Storage reserves and cellular water in mature seeds of Araucaria angustifolia

Seed tissues of Araucaria angustifolia (Bertol.) Kuntze were investigated using histochemistry, transmission electron microscopy (TEM) and energy dispersive X-ray (EDX) analysis. Moisture content and water status in tissues were also evaluated. In the embryo, TEM studies revealed the presence of one to several central vacuoles and a peripheral layer of cytoplasm in cells from different tissues of the cotyledons and axis. In the cytoplasm, lipid bodies, starch grains, mitochondria and a nucleus are evident. In most tissues, vacuoles contain proteins, indicating that the storage proteins are highly hydrated. In cells of the root cap, proteins are stored in discrete protein bodies. Both protein storage vacuoles and discrete protein bodies have inclusions of crystal globoids. EDX analysis of globoids revealed the presence of P, K and Mg as the main constituents and traces of S, Ca and Fe. In the root and shoot meristems, deposits of phytoferritin are present in the stroma of proplastids. The gametophyte consists of cells characterized by relatively thin cell walls and one to several nuclei per cell. Protein and lipid bodies are present, although starch is the most conspicuous reserve. Immediately after shedding, moisture content is approximately 145% (dry weight) for the embryo and 95% (dry weight) for the gametophyte. Calorimetric studies reveal that axes and cotyledons have a very high content of freezable water, corresponding to types 5 and 4, i.e. dilute and concentrated (or capillary) solution, respectively. The results are discussed in relation to the behaviour of the species, which has been categorized as recalcitrant.  © 2002 The Linnean Society of London . Botanical Journal of the Linnean Society , 2002, 140 , 273−281.     

Purification and biological effects of Araucaria angustifolia (Araucariaceae) seed lectin

This paper describes the purification and characterization of a new N-acetyl-d-glucosamine-specific lectin from Araucaria angustifolia (AaL) seeds (Araucariaceae) and its anti-inflammatory and antibacterial activities. AaL was purified using a combination of affinity chromatography on a chitin column and ion exchange chromatography on Sephacel-DEAE. The pure protein has 8.0kDa (SDS-PAGE) and specifically agglutinates rabbit erythrocytes, effect that was independent of the presence of divalent cations and was inhibited after incubation with glucose and N-acetyl-d-glucosamine. AaL showed antibacterial activity against Gram-negative and Gram-positive strains, shown by scanning electron microscopy. AaL, intravenously injected into rats, showed anti-inflammatory effect, via carbohydrate site interaction, in the models of paw edema and peritonitis. This lectin can be used as a tool for studying bacterial infections and inflammatory processes.     

Protein expression during seed development in Araucaria angustifolia: transient accumulation of class IV chitinases and arabinogalactan proteins

Protein accumulation and patterns during embryogenesis in the recalcitrant seeds of the gymnosperm species Araucaria angustifolia (Bert.) O. Kuntze were studied. Soluble seed proteins, chitinases, and arabinogalactan proteins (AGPs) were analyzed by means of 2-D gel electrophoresis, mass spectrometry, isoelectric focusing, Western blot, precipitation and staining with β-glucosyl Yariv reagent (β-Glc)₃Y, and gas liquid chromatography. Despite the recalcitrant nature of the seeds, the electrophoretic patterns of A . angustifolia seed proteins showed similarities with orthodox seed types. Proteins showing chitinolytic activity were observed in all seed stages analyzed, but the expression of class IV chitinases was restricted to late stages of seed development. AGPs were prominent during stages of seed development characterized by intensive cell division and differentiation, and their decrease during seed maturation might be related to cell wall modifications during the deposition of storage compounds. Gas liquid chromatographic analyzes of AGPs did not show quantitative changes in their carbohydrate moieties during seed development. A low molecular weight protein specifically expressed in mature seeds was precipitated using (β-Glc)₃Y. Amino acid sequences obtained from MS/MS analysis revealed peptides rich in valine and acidic amino acids, but devoid in amino acids normally found in AGPs polypeptides, suggesting that these peptides are not related to classical or non-classical AGPs. Possible implications of chitinases and AGPs during seed development in A . angustifolia are discussed.     

The role of gene flow in shaping genetic structures of the subtropical conifer species Araucaria angustifolia

The morphological features of pollen and seed of Araucaria angustifolia have led to the proposal of limited gene dispersal for this species. We used nuclear microsatellite and AFLP markers to assess patterns of genetic variation in six natural populations at the intra- and inter-population level, and related our findings to gene dispersal in this species. Estimates of both fine-scale spatial genetic structure (SGS) and migration rate suggest relatively short-distance gene dispersal. However, gene dispersal differed among populations, and effects of more efficient dispersal within population were observed in at least one stand. In addition, even though some seed dispersal may be aggregated in this principally barochorous species, reasonable secondary seed dispersal, presumably facilitated by animals, and overlap of seed shadows within populations is suggested. Overall, no correlation was observed between levels of SGS and inbreeding, density or age structure, except that a higher level of SGS was revealed for the population with a higher number of juvenile individuals. A low estimate for the number of migrants per generation between two neighbouring populations implies limited gene flow. We expect that stepping-stone pollen flow may have contributed to low genetic differentiation among populations observed in a previous survey. Thus, strategies for maintenance of gene flow among remnant populations should be considered in order to avoid degrading effects of population fragmentation on the evolution of A. angustifolia.     

Control of resin production in Araucaria angustifolia,an ancient South American conifer

Araucaria angustifolia is an ancient slow‐growing conifer that characterises parts of the Southern Atlantic Forest biome, currently listed as a critically endangered species. The species also produces bark resin, although the factors controlling its resinosis are largely unknown. To better understand this defence‐related process, we examined the resin exudation response of A. angustifolia upon treatment with well‐known chemical stimulators used in fast‐growing conifers producing both bark and wood resin, such as Pinus elliottii. The initial hypothesis was that A. angustifolia would display significant differences in the regulation of resinosis. The effect of Ethrel^® (ET – ethylene precursor), salicylic acid (SA), jasmonic acid (JA), sulphuric acid (SuA) and sodium nitroprusside (SNP – nitric oxide donor) on resin yield and composition in young plants of A. angustifolia was examined. In at least one of the concentrations tested, and frequently in more than one, an aqueous glycerol solution applied on fresh wound sites of the stem with one or more of the adjuvants examined promoted an increase in resin yield, as well as monoterpene concentration (α‐pinene, β‐pinene, camphene and limonene). Higher yields and longer exudation periods were observed with JA and ET, another feature shared with Pinus resinosis. The results suggest that resinosis control is similar in Araucaria and Pinus. In addition, A. angustifolia resin may be a relevant source of valuable terpene chemicals, whose production may be increased by using stimulating pastes containing the identified adjuvants.     

Spatial and temporal patterns of seed attack and germination in a large-seeded neotropical tree species

It has long been argued that seed dispersal enhances recruitment in tropical trees by allowing offspring to `escape' strong density/distance-dependent attack by insects, pathogens and rodents. Here we examined the effects of canopy openness and parent-offspring distance upon the frequency and timing of Chlorocardium rodiei seed attack and germination within a 15-ha plot of Guyanan tropical rain forest. Seeds were artificially dispersed beneath parent trees, in the understorey away from trees and in gaps. Analysing our data from an 85-week period of regular monitoring, we found that the main spatial gradients, canopy openness and distance to nearest adult conspecific, do not lead to differences in the final number of seeds attacked by infesting scolytid beetles or rodents. The timing of beetle attack, however, varied along the distance gradient and this difference affords seeds at further distances a `window' in which to germinate and produce a seedling before attack. Canopy openness was not a good predictor of rooting success, but distance was strongly associated with root and shoot formation success and the mean time to shoot formation. There was a strong negative effect of distance on the likelihood of a seed being colonised by scolytid beetles prior to removal by rodents and shoot failure was strongly associated with prior infestation. We believe these results bring a key point to bear on the well-established notion of distance-dependent attack on seeds in tropical rainforests, viz. that seed characteristics (size, germination syndrome) and the timing of attack may be more important in explaining patterns of early seedling recruitment than distance. Our studies suggest that advantages accrued through dispersal in species like Chlorocardium will depend heavily on the `race' between seed germination and attack. In the case of Chlorocardium, the `race' can be lost at considerable distances due to its prolonged dormancy and the temporal fluctuations in fruitfall and rainfall which influence attack and germination. The results presented here suggest that the lag between seed attack and germination in tropical trees can regulate the influence of parent-offspring distance on cohort recruitment at this life history stage. Received: 5 March 1998 / Accepted: 6 December 1998     

Patterns of pollen and seed dispersal in a small, fragmented population of the wind-pollinated tree Araucaria angustifolia in southern Brazil

Paternity analysis based on eight microsatellite loci was used to investigate pollen and seed dispersal patterns of the dioecious wind-pollinated tree, Araucaria angustifolia. The study sites were a 5.4 ha isolated forest fragment and a small tree group situated 1.7 km away, located in Paranalpha State, Brazil. In the forest fragment, 121 males, 99 females, 66 seedlings and 92 juveniles were mapped and genotyped, together with 210 seeds. In the tree group, nine male and two female adults were mapped and genotyped, together with 20 seeds. Paternity analysis within the forest fragment indicated that at least 4% of the seeds, 3% of the seedlings and 7% of the juveniles were fertilized by pollen from trees in the adjacent group, and 6% of the seeds were fertilized by pollen from trees outside these stands. The average pollination distance within the forest fragment was 83 m; when the tree group was included the pollination distance was 2006 m. The average number of effective pollen donors was estimated as 12.6. Mother-trees within the fragment could be assigned to all seedlings and juveniles, suggesting an absence of seed immigration. The distance of seedlings and juveniles from their assigned mother-trees ranged from 0.35 to 291 m (with an average of 83 m). Significant spatial genetic structure among adult trees, seedlings, and juveniles was detected up to 50 m, indicating seed dispersal over a short distance. The effective pollination neighborhood ranged from 0.4 to 3.3 ha. The results suggest that seed dispersal is restricted but that there is long-distance pollen dispersal between the forest fragment and the tree group; thus, the two stands of trees are not isolated.     

Integrating approaches towards the conservation of forest genetic resources: a case study of Araucaria angustifolia

In the past years the conservation of forest genetic resources has been widely recognized because of the risk of adverse effects associated with global changes in the environment. Since little effort has been made towards conservation of these resources, the assessment and the integration of information and knowledge so far generated are valuable tools in order to promote their conservation. In the present work the Brazilian conifer Araucaria angustifolia was used as a case study, integrating information, published between 1980 and 2007, about genetics, biotechnology, palaeobotany, and archeology. These studies were interpreted towards the conservation needs and strategies of the species germplasm. The main conclusions for the conservation of A. angustifolia genetic resources are: (1) despite the reduction of genetic diversity by unsustainable forest exploitation, the genetic system of A. angustifolia seems to be efficient to protect this species against rapid losses of its genetic diversity; (2) conservation measures starting now may be efficient, since the genetic structures of existing populations is more largely influenced by the past evolutionary process than by human disturbance occurred in the last century; (3) there are opportunities to save the genetic resources of the species, if simple measures are taken to safeguard the continued existence of the few remaining araucaria forests. Emerged as a general conclusion that the use of sound scientific arguments supported by different areas of the science could be a very powerful instrument in persuading decision-makers, also by governments, taking into account forest genetic resources as a primary issue for the future.     

Updating embryonic ontogenesis in Araucaria angustifolia : from Burlingame (1915) to the present

This study addresses gaps in our understanding of pre-fertilization and archegonia development and reinterprets embryonic ontogenesis from Burlingame (Bot Gaz 59:1–39, 1915) to the present based on timescale and structural features allowing us to determine functionally and developmentally accurate terminology for all these stages in A. angustifolia. Different from previous reports, only after pollination, pre-fertilization tissue development occurs (0–13 months after pollination (MAP)) and gives rise to a mature megagametophyte. During all this period, pollen is in a dormant state at the microphyla, and pollen tube germination in nucellus tissue is only observed at the stage of archegonia formation (13 MAP) and not at the free nuclei stage as reported before. For the first time, 14 months after pollination, a fertilization window was indicated, and at 15 MAP, the polyzygotic polyembryony from different archegonia was also seen. After that, subordinated proembryo regression occurs and at least three embryonic developmental stages of dominant embryo were characterized: proembryogenic, early embryogenic, and late embryogenic (15–23 MAP). Along these stages, histochemical and ultrastructural analyses suggest the occurrence of cell death in suspensor and in cap cells of dominant embryo that was not previously reported. The differentiation of meristems, procambium, pith, and cortex tissues in late embryogenic stage was detailed. The morphohistological characterization of pre-fertilization and embryonic stages, together with the timescale of megastrobili development, warranted a referential map of female reproductive structure in this species.     

Realized pollen and seed dispersal within a continuous population of the dioecious coniferous Brazilian pine [Araucaria angustifolia (Bertol.) Kuntze]

Extensive realized pollen and seed flow across populations reduces inbreeding and spatial genetic structure (SGS) and increases the genetic diversity and effective size within populations. Inbreeding, SGS and realized patterns of pollen and seed dispersal of the dioecious, wind pollinated Araucaria angustifolia were investigated based on microsatellite loci. The study was conducted in a 7.2 ha plot established within a continuous Araucaria Forest in Southern Brazil. In the plot, all 290 adult trees were mapped, measured (diameter at breast height, dbh), sexed, sampled and genotyped. We also sampled, measured (total height), mapped and genotyped 223 juveniles. A total of 86 alleles were detected in the sample (n = 513). Adults and juveniles showed a positive and significant fixation index (minimum of 0.096), suggesting inbreeding or Wahlund effect. Juveniles presented a significant aggregated distribution which was associated with a genetic aggregation (significant SGS up to 20 m), indicating that near neighbor trees may be related due the limited seed dispersal. However, the intensity of SGS was not significantly higher among juveniles (Sp = 0.0041) than adults (Sp = 0.0026). Realized pollen and seed immigration into the plot was low (pollen = 6 %; seeds = 5 %) and the patterns of pollen and seed dispersal were similar. Pollen was dispersed over long distances (343 m), but 50 % was dispersed up to 124 m. Seeds also reached long distances (318 m), with 50 % being dispersed up to 133 m. Our results are discussed in terms of auto-ecology and the genetic conservation of A. angustifolia populations.     

Seed germination in Chrysophyllum sp. nov., a large-seeded rainforest species in north Queensland: effects of seed size,litter depth and seed position

Leaf litter affects seed germination in many ways and past studies have shown greater impacts on relatively small seeds, both within and among species. In this shade-house experiment I examined the impact of forest litter on seed germination in Chrysophyllum sp. nov. (Sapotaceae), a large-seeded (2.4 g) rainforest tree from north Queensland. Seed mass varies more than 30-fold in this species, making it useful for studying the role of litter as a possible selective pressure in the evolution of seed size in large-seeded species. Seeds of varying size (small, medium, large) were sown in planting boxes containing one of three litter levels (low, medium, high) and placed either below or on top of the litter. Seed size and litter biomass had no significant impact on the number of germinating seeds or the time to germination but seeds placed below the litter germinated around twice as frequently, and 20% sooner, than seeds placed on top of the litter. There were no significant interactions between any of the three factors. This shade-house experiment suggests that leaf litter is not an important selective pressure in the evolution of seed size in this species. However, if litter disturbance under field conditions differentially affects the probability of seed germination in relation to seed size and/or litter biomass, then litter could still act as a selective pressure in the evolution of seed size in Chrysophyllum and other large-seeded species.     

Impact of monoecy in the genetic structure of a predominately dioecious conifer species, Araucaria angustifolia (Bert.) O. Kuntze

Understanding the mating system of a tree species is important in genetic conservation and tree breeding strategies because it affects the inbreeding and genetic diversity of the descendant populations. Araucaria angustifolia (Bert.) O. Kuntze is a mainly dioecious species that reproduces through outcrossing. However, some monoecious trees have been identified and they may reproduce through self-fertilization. The objective of this study was to confirm the expected relatedness of full-sibs in outcrossed hand-pollinated progenies of female seed trees, self-sibs of hand self-pollinated monoecious seed trees, and to investigate the mating system of open-pollinated progenies of female and monoecious A. angustifolia trees. To do this, eight microsatellite loci were used to genotype hand- and open-pollinated progenies. Our results show that the relatedness of outcrossed hand-pollinated progenies are true full-sibs and progenies from a selfed monoecious seed tree are self-sibs, which confirms the hand-pollination method used. Open-pollinated female seed trees reproduced only by outcrossing, generating progenies with a mixture of full- and half-sibs. Monoecious seed trees reproduced mainly by xenogamy, generating progenies with mixtures of self-sibs, full-sibs, half-sibs and self-half-sibs. We also found that an increase in the effective number of pollen donors ( $ N_{\text{ep}} $ ) would lead to an increase in the total number of alleles ( $ K $ ) within progenies. Our results also suggest that monecious trees have limited potential to modify the genetic structure through selfed seed production due to the very low estimated selfing rate in these trees and the rare occurrence of these trees in natural populations.     

Growth rate,photosynthetic activity,and leaf development of Brazil pine seedlings (Araucaria angustifolia[Bert.] O. Ktze.)

Seedlings of Brazil pine, a large-seeded South American conifer, were grown in a climate chamber to investigate vertical growth pattern and the time course of leaf development. We examined shoot growth, photosynthetic performance and markers of leaf maturation such as contents of soluble sugars and activities of sucrose-phosphate synthase (SPS), neutral invertase (nI) and sucrose synthase (Susy). The daily increment of shoot length showed an optimum curve during the first 70 days after germination. The low growth rate during the first 20 days of development correlated with net CO² emission of the seedling. Analyses of leaf maturation markers in older seedlings revealed low sucrose/hexose ratios and high activities of nI and Susy in the uppermost leaves. Although the SPS/Susy ratio was low in these leaves the extractable SPS activity did not change significantly among leaves of different age. The photosynthetic light compensation points of young leaves were about 2-fold higher than those of mature leaves and their photosynthetic capacity was only 50% as high. Our results indicate that a rapid maturation of leaves of Brazil pine seedlings may reduce the respiratory loss of carbohydrates and that the mobilisation of seed storage compounds supports initial shoot growth under light-limiting conditions which may occur in the forest-grassland succession zone.     

Desiccation studies in relation to the storage of Araucaria seed

Relationships between seed moisture content (fresh weight basis) and germination were examined for nine Araucaria species by desiccation under mild environmental conditions. The lowest safe moisture content, below which germination percentage begins to decline, was estimated in each case. Seeds can be grouped into three moisture content categories: the first group (including A. araucana, A. angustifolia, A. hunsteinii and A. bidwillii) cannot be safely dried to below 25–40%; the second group (including A. columnaris, A. rulei, A. nemorosa and A. scopulorum) cannot be dried to below about 12% without damage; the third category contains A. cunninghamii, which can be dried to 2% without damage. Seeds in the first group should be stored at 0–5 °C with moisture contents above the lowest-safe value. Provided freezing damage does not exceed 10%, seeds in the second group should be kept at - 18°C or lower with about 7% moisture content for long-term storage and at 0–5 °C with about 12% moisture content in the short term. Seed of A. cunninghamii is best retained at near 5% moisture content and in -18°C or lower. The lowest-safe moisture content was found to be associated with seed size and weight, higher moisture content values coinciding with greater size and weight of seed. Food reserve materials also differed among the groups; seeds of the first group were mainly starchy, whilst those in the other categories possessed a high lipid content.     

Drought and shade deplete nonstructural carbohydrate reserves in seedlings of five temperate tree species

Plants that store nonstructural carbohydrates (NSC) may rely on carbon reserves to survive carbon‐limiting stress, assuming that reserves can be mobilized. We asked whether carbon reserves decrease in resource stressed seedlings, and if NSC allocation is related to species' relative stress tolerances. We tested the effects of stress (shade, drought, and defoliation) on NSC in seedlings of five temperate tree species (Acer rubrum Marsh., Betula papyrifera Marsh., Fraxinus americana L., Quercus rubra L., and Quercus velutina Lam.). In a greenhouse experiment, seedlings were subjected to combinations of shade, drought, and defoliation. We harvested seedlings over 32–97 days and measured biomass and NSC concentrations in stems and roots to estimate depletion rates. For all species and treatments, except for defoliation, seedling growth and NSC accumulation ceased. Shade and drought combined caused total NSC decreases in all species. For shade or drought alone, only some species experienced decreases. Starch followed similar patterns as total NSC, but soluble sugars increased under drought for drought‐tolerant species. These results provide evidence that species deplete stored carbon in response to carbon limiting stress and that species differences in NSC response may be important for understanding carbon depletion as a buffer against shade‐ and drought‐induced mortality.     

Mineral reserves in castor beans: the dry seed

Elemental composition and distribution of the mineral reserves in the endosperm and embryo tissues of Ricinus communis cultivars Hale and Zanzibarensis were investigated. Energy dispersive x-ray analysis was used to determine the elemental composition of the globoid crystals, while atomic absorption spectrometry allowed quantification of the elements, particularly Ca, in various seed regions. No major differences were found between the two cultivars with regard to the elemental distribution in globoid crystals. While the majority of globoid crystals contained P, K, and Mg, the occasional one also contained Ca. In extremely rare instances, Fe was detected in globoid crystals. Ca-containing globoid crystals were more common in provascular cell protein bodies in the stem and radicle. Polarized light microscopy, micro-incineration, and acid solubility tests demonstrated the presence of calcium oxalate crystals in the innermost testa which adheres to the endosperm and is often mistakenly identified as endosperm. Atomic absorption spectrometry revealed that most of the calcium present in castor bean seeds is localized in the testa. On a perseed-region basis, the much larger endosperm contains more Ca than does the embryo. However, on a unit-weight basis, the radicle-plus-stem regions contain considerably more Ca than does the cotyledon or endosperm, an observation that is consistent with the observed distribution pattern for Ca-containing globoid crystals.     

Climatic signals in tree‐rings of Araucaria angustifolia in the southern Brazilian highlands

Araucaria angustifolia (Bertol.) O. Kuntze (Araucariaceae) is a Neotropical tree, widely distributed in subtropical mountain rain forests and nearby natural grasslands of Southern Brazil. This species produces annual growth rings, but its dendroclimatic potential is barely known. In the present paper, the long‐term growth patterns of A. angustifolia were investigated using annual growth ring time series and association to climate over the last century. Wood cores of A. angustifolia trees growing in forest and grassland habitats were obtained with an increment borer. The cores were surfaced, measured and cross‐dated. The dated ring‐width time series were standardized and submitted to correlation and principal component analysis to verify growth trends among sites and trees. Growth‐climate relationships were investigated using correlation and regression analyses, comparing the ordination axes scores to regional time series of precipitation and temperature. Due to anatomical irregularities, mainly partial rings, only 35 out of 60 trees were cross‐dated. The correlation and ordination analyses showed common tree‐growth trends within and between sites, indicative of a regional environmental force determining inter‐annual cambial activity variation. Despite growing in distinct habitats and disturbance regimes, A. angustifolia trees share a common long‐term growth pattern, which is significantly related to thermal conditions during the current and previous growing seasons. Moreover, site‐specific characteristics may have influenced opposite growth responses and association to climate conditions between forest and grassland trees.     

重要地下害虫东北大黑鳃金龟研究进展

东北大黑鳃金龟Holotrichia diomphalia是我国东北地区重要的蛴螬类地下害虫,主要危害麦类、玉米、豆类等大田作物,以及薯类、花生等经济作物。华北大黑鳃金龟Holotrichia oblita和江南大黑鳃金龟Holotrichia gebleri是华北及江南地区的主要蛴螬类害虫。近年来,传统分类学、杂交实验以及分子生物学的研究都证明,3种鳃金龟是同一个物种的不同地理种群,这些研究成果使东北大黑鳃金龟从一个区域性害虫转变成为分布范围从东北到长江流域的广布害虫。本文综合介绍了东北大黑鳃金龟(包括华北大黑鳃金龟、江南大黑鳃金龟)近二十年来的研究成果,包括分类学、发生规律、防治措施,以及生理生化及分子生物学等,并对东北大黑鳃金龟的综合治理研究进行了展望。     

Female parthenogenetic apomixis and androsporogenetic parthenogenesis in embryonal cells of Araucaria angustifolia: interpolation of progenesis and asexual heterospory in an artificial sporangium

Cell fate, development timing and occurrence of reproductive versus apomictic development in gymnosperms are shown to be influenced by culture conditions in vitro. In this study, female parthenogenetic apomixis (fPA), androsporogenetic parthenogenesis (mAP) and progenesis were demonstrated using embryonal initials of Araucaria angustifolia in scaled-up cell suspensions passing through a single-cell bottleneck in darkness and in an artificial sporangium (AS). Expression was based on defined nutrition, hormones and feedforward-adaptive feedback process controls at 23-25?°C and in darkness. In fPA, the nucleus of an embryonal initial undergoes endomitosis and amitosis, forming a diploid egg-equivalent and an apoptotic ventral canal nucleus in a transdifferentiated archegonial tube. Discharge of egg-equivalent cells as parthenospores and their dispersal into the aqueous culture medium were followed by free-nuclear conifer-type proembryogenesis. This replaced the plesiomorphic and central features of proembryogenesis in Araucariaceae. Protoplasmic fusions of embryonal initials were used to reconstruct heterokaryotic expressions of fPA in multiwell plates. In mAP, restitutional meiosis (automixis) was responsible for androsporogenesis and the discharge of monads, dyads, tetrads and polyads. In a display of progenesis, reproductive development was brought to an earlier ontogenetic stage and expressed by embryonal initials. Colchicine increased polyploidy, but androspore formation became aberrant and fragmented. Aberrant automixis led to the formation of chromosomal bouquets, which contributed to genomic silencing in embryonal initials, cytomixis and the formation of pycnotic micronucleated cells. Dispersal of female and male parthenospores displayed heteromorphic asexual heterospory in an aqueous environment.