reduction in pollen germination rates and a slight reduction in pollen tube length. Pollen germination is also inhibited by P4Hs inhibitors, suggesting that prolyl hydroxylation is required for pollen tube development. Plants expressing pLRX11::LRX11-GFP in the p4h4-1 p4h6-1 background show partial re-localization of LRX11-GFP from the pollen tube tip apoplast to the cytoplasm. Finally, IP-MS-MS analysis revealed a decrease in oxidized prolines (hydroxyprolines) in LRX11-GFP in the p4h4-1 p4h6-1 background compared to lrx11 plants expressing pLRX11::LRX11-GFP. Taken together, these results suggest P4H4 and P4H6 are required for pollen germination and for proper hydroxylation of LRX11 necessary for its localization at the cell wall of pollen tubes.
PubMed: 38877792
DOI: 10.1093/jxb/erae269