Demographics, pollination syndrome and conservation status of Macrozamia platyrhachis (Zamiaceae), a geographically restricted Queensland cycad

To assess the International Union for Conservation of Nature (IUCN) status of Macrozamia platyrhachis F.M.Bailey, we surveyed this central Queensland cycad for its population abundance and health and its pollinator type and pollination syndrome (thermogenesis and volatile emissions). Plants are locally abundant within the 11 discrete populations surveyed, with an estimated population of 611 315 adult plants. Plants are highly restricted to a small area of occupancy, seed dispersal is nearly non-existent and extreme fires appear to have destroyed almost all seeds and seedlings and decimated the pollinators. Of known Macrozamia pollinators, only the thrips, Cycadothrips chadwicki Mound, were found on cones, and these were found in very low numbers. The pollination syndrome for this cycad appears to be unique, based on two cone traits. For one, thermogenesis peaks in early evening, a contrast with daytime peaks of other Cycadothrips-pollinated Macrozamia, but matches that of the Tranes weevil-pollinated Macrozamia machinii. In addition, cone volatiles include both previously unreported compounds as well as those reported exclusively on either Cycadothrips- or Tranes-pollinated species. Based on its small, fragmented area of occupancy, projected population declines and the unique pollination syndrome, we recommend that M. platyrhachis retain its current status as 'Endangered'. Habitat management plans should stipulate that controlled burns be avoided during cycad coning season and that wildfires be controlled to minimise damage to seedlings and pollinators.

Pollination ecology of the Australian cycad Lepidozamia peroffskyana (Zamiaceae)

Experiments carried out to investigate the reproductive ecology of the Australian cycad Lepidozamia peroffskyana (Regal, Bull. Soc. Imp. Nat. Mosc. 1857, 1: 184) revealed that this species is pollinated exclusively by host-specific Tranes weevils (Pascoe 1875). The weevils carry out their life cycle within the tissues of the male cones but also visit the female cones in large numbers. Female cones from which insects ( but not wind) were excluded had a pollination rate that was essentially zero. In contrast, female cones from which wind ( but not insects) were excluded had a pollination rate comparable with naturally pollinated cones. Assessment of Tranes weevil pollen load indicated that they are effective pollen-carriers. No other potential insect pollinators were observed on cones of L. peroffskyana. Sampling of airborne loads of cycad pollen indicated that wind-dispersed grains were not consistently recorded beyond a 2-m radius surrounding pollen-shedding male cones. The airborne load of cycad pollen in the vicinity of pollination-receptive female cones was minimal, and the spatial distribution of the coning population indicated that receptive female cones did not usually occur close enough to pollen-shedding male cones for airborne transfer of pollen to explain observed natural rates of seed set. These multiple lines of evidence suggest that wind-once considered the only pollination vector for cycads and other gymnosperms-plays only a minimal role in the pollination of L. peroffskyana, if any at all. The global diversity of insects associated with cycads suggests that some lineages of pollinating beetles may have been associated with cycad cones since Mesozoic times.

Pollination of Australian Macrozamia cycads (Zamiaceae): Effectiveness and behavior of specialist vectors in a dependent mutualism

Complementary field and laboratory tests confirmed and quantified the pollination abilities of Tranes sp. weevils and Cycadothrips chadwicki thrips, specialist insects of their respective cycad hosts, Macrozamia machinii and M. lucida. No agamospermous seeds were produced when both wind and insects were excluded from female cones; and the exclusion of wind-vectored pollen alone did not eliminate seed set, because insects were able to reach the cone. Based on enclosure pollination tests, each weevil pollinates an average 26.2 ovules per cone and each thrips 2.4 ovules per cone. These pollinators visited similar numbers of ovules per cone in fluorescent dye tests that traced insect movement through cones. Fluorescent dye granules deposited by Cycadothrips were concentrated around the micropyle of each visited ovule, the site of pollen droplet release, where pollen must be deposited to achieve pollination. In contrast, Tranes weevils left dye scattered on different areas of each visited ovule, indicating that chance plays a greater role in this system. Each weevil and 25 thrips delivered 6.2 and 5.2 pollen grains, respectively, on average, to each visited ovule per cone, based on examination of dissected pollen canals. In sum, the pollination potential of 25 Cycadothrips approximates that of one Tranes weevil.

Dioon Lindl. (Zamiaceae): Perspectives from phylogeny and a population genetic study of Dioon edule

Dioon Lindl. (Zamiaceae) is a small genus restricted to Mexico (12 species) and Honduras (one species). Previous systematic studies have been unable to fully resolve species relationships within the genus. Phylogenetic analyses were conducted with data from several sources, including Restriction Fragment Length Polymorphisms from the chloroplast genome, morphology, two introns of the low copy nuclear gene S-adenosyl-L-homocysteine hydrolase (SAHH) and the 5.8S/ITS2 regions of the nuclear ribosomal DNA. The goals of the study were to construct a total evidence species level phylogeny and to explore current biogeographical hypotheses. None of the analyses performed produced a fully resolved topology. Dioon is comprised of two main lineages (the Edule and Spinulosum Clades), which represents an ancient divergence within the genus. The two introns of the nuclear gene SAHH offer additional evidence for the split into two lineages. Intron 2 contains a 18 bp deletion in the Spinulosum Clade, providing a synapomorphy for that group. The 5.8S/ITS2 regions were highly polymorphic and subsequently omitted from the combined analyses. In order to visualize congruence between morphology and molecular data, morphological characters were mapped onto the combined molecular tree. Current biogeographical hypotheses of a general northward pattern of migration and speciation are supported here. However, sister relationships within the Edule Clade are not fully resolved. Seven DNA microsatellite markers were developed to investigate patterns of genetic variation of seven populations of D. edule, a species restricted to Eastern Mexico. We found that most of the genetic variation lies within populations (Ho = 0.2166–0.3657) and that levels of population differentiation are low (Fst = 0.088); this finding is congruent with the breeding system of this species, dioicy. Four of the populations deviate from Hardy Weinberg Equilibrium and have a high number of identical genotypes, we suggest that this unexpected pattern is due to the life-history strategy of the species coupled with the few number of polymorphic loci detected in these populations. Our results are not congruent with earlier evidence from morphology and allozyme markers that suggest that the two northernmost populations represent a distinct entity that is recognized by some taxonomists as D. angustifolium.

苏铁叶绿体的超微结构和功能及对CO2浓度倍增的响应

本研究首次揭示了七种苏铁类植物叶绿体的超微结构。即苏铁科(Cycadaceae)的攀枝花苏铁(C. panzhihuaensis).苏铁(C. revoluta)、叉叶苏铁(C.micholitzii)、 刺叶苏铁(C.rumphii和多歧苏铁(C.multipinnata)，蕨铁科(Stangeriaceae)的蕨铁(Stangeria eriopus)和泽米科(Zamiaceae)的米德尔堡大苏铁(Encephalortos middelburgensis).根据它们叶绿体内膜结构的不同将其大体分为两种类型：1、阴生型叶绿体：多歧苏铁和刺叶苏铁为此类型，它们的叶绿体类囊体垛叠程度高，基粒垛较宽，单个基粒中类囊体的数量很多，有的甚至上百;2、阳生型叶绿体：攀枝花苏铁、米德尔堡大苏铁、苏铁、蕨铁和刺叶苏铁以及外类群的凤尾蕨(Pteris vittata)的叶绿体均有阳生型叶绿体的特征：类囊体膜垛叠程度低，基粒较小。根据它们叶绿体的结构特征，又将其分为两组： ( 1)攀枝花苏铁，米德尔堡大苏铁和叉叶苏铁叶绿体中均有类囊体膜膨大的现象： (2)苏铁和蕨铁在苏铁类中是比较原始的种类，它们的叶绿体与在系统进化上较苏铁类低等的凤尾蕨的叶绿体中都有几个基粒聚集成簇的现象，说明苏铁类在进化的同时一些较原始的性状仍保留了下来。这些苏铁都在温室相同的条件下生长了两年多，但它们的叶绿体结构仍然能够反应原产地生境特点，说明在长期进化中，叶绿体对环境的适应方式已在基因水平上稳定下来，苏铁类植物不同叶绿体结构的形成有其遗传基础。 多歧苏铁，攀枝花苏铁，叉叶苏铁的叶绿体膜垛叠程度依次降低，与此相应，它们的chla/b和F730/F684依次升高，反应了结构与功能的一致性。 选用具有阳生型叶绿体的攀枝花苏铁和有阴生型叶绿体的多歧苏铁用不同的C02浓度（350 umol mol-l和700umol mol-l）处理后观察其叶绿体结构的变化，结果发现C02浓度倍增对它们的叶绿体影响甚微，而作为对照的无论是C3植物小麦(Triticum italica)还是C4植物谷子(Setaria italica)在C02倍增的条件下叶绿体内均有大量淀粉粒积累，并有膜结构改变。这说明苏铁的叶绿体结构有保守性，这有可能是苏铁类能历经亿万年的沧桑而生存下来的结构基础之一。

Association of cone thermogenesis and volatiles with pollinator specificity in Macrozamia cycads

Cone traits (volatile components and thermogenesis) of three cycad species in the genus Macrozamia were examined for differences related to their specific insect pollinators, the weevil, Tranes spp., or the thrips, Cycadothrips chadwicki. Linalool (>80% of emissions) dominated cone volatile components of M. machinii (Tranes-pollinated) and beta-myrcene was a minor component (