Aulacaspis yasumatsui Invasion Reduced Cycas micronesica Microstrobilus Size and Pollinator Brood Site Competence.

Aulacaspis yasumatsui Takagi invaded Guam in 2003, and the influence on survival and demography of the host Cycas micronesica K.D. Hill population has been well-studied. To more fully understand how A. yasumatsui has threatened the host cycad species, we determined the microstrobilus size and number of pollinators per microstrobilus from 2001 to 2021. The microstrobilus height and diameter were measured directly, and the volume was calculated. Microstrobili were 58 cm in height, 13 cm in diameter, and 4740 cm3 in volume prior to direct A. yasumatsui infestations. Microstrobili decreased in size immediately after direct infestations by A. yasumatsui, and then began to slowly increase in size until 2021. For example, the volume was 24% of pre-invasion volume in 2007, and was 57% of pre-invasion volume in 2021. Microstrobili were harvested; then, the number of pollinator pupae were counted after an incubation period. Pollinator pupae counts per microstrobilus declined to 66% of pre-invasion levels by 2007 and have remained similarly constrained through 2021. Our results revealed that A. yasumatsui damage to the host C. micronesica population is not limited to attrition of the extant plant population, but also includes a loss in male reproductive effort and the risk of coextinction of the insular pollinator.

Unique chemistry associated with diversification in a tightly coupled cycad-thrips obligate pollination mutualism.

Cycad cone thermogenesis and its associated volatiles are intimately involved in mediating the behavior of their obligate specialist pollinators. In eastern Australia, thrips in the Cycadothrips chadwicki species complex are the sole pollinators of many Macrozamia cycads. Further, they feed and reproduce entirely in the pollen cones. M. miquelii, found only in the northern range of this genus, is pollinated only by a C. chadwicki cryptic species that is the most distantly related to others in the complex. We examined the volatile profile from M. miquelii pollen and ovulate (receptive and non-receptive) cones to determine how this mediates pollination mechanistically, using GC-MS (gas chromatography-mass spectrometry) and behavioral tests. Monoterpenes comprise the bulk of M. miquelii volatile emissions, as in other Macrozamia species, but we also identified compounds not reported previously in any cycad, including three aliphatic esters (prenyl acetate and two of uncertain identity) and two aliphatic alcohols. The two unknown esters were confirmed as prenyl (3-methylbut-2-enyl) esters of butyric and crotonic ((E))-but-2-enoic) acids after chemical synthesis. Prenyl crotonate is a major component in emissions from pollen and receptive ovulate cones, is essentially absent from non-receptive cones, and has not been reported from any other natural source. In field bioassays, Cycadothrips were attracted only to those volatile treatments containing prenyl crotonate. We discuss M. miquelii cone odorants relative to those of other cycads, especially with respect to prenyl crotonate being a species-specific signal to this northern C. chadwicki cryptic species, and how this system may have diversified.

Biotic Threats to Cycas micronesica Continue to Expand to Complicate Conservation Decisions.

Invasions of non-native species can threaten native biodiversity, and island ecosystems are ideal for studying these phenomena. In this article, first, we report on the invasive species that combine to threaten the island cycad Cycas micronesica by reviewing the history of previously reported invasions and providing an update of recent invasions. Then, we prioritize the threat status of each herbivore and the interactions among them. Plant damage was initiated in 2003─2005 by the non-native Aulacaspis yasumatsui Takagi armored scale, Erechthias sp. Meyrick leaf miner, and Luthrodes pandava Horsfield butterfly, which elicited unprecedented irruptions of the native Acalolepta marianarum Aurivillius stem borer and increased herbivory by feral pigs (Sus scrofa L.). The combined impact of these five consumers represents the greatest sustained threat to the cycad tree species. Mitigation of the damage caused by phytophagous non-native species is urgently needed to conserve this unique gymnosperm tree.

Burrowing activity of coconut rhinoceros beetle on Guam cycads.

Guam's established population of non-native coconut rhinoceros beetle (CRB, Oryctes rhinoceros L.) began creating burrows in stem apices of several cycad species in a managed garden. We conducted an island-wide survey to determine the spatial patterns of CRB burrowing of stems of in situ Cycas micronesica. We also measured starch of healthy and unhealthy coconut leaf tissue and compared this with starch of cycad stem tissue. The starch concentration of the central unexpanded leaf in healthy Philippine coconut trees was ≈90 mg·g-1, and that of unhealthy Guam coconut trees was ≈40 mg·g-1. The starch concentration of the tissue within the CRB burrow locations on C. micronesica trees was 145 mg·g-1. Burrowing of C. micronesica was restricted to female CRB adults and was found throughout the full latitudinal gradient of Guam. Our findings indicate Guam's unhealthy coconut trees are no longer nutrient-dense, and the female CRB population may have exhibited a phylogenetically distant host shift to the abundant C. micronesica plants for a starch-rich diet within the concepts of the 'ecological fitting' hypothesis. We add proximity to coconut tree habitats as a new threat to Guam's endangered C. micronesica population.

Thermogenic respiratory processes drive the exponential increase of volatile organic compound emissions in Macrozamia cycad cones.

An important outcome of plant thermogenesis is increased emissions of volatiles that mediate pollinator behaviour. We investigated whether the large increase in emissions, mainly the monoterpene ß-myrcene (>90%), during daily thermogenic events of Macrozamia macleayi and lucida cycad cones are due solely to the influence of high cone temperatures or are, instead, a result of increased respiratory rates during thermogenesis. We concurrently measured temperature, oxygen consumption and ß-myrcene emission profiles during thermogenesis of pollen cones under typical environmental temperatures and during experimental manipulations of cone temperatures and aerobic conditions, all in the dark. The exponential rise in ß-myrcene emissions never occurred without a prior, large increase in respiration, whereas an increase in cone temperature alone did not increase emissions. When respiration during thermogenesis was interrupted by anoxic conditions, ß-myrcene emissions decreased. The increased emission rates are not a result of increased cone temperature per se (through increased enzyme activity or volatilization of stored volatiles) but are dependent on biosynthetic pathways associated with increased respiration during thermogenesis that provide the carbon, energy (ATP) and reducing compounds (NADPH) required for ß-myrcene production through the methylerythritol phosphate (MEP) pathway. These findings establish the significant contribution of respiration to volatile production during thermogenesis.

Cone thermogenesis and its limits in the tropical Cycas micronesica (Cycadaceae): association with cone growth, dehiscence, and post-dehiscence phases.

PREMISE OF THE STUDY: Thermogenesis is a prominent pollination-related feature of cycad cones and is generally assumed to play a role in pollination. Although typically studied just before, during, and immediately after the cones' pollination phase, thermogenesis may be present in other cone developmental phases. • METHODS: We assayed thermogenesis in Cycas micronesica, Guam's endangered cycad, over successive cone developmental phases by measuring temperatures in shaded and unshaded in situ cones for up to 7 wk. We also studied the effect of ambient conditions on cone thermogenesis in laboratory experiments and estimated the cones' metabolic heating rates. • KEY RESULTS: Pollen cones exhibit a continuous, but small, metabolically generated thermogenesis for multiple weeks, including a single thermogenic peak temperature greater than peak ambient each day. The magnitudes of those daily peak temperature elevations above ambient reach maxima twice during cone development: a few days before dehiscence and approximately 1 wk post-dehiscence. Excised cones in dark, fixed temperature environments generated multiple thermogenic events (∼24 h period) over ∼10 d. Cones appear to initiate a protective temperature regulatory response at temperatures ≥∼38°C. • CONCLUSIONS: Cycas micronesica pollen cones exhibit several thermogenic attributes not reported in other cycads, including continuous thermogenesis for many weeks. These cones grow in a hot tropical environment that likely confines their metabolically generated temperature increases to a small thermogenic window beyond which they encounter heat stress. These findings suggest the presence of thermogenic functions not strictly related to pollination and a potential vulnerability to warming climates.

Information-based or resource-based systems may mediate Cycas-herbivore interactions.

Invasive arthropod herbivores comprise one of the greatest threats to cycad conservation both in situ and ex situ. We discuss two mechanisms, not necessarily mutually exclusive, that may underlie the disparity in Chilades pandava damage among Cycas species. In an information-based system, plant infochemicals may differentially influence oviposition behavior of Ch. pandava adults or host finding behavior of this butterfly's natural enemies. Alternatively, heterogeneity in damage may be mediated by a resource-based system whereby plant substrate is more palatable to larvae for susceptible species or more defended by less damaged species.

Arthropod invasion disrupts Cycas micronesica seedling recruitment.

We recently described characteristics of reproductive effort for the cycad Cycas micronesica on the island of Guam. The data were serendipitously recorded just prior to the devastating invasion of the armored scale Aulacaspis yasumatsui. This invasion decimated the cycad population and after six years of infestation no recruitment is occurring among the survivors. We describe various underlying mechanisms that may explain how this homopteran insect has eliminated host recruitment among categories including plant-pollinator mutualism disruptions, direct damage to reproductive structures, population level responses to declining plant health, and failures of seedlings to establish. Our pre-invasion data on reproductive effort will serve as the benchmark for quantifying how this alien pest is endangering the endemic cycad.

Unstable, self-limiting thermochemical temperature oscillations in Macrozamia cycads.

Field measurements and laboratory experiments on the Australian cycads Macrozamia lucida and Macrozamia macleayi demonstrate that their cones' diel peak thermogenic temperature increase varies systematically with cone stage, with single thermogenic temperature peaks occurring daily for up to 2 weeks and reaching 12 degrees C above ambient at midstage. The initiation, magnitude and timing of those peaks are strongly modulated by ambient temperature; the period between successive thermogenic temperature peaks is not circadian, and light is neither necessary nor sufficient to initiate a thermogenic event. A mathematical analysis is developed that provides a unified explanation of the experimental results. It describes these unstable, self-limiting thermogenic events in terms of conservation of energy and a first-order chemical reaction rate model that includes an Arrhenius equation dependence of the cone's metabolic heating rate on the cone temperature.

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.