INSECT POLLINATION IN THE CYCAD ZAMIA PUMILA (ZAMIACEAE)

A wind and insect exclusion pollination experiment was conducted in a wild population of the cycad Zamia pumila L. in Florida. Cones from which insects but not wind were excluded produced no viable seeds, while cones from which wind but not insects were excluded produced abundant viable seeds. Two beetle species have been identified which may be effecting pollination. Adults and larvae of both beetles are found abundantly on the male cones, and adults of both species and larvae of one species are found occasionally on female cones. Z. pumila produces sugar and amino acid-rich micropyle droplets which may serve as pollinator rewards. Mimicry of resource-rich male cones by female cones and the use of cones as refuges between mating and feeding bouts may account for beetle movement from male to female cones.     

Two Global Conformation States of a Novel NAD(P) Reductase Like Protein of the Thermogenic Appendix of the Sauromatum guttatum Inflorescence

A novel NAD(P) reductase like protein (RL) belonging to a class of reductases involved in phenylpropanoid synthesis was previously purified to homogeneity from the Sauromatum guttatum appendix. The Sauromatum appendix raises its temperature above ambient temperature to ~30 °C on the day of inflorescence opening (D-day). Changes in the charge state distribution of the protein in electrospray ionization–mass spectrometry spectra were observed during the development of the appendix. RL adopted two conformations, state A (an extended state) that appeared before heat-production (D ? 4 to D ? 2), and state B (a compact state) that began appearing on D ? 1 and reached a maximum on D-day. RL in healthy leaves of Arabidopsis is present in state A, whereas in thermogenic sporophylls of male cones of Encephalartos ferox is present in state B. These conformational changes strongly suggest an involvement of RL in heat-production. The biophysical properties of this protein are remarkable. It is self-assembled in aqueous solutions into micrometer sizes of organized morphologies. The assembly produces a broad range of cyclic and linear morphologies that resemble micelles, rods, lamellar micelles, as well as vesicles. The assemblies could also form network structures. RL molecules entangle with each other and formed branched, interconnected networks. These unusual assemblies suggest that RL is an oligomer, and its oligomerization can provide additional information needed for thermoregulation. We hypothesize that state A controls the plant basal temperature and state B allows a shift in the temperature set point to above ambient temperature.     

Salicylic Acid Levels in Thermogenic and Non-Thermogenic Plants

The natural trigger for heat production in the thermogenic inflorescencesof Sauromatum guttatum Schott (voodoo lily) was recently identifiedas salicylic acid (SA), which induced heat production at levelsas low as 13 ng g f. wt^–1. Since then the levels of SAwere determined in other thermogenic and non-thermogenic plantspecies. In thermogenic inflorescences of five aroid species,and in male cones of at least four thermogenic cycads SA levelsduring heat production exceeded 1 µg g f. wt^–1.SA was not detected in the thermogenic flowers of a water lily,Victoria regia Lindl. (Nymphaeaceae), and Bactris major Jacq.(Palmae). Levels of salicylic acid varied substantially in thefloral parts of seven non-thermogenic species and in the leavesof 27 non-thermogenic species. Amorphophallus campanulatus Blume ex Decne, Arum italicum Mill., Arum dioscoridis Sibth. & Son., Philodendron selloum Koch, Monstera deliciosa Liebm., Encephalartosferox Bertol. f., Encephalartos hildebrandtii A. Br. & Bouché, Encephalartos gratus Prain, Dioon edule Lindl. cv. edule, Dioon edule Lindl. cv angustifolium, Sauromatum guttatum Schott, voodoo lily, Victoria regia Lindl., Bactris major Jack, salicylic acid, thermogenicity, heat production     

Energy Loss in Tissue 2Sauromatum guttatum (Schott) Analysed by Microcalorimetry

The heat evolved by 1 mm-thick tissue slices of the appendixof the Sauromatum guttatum inflorescence was measured calorimetricallyduring development. From D–5 (5 d before inflorescence-opening,designated as D-day) to D–2 about 8 µW mg^–1fresh wt. was observed, and during D–1 an increase inheat evolution to 14 µW mg^–1 fresh wt. was monitored.The heat was produced through oxidative metabolism, since replacingthe air in the microcalorimeter with nitrogen blocked heatproduction.Addition of salicylic acid to tissue slices of thermogenic organs(appendix, lowest part of the spadix and male flowers) and ofnon-thermogenic organs (female flowers, club-shaped organs andlower part of the spadix) of Sauromatum inflorescences revealedthat the acid boosted heat-production only in the thermogenicorgans. The effect of the acid manifested itself with no appreciablelag time, and it generated non-linearity in the rate of heat-productionby tissue slices of the appendix. In the appendices of the highlythermogenic Arum italicum and of the weakly thermogenic Amorphophallusrivieri, salicylic acid selectively boosted the rate of heat-productionin the appendix of A. italicum. A Q₁₀ of 24 was found between15C to 25 C for 1 mm-thick slices of D–4 appendicesof S guttatum. Addition of digitonin or deoxycholate to pre-D-daytissue slices of the appendix increased the rate of heat-production. Key words: Amorphophallus rivieri, Arum italicum, microcalorimetry, salicylic acid, Sauromatum guttatum     

Oleic Acid and Its Positional Isomer,cis-Vaccinic Acid,in the Appendix of Sauromatum guttatum during Anthesis

The fatty acid profiles of various organs of the thermogenic inflorescence of Sauromatum guttatum and of the sporophylls of thermogenic male cones of two cycad species (Encephalartos ferox and Dioon edule var edule and var angustifolium) were determined by gas chromatography. During anthesis, palmitate (16:0), oleate [18:1 (9)], cis-vaccinate [18:1 (11)], and linoleate [18:2 (9, 12)] were the most abundant fatty acids in the Sauromatum appendix. cis-Vaccinic acid, a positional isomer of oleic acid, was identified by comparing its retention time on a gas chromatography column and its mass spectrum to an authentic compound. The percentage of oleic acid from total fatty acids dropped from about 9 in the morning 3 d before heat production to 6 in the morning 2 d before heat production. At this time, the percentage of cis-vaccinic acid increased from 3 to 11%, and then remained at this level until the inflorescence dried and died. Palmitoleic acid [16:1 (9)], the common precursor of cis-vaccinic acid, is a minor component of total fatty acids. In six other organs of the Sauromatum inflorescence including thermogenic organs, such as male flowers and lower spadix, palmitate, oleate, and linoleate were prevalent but cis-vaccinate was not. The thermogenic male cones of the two cycad species were rich in palmitic, oleic, and linolenic acids. The level of cis-vaccinic acid in these organs was less than 0.5%.     

METABOLIC ASPECTS OF THERMOGENESIS IN MALE CONES OF FIVE CYCAD SPECIES

High starch concentrations in the sporophylls of male cones of five species of cycad and their depletion during their heat-producing phase of development suggest that starch is a major fuel source for thermogenesis in these plants. Carbon-13/carbon-12 ratio analyses of respired CO₂, starch, and lipids in a male cone of one of these species, Macrozamia moorei F. Muell., indicate that although starch is a major contributor to respiration, lipids are also an important respiratory substrate.     

INSECT-CYCAD SYMBIOSIS AND ITS RELATION TO THE POLLINATION OF ZAMIA FURFURACEA (ZAMIACEAE) BY RHOPALOTRIA MOLLIS (CURCULIONIDAE)

Zamia furfuracea, a cycad of Mexican origin, in cultivation is pollinated by the snout weevil, Rhopalotria mollis, also of Mexican origin. This weevil apparently is host specific, swarming upon male cones of the cycad, where mating, feeding, and oviposition occur. Sporophylls of male cones are rich in starch; those of female cones are poor in starch, and weevils feed upon male cones and are visitors to but not feeders upon or within female cones. Pollen transport to female cones occurs during such visitation. All stages of metamorphosis of R. mollis occur within male cones; larvae feed exclusively on parenchyma of microsporophylls, pupate within stalks of microsporophylls, and emerge as adults from the outer ends of microsporophylls. They do not feed on pollen and do not damage microsporangia or pollen. Toward the end of the breeding season of the weevil (and the cycad), some larvae enter diapause in thick-walled pupal cases within microsporangial stalks of pollen-spent cones. These remain in diapause until the next reproductive season of the cycad.     

Cyanide-Insensitive Respiration in Thermogenic Flowers of Victoria and Nelumbo

The thermogenic carpellary appendages of the flowers of Victoriacruziana d'Orb. and the thermogenic staminal appendages of Nelumbolutea (Willd.) Pers. possess the cyanide-insensitive, ‘alternative’respiratory pathway. The presence of this pathway was demonstratedin tissue slices as well as in mitochondria. The thermogenicactivity was accompanied by ultrastructural changes in the Victoriamitochondria. Before anthesis, mitochondria with well-developedcristae were present in appendage tissue in large numbers. Duringanthesis, lamelliform cristae appeared in a different orientation. Key words: Victoria, Nelumbo, flowers, thermogenicity, mitochondria     

Scent chemistry and patterns of thermogenesis in male and female cones of the African cycad Encephalartos natalensis (Zamiaceae)

Most, if not all, extant cycads are pollinated by insects which use the cones as larval brood sites. These interactions appear to be mediated by cone volatiles, and, in some species, by patterns of thermogenesis. We investigated the chemical composition of volatile emissions and patterns of thermogenesis in cones of the South African cycad Encephalartos natalensis, using a gas chromatograph–mass spectrometer (GC–MS) and miniature temperature data loggers (ibuttons), respectively. This was done during various developmental stages (before and during receptivity and pollen release) for both female and male cones. A total of 31 compounds were identified in headspace samples; 17 of which were common to both sexes, 12 found only in male cones, and two found only in female cones. The major volatiles in pollen and female cones are (3E)-1,3-octadiene (averaging 54.25% and 15.82% of total emissions), (3E,5Z)-1,3,5-octatriene (averaging 13.37% and 47.66%), and α-pinene (averaging 16.29% and 12.24%). Female cones were not thermogenic before and during receptivity whereas pollen cones were thermogenic during pollen shedding. Thermogenesis of male cones occurred between 1400 h and 1530 h on successive afternoons, reaching an average of c. 10.5 °C above ambient temperature. Volatile emissions and thermogenesis occurred in association with insect activity on the cones suggesting that they both play a role in regulating insect behaviour.     

Pollination of the cycad <Emphasis Type="Italic">Zamia incognita</Emphasis> A. Lindstr. &amp; Idárraga by <Emphasis Type="Italic">Pharaxonotha</Emphasis> beetles in the Magdalena Medio Valley,Colombia: a mutualism dependent on a specific pollinator and its significance for conservation

The genus Zamia (Zamiaceae: Cycadales) exhibits its greatest diversity in Colombia and is highly threatened by habitat loss, extraction for ornamental plant trade, and mining, among other factors. One of the most important considerations for the effective conservation of Zamia is its highly specialized reproductive biology. Despite the importance of pollination for the populations’ viability, no studies have examined the pollination process of cycads in Colombia. Herein, we describe the pollination process of Zamia incognita A. Lindstr. & Idárraga, in a natural population. Exclusion experiments were performed by selectively excluding wind, beetles, both, or neither, which demonstrated that Pharaxonotha beetles are effective pollinators of Zamia incognita and that wind does not play any role as pollen vector. By following beetles marked with fluorescent dyes and directly observing beetle movements on and into female cones and micropyles, we confirmed that Pharaxonotha sp. is the effective pollinator of Z. incognita. The beetles traveled a maximum dispersal distance from a male to female cone of nearly 22 m and a minimum distance of 5 m. We found Pharaxonotha beetles in male cones, where they complete their life cycle. Cones produce heat in a circadian pattern associated with the elongation of the cones and pollen shedding. The increase in cones’ temperature appears to play an important role in beetle attraction. We suggest that pollination droplets on the micropyles would be a reward to pollinators. We also discuss the relationship of this Zamia species with other insects, which have important consequences for the conservation of web interactions.     

MORPHOLOGICAL STUDIES OF LYMNAEID SNAILS FROM THE HUMAN FASCIOLIASIS ENDEMIC ZONE OF BOLIVIA

The taxonomic position of the Lymnaeidae from the BolivianAltiplano has been unclear. On the basis of conchological characters,some authors reported two species from this area, Lymnaea viatrix andL. cubensis while others, considering also anatomical characters,considered L. viatrix as a synonym of L. cubensis. More recentstudies demonstrated genetic identity between the Bolivian lymnaeidsand L. truncatula from the Iberian Peninsula. Populations recognizedas L. cubensis correspond to a distinct genetic group, but geneticinformation was not available for L. viatrix. In the light ofthese genetic results, a morphometric study of both the shell (usingRaupian parameters) and male reproductive system was carried outof L. cubensis from Cuba (type locality), Dominican Republic,Guadeloupe and Venezuela, and of L. truncatula from Bolivia,France, Portugal, Spain and Morocco. Syntypes of L. viatrixfrom Argentina (var. A. ventricosa) and specimens of L. viatrix(var. B. elongata) from the type locality, Peru, were also studied(conchological characters only). The conchological study showedthe presence of a large amount of variability between populations.This variability was not congruent with genetic results. Alarge amount of variability was also found using anatomicalcharacters of the male reproductive system and all of them clearlyseparate L. cubensis from L. truncatula independently from geographicalorigin. Thus anatomical characters, unlike conchological parameters,do differentiate taxonomic species inferred from genetic studies. (Received 11 September 1997; accepted 5 March 1999)     

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.     

Arboreal camera trapping sheds light on seed dispersal of the world’s only epiphytic gymnosperm: Zamia pseudoparasitica

Epiphytic lifestyles have evolved independently in ecologically, morphologically, and taxonomically diverse plant species. Although this adaptation is widespread among angiosperms, it is only known to have arisen in a single gymnosperm species, Zamia pseudoparasitica (Cycadophyta). Zamia pseudoparasitica is endemic to the mountains of Western Panama, and little is known about the ecology of this unusual cycad. Here, we provide the first report of a potential seed disperser of Z. pseudoparasitica. Between late October 2019 and March 2020, we conducted arboreal camera trapping at three sites along the Talamanca Cordillera in Western Panama, yielding an accumulated survey effort of 271 camera days. Weekly direct observations were also performed using handheld binoculars at one site. Arboreal camera trapping revealed at least seven mammal species that visit this epiphytic cycad. At all three sites, the Northern olingo (Bassaricyon gabbii) was seen visiting individuals of Z. pseudoparasitica repeatedly, both while cones were closed and after they had opened. We estimated the time‐varying intensity of the visits throughout our sampling and used mixed models to compare the length of visits when cones were closed versus when they were open. Both duration and time‐varying intensity of visits increased after cones had opened and we documented Northern olingo removing and carrying away seeds. We also observed predation by the yellow‐eared toucanet (Selenidera spectabilis) which picked and destroyed mature Z. pseudoparasitica seeds. These results suggest that the Northern olingo could be an important seed dispersal agent for this rare epiphytic gymnosperm.     

危害黄杉球果的实小卷蛾属一新种(鳞翅目：卷蛾科)

记述了实小卷蛾属 (小卷蛾亚科,花小卷蛾族)的一个新种,黄杉实小卷蛾Retinia pseudotsugaicola Liu et Wu, sp. nov.。其幼虫危害黄杉Pseudotsuga sinensis Dode的球果,在云南省禄劝县云龙乡球果受害率达80%～90%。该新种雄性外生殖器的抱器瓣在腹面中央凹陷很深,抱器腹拐角几乎呈直角,抱器端的腹角尖,可与该属其它种相区别。该新种的雌性外生殖器与松实小卷蛾R.cristata很相似,但成虫的花纹完全不同,该新种前翅的中横带很宽,且布满了灰绿色和灰黄色的云状纹,而松实小卷蛾前翅的中横带则较窄,翅端有明显的肛上纹。在昆明禄劝县一年发生1代,以蛹在球果中越冬。越冬蛹于翌年3月下旬开始羽化。4月上旬至7月下旬是幼虫危害期。老熟幼虫7月下旬开始陆续化蛹越冬。提供了成虫外形、翅脉、雌雄外生殖器解剖图及幼虫的毛序图。模式标本保存在中国科学院动物研究所。     

Human Ortholog of a Plant Salicylic Acid Receptor Found in SK-N-SH Cell Line

Our previous studies have described the purification and characterization of a novel plant NAD(P)-reductase like protein (RL) from the thermogenic appendix of the Sauromatum guttatum inflorescence. RL is mainly located in cytoplasm of thermogenic plants and it can act like a bistable switch. It adopts a compact conformation during heat-production and a more expanded conformation when heat is not generated. Addition of salicylic acid, a natural thermogenic inducer, at picomolar concentration to a solution of purified RL induced a discontinuous volume phase transition in which the volume of RL in the oligomeric form expanded and shrunk repeatedly every 4–5 min. In the present study using ESI–MS analysis we have demonstrated the existence of RL in the human SK-N-SH cell line and in mouse brain tissue. The molecular mass of human RL is in the same range as of its plant counterpart, 34,140 ± 34 Da. The charge state distribution of the human RL is identical to its plant counterpart from the Sauromatum appendix during heat-production. Human RL was present in the compact state when it was purified from the SK-N-SH cell line When these cells were treated with salicylic acid (10 μM) a shift to a much more compact conformation was observed. It seems that the potential of RL to respond to salicylic acid was conserved. These results may reveal the existence of a thermoregulation system that is evolutionarily conserved and is operating by conformational changes. This discovery may also represent an opportunity for a better understanding of some of the diverse functions of salicylic acid and aspirin in plants and humans.     

Synchronicity of thermogenic activity, alternative pathway respiratory flux, AOX protein content, and carbohydrates in receptacle tissues of sacred lotus during floral development

The relationships between heat production, alternative oxidase(AOX) pathway flux, AOX protein, and carbohydrates during floraldevelopment in Nelumbo nucifera (Gaertn.) were investigated.Three distinct physiological phases were identified: pre-thermogenic,thermogenic, and post-thermogenic. The shift to thermogenicactivity was associated with a rapid, 10-fold increase in AOXprotein. Similarly, a rapid decrease in AOX protein occurredpost-thermogenesis. This synchronicity between AOX protein andthermogenic activity contrasts with other thermogenic plantswhere AOX protein increases some days prior to heating. AOXprotein in thermogenic receptacles was significantly higherthan in post-thermogenic and leaf tissues. Stable oxygen isotopemeasurements confirmed that the increased respiratory flux supportingthermogenesis was largely via the AOX, with little or no contributionfrom the cytochrome oxidase pathway. During the thermogenicphase, no significant relationship was found between AOX proteincontent and either heating or AOX flux, suggesting that regulationis likely to be post-translational. Further, no evidence ofsubstrate limitation was found; starch accumulated during theearly stages of floral development, peaking in thermogenic receptacles,before declining by 89% in post-thermogenic receptacles. Whilstcoarse regulation of AOX flux occurs via protein synthesis,the ability to thermoregulate probably involves precise regulationof AOX protein, most probably by effectors such as -keto acids. Key words: Alternative oxidase, alternative pathway respiration, Nelumbo nucifera, plant thermogenesis, starch Received 11 November 2007; Accepted 28 November 2007     

A Comparison of the Flowering and Branching Control Systems in Lathyrus odoratus L. and Pisum sativum L.

In the sweet pea (Lathyrus odoratus L.) the difference in floweringbehaviourbetween photoperiodic (long-day) and day-neutral cultivars appearsto be due to a difference in their ability to produce a graft-transmissiblefloral inhibitor. The flowering control systems in the sweetpea and the garden pea (Pisum sativum L.) appear to be verysimilar on the basis of inter-generic graft results. It is suggestedthat the major flowering genes Dn* in L. odoratus and Sn andDne in P. sativum control steps in a biochemical pathway commonto these two species (which are related at the tribal level)and that the product of this pathway inhibits flowering andpromotes outgrowth of basal laterals in both species.     

PETRIFIED PLANTS FROM THE UPPER MISSISSIPPIAN OF NORTH AMERICA. II. LEPIDOSTROBUS FAYETTEVILLENSE SP. N.

Structurally preserved arborescent lycopsid fructifications are described from the Fayetteville Shale (Upper Mississippian) of northwestern Arkansas. Specimens of Lepidostrobus fayettevillense sp. n. range from complete cones 22.5 cm long and approximately 1.0 cm in diameter to smaller water-worn fragments. The cones consist of a central axis bearing closely spaced, spirally arranged sporophylls which extend from the cone axis at right angles. Each sporophyll consists of a pedicel which is turned up at its end to form a distal lamina. Sporangia are large and attached to the adaxial surface of each sporophyll. The vascular cylinder consists of a centrally located exarch protostele at least 1.1 mm in diameter. The new species is compared with morphologically similar lycopsid fructifications of equivalent age.     

五种棺头蟋核型的比较研究

报道了中国5种棺头蟋的核型：石首棺头蟋Loxoblemmus equestris, 2n=17, XO(♂);小棺头蟋L. aomoriensis, 2n=11, XO(♂); 哈尼棺头蟋L. haani, 2n=11, XO(♂);多伊棺头蟋L. doenitzi, 2n=11, XO(♂)和窃棺头蟋L. detectus, 2n=11,XO(♂)。并应用核型似近系数及进化距离对这5种蟋蟀作了聚类分析,得出5种蟋蟀的演化方向为石首棺头蟋→小棺头蟋→哈尼棺头蟋→多伊棺头蟋→窃棺头蟋。     

Histological changes during maturation in male and female cones of the cycad Zamia furfuracea and their significance in relation to pollination biology

The cycad Zamia furfuracea L.Fil. Is pollinated by a curculionid beetle, Rhopalotria mollis Sharp which completes its life cycle in male cones of the cycad, and effectively pollinates female coneS. Idioblasts within parenchyma in both male and female cones appear to contain toxic compounds, including at least one neurotoxin, 2-amino-3-(methylamino) propanoic acid (BMAA), and a toxic glycoside, methylazoxymethanol-/!-primeveroside (macrozamin). Idioblasts appear structurally unmodified in male cones throughout the period of pollen maturation, and feeding weevils consume much of the starch-rich microsporophyll parenchyma tissue, including idioblastS. During this activity no appreciable change in morphology or staining reactions of male-cone idioblasts is detectable. Prior to pollen receptivity, female-cone idioblasts resemble those of male cones. Thereafter, many female-cone idioblasts show marked changes in morphology and content not caused by the weevils themselveS. Idioblast changes in female cones are probably associated with the defence of female-cone resources against predation by animals, including pollinating weevils, and may relate to mobilization of toxinS. Absence of similar morphological changes in male-cone idioblasts is correlated with toxin sequestration, enabling the pollinator to breed and feed without intoxication.