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The complexity of citrus phylogeny and taxonomy, influenced by polyembryony, genetic diversity, and extended generation cycles, necessitates assessing genetic relationships for improved production and disease resilience. DNA polymorphism markers are pivotal in genotyping and resolving phenotypic varietal ambiguities through accurate and objective genetic relatedness and distinctiveness assessments. In this study, 27 accessions of kagzi and ornamental lime were evaluated at morphological (based on IPGRI citrus descriptor), biochemical, and molecular levels using DNA-based markers, namely, random amplified polymorphic DNA (RAPD), start codon targeted (SCoT) polymorphism, and simple sequence repeats (SSR). Morphological analysis unveiled significant variation in leaf lamina attachment, shape, margins, fruit characteristics, and texture across all parameters. Biochemical research noted minimal differences, with total soluble solids (TSS) ranging from 5.7° to 8.0° Brix and titratable acidity (TA) from 0.36 to 0.50%. Notably, ornamental lime accessions exhibited higher TSS:TA ratios of 16.00. Molecular markers (RAPD, SCoT, and SSR) corroborated these findings, yielding 115, 74, and 64 amplicons, respectively, with polymorphic percentages varying from 33.33 to 100%. Polymorphic information content ranged from 0.20 to 0.90 for RAPD, 0.41 to 0.99 for SCoT, and 0.07 to 0.67 for SSR markers. Analysis of molecular variance (AMOVA) substantiated significant genetic differences among the kagzi and ornamental lime accessions (RAPD PhiPT = 0.267, SCoT PhiPT = 0.351, SSR PhiPT = 0.107). Furthermore, STRUCTURE software 2.3.4 revealed a ΔK value of two (K = 2), indicating two distinct genetic pools within the accessions, possibly attributable to bud mutations generating varying morphological traits within the same plant. Molecular marker-based germplasm characterization can mitigate confusion linked to morphological characteristics and multiple cultivar registrations. [ABSTRACT FROM AUTHOR]

Rootstocks/scions with potential for commercial growing, either for the industry or fresh market, have been one of the main objectives of the fruit breeding programs. Fruit crop breeding has impediments like polyembryony (PE) which is otherwise desirable for clonal plant propagation. In nature PE exist in some fruit crops like citrus, mango, jamun, rose apple, almond etc. In polyembryonic crops, there is an occurrence of the more than one embryo in seed and subsequently numerous seedlings emerge from that single seed. The mother type seedlings are considered nucellar are often vigorous in nature as compared to zygotic ones. In this review, the classification, genetic basis and mechanism of PE has been discussed. In current review, morphogenetic authentication of seedling from polyembronic seed has been discussed in the light of recent research. Transcriptional pathway along with the inheritance pattern and molecular breeding aspects of PE has also been summarized. [ABSTRACT FROM AUTHOR]

n the Neotropics, the focus of apomictic studies predominantly centres on trees within the Brazilian savanna, characterized, mostly as sporophytic and facultative, associated with polyploidy and polyembryony. To enhance our understanding of the mechanisms governing apomixis and sexual reproduction in tropical herbaceous plants, we clarify the relationship between apomixis, chromosome counts, and polyembryony in the epiphytic orchid Zygopetalum mackayi , which forms a polyploid complex within rocky outcrops in both the Brazilian savanna and the Atlantic forest. To define embryo origins and describe megasporogenesis and megagametogenesis, we performed manual self-pollinations in first-day flowers of cultivated plants, considering all three cytotypes (2 x , 3 x , 4 x) of this species. Flowers and fruits at different stages were collected to describe the development and morphology of ovules and seeds considering sexual and apomictic processes. As self-pollination treatments resulted in high fruit abortion in diploids, we also examined pollen tube development in aborted flowers and fruits to search for putative anomalies. Megasporogenesis and megagametogenesis occur regularly in all cytotypes. Apomixis is facultative and sporophytic, and associated with polyploid cytotypes, while diploid individuals exclusively engage in sexual reproduction. Polyembryony is caused mainly by the production of adventitious embryos from nucellar cells of triploids and tetraploids, but also by the development of multiple archesporia in all cytotypes. Like other apomictic angiosperms within the Brazilian savanna, our findings demonstrate that apomixis in Z. mackayi relies on pollinators for seed production. We also consider the ecological implications of these apomictic patterns in Z. mackayi within the context of habitat loss and its dependence on pollinators. [ABSTRACT FROM AUTHOR]

Rough lemon is indigenous and has wide adaptability in Indian citriculture. Rough lemon and Volkamer lemon are comparable in horticultural performance and their resistance against biotic and abiotic stresses are almost similar but Volkamer lemon has better adaptability in different agroclimatic conditions. To improve Volkamer lemon hybridization has been attempted with Kinkoji (citrus greening tolerance), Swingle citrumelo (Phytophthora resistance), and Cleopatra (salinity tolerance). Polyembryony in Volkamer lemon and absence of the heritable morphological markers (Cleopatra) are limiting factors for hybrid identification and morphological characters are not being segregated in Kinkoji and Swingle citrumelo at early stage, then molecular markers are employed for detection of putative zygotic seedlings. We performed hybridization between Volkamer lemon (♀), Kinkoji (B cross) (58 F1) (♂), Swingle citrumelo (D cross) (71 F1) (♂), Cleopatra (N cross) (83 F1) (♂) and discriminate the hybrids by using 25 SSR markers. The fifteen markers were found efficient for hybrid identification (> 71% hybrids detected). CCSME23 (88.21%) marker has detected 187 hybrids in all crosses thereafter, 184 hybrids detected each by HvSSRCS-2, HvSSRCS-135, HvSSRCS-237, HvSSRCM-92 and HvSSRCM-466 with 86.79%. A total of 44 qualitative and 18 quantitative characters were analysed. Fifteen qualitative and six quantitative characters were found significant variability in leaf, spine, branch, and stem. Sufficient amount of genetic variability was detected through phylogenetic analysis and population structure analysis showed two subpopulations in B and N cross but in D cross five subpopulations were detected. After primary screening, nine F1 hybrids for citrus greening tolerance and twelve F1 hybrids for Phytophthora tolerance were identified. The newly developed F1 hybrids exhibited phenotypic and genetic diversity along with disease tolerance. These potential hybrids may be recommended for commercial citriculture after thorough horticultural evaluation. [ABSTRACT FROM AUTHOR]

Background: Genetic variation for salt tolerance remains elusive in jamun (Syzygium cumini). Methods: Effects of gradually increased salinity (2.0–12.0 dS/m) were examined in 20 monoembryonic and 28 polyembryonic genotypes of jamun. Six genotypes were additionally assessed for understanding salt-induced changes in gas exchange attributes and antioxidant enzymes. Results: Salt-induced reductions in leaf, stem, root and plant dry mass (PDM) were relatively greater in mono- than in poly-embryonic types. Reductions in PDM relative to control implied more adverse impacts of salinity on genotypes CSJ-28, CSJ-31, CSJ-43 and CSJ-47 (mono) and CSJ-1, CSJ-24, CSJ-26 and CSJ-27 (poly). Comparably, some mono- (CSJ-5, CSJ-18) and poly-embryonic (CSJ-7, CSJ-8, CSJ-14, CSJ-19) genotypes exhibited least reductions in PDM following salt treatment. Most polyembryonic genotypes showed lower reductions in root than in shoot mass, indicating that they may be more adept at absorbing water and nutrients when exposed to salt. The majority of genotypes did not exhibit leaf tip burn and marginal scorch despite significant increases in Na+ and Cl−, suggesting that tissue tolerance existed for storing excess Na+ and Cl− in vacuoles. Jamun genotypes were likely more efficient in Cl− exclusion because leaf, stem and root Cl− levels were consistently lower than those of Na+ under salt treatment. Leaf K+ was particularly little affected in genotypes with high leaf Na+. Lack of discernible differences in leaf, stem and root Ca2+ and Mg2+ contents between control and salt treatments was likely due to their preferential uptake. Correlation analysis suggested that Na+ probably had a greater inhibitory effect on biomass in both mono- and poly-embryonic types. Discriminant analysis revealed that while stem and root Cl− probably accounted for shared responses, root Na+, leaf K+ and leaf Cl− explained divergent responses to salt stress of mono- and poly-embryonic types. Genotypes CSJ-18 and CSJ-19 seemed efficient in fending off oxidative damage caused by salt because of their stronger antioxidant defences. Conclusions: Polyembryonic genotypes CSJ-7, CSJ-8, CSJ-14 and CSJ-19, which showed least reductions in biomass even after prolonged exposure to salinity stress, may be used as salt-tolerant rootstocks. The biochemical and molecular underpinnings of tissue tolerance to excess Na+ and Cl− as well as preferential uptake of K+, Ca2+, and Mg2+ need to be elucidated. [ABSTRACT FROM AUTHOR]

Background Genetic variation for salt tolerance remains elusive in jamun (Syzygium cumini). Methods Effects of gradually increased salinity (2.0–12.0 dS/m) were examined in 20 monoembryonic and 28 polyembryonic genotypes of jamun. Six genotypes were additionally assessed for understanding salt-induced changes in gas exchange attributes and antioxidant enzymes. Results Salt-induced reductions in leaf, stem, root and plant dry mass (PDM) were relatively greater in mono- than in poly-embryonic types. Reductions in PDM relative to control implied more adverse impacts of salinity on genotypes CSJ-28, CSJ-31, CSJ-43 and CSJ-47 (mono) and CSJ-1, CSJ-24, CSJ-26 and CSJ-27 (poly). Comparably, some mono- (CSJ-5, CSJ-18) and poly-embryonic (CSJ-7, CSJ-8, CSJ-14, CSJ-19) genotypes exhibited least reductions in PDM following salt treatment. Most polyembryonic genotypes showed lower reductions in root than in shoot mass, indicating that they may be more adept at absorbing water and nutrients when exposed to salt. The majority of genotypes did not exhibit leaf tip burn and marginal scorch despite significant increases in Na+ and Cl−, suggesting that tissue tolerance existed for storing excess Na+ and Cl− in vacuoles. Jamun genotypes were likely more efficient in Cl− exclusion because leaf, stem and root Cl− levels were consistently lower than those of Na+ under salt treatment. Leaf K+ was particularly little affected in genotypes with high leaf Na+. Lack of discernible differences in leaf, stem and root Ca2+ and Mg2+ contents between control and salt treatments was likely due to their preferential uptake. Correlation analysis suggested that Na+ probably had a greater inhibitory effect on biomass in both mono- and poly-embryonic types. Discriminant analysis revealed that while stem and root Cl− probably accounted for shared responses, root Na+, leaf K+ and leaf Cl− explained divergent responses to salt stress of mono- and poly-embryonic types. Genotypes CSJ-18 and CSJ-19 seemed efficient in fending off oxidative damage caused by salt because of their stronger antioxidant defences. Conclusions Polyembryonic genotypes CSJ-7, CSJ-8, CSJ-14 and CSJ-19, which showed least reductions in biomass even after prolonged exposure to salinity stress, may be used as salt-tolerant rootstocks. The biochemical and molecular underpinnings of tissue tolerance to excess Na+ and Cl− as well as preferential uptake of K+, Ca2+, and Mg2+ need to be elucidated.

The use of doubled-haploids in plant breeding programs enables accelerating the release of new varieties adapted to climate change. The durum wheat x maize crosses technique is a method of choice for producing durum wheat haploid plants. The haploid embryos produced by this method develop without albumen and their survival is ensured by post-pollination hormonal treatments. In this study, nine post-pollination treatments with 2,4-Dichlorophenoxyacetic acid (2,4-D), Picloram and Dicamba at the concentrations of 10, 50 and 100 mg.L-1 were applied to 7 durum wheat genotypes. The effects of genotype and post-pollination treatment on durum wheat haploid embryos produced by durum wheat x maize crosses and the use of the endosperm nursing technique for haploid plantlets regeneration were investigated. The haploid induction parameters varied with the durum wheat genotypes as well as the post-pollination treatments. The phenomenon of polyembryony resulting from durum wheat x maize crosses is reported for the first time in this article. The durum wheat genotypes showed different abilities to produce monoembryo and polyembryos. The post-pollination treatments with 2,4-D (10 mg.L-1) and Picloram (10 and 100 mg.L-1) gave a higher embryo formation frequency than the treatments with Dicamba. The embryo conversion to plantlet was greatly improved, especially in recalcitrant genotypes using the durum wheat endosperm as supplemental nourishment in combination with the Gamborg B5 regeneration medium. [ABSTRACT FROM AUTHOR]

Introduction: Putative mutants were generated through gamma irradiation in the polyembryonic mango genotype Nekkare. The putative mutant progenies along with control seedlings and mother plants were evaluated by comparing the compositions and relative proportions of their major volatile compounds. Methods: Volatile profiling was done using headspace-solid phase microextraction (HS SPME) method coupled with gas chromatography-mass spectrometry (GC-MS MS). Furthermore, characterisation of putative mutants and control seedlings was carried out using simple sequence repeat (SSR) markers to ascertain the genetic diversity present in the samples under study. Results: Monoterpenes were the most abundant volatile compound in all the studied samples (ranging from 34.76% to 91.41%) out of which I-Phellandrene and cis-Ocimene formed the major fraction in mother plants (20.45%-21.86% and 16.17%-21.27%, respectively) and control seedlings (23.32%-24.95% and 18.95%-20.81%, respectively), while beta-Phellandrene was dominant in the selected putative mutant samples (2.34%-29.53%). Among sesquiterpenes, trans-Caryophyllene was detected only in the putative mutant samples (0.10%-30.18%). Grouping together of mother plants and control seedlings was seen in the cluster analysis, while the putative mutants grouped apart from them suggesting genetic diversity. Genetic distance between the mother plants and control seedlings ranged from 0.97 to 2.73, while between putative mutants, control seedlings, and mother plants, it ranged from 6.54 to 9.82. SSRbased characterisation of putative mutant seedlings showed that mutation caused variability in the treated population. This was evident from the high allelic richness ranging from 4 to 12 with a mean of 7 and a higher mean Shannon's Information Index (1.50) of the putative mutant population. Discussion: The study demonstrates that volatile profiling and molecular characterisation using SSR markers could be used as a tool to detect variation in a mutated population. In addition, volatile profiling can be used to validate putative mutants in polyembryonic mango genotypes where the seedlings of nucellar origin are similar to mother plants. [ABSTRACT FROM AUTHOR]

The occurrence of more than one embryo per seed (polyembryony) is common among angiosperms; however, there are gaps in the knowledge of its effects on the early stages of plant development. In this context, we study the effects of polyembryony and intraspecific variability in gemellar competition during the seed–seedling transition in Neotropical Handroanthus chrysotricus (Bignoniaceae). We used seeds from five cultivated trees in an urban environment inserted in a biodiversity hotspot (Cerrado). Embryo mass, seed germination, seedling emergence and seedling morphometry were evaluated. We did not find intraspecific variability in seed germination, seedling emergence or the mean number of embryos and seedlings per seed. On the other hand, intraspecific variability was observed during the transition from embryo to seedling. When only one seedling emerged from a seed, the seed–seedling transition was more asynchronous than when more seedlings emerged from one seed (with higher uncertainty and a longer time to emergence of the last seedling). The mass of embryos and seedlings decreased with the increase in the number of embryos in a seed, reinforcing the occurrence of gemellar competition. However, the total mass of embryos per seed was similar. The increase in seedlings per seed also decreased the morphometric measurements of each one. A positive morphometric aspect of the emergence of two seedlings per seed was that they had the highest total seedling mass, evidencing the positive Allee effect. Polyembryony had both positive and negative effects on seed germination and seedling morphology in the species, which helps to understand how this phenomenon acts on seed biology and plant establishment.

IntroductionPutative mutants were generated through gamma irradiation in the polyembryonic mango genotype Nekkare. The putative mutant progenies along with control seedlings and mother plants were evaluated by comparing the compositions and relative proportions of their major volatile compounds.MethodsVolatile profiling was done using headspace-solid phase micro-extraction (HS SPME) method coupled with gas chromatography–mass spectrometry (GC-MS MS). Furthermore, characterisation of putative mutants and control seedlings was carried out using simple sequence repeat (SSR) markers to ascertain the genetic diversity present in the samples under study.ResultsMonoterpenes were the most abundant volatile compound in all the studied samples (ranging from 34.76% to 91.41%) out of which I-Phellandrene and cis-Ocimene formed the major fraction in mother plants (20.45%–21.86% and 16.17%–21.27%, respectively) and control seedlings (23.32%–24.95% and 18.95%–20.81%, respectively), while beta-Phellandrene was dominant in the selected putative mutant samples (2.34%–29.53%). Among sesquiterpenes, trans-Caryophyllene was detected only in the putative mutant samples (0.10%–30.18%). Grouping together of mother plants and control seedlings was seen in the cluster analysis, while the putative mutants grouped apart from them suggesting genetic diversity. Genetic distance between the mother plants and control seedlings ranged from 0.97 to 2.73, while between putative mutants, control seedlings, and mother plants, it ranged from 6.54 to 9.82. SSR-based characterisation of putative mutant seedlings showed that mutation caused variability in the treated population. This was evident from the high allelic richness ranging from 4 to 12 with a mean of 7 and a higher mean Shannon’s Information Index (1.50) of the putative mutant population.DiscussionThe study demonstrates that volatile profiling and molecular characterisation using SSR markers could be used as a tool to detect variation in a mutated population. In addition, volatile profiling can be used to validate putative mutants in polyembryonic mango genotypes where the seedlings of nucellar origin are similar to mother plants.

Protoplast has been widely used in biotechnologies to circumvent the breeding obstacles in citrus, including long juvenility, polyembryony, and male/female sterility. The protoplast-based transient gene expression system is a powerful tool for gene functional characterization and CRISPR/Cas9 genome editing in higher plants, but it has not been widely used in citrus. In this study, the polyethylene glycol (PEG)-mediated method was optimized for citrus callus protoplast transfection, with an improved transfection efficiency of 68.4%. Consequently, the efficiency of protein subcellular localization assay was increased to 65.8%, through transient expression of the target gene in protoplasts that stably express the fluorescent organelle marker protein. The gene editing frequencies in citrus callus protoplasts reached 14.2% after transient expression of CRISPR/Cas9 constructs. We demonstrated that the intronic polycistronic tRNAgRNA (inPTG) genome editing construct was functional in both the protoplast transient expression system and epicotyl stable transformation system in citrus.With this optimized protoplast transient expression system, we improved the efficiency of protein subcellular localization assay and developed the genome editing system in callus protoplasts, which provides an approach for prompt test of CRISPR vectors. [ABSTRACT FROM AUTHOR]

The occurrence of more than one embryo per seed (polyembryony) is common among angiosperms; however, there are gaps in the knowledge of its effects on the early stages of plant development. In this context, we study the effects of polyembryony and intraspecific variability in gemellar competition during the seed–seedling transition in Neotropical Handroanthus chrysotricus (Bignoniaceae). We used seeds from five cultivated trees in an urban environment inserted in a biodiversity hotspot (Cerrado). Embryo mass, seed germination, seedling emergence and seedling morphometry were evaluated. We did not find intraspecific variability in seed germination, seedling emergence or the mean number of embryos and seedlings per seed. On the other hand, intraspecific variability was observed during the transition from embryo to seedling. When only one seedling emerged from a seed, the seed–seedling transition was more asynchronous than when more seedlings emerged from one seed (with higher uncertainty and a longer time to emergence of the last seedling). The mass of embryos and seedlings decreased with the increase in the number of embryos in a seed, reinforcing the occurrence of gemellar competition. However, the total mass of embryos per seed was similar. The increase in seedlings per seed also decreased the morphometric measurements of each one. A positive morphometric aspect of the emergence of two seedlings per seed was that they had the highest total seedling mass, evidencing the positive Allee effect. Polyembryony had both positive and negative effects on seed germination and seedling morphology in the species, which helps to understand how this phenomenon acts on seed biology and plant establishment. [ABSTRACT FROM AUTHOR]

Simple polyembryony, where one gametophyte produces multiple embryos with different sires but the same maternal haplotype, is common among vascular plants. We develop an infinite-sites, forward population genetics model showing that together polyembryony's two benefits—"reproductive compensation" achieved by providing a backup for inviable embryos and the opportunity to favor the fitter of surviving embryos—can favor its evolution. Our model tests how these factors can favor the evolution of polyembryony and how these underlying benefits of polyembryony shape the genetic load under a range of biological parameters. While these two benefits are difficult to disentangle in nature, we construct variant models of polyembryony that either only include or only exclude the opportunity for reproductive compensation. We find that reproductive compensation strongly favors the evolution of polyembryony and that polyembryony is favored much more weekly in its absence, suggesting that the benefit of a backup embryo is the major force favoring polyembryony. Remarkably, we find nearly identical results in cases in which mutations impact either embryonic or postembryonic fitness (no pleiotropy) and in cases in which mutations have identical effects on embryonic and postembryonic fitness (extreme pleiotropy). Finally, we find that the consequences of polyembryony depend on its function—polyembryony results in a decrease in mean embryonic fitness when acting as a mechanism of embryo compensation and ultimately increases mean embryonic fitness when we exclude this potential benefit. [ABSTRACT FROM AUTHOR]

Introduction: Increased soil salinity in the recent years has adversely affected the productivity of mango globally. Extending the cultivation of mango in salt affected regions warrants the use of salinity tolerant/resistant rootstocks. However, the lack of sufficient genomic and transcriptomic information impedes comprehensive research at the molecular level. Method: We employed RNA sequencing-based transcriptome analysis to gain insight into molecular response to salt stress by using two polyembryonic mango genotypes with contrasting response to salt stress viz., salt tolerant Turpentine and salt susceptible Mylepelian. Results: RNA sequencing by Novaseq6000 resulted in a total of 2795088, 17535948, 7813704 and 5544894 clean reads in Mylepelian treated (MT), Mylepelian control (MC), Turpentine treated (TT) and Turpentine control (TC) respectively. In total, 7169 unigenes annotated against all the five public databases, including NR, NT, PFAM, KOG, Swissport, KEGG and GO. Further, maximum number of differentially expressed genes were found between MT and MC (2106) followed by MT vs TT (1158) and TT and TC (587). The differentially expressed genes under different treatment levels included transcription factors (bZIP, NAC, bHLH), genes involved in Calcium-dependent protein kinases (CDPKs), ABA biosynthesis, Photosynthesis etc. Expression of few of these genes was experimentally validated through quantitative real-time PCR (qRTPCR) and contrasting expression pattern of Auxin Response Factor 2 (ARF2), Late Embryogenesis Abundant (LEA) and CDPK genes were observed between Turpentine and Mylepelian. Discussion: The results of this study will be useful in understanding the molecular mechanism underlying salt tolerance in mango which can serve as valuable baseline information to generate new targets in mango breeding for salt tolerance. [ABSTRACT FROM AUTHOR]

Seeds may differ in terms of dormancy, longevity, sensitivity to desiccation and dry mass, according to the timing (dry season/rainy season) of diaspore dispersal. In addition, seasonal variations in temperature and water availability can act as signals of the season during seed development, influencing germination responses and root growth. We evaluated the effects of temperature variations and water availability on germination parameters, root growth and seed traits of four coexisting Piper species in seasonal vegetation that differed in diaspore dispersal timing.Eight temperature treatments (15, 20, 23, 25, 28, 30, 35 °C, and alternate 30 °C–20 °C) and four induced water potentials (0, −0.3, −0.6 and −1.2 MPa) were used. The parameters germination onset, germination percentage (G%), mean germination time (MGT), root elongation, seed longevity during ex situ storage and dry mass of seeds were evaluated.Germination responses observed were independent of the diaspore dispersal timing, such as variations in germination onset, G% and MGT, both in temperature and water availability treatments. In contrast, root elongation, longevity and dry mass of seeds varied according to the time of diaspore dispersal.Our results corroborate the hypothesis that the timing of diaspore dispersal is an important factor in controlling the initial development of seedlings in seasonal vegetation, but not in germination responses. The predominance of negative effects of temperature increases and water deficit on root growth shows that the initial stages of plant development can be strongly impacted by these environmental factors. [ABSTRACT FROM AUTHOR]

IntroductionIncreased soil salinity in the recent years has adversely affected the productivity of mango globally. Extending the cultivation of mango in salt affected regions warrants the use of salinity tolerant/resistant rootstocks. However, the lack of sufficient genomic and transcriptomic information impedes comprehensive research at the molecular level. MethodWe employed RNA sequencing-based transcriptome analysis to gain insight into molecular response to salt stress by using two polyembryonic mango genotypes with contrasting response to salt stress viz., salt tolerant Turpentine and salt susceptible Mylepelian.ResultsRNA sequencing by Novaseq6000 resulted in a total of 2795088, 17535948, 7813704 and 5544894 clean reads in Mylepelian treated (MT), Mylepelian control (MC), Turpentine treated (TT) and Turpentine control (TC) respectively. In total, 7169 unigenes annotated against all the five public databases, including NR, NT, PFAM, KOG, Swissport, KEGG and GO. Further, maximum number of differentially expressed genes were found between MT and MC (2106) followed by MT vs TT (1158) and TT and TC (587). The differentially expressed genes under different treatment levels included transcription factors (bZIP, NAC, bHLH), genes involved in Calcium-dependent protein kinases (CDPKs), ABA biosynthesis, Photosynthesis etc. Expression of few of these genes was experimentally validated through quantitative real-time PCR (qRT-PCR) and contrasting expression pattern of Auxin Response Factor 2 (ARF2), Late Embryogenesis Abundant (LEA) and CDPK genes were observed between Turpentine and Mylepelian.DiscussionThe results of this study will be useful in understanding the molecular mechanism underlying salt tolerance in mango which can serve as valuable baseline information to generate new targets in mango breeding for salt tolerance.

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Tillandsia subgenera Diaphoranthema and Phytarrhiza s.s. are closely related epiphytic and epilithic plants adapted to xeric habitats in the Americas. Several studies suggested that subgenus Diaphoranthema and the xerophytic species of subgenus Phytarrhiza should not be segregated into different subgenera. In recent phylogenetic analyses of Tillandsioideae, subgenus Phytarrhiza s.l. is revealed as highly polyphyletic, showing a xerophytic lineage closely related to subgenus Diaphoranthema. Moreover, the evolution of neotenic traits, a selfing breeding system and polyembryony have yet to be fully investigated in a phylogenetic context. Infrageneric classifications and phylogenetic relationships were inferred using the plastid matK - trnK and nuclear ETS markers and morphology. Phylogenetic analyses of individual and combined molecular data sets were performed under parsimony and Bayesian inference. Our results confirm subgenus Diaphoranthema as monophyletic if three xeric species of subgenus Phytarrhiza are included. In subgenus Diaphoranthema, aggregates Loliacea and Rectangula were recovered as monophyletic, whereas Myosura was paraphyletic and Caliginosa, Capillaris and Recurvata were polyphyletic. Alternative groupings and morphological trends are discussed. Analysis of morphological characters associated with heterochrony indicates that neoteny may not be the only paedomorphic process in the evolution of this group. Furthermore, paedomorphosis would occur in some vegetative structures, inflorescences and flowers, whereas some fruit and seed traits could be modified by peramorphosis. [ABSTRACT FROM AUTHOR]

In mango, leaf and fruit volatile aroma profiles are variety specific which can be used as fingerprint of a variety. Such biochemical markers can also discriminate the nucellar and zygotic seedlings in polyembryonic mango varieties. In order to validate the applicability of volatile as well as phenolic acid profiles as biomarkers, the open pollinated seedlings of three polyembryonic varieties of mango were compared with their mother trees. Leaf volatile and phenol acid profiling were done using Gas Chromatography/Mass Spectrometry (GCMS) and Liquid Chromatography/Mass Spectrometry (LCMS) methods respectively. The sesquiterpene hydrocarbons were the most abundant in all the genotypes studied. Monoterpenoids were the major compounds in cultivars Vellaikolumban and Olour, while the sesquiterpenoids were the major compounds in cv. Turpentine. While terpinolene was the major monoterpenoid compound in Vellaikolumban and limonene in cv. Olour, the sesquiterpene á-gurjunene was the major compound in cv. Turpentine. Volatile profiling showed clear differences between the varieties but was similar within a variety. Among the 15 phenolic acids quantified in the leaves, P-coumaric acid, gallic acid, and ferulic acids were predominant whereas, vanillic acid, syringic acid, gentisic acid, benzoic acid, and sinapic acids were low in quantity. Phenolic acid profile did not show significant diversity among the varieties and therefore cannot be used for identification of varieties. The volatile profiling can be used for the identification and differentiation of polyembryonic mango genotypes. [ABSTRACT FROM AUTHOR]

Background: Floral sterility and nucellar embryony are peculiar traits of several Citrus species and represent an obstacle to traditional breeding. Morphological sterility mainly results in pistil abortion and anther atrophy, while polyembryony is due to a mechanism known as sporophytic apomixis, which consists of the presence of embryos in the seed generated from the nucellar (maternal) tissue alongside the zygotic embryo (sexual origin). Considering the growing interest in lemon [Citrus limon (L.) Burm. f.] breeding, and the lack of information on floral sterility and the polyembryony trait among different lemon cultivars, a morphological and molecular characterization of these traits of interest was performed on forty Sicilian and international lemon cultivars available in the citrus germplasm collection of Catania University (Italy). Methods: Eight traits related to the reproductive biology were assessed on the selected lemon cultivars, namely: pistil abortion and anther atrophy, number of seeds per fruit, number of embryos per seed, percentage of seeds showing polyembryony, germination, percentage of seeds resulting in more than one plantlet, and average seed weight. Moreover, seedlings recovered after the germination assay were genotyped with SNP and SSR markers for ascertaining their nucellar or zygotic origin. In addition, PCR analysis were performed to assess the allele combination of the miniature inverted-repeat transposable element (MITE) insertion in CitRKD1, a gene associated with the occurrence of apomixis in citrus. Results: All traits showed high variability among the accessions analyzed. As for polyembryony, lemon 'Adamopoulos' scored the highest percentage of polyembryonic seeds (67.6%), whilst lemon 'Lunario' showed the lowest value (8.7%). Conclusions: Insights on the level of polyembryony within lemon varieties will represent a valuable tool for breeders for the set-up of novel mating schemes. In fact, when a polyembryonic female parent is used in cross breeding, the selection of the zygotic individual is hampered by the presence of a nucellar one. [ABSTRACT FROM AUTHOR]

In mango, leaf and fruit volatile aroma profiles are variety specific which can be used as fingerprint of a variety. Such biochemical markers can also discriminate the nucellar and zygotic seedlings in polyembryonic mango varieties. In order to validate the applicability of volatile as well as phenolic acid profiles as biomarkers, the open pollinated seedlings of three polyembryonic varieties of mango were compared with their mother trees. Leaf volatile and phenol acid profiling were done using Gas Chromatography/Mass Spectrometry (GCMS) and Liquid Chromatography/Mass Spectrometry (LCMS) methods respectively. The sesquiterpene hydrocarbons were the most abundant in all the genotypes studied. Monoterpenoids were the major compounds in cultivars Vellaikolumban and Olour, while the sesquiterpenoids were the major compounds in cv. Turpentine. While terpinolene was the major monoterpenoid compound in Vellaikolumban and limonene in cv. Olour, the sesquiterpene á-gurjunene was the major compound in cv. Turpentine. Volatile profiling showed clear differences between the varieties but was similar within a variety. Among the 15 phenolic acids quantified in the leaves, P-coumaric acid, gallic acid, and ferulic acids were predominant whereas, vanillic acid, syringic acid, gentisic acid, benzoic acid, and sinapic acids were low in quantity. Phenolic acid profile did not show significant diversity among the varieties and therefore cannot be used for identification of varieties. The volatile profiling can be used for the identification and differentiation of polyembryonic mango genotypes.

Apomixis can fix the heterosis of Hybrid F1, by maintaining its heterozygous genotype, and is an ideal way for the development of hybrid rice. In this paper, we designed an engineering strategy for realizing apomictic reproduction of hybrid rice in the way of induce adventitious embryos. An embryogenesis gene, AtWUS, controlled by the ovule-specific promoter, a ribonuclease gene Barnase driven by the egg cell-specific promoter pDD45, and an inactivation gene ZmAA1 driven by the pollen-specific promoter pG47 were simultaneously integrated into one T-DNA, and co-transformed with the second T-DNA carrying a Barstar gene. Double-seedlings were observed in transgenic line. Whole-genome sequencing and ploidy levels confirmed by flow cytometry showed that one of the double-seedlings was heterozygous diploid and the other seedling was homozygous haploid, which confirmed that embryogenesis in one of the double-seedlings arises from the zygote after fertilization and the other derived from an unfertilized gamete. Meanwhile we obtained embryofree seeds at frequencies of 2.6% to 3.8% in T1 generation, and 0.75% to 3% in T2 generation. Though we did not obtained adventitious embryos in hybrid rice in this study, the phenomenon of double-seedlings and embryo-free seeds in transgenic line was informative and strongly suggested that endosperm development is an autonomously organized process in rice, independent of egg cell fertilization and embryo-endosperm communication. This provides novel insights into the induction of haploid embryos and lends theoretical support to successful clonal propagation using synthetic apomixis. [ABSTRACT FROM AUTHOR]

We provide the first documented case of the black mangrove, Avicennia germinans, producing multiple seedlings from a single propagule, a phenomenon called polyembryony, on several occasions. There is a lack of knowledge about polyembryony in mangroves, as it is an extremely rare phenomenon previously observed in only three other mangrove species. A higher weight of mature propagules may indicate polyembryony, thus enabling early detection. A. germinans may make use of mixed reproductive strategies to ensure the continued survival of the species, or to increase its expansion under favourable environmental conditions. Potential causes and implications of polyembryony in A. germinans are discussed. [ABSTRACT FROM AUTHOR]

Because of the growing worldwide demand for maize grain, new alternatives have been sought for breeding of this cereal, e.g., development of polyembryonic varieties, which agronomic performance could positively impact the grain yield per unit area, and nutritional quality. The objectives of this study were to (1) determine the phytochemicals present in the embryo and endosperm of grain from maize families with high, low, and null polyembryony frequency, which were planted at different locations, and (2) state the relationship between these compounds and seed germination. The extracted phytochemicals from corn were identified by HPLC-MS. The results showed that the genotype with the highest presence of phytochemicals was the brachytic population with high polyembryony called "BAP", which also required less water during the germination process. The number of phytochemicals in both embryo and endosperm tissues was not related to the sowing location where they came from or the type of polyembryony. The number of different phytochemicals depended on the grain tissue from where they were extracted. The chemical compounds found in the different maize tissues were related to the development of the plant, either in roots or nibs because these are mainly associated with the lignin synthesis. [ABSTRACT FROM AUTHOR]

Abstract Background Placentation has evolved multiple times among both chordates and invertebrates. Although they are structurally less complex, invertebrate placentae are much more diverse in their origin, development and position. Aquatic colonial suspension-feeders from the phylum Bryozoa acquired placental analogues multiple times, representing an outstanding example of their structural diversity and evolution. Among them, the clade Cyclostomata is the only one in which placentation is associated with viviparity and polyembryony—a unique combination not present in any other invertebrate group. Results The histological and ultrastructural study of the sexual polymorphic zooids (gonozooids) in two cyclostome species, Crisia eburnea and Crisiella producta, revealed embryos embedded in a placental analogue (nutritive tissue) with a unique structure—comprising coenocytes and solitary cells—previously unknown in animals. Coenocytes originate via nuclear multiplication and cytoplasmic growth among the cells surrounding the early embryo. This process also affects cells of the membranous sac, which initially serves as a hydrostatic system but later becomes main part of the placenta. The nutritive tissue is both highly dynamic, permanently rearranging its structure, and highly integrated with its coenocytic ‘elements’ being interconnected via cytoplasmic bridges and various cell contacts. This tissue shows evidence of both nutrient synthesis and transport (bidirectional transcytosis), supporting the enclosed multiple progeny. Growing primary embryo produces secondary embryos (via fission) that develop into larvae; both the secondary embyos and larvae show signs of endocytosis. Interzooidal communication pores are occupied by 1‒2 specialized pore-cells probably involved in the transport of nutrients between zooids. Conclusions Cyclostome nutritive tissue is currently the only known example of a coenocytic placental analogue, although syncytial ‘elements’ could potentially be formed in them too. Structurally and functionally (but not developmentally) the nutritive tissue can be compared with the syncytial placental analogues of certain invertebrates and chordates. Evolution of the cyclostome placenta, involving transformation of the hydrostatic apparatus (membranous sac) and change of its function to embryonic nourishment, is an example of exaptation that is rather widespread among matrotrophic bryozoans. We speculate that the acquisition of a highly advanced placenta providing massive nourishment might support the evolution of polyembryony in cyclostomes. In turn, massive and continuous embryonic production led to the evolution of enlarged incubating polymorphic gonozooids hosting multiple progeny.

In the present work, we study the ovule, seed, and fruit development in six Bulbostylis species in order to characterize the genus in a comparative approach and to identify the characteristics that can be used in taxonomy and phylogeny. Flowers and fruits at different developmental stages were analyzed using LM and SEM after processing according to standard techniques. The species studied have the following: anatropous and bitegmic ovules, weak crassinucellar ovules, obturator of integumentary origin, monosporic embryo sac of the Polygonum type, nuclear endosperm, hypostase formation, seed coat formed by tanniferous endotegmen and exotesta, and Bulbostylis-type embryo. On the other hand, the pericarp development constitutes the main variation within Bulbostylis since the cells of the exocarp may or may not present starch grains, and their inner periclinal walls may be slightly or deeply concave depending on the degree of development of the mesocarp sclereids. In a taxonomic context, the results herein obtained are in conflict with studies which suggest infrageneric groupings based on fruit micromorphology, and also with the relationship among the Bulbostylis species based on molecular analysis. This work contributes to a better understanding of the reproductive anatomy and embryology in Bulbostylis, and reveals the first insights about the origin of multiple embryos in Cyperaceae. Given the frequent presence of polyembryony in Bulbostylis, and the poor mention of this condition in the family, this work highlights an aspect in the anatomy of Cyperaceae that must be re-explored. [ABSTRACT FROM AUTHOR]

Polyembryony has been reported in maize (Zea mays L.) since the 20th century. This trait is defined as the appearance of two or more seedlings from a germinated seed and has been associated with improvements in the nutritional content of the grain. Maize kernel traits are affected by different factors among them the growing location. Thus, it is important to determine the environmental (location) effect on polyembryonic kernel traits. The purpose of this study was to determine the environmental effects on the physical and chemical properties of polyembryonic maize kernels. Thirty-two maize genotypes with different levels of polyembryony (high, low and non-polyembryonic) were planted in two different locations with distinct climate (Buenavista, Coahuila and Río Bravo, Tamaulipas, Mexico). Results showed that at the Buenavista location, maize lines had a greater weight of one thousand grains (272 - 367 g), the non-polyembryonic lines showed the highest weight. Besides, the low polyembryonic maize lines promoted the largest grain dimensions. The polyembryonic maize had the biggest embryos, which occupied 11.37 -11.59% of the grain. At the Río Bravo, in comparison to Buenavista, the polyembryonic maize lines had the highest protein (4.56%) and fat (5%) contents, while the raw fiber (1.2%) was highest in non-polyembryonic maize, meanwhile at, the Buenavista location, the starch content (59.2%) was higher in non-polyembryonic maize, while ashes (1.4%) and moisture (13.7%) were higher in polyembryonic maize. Growing polyembryonic maize genotypes in different environments, it was observed that the bromatological profile is affected by environmental conditions. In this study, a better nutritional content was observed in maize genotypes grown at Río Bravo in comparison to those maize genotypes grown at Buenavista. So, this study suggests that it is possible to produce polyembryonic maize genotypes in different environments which will have a different bromatological profile and can be employed for specific industrial uses.

Abstract Background Polyembryony is defined as the formation of several embryos from a single egg. This phenomenon can occur in humans, armadillo, and some endoparasitoid insects. However, the mechanism underlying polyembryogenesis in animals remains to be elucidated. The polyembryonic parasitoid wasp Copidosoma floridanum oviposits its egg into an egg of the host insect; eventually, over 2000 individuals will arise from one egg. Previously, we reported that polyembryogenesis is enhanced when the juvenile hormone (JH) added to the culture medium in the embryo culture. Hence, in the present study, we performed RNA sequencing (RNA-Seq) analysis to investigate the molecular mechanisms controlling polyembryogenesis of C. floridanum. Functional annotation of genes is not fully available for C.floridanum; however, whole genome assembly has been archived. Hence, we constructed a pipeline for gene functional annotation in C. floridanum and performed molecular network analysis. We analyzed differentially expressed genes between control and JH-treated molura after 48 h of culture, then used the tblastx program to assign whole C. floridanum transcripts to human gene. Results We obtained 11,117 transcripts in the JH treatment group and identified 217 differentially expressed genes compared with the control group. As a result, 76% of C. floridanum transcripts were assigned to human genes. Gene enrichment analysis revealed genes associated with platelet degranulation, fatty acid biosynthesis, cell morphogenesis in the differentiation and integrin signaling pathways were fluctuated following JH treatment. Furthermore, Cytoscape analysis revealed a molecular interaction that was possibly associated with polyembryogenesis . Conclusions We have constructed a pipeline for gene functional annotation of C. floridanum, and identified transcripts with high similarity to human genes during early embryo developmental. Additionally, this study reveals new molecular interactions associated with polyembryogenesis; these interactions could indicate the molecular mechanisms underlying polyembryony. Our results highlight the potential utility of molecular interaction analysis in human twins.

ABSTRACT: A generalized concept in the Citrus genus is that highly polyembryonic varieties produce only a small number of hybrids. Small zygotic embryos congregate primarily near the micropyle, while nucellar embryos organize near or away from the micropyle. In the present study, the authors determined the number of polyembryonic seeds, embryos per seed, and the largest embryo (LE) position in five citrus cultivars: C35 citrange, Volkamer lemon, Amblycarpa mandarin, Minneola tangelo, and Valencia orange. The percentage of nucellar seedlings obtained exclusively from the LE per seed was then calculated. The polyembryony percentage varied largely between genotypes, from 65 to 98 %, and the mean number of embryos ranged from 2.9 to 4.6. The chalaza contained up to 87 % of the LE. Out of 30 primers, 17 Simple sequence repeats [SSRs] (AG14, ATC09, CAT01, CCSM147, CCSM18, CCSM13, CCSM4, F2, F4, F6, GT03, TAA41, TAA45, TAA1, F7, F11 and TAA52) identified nucellar plants identical to the female parent (genetic similarity index [GSI] value ≥ 0.95). This study establishes for the first time the relationship between the sexual or asexual origin of seedlings derived from LE embryos isolated from seeds and the SSR primers described above. While the five citrus cultivars had high polyembryony levels, 30 % of the resulting plants differed from the female parent in C35 Citrange, 45 % in Volkamer lemon, 15 % in Amblycarpa mandarin, 15 % in Valencia orange, and 45 % in Minneola tangelo. The largest seedling is not always nucellar: in the five citrus species studied, the LE produces 55 to 85 % of the nucellar embryos.

n the Neotropics, the focus of apomictic studies predominantly centres on trees within the Brazilian savanna, characterized, mostly as sporophytic and facultative, associated with polyploidy and polyembryony. To enhance our understanding of the mechanisms governing apomixis and sexual reproduction in tropical herbaceous plants, we clarify the relationship between apomixis, chromosome counts, and polyembryony in the epiphytic orchid Zygopetalum mackayi , which forms a polyploid complex within rocky outcrops in both the Brazilian savanna and the Atlantic forest. To define embryo origins and describe megasporogenesis and megagametogenesis, we performed manual self-pollinations in first-day flowers of cultivated plants, considering all three cytotypes (2 x , 3 x , 4 x ) of this species. Flowers and fruits at different stages were collected to describe the development and morphology of ovules and seeds considering sexual and apomictic processes. As self-pollination treatments resulted in high fruit abortion in diploids, we also examined pollen tube development in aborted flowers and fruits to search for putative anomalies. Megasporogenesis and megagametogenesis occur regularly in all cytotypes. Apomixis is facultative and sporophytic, and associated with polyploid cytotypes, while diploid individuals exclusively engage in sexual reproduction. Polyembryony is caused mainly by the production of adventitious embryos from nucellar cells of triploids and tetraploids, but also by the development of multiple archesporia in all cytotypes. Like other apomictic angiosperms within the Brazilian savanna, our findings demonstrate that apomixis in Z. mackayi relies on pollinators for seed production. We also consider the ecological implications of these apomictic patterns in Z. mackayi within the context of habitat loss and its dependence on pollinators., Competing Interests: None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Annals of Botany Company.)

Background: Floral sterility and nucellar embryony are peculiar traits of several Citrus species and represent an obstacle to traditional breeding. Morphological sterility mainly results in pistil abortion and anther atrophy, while polyembryony is due to a mechanism known as sporophytic apomixis, which consists of the presence of embryos in the seed generated from the nucellar (maternal) tissue alongside the zygotic embryo (sexual origin). Considering the growing interest in lemon [Citrus limon (L.) Burm. f.] breeding, and the lack of information on floral sterility and the polyembryony trait among different lemon cultivars, a morphological and molecular characterization of these traits of interest was performed on forty Sicilian and international lemon cultivars available in the citrus germplasm collection of Catania University (Italy). Methods: Eight traits related to the reproductive biology were assessed on the selected lemon cultivars, namely: pistil abortion and anther atrophy, number of seeds per fruit, number of embryos per seed, percentage of seeds showing polyembryony, germination, percentage of seeds resulting in more than one plantlet, and average seed weight. Moreover, seedlings recovered after the germination assay were genotyped with SNP and SSR markers for ascertaining their nucellar or zygotic origin. In addition, PCR analysis were performed to assess the allele combination of the miniature inverted-repeat transposable element (MITE) insertion in CitRKD1, a gene associated with the occurrence of apomixis in citrus. Results: All traits showed high variability among the accessions analyzed. As for polyembryony, lemon ‘Adamopoulos’ scored the highest percentage of polyembryonic seeds (67.6%), whilst lemon ‘Lunario’ showed the lowest value (8.7%). Conclusions: Insights on the level of polyembryony within lemon varieties will represent a valuable tool for breeders for the set-up of novel mating schemes. In fact, when a polyembryonic female parent is used in cross breeding, the selection of the zygotic individual is hampered by the presence of a nucellar one.

Plant sexual and asexual reproduction through seeds (apomixis) is tightly controlled by complex gene regulatory programs, which are not yet fully understood. Recent findings suggest that RNA helicases are required for plant germline development. This resembles their crucial roles in animals, where they are involved in controlling gene activity and the maintenance of genome integrity. Here, we identified previously unknown roles of Arabidopsis RH17 during reproductive development. Interestingly, RH17 is involved in repression of reproductive fate and of elements of seed development in the absence of fertilization. In lines carrying a mutant rh17 allele, development of supernumerary reproductive cell lineages in the female flower tissues (ovules) was observed, occasionally leading to formation of two embryos per seed. Furthermore, seed coat, and putatively also endosperm development, frequently initiated autonomously. Such induction of several features phenocopying distinct elements of apomixis by a single mutation is unusual and suggests that RH17 acts in regulatory control of plant reproductive development. Furthermore, an in-depth understanding of its action might be of use for agricultural applications. [ABSTRACT FROM AUTHOR]

A synopsis is provided of different expressions of whole-animal vertebrate clonality (asexual organismal-level reproduction), both in the laboratory and in nature. For vertebrate taxa, such clonal phenomena include the following: human-mediated cloning via artificial nuclear transfer; intergenerational clonality in nature via parthenogenesis and gynogenesis; intergenerational hemiclonality via hybridogenesis and kleptogenesis; intragenerational clonality via polyembryony; and what in effect qualifies as clonal replication via self-fertilization and intense inbreeding by simultaneous hermaphrodites. Each of these clonal or quasi-clonal mechanisms is described, and its evolutionary genetic ramifications are addressed. By affording an atypical vantage on standard vertebrate reproduction, clonality offers fresh perspectives on the evolutionary and ecological significance of recombination-derived genetic variety.

The results of the investigation proved the gametophytic origin of diploids derived from two-embryonic seeds of pepper C. annuum L. hybrid. During the germination of seeds harvested from the red fruited hybrid (C. annuum L. ATZ × C. annuum L.`Sono`) F1, additional embryos have been found. Four of diploid twins F2 generation were different in their phenotype within the pair and/or from the F1 mother plant. Plants of three pairs: 1A-1B, 2A-2B, 4A-4B were significantly different with regard to the average fruit weight, length and seeds number. Yellow colour of ripe fruit was characteristic for 2A, 3A and 3B plants. In RAPD molecular analysis, twenty-three primers were used and six of them enabled polymorphic products to be obtained in reactions. The results of the analysis confirmed phenotypic differentiation of the twins and their parental forms. The phenotypic and molecular analyses proved that spontaneous diploids from a two-embryonic seeds are ready for the production of genetically stable, sexual progeny.

Apomixis, the asexual formation of embryos and seeds, occurs in approximately 18% of angiosperm families. Melastomataceae exhibits a remarkable number of apomictic species, distributed among different tribes. This mode of reproduction has been elucidated in Miconieae, but remains unclarified for other groups, such as Microlicieae. Although apomixis has been previously described for Microlicieae species, the cytological basis for this phenomenon is entirely unknown in this group. Thus, populations of Microlicia fasciculata and M. polystemma were used in order to (a) investigate the presence of autonomous apomixis; (b) verify if this mode of reproduction leads to polyembryony; and (c) investigate whether apomixis may occur in parallel with the sexual process. We tested these species for autonomous fruit set and polyembryony, and pollen viability, and analyzed pollen tube growth. Anatomical techniques were used to elucidate the micro‐ and megasporogenesis and gametogenesis. The species showed autonomous fruit and seed formation and exhibited polyembryony. Apospory and adventitious embryony were the developmental mechanisms of apomixis in M. fasciculata and M. polystemma, respectively. Both species exhibited low pollen viability. However, some viable pollen, reduced embryo sac formation, natural pollination and pollen tube growth enable sexual reproduction and characterize these species as facultative apomicts. The independence of pollinators for fruit set, uniparental reproduction and the possibility of sexual reproduction, confer reproductive assurance and flexibility, bringing together advantages of sexual and asexual reproduction. In this sense, apomixis may have played an important role in the evolution and diversification of Microlicia, a widely distributed genus in the Brazilian Cerrado. [ABSTRACT FROM AUTHOR]

The results of the investigation proved the gametophytic origin of diploids derived from two-embryonic seeds of pepper C. annuum L. hybrid. During the germination of seeds harvested from the red fruited hybrid (C. annuum L. ATZ × C. annuum L. 'Sono') F1, additional embryos have been found. Four of diploid twins F2 generation were different in their phenotype within the pair and/or from the F1 mother plant. Plants of three pairs: 1A-1B, 2A-2B, 4A-4B were significantly different with regard to the average fruit weight, length and seeds number. Yellow colour of ripe fruit was characteristic for 2A, 3A and 3B plants. In RAPD molecular analysis, twenty three primers were used and six of them enabled polymorphic products to be obtained in reactions. The results of the analysis confirmed phenotypic differentiation of the twins and their parental forms. The phenotypic and molecular analyses proved that spontaneous diploids from a two-embryonic seeds are ready for the production of genetically stable, sexual progeny. [ABSTRACT FROM AUTHOR]

• We report for the first time comprehensive evaluation of salinity tolerance in jamun (Syzygium cumini L. Skeels). • The evaluation procedure used in this work closely mimics salinity under field conditions, and may be used to evaluate the responses of other fruit crops to a prolonged, gradually increased salinity. • Membership function analysis ranked three genotypes as highly salt tolerant. • Na+ exclusion was found to be a major trait underlying salt tolerance. We assessed the effects of salinity stress on 48 genotypes of jamun (Syzygium cumini L. Skeels) including 20 monoembryonic and 28 polyembryonic types to identify the salt tolerant genotypes and the morpho-physiological traits that underpin salt tolerance. Salinity of irrigation water was gradually raised over a period of 21 weeks, from 2.0 to 12.0 dS/m. Control plants were irrigated using fresh water (0.70 dS/m). The experiment was conducted in a Completely Randomized Block Design with four replications. Salinity had a more repressive effect on shoot than on root biomass, regardless of the seedling type, reflecting that roots might tolerate salt concentrations high enough to inhibit the growth of shoots. Salinity had little effects on the maximum quantum yield of photosystem II (F v /F m), and the relative leaf chlorophyll in most genotypes evaluated. Salt-induced decreases in photosynthetic assimilation were generally greater in genotypes with inherently high photosynthetic rates. Genotypes with strong capacities for Na+ exclusion (e.g., CSJ-18, and CSJ-19) were generally highly tolerant to salinity stress. Leaf Cl−concentrations under saline conditions were lower than Na+ concentrations, suggesting a more effective mechanism for Cl− exclusion. Leaf K+, Ca2+ and Mg2+ levels were not significantly different between control and salt treatments in both mono- and poly-embryonic types. The mean membership function value (MFV) was hardly different between mono- (0.5052) and poly-embryonic (0.5254) types, given quite similar reductions in plant height (∼22.0 %), leaf area (≤ 14.0 %), leaf (35–40 %), stem (∼40 %), and root (24–29 %) biomass, and net photosynthesis (∼64 %) as well as increases in leaf Na+ (79–85 %) and Cl− (70–76 %) under saline conditions. Based on mean MFV for all the traits, genotypes CSJ-19, CSJ-18 and CSJ-5 were found to be highly salt tolerant, whereas CSJ-1 and CSJ-31 were ranked as highly salt sensitive. We deduce that maintenance of leaf and root growth, reduced uptake of Na+, and preferential accumulation of K+, Ca2+ and Mg2+ could be the major traits underlying salinity tolerance in jamun. [Display omitted] [ABSTRACT FROM AUTHOR]

In this paper we report the fruit characteristics and seed polyembryony in langsat, Lansium parasiticum (syn. Lansium domesticum Corr.). This fruit tree belongs to the family Meliaceae in order Sapindales and is considered to be native of western South East Asia and is common in both wild and cultivated forms throughout Malaysia and Philippines where the fruits are very popular and the tree is being utilized in reforestation efforts. It is also grown in southern Thailand and Vietnam and flourishes in the Nilgiris and other humid areas of South India. In the present investigation we report the morphological and biochemical parameters of the plants and fruits obtained from State Horticultural Farm, Buliar (latitude 11.34; longitude 76.79) in Tamil Nadu, at elevation of 360 m MSL and receiving average annual rainfall of 125.14 cm. The plantation was established in the year 1900 and consists of various tropical trees like mangosteen, langsat, arecanut, coffee, silveroak, pepper, cinnamon in tier system of planting.

ABSTRACT Polyembryony is the differentiation and development of multiple embryos in a single seed. This characteristic can provide advantages, as more than one embryo is produced with the same amount of resources, and the probability of establishment of at least one seedling increases. However, sibling seedlings may also increase competition, affecting development and survival. In the present study, the possible advantages and disadvantages of polyembryony were analyzed in the initial establishment of seedlings of Carapa surinamensis (Meliaceae), a tree species that produces monoembryonic or polyembryonic seeds. In this regard, the development of single seedlings was compared with a pair of seedlings emerging from polyembryonic seeds. We compared the development of seedlings attached to or detached from each other and to the seed resources. We observed two levels of competition: (a) for the seed reserves during germination and initial development, as multiple embryos of C. surinamensis share the same reserves, and (b) for external factors, mostly space for root and shoot development, and also for light. Reducing the competition for external factors by separating the siblings was not enough to reduce the effects of competition for seed reserves in the first six months of development. Nevertheless, viable seedlings were produced in all treatments. Thus, depending on sprout management in the nursery, the number of seedlings per seed can be significantly increased by detaching the seedlings, or more vigorous seedlings can be obtained when only one seedling is maintained.

Background: Placentation has evolved multiple times among both chordates and invertebrates. Although they are structurally less complex, invertebrate placentae are much more diverse in their origin, development and position. Aquatic colonial suspension-feeders from the phylum Bryozoa acquired placental analogues multiple times, representing an outstanding example of their structural diversity and evolution. Among them, the clade Cyclostomata is the only one in which placentation is associated with viviparity and polyembryony-a unique combination not present in any other invertebrate group. Results: The histological and ultrastructural study of the sexual polymorphic zooids (gonozooids) in two cyclostome species, Crisia eburnea and Crisiella producta, revealed embryos embedded in a placental analogue (nutritive tissue) with a unique structure-comprising coenocytes and solitary cells-previously unknown in animals. Coenocytes originate via nuclear multiplication and cytoplasmic growth among the cells surrounding the early embryo. This process also affects cells of the membranous sac, which initially serves as a hydrostatic system but later becomes main part of the placenta. The nutritive tissue is both highly dynamic, permanently rearranging its structure, and highly integrated with its coenocytic 'elements' being interconnected via cytoplasmic bridges and various cell contacts. This tissue shows evidence of both nutrient synthesis and transport (bidirectional transcytosis), supporting the enclosed multiple progeny. Growing primary embryo produces secondary embryos (via fission) that develop into larvae; both the secondary embyos and larvae show signs of endocytosis. Interzooidal communication pores are occupied by 1-2 specialized pore-cells probably involved in the transport of nutrients between zooids. Conclusions: Cyclostome nutritive tissue is currently the only known example of a coenocytic placental analogue, although syncytial 'elements' could potentially be formed in them too. Structurally and functionally (but not developmentally) the nutritive tissue can be compared with the syncytial placental analogues of certain invertebrates and chordates. Evolution of the cyclostome placenta, involving transformation of the hydrostatic apparatus (membranous sac) and change of its function to embryonic nourishment, is an example of exaptation that is rather widespread among matrotrophic bryozoans. We speculate that the acquisition of a highly advanced placenta providing massive nourishment might support the evolution of polyembryony in cyclostomes. In turn, massive and continuous embryonic production led to the evolution of enlarged incubating polymorphic gonozooids hosting multiple progeny. [ABSTRACT FROM AUTHOR]