Reproductive biology of the South American cycad Zamia boliviana,involving brood-site pollination

Brood-site pollination mutualisms represent extreme levels of reciprocal specialization between plants and insects. Several studies have revealed intimate associations between cycads and their insect pollinators across different continents. However, the reproductive biology of South American cycads is still poorly understood, with limited empirical data available. Here, we investigated the plant–pollinator interactions of Zamia boliviana (Cycadales, Zamiaceae), which is distributed on savanna patches in Brazil and Bolivia. Our aim was to examine reproductive biology (strobili development and their visitors) to identify potential pollinators from the perspective of dioecy and expectations of obligatory mutualism. We monitored reproductive cohorts and registered the strobiliʼs visitors. We classified the visitorsʼ activities and described the behavior of the most active and recurring ones in both reproductive structures to ascertain the potential pollinators and their relationship with the Z. boliviana. In situ experiments were used to demonstrate efficacy in seed production. Although other insects visit the strobili, only one Coleoptera (Erotylidae: Pharaxonotha) was present seasonally. Pharaxonotha cerradensis was the single species capable of transporting pollen and developing a life cycle associated with polleniferous strobili, while concomitantly visiting ovuliferous strobili. Experimental tests associated with observational evidence in the cycad indicated brood-site pollination mutualism with the Erotylidae beetle, a potential pollinator of Z. boliviana. Our investigation confirmed a new case of obligatory mutualism between cycads and P. cerradensis in the New World. Future research on Zamia from South America should address pollination systems of this genus to better understand the ecology and evolution of plant–pollinator interactions.     

When North and South don’t mix: genetic connectivity of a recently endangered oceanic cycad,Cycas micronesica,in Guam using EST‐microsatellites

Subject to environmental changes and recurrent isolation in the last ca. 250 Ma, cycads are often described as relicts of a previously common lineage, with populations characterized by low genetic variation and restricted gene flow. We found that on the island of Guam, the endemic Cycas micronesica has most of the genetic variation of 14 EST‐microsatellites distributed within each of 18 genetic populations, from 24 original sampling sites. There were high levels of genetic variation in terms of total number of alleles and private alleles, and moderate levels of inbreeding. Restricted but ongoing gene flow among populations within Guam reveals a genetic mosaic, probably more typical of cycads than previously assumed. Contiguous cycad populations in the north of Guam had higher self‐recruitment rates compared to fragmented populations in the south, with no substantial connection between them except for one population. Guam’s genetic mosaic may be explained by the influence of forest continuity, seed size, edaphic differences, and human transport of cycads. Also important are the extent of synchrony among flushes of reproductive female seed‐bearing sporophylls and restricted pollen movement by an obligate mutualist and generalist insects. An NADH EST‐locus under positive selection may reflect pressure from edaphic differences across Guam. This and three other loci are ideal candidates for ecological genomic studies. Given this species’ vulnerability due to the recent introduction of the cycad aulacaspis scale, we also identify priority populations for ex situ conservation, and provide a genetic baseline for understanding the effects of invasive species on cycads in the Western Pacific, and islands in general.     

Repeat photography confirms alarming decline in South African cycads

Although South African cycads are known to be declining in the wild, there is at present no broad-scale, quantitative analysis to support this view. In this study the fate of 626 individual cycads was assessed from 107 repeat photographs taken at 53 locations over three time-steps (broadly 1940s, mid-1990s and 2014). Of the cycads photographed in the 1940s, 78 % survived to the mid-1990s. By 2014, however, only 16 % of the original cycads remained. The Limpopo Province had the highest rate of cycad loss over the recorded period, followed by the Eastern Cape and KwaZulu-Natal. Gauteng and Mpumalanga had the lowest rate of cycad loss. In general, cycad losses were greatest on land under private ownership when compared to communal land and conservation areas. However, cycad loss as a result of damage sustained due to traditional medicine collection was highest on communal lands. Continued declines in most of the studied cycad populations are a concern due to the potential Allee effects this may introduce. While legislation to protect cycads is in place, enforcement is difficult given the spatially extensive and often remote and patchy distribution of cycad populations. Conservation approaches that work actively with farmers to protect cycads on their property, as well as education about sustainable traditional medicine collection practices in communal areas have the best chance of ensuring the viability of wild cycad populations.     

PATTERNS OF ALLOZYME DIVERSITY IN THE WEST INDIES CYCAD ZAMIA PUMILA (ZAMIACEAE)

The West Indies cycad, Zamia pumila, is restricted to the Greater Antilles, northern Bahama Islands, Florida, and the southeastern coast of Georgia. An electrophoretic study based on nine enzymes compared 21 accessions from throughout the range of the species. Lower levels of intrapopulation variation than those reported for ferns, other gymnosperms, and angiosperms were discovered for the two more extensively sampled populations. However, this variation was similar to that found in other island taxa and in the endemic Australian cycad, Macrozamia communis. In contrast, allozyme divergence among accessions of Z. pumila appeared relatively high, mostly as a result of rare alleles restricted in geographic distribution. The age and biogeography of Z. pumila may gave contributed to population differentiation. Also, mean number of alleles per locus was low for the species (1.75). Finally, the time-since-divergence value (10.8 million years ago) between Z. pumila and its closest extant relative, Z. splendens, was much smaller than the age of Z. pumila suggested by the fossil record and historical geology of the Caribbean (30–60 million years ago). Together, these data indicate that biochemical evolution within this species, and perhaps in all cycads, is slow when compared to that of noncycad seed plants.     

苏铁类植物传粉生物学研究进展(综述)

苏铁类植物是现存最古老的种子植物,研究其传粉特点,对于研究种子植物的起源与演化、植物与动物的协同进化以及苏铁类植物的繁殖机制、濒危机制有重要意义。本文从苏铁雌雄株开花物候学、传粉媒介及传粉机制等方面,对苏铁类植物传粉生物学相关研究进行综述,并提出今后该类植物传粉生物学研究的建议：对更多苏铁属种类未知的传粉机制进行研究;更广泛地应用排除法研究苏铁类特别是苏铁属植物的传粉媒介,进一步探讨传粉昆虫与苏铁类的共生关系问题。     

Comparison of DNA restriction fragment length polymorphisms of Nostoc strains in and from cycads

DNA was prepared from cyanobacteria freshly isolated from coralloid roots of natural populations of five cycad species: Ceratozamia mexicana mexicana (Mexico), C. mexicana robusta (Mexico), Dioon spinulosum (Mexico), Zamia furfuraceae (Mexico) and Z. skinneri (Costa Rica). Using the Southern blot technique and cloned Anabaena PCC 7120 nifK and glnA genes as probes, restriction fragment length polymorphisms of these cyanobacterial symbionts were compared. The five cyanobacterial preparations showed differences in the sizes of their DNA fragments hybridizing with both probes, indicating that different cyanobacterial species and/or strains were in the symbiotic associations. On the other hand, a similar comparison of cyanobacteria freshly collected from a single Encephalartos altensteinii coralloid root and from three independently subcultured isolates from the same coralloid root revealed that these were likely to be one and the same organism. Moreover, the complexity of restriction patterns shows that a mixture of Nostoc strains can associate with a single cycad species although a single cyanobacterial strain can predominate in the root of a single cycad plant. Thus, a wide range of Nostoc strains appear to associate with the coralloid roots of cycads.Non-standard abbreviations bp base pairs - kbp kilobase pairs - RFLP's restriction fragment length polymorphisms     

Is lack of cycad (Cycadales) diversity a result of a lack of polyploidy?

Many hypotheses exist for the relative diversity of seed plant taxa. We discuss one of them: how the relative dearth of cycad diversity throughout their 300 million year history may be a result of a lack of duplication of their entire nuclear genome, often termed polyploidy. We show theoretically how polyploidy causes speciation via cryptic reproductive isolation. Polyploidy can also cause radiations via epigenetically induced heterochrony and plasticity. Most cycads have only a few large chromosomes because of a lack of whole genome duplication or, except possibly in the genus Zamia, chromosomal fission. Large chromosomes and extremely small effective population sizes result in substantial linkage disequilibrium, genetic hitchhiking and genetic drift in cycads. By contrast, other seed plants have higher incidences of polyploidy and may therefore have been more prone to radiations. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 165 , 156–167.     

Variation in reproductive output of Ficus superba despite aseasonal reproduction

Studies of Ficus reproductive phenology have focused on documenting its seasonality and dependence on meteorological factors. While there have been reports that duration of receptivity of syconia can be prolonged in response to pollinator limitation, the effect of pollination rate on the duration of phenological phases over a year has not been examined. Percentage of unpollinated syconia, number of foundresses per pollinated syconium, duration of receptivity, frequency of single-sex broods, crop size, frequency of parasitisation, persistence of ripe syconia, and mass abscission were recorded for Ficus superba at the crop level. Percentage of unpollinated syconia is significantly negatively correlated with the number of foundresses per syconium, and positively correlated with the duration of receptivity, and the persistence of ripe syconia. Despite the absence of sesonality in reproduction, the occurrences of receptive syconia and ripe syconia showed unimodal and bimodal peaks, respectively, owing to prolongment of these phases. This is attributable to meteorological factors, which are hypothesized to influence pollinator dispersal, or population dynamics, and thus pollination rate. This highlights the overlooked significance of pollinator, and possibly frugivore phenology in accounting for Ficus reproduction, and suggests that while reproduction may be aseasonal, reproductive potential may not be. Furthermore, crop size was shown to affect the number of foundresses per syconium, duration of receptivity, and persistence of ripe syconia. Possible adaptive value of producing crops of different sizes is discussed.     

Mast-seeding in the cycad genus Encephalartos: a test of the predator satiation hypothesis

Mast-seeding behaviour was monitored in 18 populations of eight species of the African cycad genus Encephalartos between 1988 and 1991. The coefficient of variation (V) in annual cone production for each population ranged between 88 and 200, indicating large fluctuations in reproductive effort between years. Data were collected to determine whether mast-seeding reduced levels of predispersal seed predation by satiating seed predators in mast years and whether it resulted in a reproductive advantage over plants which reproduced more frequently. Masting intensity was greatest in those populations in which individual plants suffered the highest levels of predispersal seed predation in years when only a few plants produced seeds. The principal seed predators were two congeneric weevil species, Antliarhinus zamiae and A. signatus, which develop exclusively on cycad seeds. The lowest intensity of mast-seeding was recorded for cycad populations with low levels of seed predation and in which A. zamiae and A. signatus occurred only in low numbers or were entirely absent. Larger seed crops appeared to result in lower levels of seed predation by A. zamiae and A. signatus in four populations of E. altensteinii, and differences in seed crop size accounted for 48–66% of variation in levels of seed predation in populations of five cycad species. In one population of E. altensteinii, lower levels of seed predation in plants reproducing periodically resulted in a reproductive advantage over plants reproducing more frequently. These results are consistent with the predator satiation hypothesis. However, in most cycad populations, numbers of seed predators did not appear to decrease significantly after a period of 2–8 years between reproductive episodes and, in two of three populations examined, periodic reproduction did not increase the number of seeds surviving to dispersal over a 4-year period. These results are interpreted to mean that periodic reproduction has not evolved in response to selection imposed by seed predators, but that selection may favour those plants which experience lower levels of seed predation by coning in synchrony with the majority of plants in the population.     

Exploring cycad foliage as an archive of the isotopic composition of atmospheric nitrogen

Molecular nitrogen (N₂) constitutes the majority of Earth's modern atmosphere, contributing ~0.79 bar of partial pressure (pN₂). However, fluctuations in pN₂ may have occurred on 10⁷–10⁹ year timescales in Earth's past, perhaps altering the isotopic composition of atmospheric nitrogen. Here, we explore an archive that may record the isotopic composition of atmospheric N₂ in deep time: the foliage of cycads. Cycads are ancient gymnosperms that host symbiotic N₂‐fixing cyanobacteria in modified root structures known as coralloid roots. All extant species of cycads are known to host symbionts, suggesting that this N₂‐fixing capacity is perhaps ancestral, reaching back to the early history of cycads in the late Paleozoic. Therefore, if the process of microbial N₂ fixation records the δ¹⁵N value of atmospheric N₂ in cycad foliage, the fossil record of cycads may provide an archive of atmospheric δ¹⁵N values. To explore this potential proxy, we conducted a survey of wild cycads growing in a range of modern environments to determine whether cycad foliage reliably records the isotopic composition of atmospheric N₂. We find that neither biological nor environmental factors significantly influence the δ¹⁵N values of cycad foliage, suggesting that they provide a reasonably robust record of the δ¹⁵N of atmospheric N₂. Application of this proxy to the record of carbonaceous cycad fossils may not only help to constrain changes in atmospheric nitrogen isotope ratios since the late Paleozoic, but also could shed light on the antiquity of the N₂‐fixing symbiosis between cycads and cyanobacteria.     

An ecological study of the relationship between two living fossils in Malawi: the Mulanje Tiger Moth (Callioratis grandis) and the Mulanje Cycad (Encephalartos gratus)

The Mulanje tiger moth (Callioratis grandis) has recently been discovered feeding solely on the Mulanje Cycad, Encephalartos gratus. In its type locality Encephalartos gratus is severely threatened through habitat destruction and human population growth. As a result the first detailed demographic survey of Encephalartos gratus on Mount Mulanje was undertaken with particular emphasis on the relationship between E. gratus and Callioratis grandis. What is unusual about C. grandis is that the males exhibit lekking behaviour, for which aposematism may be a prerequisite. Cycads were surveyed in a systematic manner. Both the biotic and abiotic parameters were recorded from 532 wild Encephalartos gratus. Results showed the morphological range of measurements differed from that of previously published literature on this species. Results also showed that the incidence of Callioratis larval damage was related to the maturity of the wild cycads – moth damage was greater on more mature plants. Approximately 50% of surveyed cycads showed evidence of Callioratis moth damage, however, there was no apparent preference from the moth to select either male or female cycads as food plants for the larvae. In some cases there was severe damage to cycad fronds, but this did not seem to affect the production of reproductive cones or even the survival of individual cycads.     

Zamia grijalvensis sp. nov. (Zamiaceae,Cycadales) from Chiapas,Mexico with notes on hybridization and karyology

Zamia grijalvensis, a new cycad species from Chiapas, México, is described and illustrated. This species is similar to Z. lacandona with respect to leaves, but differs in habit, morphology and colour of the female and male cones. The female cone of Z. grijalvensis is reddish brown to orange brown, whereas that of Z. lacandona is dark brown with lighter beige tomentum. The male cone of Z. grijalvensis is erect, whereas that of Z. lacandona is decumbent. The new species may be of natural hybrid origin and is discussed in the light of other findings among cycads. Preliminary diploid chromosome counts for Z. grijalvensis 2n = 19, 20 and karyotypes appear variable, as previously found in its congeners with wide distribution, such as Z. paucijuga and Z. prasina, both of which are characterised by high diploid numbers with a high number of telocentric and few metacentric chromosomes. These changes in chromosome number are probably the result of stressful environmental factors.     

珍稀濒危植物距瓣尾囊草(Urophysa rochkii)的开花物候和繁育系统特性

通过野外观察,运用套袋授粉和联苯胺-过氧化氢法等实验方法对距瓣尾囊草开花物候以及种群的繁育系统特性进行研究。研究结果:1)距瓣尾囊草为两性花,聚伞状花序,花期较长,从头年12月持续到次年4月,单花花期持续8—15 d,种群花期可持续4个月左右。2)距瓣尾囊草在花朵未开放前花粉没有活力,花药开裂当天花粉活力达到94.8%,第2天为90.3%,第3天为81.6%,第4天为62.9%,此后其花粉活力明显减弱;柱头可授性于散粉后第3天开始出现,散粉后4—5天最强,散粉后第8天柱头失去可授性;柱头可授性和花粉活力有5 d左右的重叠期。3)其花粉胚珠比P/O为938.5±250,按照Cruden标准划分,其有性繁育系统为兼性异交。4)按照Dafni的标准,距瓣尾囊草的OCI=4,即繁育系统为异交、部分自交亲和、需要传粉者。5)人工异花授粉结实率达80%以上,略高于自然结实率78.94%;直接套袋结实率为5.71%;去雄套袋和去雌套袋均不结实。以上结果表明,距瓣尾囊草繁育系统表现为异交为主、部分自交亲和并且不存在无融合生殖现象,其开花物候、花部特征和繁育系统为适应特殊的环境提供了一定的生殖保障,本次研究结果为距瓣尾囊草的迁地保护和种群的繁衍复壮提供理论依据。     

Cycad cone and angiosperm floral volatiles: Inferences for the evolution of insect pollination

The chemistry of the volatile emissions of male cones of four putatively insect-pollinated cycad (Cycadales) species and cones of both sexes of one wind-pollinated cycad species was analysed using GC-MS Zamia furfuracea, Macrozamia moorei, and Encephalartos altensteinii had blends consisting primarily of monoterpenoids, benzenoids, and in two causes, a di-unsaturated aliphatic hydrocarbon, while that of Zamia pumila was dominated by methyl salicylate, with sesquiterpenes and aliphatic hydrocarbons also present. In contrast, the wind-pollinated Cycas rumphii contained a series of highly alcohols, ketones and esters. The compounds classes found in the cycads are potent herbivore deterrents, and there is no apparent difference in this regard between insect- and wind-pollinated species. The results suggest convergent evolution in the gymnospermous cycads and magnoliid angiosperms of the olfactory cues that attract pollinating insects; the data are consistent with the hypothesis that early chemical attractants for pollinators evolved from herbivore deterrents.     

Population genetics and conservation of critically small cycad populations: a case study of the Albany Cycad,Encephalartos latifrons (Lehmann)

Declining populations of less than 250 mature individuals are symptomatic of many Critically Endangered cycads, which, globally, comprise the most threatened group of organisms as a result of collecting and habitat loss. Survival plans focus on law enforcement, reintroduction, and augmentation programmes using plants from the wild and botanical gardens. Augmentation is one of the few remaining options for cycad populations, although the assumed benefits remain untested and there is a possibility that augmentation from different sources could compromise the genetic integrity of existing populations, especially when garden plants have no provenance data. We studied Encephalartos latifrons, a South African endemic, which is a typical Critically Endangered cycad. We studied the extent and structure of genetic diversity in wild and ex situ populations to assess the potential benefits and risks associated with augmentation programmes. We examined 86 plants using amplified fragment length polymorphisms (AFLPs). The 417 AFLP markers thus generated yielded a unique DNA ‘fingerprint’ for each plant. Wild populations retain high levels of genetic diversity and this is reflected among the ex situ holdings at the Kirstenbosch Botanical Garden. No population differentiation is evident, indicating a single panmictic population, consistent with moderately high levels of gene flow between subpopulations and a sexual mode of reproduction. Bayesian clustering identified four genotype groups in the wild, as well as a genotype group only found in ex situ collections. Our results indicate that E. latifrons would benefit from augmentation programmes, including the use of undocumented collections, and careful management of breeding plants would increase the heterogeneity of propagules. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 293–308.     

The dissimilar costs of love and war: age-specific mortality as a function of the operational sex ratio

Lifespan and ageing are strongly affected by many environmental factors, but the effects of social environment on these life-history traits are not well understood. We examined effects of social interaction on age-specific mortality rate in the sexually dimorphic neriid fly Telostylinus angusticollis. We found that although interaction with other individuals reduced longevity of both sexes, the costs associated with variation in operational sex ratio were sex specific: males' early-life mortality rate increased, and lifespan decreased, with increasing male bias in the sex ratio, whereas surprisingly, the presence of males had no effect on early-life mortality or lifespan of females. Intriguingly, early-life (immediate) mortality costs did not covary with late-life (latent) costs. Rather, both sexes aged most rapidly in a social environment dominated by the opposite sex. Our findings suggest that distinct reproductive activities, such as mating and fighting, impose different age-specific patterns of mortality, and that such costs are strongly sex specific.     

GEOGRAPHIC VARIATION IN REPRODUCTIVE BEHAVIOR AND SIZE STRUCTURE OF THE AUSTRALIAN CYCAD MACROZAMIA COMMUNIS (ZAMIACEAE)

Macrozamia communis is a common cycad that occurs in eastern New South Wales, Australia, where it forms an understory in various forest or woodland communities. Leaf number of reproductive individuals as well as of nonreproductive ones, taken as an indication of size class, varies clinally from north to south, with reproductive individuals in southern populations having mean leaf numbers about 12 times greater than those at the northern end of the range of this species. Female cone size and mature ovule weight appear to vary also along this gradient. Using leaf number as a predictor of sexual maturity, from 37 to 100% of the individuals present in a population were judged to be mature. The percent of mature individuals participating in the 1987 coning episode ranged from none in three southern populations to 68% elsewhere. In 9 of 10 samples more males than females participated in the 1987 coning episode, with a collective sex ratio slightly male biased for all populations sampled. In three population samples average cone numbers for male and female plants were equal. In two samples males produced more cones per plant than females. Leaf numbers of males and females in individual populations were equal. Few individuals producing cones in 1987 showed evidence of recent prior cone production, suggesting that successive cone production by both sexes is separated by an interval of several years. The periodicity with which fire is used as a fire suppression method may affect the frequency of cone production in some populations, perhaps preventing it entirely. The apparent absence of recruitment in some populations may result from long-term seed collection by humans for horticultural purposes as well as the very poor seed dispersal that appears to characterize this cycad throughout its range. The weevil Tranes internatus was observed occupying the internal tissues of female cones; this insect may be involved in the pollination of M. communis.     

Genetic diversity and structure of the cycad Zamia loddigesii Miq. (Zamiaceae): implications for evolution and conservation

The genetic diversity and structure of four populations of the cycad Zamia loddigesii were studied throughout its range in Mexico. Allozyme electrophoresis of 15 loci was conducted. The mean number of alleles per locus was 1.80 ± 0.09, the percentage of polymorphic loci was 66.6 ± 5.4, and the expected heterozygosity was 0.266 ± 0.02. The results indicated that the genetic diversity was relatively higher, with respect to tropical tree species and other cycads. The genetic variation explained by differences among populations was 18%. On average, gene flow between paired populations was similar ( Nm  = 1.6) to other tropical forest trees and cycad species. Our results indicated that the geographical isolation among populations of Z. loddigesii generated allele loss, as well as a clinal variation in the frequencies of two loci ( MDH and MNR2 ), in relation to the latitudinal distribution of populations. The populations have become fragmented due to increasingly higher pressure of habitat conversion and disturbance. The importance of the establishment of sanctuaries and protected areas and a reduction in deforestation is highlighted in this research as a way of preserving the high genetic diversity of this and other endemic species.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 152 , 533–544.