Embryo Sac Development Research Articles

Unlocking fertility in the female gametophyte: a DEAD-box RNA helicase is essential for embryo sac development and seed setting.

This article comments on: Huang J, Qiao Z, Yu H, Lu Z, Chen W, Lu J, Wu J, Bao Y, Shahid MQ, Liu X. 2024. OsRH52A, a DEAD-box protein, regulates functional megaspore specification and is required for embryo sac development in rice. Journal of Experimental Botany 75, 4802–4821. https://doi.org/10.1093/jxb/erae180

Morphogenesis, megagametogenesis, and microgametogenesis in Actinidia arguta flower buds

In dioecious plants, understanding sexual differentiation is important for improving cultivation techniques. Flower bud morphogenesis and gametophyte formation underlie the mechanism of sexual differentiation. Actinidia arguta (hardy kiwi) is a unisexual plant, and there have been few studies on its flower bud differentiation or gametophyte formation. We studied flower bud morphogenesis and gametophyte formation in A. arguta using paraffin sections and found that the morphogenesis of male and female flower buds was divided into six stages: the undifferentiated stage, inflorescence initials differentiation stage, calyx primordium differentiation stage, petal primordium differentiation stage, stamen primordium differentiation stage, and pistil primordium differentiation stage. Morphogenesis in the male inflorescence can reach quaternary flower primordia, and morphogenesis in the female inflorescence can reach the tertiary flower primordia, however, no Stage 4 lateral flowers in the male inflorescence or Stage 3 lateral flowers in the female inflorescence were found in natural situation. The embryo sac in female plants was a monocellular Polygonum type, and the embryo sac development in male plants was incomplete; abortion occurred before the archesporial cells developed. The male gametophyte differentiation process in the male plant was normal, with pollen grains similar in shape to a rugby ball and arranged neatly in the anther. The pollen grain differentiation in female plants was abnormal; abortion occurred at the uninucleate microspore stage, and mature pollen grains were irregularly spherical.

Lectin Receptor-Like Protein Kinase OsNRFG6 is Required for Embryo Sac Development and Fertilization in Neo-Tetraploid Rice

Great yield-enhancing prospects of autotetraploid rice was restricted by various polyploidy-induced reproductive dysfunction. To surmount these challenges, our group has generated a series of valuable fertile tetraploid lines (denoted as neo-tetraploid rice) through 20-year efforts. With this context, a G-type lectin receptor-like kinase, OsNRFG6, was identified as a pivotal factor associated with reproductive regulation in neo-tetraploid rice. Nevertheless, it is still elusive about a comprehensive understanding of its precise functional roles and underlying molecular mechanisms during reproduction of neo-tetraploid rice. Here, we demonstrated that OsNRFG6 executed a constitutive expression pattern and encoded proteins localizing in perinucleus and endoplasmic reticulum. Subsequently, four independent mutant lines of OsNRFG6 within neo-tetraploid rice background were further identified, all displaying low seed-setting rate due to abortive embryo sacs and defective double fertilization. RNA-seq and RT-qPCR revealed a significant down-regulation of OsNRFG6 and female reproductive genes such as OsMEL1 and LOG in ovaries prior to and post-fertilization, attributing this effect to OsNRFG6 mutation. Furthermore, through yeast-two hybrids, bimolecular fluorescence complementation assays, and luciferase complementation imaging assays, it was determined that OsNRFG6 could interact with itself and two female reproductive proteins (LOG and OsDES1) to form protein complexes. These results elucidate the reproductive functions and molecular pathway governed by OsNRFG6 in regulating fertility of neo-tetraploid rice, offering insights into molecular understanding of fertility improvement in polyploid rice.

Contribution to the Embryology, Sporoderm and Spermoderm Pattern of <I>Bothriochloa pertusa</I> (L.) A. Camus, <I>Capillipedium filiculme</I> (Hook.F.) Stapf and <I>Dichanthium annulatum</I> (Forssk.) Stapf with a Brief discussion on its Taxonomic Status

Embryological features in Bothriochloa pertusa, Capillipedium filiculme and Dichanthium annulatum have been studied. The male reproductive characters are constant as evidenced by anther wall development and it corresponds to Monocot type. At maturity the pollen grains are 3-celled. Embryo sac development and embryogeny conforms to Polygonum type and Asterad type respectively. Endosperm, seed coat and pericarp have been studied and data on SEM investigation on pollen wall and seed coat of these taxa are added. The three taxa viz., Bothriochloa, Capillipedium and Dichanthium were considered under Andropogoneae of Panicoideae as separate genera by Bor (1960). De Wet and Harlan (1970) merged them in Dichanthium . The evidences given by them are intergeneric hybridization and production of fertile hybrids between these three genera. Clayton (1977) also followed the concept of merger taking priority. The authors assessed the possibility of merger into Dichanthium on the basis of embryology, sporoderm and spermoderm pattern

OsRH52A, a DEAD-box protein, regulates functional megaspore specification and is required for embryo sac development in rice.

The development of the embryo sac is an important factor that affects seed setting in rice. Numerous genes associated with embryo sac (ES) development have been identified in plants; however, the function of the DEAD-box RNA helicase family genes is poorly known in rice. Here, we characterized a rice DEAD-box protein, RH52A, which is localized in the nucleus and cytoplasm and highly expressed in the floral organs. The knockout mutant rh52a displayed partial ES sterility, including degeneration of the ES (21%) and the presence of a double-female-gametophyte (DFG) structure (11.8%). The DFG developed from two functional megaspores near the chalazal end in one ovule, and 3.4% of DFGs were able to fertilize via the sac near the micropylar pole in rh52a. RH52A was found to interact with MFS1 and ZIP4, both of which play a role in homologous recombination in rice meiosis. RNA-sequencing identified 234 down-regulated differentially expressed genes associated with reproductive development, including two, MSP1 and HSA1b, required for female germline cell specification. Taken together, our study demonstrates that RH52A is essential for the development of the rice embryo sac and provides cytological details regarding the formation of DFGs.

Zygotic quiescence prolongs the reproductive cycle in Berchemia sinica (Rhamnaceae)

More than two-thirds of the species in Berchemia exhibit remarkable reproductive phenology, characterized by prolonged fruit ripening and the occurrence of overlapping fruit ripening and flowering. However, the underlying reasons for these phenomena remain unknown. Therefore, this study employed routine paraffin section technology, histochemical technique and scanning electron microscopy to investigate embryo sac development, differences between male and hermaphroditic flowers, as well as the overwintering strategy in Berchemia sinica. The findings of this study revealed a significant reduction in pollen viability and the number of pollen grains per flower in hermaphroditic compared to male flowers. Following fertilization, the ovary of hermaphroditic flowers does not undergo significant enlargement but gradually enters the state of zygotic quiescence. Zygotic quiescence prolongation is the main reason for the long reproductive cycle of B. sinica, and it takes approximately 14 months for the entire cycle from flower bud differentiation to fruit ripening to complete. Long reproductive cycle and complex overwintering mechanism exhibited by B. sinica resemble those seen in species belonging to Betulaceae, Fagaceae, Tapisciaceae, and Theaceae, suggesting convergent evolution during Earth's glacial period resulted in similar adaptive structures among these groups delayed fruit ripening leads to overlap between current year's flowers with previous year's fruits. The trade-off between flower and fruit for reproductive resources may have driven evolutionary transition from ancestral hermaphroditism towards androdioecy in Berchemia.

Morphology of flower bud differentiation and floral organ specialization in Caucasian clover

AbstractFlower bud differentiation is the commencement of plant reproductive development. Flower organ specialization follows flower bud differentiation. This biological mechanism controls whether plants reproduce successfully via seed. Caucasian clover (Trifolium ambiguum) is used in mixed temperate pastures. However, its low natural seed setting rate contributes to a low seed yield. This study aimed to elucidate how the reproductive biology of Caucasian clover could affect seed production. Caucasian clover flower bud differentiation and flower organ development were systematically examined. Flower bud differentiation has six consecutive stages: pre‐differentiation, differentiation of the inflorescence, the floret, the perianth, the stamen, and finally the pistil primordium. The anther has two chambers. Pollen grains have six development phases: the tetraspore, early‐uninucleate, late‐uninucleate, vegetative cell and germ‐cell form, binucleate, and maturation phase. The microspore tetrad is tetrahedral, and the mature pollen grains are bicellular. Pollen grain viability was found 35%. The Caucasian clover monocarpelate ovary contains 1–2 ovules that are covered with a 2‐layer integument. The ovule is anatropous. The embryo sac development type is a polygonum. The floral formula is “⚥↑K(5) C1+1+2 A(9)+1 G(1:1:1−2)”. These genetic traits of two pollen sacs and low viability pollen grains may be responsible for the low natural seed setting. Poor seed setting may also be due to embryo sac dysplasia that leads to ovule sterility. Therefore, the breeding aim should be to improve pollen viability, to decrease embryo sac dysplasia and to reduce embryo abortion that could be targeted for genetic improvement to enhance seed yields.

DEVELOPMENT OF FEMALE REPRODUCTIVE STRUCTURES IN <i>BOECHERA</i> (BRASSICACEAE) SPECIES UNDER SEXUAL AND APOMICTIC REPRODUCTIVE MODES

The results of comparative embryological study of the two species from the genus Boechera (B. falcata and B. stricta), which are characterized by a sexual mode of reproduction, and the apomictic M4B accession, an ancient natural hybrid of B. stricta × B. retrofracta, are presented. A detailed description of the ovule development of the studied plants is described. A significant similarity of the ovule development has been shown; a general plan of the ovules structure is ortho-campylotropic, bitegmal, medionucellate. The development of the embryo sac in B. falcata and B. stricta corresponds to the monosporic Polygonum type; in the ovules of the M4B accession, an apomictic unreduced female gametophyte is predominantly formed as a result of Taraxacum-type diplospory. However, in this accession apomixis is facultative, since a small amount of the ovules in the ovary forms a reduced embryo sac with participation of meiosis, but unlike B. falcata and B. stricta, after the second meiotic division, a triad of megaspores is formed, of which the chalazal one is functional. In the ovules of the apomictic M4B accession, underdevelopment of the inner integument was often observed, which, apparently, correlates with arrest of the megasporocyte and embryo sac development, since destruction was often observed in ovules with a short inner integument. This abnormality is likely due to the hybrid nature of the M4B accession rather than apomixis. In this accession, the embryo is formed parthenogenetically, but its development requires endosperm, which apparently develops by pseudogamy, only after triple fusion (fertilization of the fused polar nuclei by sperm). In the absence of endosperm, the embryo perishes, and seed development is arrested.

ClBBM and ClPLT2 function redundantly during both male and female gametophytes development in watermelon

As a prerequisite for sexual reproduction, gametophyte development is an interesting process involving cell proliferation, differentiation and specialization. Gametogenesis has been extensively explored in model plants, but the regulatory mechanism of gametophyte development largely remains unknown in Cucurbitaceae species. In present study, we have shown that watermelon ClBBM and ClPLT2, two AP2/ERF transcription factors, participated in both male and female gametophyte development. Clbbm and Clplt2 single mutants resembled wild-type phenotypes in both vegetative and reproductive development. But Clbbm/Clplt2 double mutant showed partial pollen abortion and bore less seeds comparing to WT. Our results indicated that the abnormal pollen grains were caused by premature tapetum degeneration, and reduced seed-set was due to faulty embryo sac development. ClBBM and ClPLT2 were expressed in FG4 embryo sacs, and their transcripts were also detectable in the tapetum and microspore of stage 9 anthers, which was consistent with developmental stages of defective phenotypes observed in double mutant. The expression of genes essential for tapetum development, ClATM1, ClAMS, ClMS1 and ClMS188, was decreased in Clbbm/Clplt2 double mutants. Moreover, the transcriptome analysis indicated that ClBBM and ClPLT2 participated in tapetum and pollen wall development by regulate cell cycle, transmembrane transport, glucan and cellulose metabolic process. Collectively, ClBBM and ClPLT2 were functionally redundant in regulating gametophyte development in watermelon, and their functions differ from their homologous genes in model plant Arabidopsis.

OsmiRNA5488 Regulates the Development of Embryo Sacs and Targets OsARF25 in Rice (Oryza sativa L.).

Small RNAs are a class of non-coding RNAs that typically range from 20 to 24 nucleotides in length. Among them, microRNAs (miRNAs) are particularly important regulators for plant development. The biological function of the conserved miRNAs has been studied extensively in plants, while that of the species-specific miRNAs has been studied in-depth. In this study, the regulatory role of a rice-specific OsmiRNA5488 (OsmiR5488) was characterized with the miR5488-overexpressed line (miR5488-OE) and miR5488-silenced line (STTM-5488). The seed-setting rate was notably reduced in miR5488-OE lines, but not in STTM-5488 lines. Cytological observation demonstrated the different types of abnormal mature embryo sacs, including the degeneration of embryo sacs and other variant types, in miR5488-OE lines. The percentage of the abnormal mature embryo sacs accounted for the reduced value of the seed-setting rate. Furthermore, OsARF25 was identified as a target of OsmiR5488 via RNA ligase-mediated 3'-amplifification of cDNA ends, dual luciferase assays, and transient expression assays. The primary root length was decreased with the increases in auxin concentrations in miR5488-OE lines compared to wild-type rice. Summarily, our results suggested that OsmiR5488 regulates the seed-setting rate and down-regulates the targeted gene OsARF25.

Embryo sac development relies on symplastic signals from ovular integuments in Arabidopsis.

Ovules are female reproductive organs of angiosperms, consisting of sporophytic integuments surrounding female gametophytes, that is, embryo sacs. Synchronization between integument growth and embryo sac development requires intracellular communication. However, signaling routes through which cells of the two generations communicate are unclear. We report that symplastic signals through plasmodesmata (PDs) of integuments are critical for the development of female gametophytes. Genetic interferences of PD biogenesis either by functional loss of CHOLINE TRANSPORTER-LIKE1 (CTL1) or by integument-specific expression of a mutated CALLOSE SYNTHASE 3 (cals3m) compromised PD formation in integuments and reduced fertility. Close examination of pINO:cals3m or ctl1 ovules indicated that female gametophytic development was either arrested at various stages after the formation of functional megaspores. In both cases, defective ovules could not attract pollen tubes, leading to the failure of fertilization. Results presented here demonstrate a key role of the symplastic route in sporophytic control of female gametophytic development.

The mutation of CaCKI1 causes seedless fruits in chili pepper (Capsicum annuum)

The seedless mutant tn-1 in chili pepper is caused by a mutation in CaCKI1 (CA12g21620), which encodes histidine kinase involving female gametophyte development. An amino acid insertion in the receiver domain of CaCKI1 may be the mutation responsible for tn-1. Seedlessness is a desirable trait in fruit crops because the removal of seeds is a troublesome step for consumers and processing industries. However, little knowledge is available to develop seedless chili peppers. In a previous study, a chili pepper mutant tn-1, which stably produces seedless fruits, was isolated. In this study, we report characterization of tn-1 and identification of the causative gene. Although pollen germination was normal, confocal laser microscopy observations revealed deficiency in embryo sac development in tn-1. By marker analysis, the tn-1 locus was narrowed down to a 313kb region on chromosome 12. Further analysis combined with mapping-by-sequencing identified CA12g21620, which encodes histidine kinase as a candidate gene. Phylogenetic analysis revealed CA12g21620 was the homolog of Arabidopsis CKI1 (Cytokinin Independent 1), which plays an important role in female gametophyte development, and CA12g21620 was designated as CaCKI1. Sequence analysis revealed that tn-1 has a 3-bp insertion in the 6th exon resulting in one lysine (K) residue insertion in receiver domain of CaCKI1, and the sequence nearby the insertion is widely conserved among CKI1 orthologs in various plants. This suggested that one K residue insertion may reduce the phosphorylation relay downstream of CaCKI1 and impair normal development of female gametophyte, resulting in seedless fruits production in tn-1. Furthermore, we demonstrated that virus-induced gene silencing of CaCKI1 reducednormally developed female gametophyte in chili pepper. This study describes the significant role of CaCKI1 in seed development in chili pepper and the possibility of developing seedless cultivars using its mutation.

Mutation of DEFECTIVE EMBRYO SAC1 results in a low seed-setting rate in rice by regulating embryo sac development.

The seed-setting rate has a significant effect on grain yield in rice (Oryza sativa L.). Embryo sac development is essential for seed setting; however, the molecular mechanism underlying this process remains unclear. Here, we isolated defective embryo sac1 (des1), a rice mutant with a low seed-setting rate. Cytological examination showed degenerated embryo sacs and reduced fertilization capacity in des1. Map-based cloning revealed a nonsense mutation in OsDES1, a gene that encodes a putative nuclear envelope membrane protein (NEMP)-domain-containing protein that is preferentially expressed in pistils. The OsDES1 mutation disrupts the normal formation of functional megaspores, which ultimately results in a degenerated embryo sac in des1. Reciprocal crosses showed that fertilization is abnormal and that the female reproductive organ is defective in des1. OsDES1 interacts with LONELY GUY (LOG), a cytokinin-activating enzyme that acts in the final step of cytokinin synthesis; mutation of LOG led to defective female reproductive organ development. These results demonstrate that OsDES1 functions in determining the rice seed-setting rate by regulating embryo sac development and fertilization. Our study sheds light on the function of NEMP-type proteins in rice reproductive development.

OsPHS1 is required for both male and female gamete development in rice

Meiosis plays an essential role in the production of male and female gametes. Extensive studies have elucidated that homologous chromosome association and pairing are essential for crossing-over and recombination of chromosomal segments. However, the molecular mechanism of chromosome recognition and pairing remains elusive. Here, we identified a rice male–female sterility mutant plant. Cytological observations showed that the development of both pollen and embryo sacs of the mutant were abnormal due to defects in homologous chromosome recognition and pairing during prophase I. Map-based cloning revealed that Os06g0473000 encoding a poor homologous synapsis 1 (PHS1) protein is the candidate target gene, which was confirmed by knockout using CRISPR/Cas9 technology. Sequence analysis revealed a single base mutation (G > A) involving the junction of the fourth exon and intron of OsPHS1, which is predicted to alter splicing, resulting in an Osphs1 mutant. Expression pattern analysis indicated that OsPHS1 expression levels were mainly expressed in panicles at the beginning of meiosis. Subcellular localization analysis demonstrated that the OsPHS1 protein is situated in the nucleus and cytoplasm. Taken together, our results suggest an important role for OsPHS1 in homologous chromosome pairing in both male and female gametogenesis in rice.

Differential proMMP-2 and proMMP-9 secretion in human pre-implantation embryos at day 5 of development.

Morphological development is the most common non-invasive criterion used to select in vitro human embryos for implantation. With this criterion, however, embryos in cellular arrest go unnoticed. A more accurate criterion is needed to improve the success rate of implantation. Extracellular matrix metalloproteases type 2 (MMP-2) and MMP-9 are key markers of embryonic development and the implantation process, according to various animal studies. The first objective of this study was to examine proMMP-2 and proMMP-9 activity in the culture media of human embryos with good morphological development. Secondly, the results of proMMP-2 and proMMP-9 activity in the culture medium were compared between pregnant and non-pregnant. Forty-two patients were approved by the Ethics and Research Committees of the Instituto Nacional de Perinatología in México City hospital, based on institutional inclusion criteria for in vitro fertilization. On day 5 of development, embryos were transferred to patients, and the culture media secretion profile of proMMP-2 and proMMP-9 activity was determined by substrate gel zymography. After analysis of embryo sac development, each patient was assigned to the pregnant (n=17) or non-pregnant (n=25) group. Our results demonstrate that proMMP-2 was active in the culture media corresponding to all 17 women achieving full-term pregnancy and proMMP-9 in the media corresponding to 11 of these women. Contrarily proMMP-2 and proMMP-9 were active in the culture media corresponding to 3 and 11 of the 25 non-pregnant patients, respectively. The clinical implications of this study suggest the activity evaluation of proMMP-2 and proMMP-9 in embryonic culture media on day 5 of development appears to be a reliable indicator of the quality of embryos and their capacity to establish a pregnancy.

Cytokinin regulates female gametophyte development by cell cycle modulation in Arabidopsis thaliana

Male and female gametophyte development, double fertilization, and embryogenesis are key to alternating generations in angiosperms. The female gametophyte of Arabidopsis is an eight-nucleate haploid structure developed from functional megaspores (FMs) through three flawless mitoses regulated by a series of cell cycle genes. Cytokinin, an important phytohormone, plays a critical role in the regulation of plant growth and development. However, the mechanisms by which cytokinins regulate female gametophyte development remain largely unknown. In this study, we constructed transgenic plants (pES1::CKX1) with low cytokinin levels in the embryo sac. Phenotypic analysis showed that pES1::CKX1 inhibits female gametophyte development. Microscopic observation revealed that female gametophyte development of pES1::CKX1 was delayed. The promoters of all cell cycle genes were cloned and transformed into wild-type (WT). We crossed these transgenic plants of cell cycle genes expressed in ovules with pES1::CKX1 and compared the expression level of β-glucuronidase (GUS) in pES1::CKX1 and WT. Many cell cycle-regulated genes were up or downregulated in pES1::CKX1 compared with WT, and the embryo sac development cell cycle in cycd2;1/+ cycd3;3 was defective. Our results demonstrated that cytokinin affects cell division in the female gametophyte by affecting the expression of cell cycle genes.

A Preliminary Study of Apomictic Characteristics of ‘Mianli’ (Pyrus sinkiangensis Yü)

This study investigated the ploidy of ‘Mianli’ with flow cytometry and the traditional chromosome squash technique. Its pollination biology and the occurrence and formation of embryo sacs before and after flowering were observed in paraffin sections to characterize its embryo sacs. The intersimple sequence repeat (ISSR) marker technique was used to test the uniformity of progeny of ‘Mianli’ treatments. The chromosome number of ‘Mianli’ is 2n = 2x = 34. The ploidy results were consistent with those identified by flow cytometry. ‘Mianli’ is male-sterile, and the anatropous ovule has double integuments. ‘Mianli’ can bear fruit normally and produce fertile seeds under the treatments of emasculation with bagging or no emasculation with bagging, but the seed yield is very low and significantly lower than that under artificial pollination or natural pollination. The developmental process of embryo sacs under natural pollination showed that most megasporocytes develop into mature sexual embryo sacs through meiosis and a few megasporocytes degenerate. Some sexual embryo sacs continue to develop into embryos after fertilization, and some sexual embryo sacs are aborted. In addition, new aposporous initial cells are generated irregularly at each stage from the emergence of megasporocyte to the end of sexual reproduction or abortion. The observation of the development of embryo sacs under emasculation with bagging showed that after pollination is blocked, mature sexual embryo sacs degenerate, and aposporous mononucleate embryo sacs appear around the degenerated sexual embryo sacs or in the peripheral tissues. Then, the process of proembryonic masses developing into spherical embryo was observed. A genetic uniformity analysis of progeny of ‘Mianli’ using ISSR was performed. The results showed that the progeny population under emasculation with bagging has high consistency at the molecular level, with some plants having full consistency with the female parent’s banding pattern, demonstrating consistency with the maternal genetic characteristics. The progeny under artificial pollination or natural pollination do not have the same banding pattern as the female parent. Because there is no pseudogamy, all of the progeny are true hybrids. In summary, it seems that ‘Mianli’ only has sexual reproduction in the presence of pollen, and only a few ovules are stimulated to undergo apomixis after pollination is blocked.

Cytological Observation and Candidate Gene Identification of a Female Semi-Sterile Rice Mutant

Rice is an important staple crop, and fertilization has a very important role in the production of high-yield and well-quality hybrid of rice. The abnormal development of female gametes will lead to a significant decrease in rice yields. Due to the complex mechanism of female gamete development, the molecular mechanism of female sterility has not made a breakthrough. In this study, one semi-sterile mutant m21 and its fifth generation of backcross material (WZN58 and WZN68) were used as materials, and the phenotypic observation and cytological observation of embryo sac development were performed. Moreover, the pollen viability was evaluated by iodine potassium iodide (I2-KI) staining. Bulked-segregant analysis (BSA) re-sequencing and transgenic analysis were further applied to investigate the molecular mechanism of semi-sterile phenotype. These results demonstrated that male gamete development was normal, while the abnormality of the mutant was caused by the abnormal development of female gamete. Among 112 candidate genes detected by BSA, one gene Os10g0506000 was identified responsible for the semi-sterile phenotype of m21 mutant. This study provides an ideal material and candidate gene for rice female fertility analysis.

Flower differentiation and fruiting dynamics in olive trees (Olea europaea): Eco-physiological analysis in the Mediterranean basin

The formation of flowers in sufficient number and quality is a prerequisite for a successful subsequent fruit set. Despite the abundant flowering, olive trees (Olea europea) are characterized by a very low fruit set, and a very severe yield alteration leading to market fluctuation over time. The goal of this paper is to explore and analyze eco-physiological driving factors behind the poor fruit set in Mediterranean olive groves. The key mechanisms causing floral differentiation and extreme yield alternate are functional of plant genetic variability, nutrient competition, and some ecological aspects as a response to climate change. Additionally, olive inflorescence architecture appears to be complex and can vary between cultivars; the olive flower differentiation results in a variable proportion of hermaphrodite, pistillate and staminate flowers among olive cultivars as well as across canopy positions and branches, enhancing nutrient competition between flowers. Self-pollination could be one of the limiting factors for increasing early fruit abscission and extreme alternate fruit-bearing. Hormonal treatments to reduce alternate production in olive trees should be explored. The current review analysis shall help to improve olive grove management, but also for breeding new cultivars more suitable for Mediteranean agro-ecological constraints. Ovule viability and fertilisation, and embryo sac development abnormalities should all be further investigated.

Viability of Embryo Sacs and Fruit Set in Different Plum (Prunus domestica L.) Cultivars Grown under Norwegian Climatic Conditions.

Compatibility and synchrony between specialized tissues of the pistil, female gametophytes and male gametophytes, are necessary for successful pollination, fertilization, and fruit set in angiosperms. The aim of the present work was to study the development and viability of embryo sacs, as well as fertilization success, in relation to the fruit set of the cultivars ‘Mallard’, ‘Edda’, ‘Jubileum’, and ‘Reeves’, under specific Norwegian climatic conditions. Emasculated, unpollinated, and open-pollinated flowers were collected at the beginning of flowering, and on the 3rd, 6th, 9th, and 12th days after flowering, from all four plum cultivars over two years (2018/2019). Ovaries were dehydrated, embedded in paraffin wax, sectioned, stained, and observed under a light microscope. Results showed the existence of synchronization between successive phases in the development of the embryo sac and individual phases of flowering. All plum cultivars had higher percentages of viable embryo sacs, fertilized embryo sacs, and fruit set in 2018 than in 2019. These differences may be related to the very low temperatures during the post-full-flowering period in 2019, and to the low adaptation of some studied cultivars to unfavorable conditions. In our study, the cultivar ‘Jubileum’ showed the highest percentage of viable embryo sacs, fertilized embryo sacs, and fruit set compared to other cultivars, i.e., the best low-temperature adaptation.