Is there a floral parasite mutualism in cycad pollination? the pollination biology of Encephalartos villosus (Zamiaceae)

Observations and experiments were carried out over 5 yr to distinguish between wind and insect pollination in the cycad Encephalartos villosus Lemaire (Zamiaceae). They were also designed to determine whether a pollination mutualism exists between E. villosus and Antliarhinus zamiae (Thunberg) (Coleoptera: Brentidae), an obligate ovule parasite that routinely parasitizes a large proportion of the ovules. The percentage of fertilized ovules dropped slightly when wind was excluded from the megastrobilus. However, when insects were excluded by either net bags or insecticide there was a substantial decrease in the proportion of fertilized ovules. Five beetle species belonging to four families were found on the strobili at the time of pollination. Using data on the effectiveness of pollen transfer to the receptive ovule, as well as data on abundance and pollen loads, a pollinator importance value (PIV) was determined for each beetle species and a pollinator importance index (PII) was determined for each population. PII values showed that an undescribed weevil (Porthetes sp., Curculionidae) was consistently the most important pollinator. A. zamiae and an undescribed beetle species within the Xenoscelinae (Languriidae) played a minor role in pollination, and their contributions varied from year to year and between populations. Two additional beetle species, Metacucujus goodei Endrödy-Younga (Boganiidae) and a second species of Xenoscelinae, had very low PII values and probably had little or no effect on pollination. Low PIV scores for A. zamiae were a result of its low numbers on the microstrobilus and the tendency of the beetles to remain on the outside of the megastrobilus. In the interaction between E. villosus and A. zamiae, the cycad does not appear to benefit significantly from a pollination service and I interpret this to mean that the relationship is antagonistic rather than mutualistic. There is, however, a possible mutualism between Porthetes sp. and E. villosus.     

合意非洲铁羽片表皮及气孔器的发育研究

用光学显微镜和扫描电镜对合意非洲铁羽片表皮及气孔器的发育过程进行观察研究。结果显示,(1)在羽片发育过程中表皮细胞变化不明显,主要是细胞伸长和壁增厚的过程;表皮的角质层连续出现。(2)气孔器发育可大致分3个阶段:保卫细胞母细胞阶段、幼保卫细胞阶段和成熟保卫细胞阶段;气孔器发育方式为单唇型或周源型;成熟气孔器属单环型。(3)在羽片发育过程中,气孔密度起初很低,后急剧升高并达到峰值,之后逐渐下降并趋于稳定。(4)有早熟气孔器。(5)羽片表皮发育成熟的方式是由基部向顶部逐步推进。     

Molecular relationships in Encephalartos (Zamiaceae, Cycadales) based on nucleotide sequences of nuclear ITS 1&2, rbcL, and genomic ISSR fingerprinting

The cycad genus Encephalartos is restricted to Africa and is threatened with extinction in most of its range. Total DNA was extracted from 51, i.e., 78 %, of the described species of Encephalartos. The accessions were sampled from the furthest western occurrence of the genus in Nigeria, via Sudan and Uganda, to southern South Africa. The sequences of nuclear ribosomal internal transcribed spacer regions 1 and 2 (ITS 1&2), the chloroplast encoded rbcL gene, and ISSR genomic fingerprinting were employed to resolve the molecular history and the relationships within the genus. Sequence alignment, as well as ISSR fingerprinting, data show low genetic variation among all analysed accessions, indicating diversification within the Pliocene/Pleistocene. ITS 1&2 data agree well with morphological and geographical characters and resolved three major genetic clusters with overlapping distribution ranges in eastern South Africa. This area, that contains the largest diversity of genotypes of Encephalartos, may have served as a Pliocene/Pleistocene refugium.     

Geographic variation in cone volatiles and pollinators in the thermogenic African cycad Encephalartos ghellinckii Lem

• Heat and odour production can have profound effects on pollination in cycads. It is therefore expected that these traits would co‐vary geographically with pollinator assemblages. Such intraspecific variation, may lead to the evolution of pollination ecotypes, which can be an early stage of pollinator‐mediated speciation.
• We measured cone temperatures using miniature temperature data loggers and examined the composition of cone volatile odours using headspace sampling and analysis with gas chromatography‐mass spectrometry in four populations spanning the range of the African cycad Encephalartos ghellinckii. Pollinator assemblages were also investigated in three populations.
• Male and female cones were thermogenic at pollen shed and receptive stages, respectively, but patterns of thermogenesis did not vary among populations. Scent emissions from cones in populations in the Drakensberg Mountains were characterised by cis‐β‐ocimene, β‐myrcene and (3E)‐1,3‐octadiene, while camphene and α‐pinene were characteristic of scent emissions from cones in populations closer to the coast. These differences in volatile blends corresponded with differences in insect assemblages.
• These results confirm intraspecific variation in volatile emissions of E. ghellinckii and support the predictions that intraspecific variation in volatile emissions will be associated with shifts in pollinator assemblages. While further work needs to be done to test for local adaptation in this system, this preliminary evidence is consistent with the formation of pollination ecotypes in the E. ghellinckii species complex.

     

Exploitation of Encephalartos transvenosus outside and inside Mphaphuli Cycads Nature Reserve, Limpopo Province, South Africa

This study has investigated the use and threats of Encephalartos transvenosus outside and inside Mphaphuli Cycads Nature Reserve, Limpopo Province, South Africa. Data were collected from 2005 to 2006 through social and ecological surveys. Semi-structured interviews were conducted with a total of 40 people per household, sampled from four selected villages around the reserve. E. transvenosus is used for medicinal purposes, decorations, cultural activities and as food. Youth and adult persons harvest E. transvenosus for income generation, while children and aged people mainly use it for subsistence purposes. The survival of E. transvenosus in the wild is uncertain due to various threats, such as illegal collection, habitat destruction, fire and grazing. Some conservation measures are also discussed.     

Patterns of odour emission,thermogenesis and pollinator activity in cones of an African cycad: what mechanisms apply?

Background and Aims

Ontogenetic patterns of odour emissions and heating associated with plant reproductive structures may have profound effects on insect behaviour, and consequently on pollination. In some cycads, notably Macrozamia, temporal changes in emission of specific odour compounds and temperature have been interpreted as a ‘push–pull’ interaction in which pollinators are either attracted or repelled according to the concentration of the emitted volatiles. To establish which mechanisms occur in the large Encephalartos cycad clade, the temporal patterns of volatile emissions, heating and pollinator activity of cones of Encephalartos villosus in the Eastern Cape (EC) and KwaZulu Natal (KZN) of South Africa were investigated.

Methods and Key Results

Gas chromatography–mass spectrometry (GC-MS) analyses of Encephalartos villosus cone volatiles showed that emissions, dominated by eucalyptol and 2-isopropyl-3-methoxypyrazine in EC populations and (3E)-1,3-octadiene and (3E,5Z)-1,3,5-octatriene in the KZN populations, varied across developmental stages but did not vary significantly on a daily cycle. Heating in male cones was higher at dehiscence than during pre- and post-dehiscence, and reached a maximum at about 1830 h when temperatures were between 7·0 and 12·0 °C above ambient. Daily heating of female cones was less pronounced and reached a maximum at about 1345 h when it was on average between 0·9 and 3·0 °C above ambient. Insect abundance on male cones was higher at dehiscence than at the other stages and significantly higher in the afternoon than in the morning and evening.

Conclusions

There are pronounced developmental changes in volatile emissions and heating in E. villosus cones, as well as strong daily changes in thermogenesis. Daily patterns of volatile emissions and pollinator abundance in E. villosus are different from those observed in some Macrozamia cycads and not consistent with the push–pull pattern as periods of peak odour emission do not coincide with mass exodus of insects from male cones.     

Some observations on the cycads of Central Africa

The results of two expeditions to Central Africa to collect and study cycads, mainly Encephalartos species, are presented. Further morphological information on the living plants growing in their natural habitats is given, together with new localities. Species delimitation is reassessed.     

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.     

Adaptation for oviposition into concealed cycad ovules in the cycad weevils Antliarhinus zamiae and A. signatus (Goleoptera: Gurculionoidea)

Antliarhinus zamiae (Thunberg) and A. signatus Gyllenhal are the only insects known in which the larvae feed exclusively on the cycad gametophyte. In their host plants, species of the cycad genus Encephalartos (Zamiaceae), the gametophyte is protected by a three-layered integument and substantial sporophyll tissues. Adaptation for oviposition into the gametophyte may, therefore, be crucial to this feeding specialization. Ovipositional behaviour and ovipositor structure in four species of Antliarhinus suggest that ovipositional traits suited to ovule predation preceded the evolution of this habit and that ovipositional traits in ancestral species provided the right environment for the evolution of ovule predation. Current ovipositional traits in A. zamiae and A. signatus probably represent responses to changes in cone structure after the evolution of ovule predation.     

Somatic embryogenesis in Encephalartos cycadifolius

Callus cultures of Encephalartos cycadifolius were established from zygotic embryo explants on a modified B5 medium containing 1 mg l^–1 2,4-D and 1 mg l^–1 kinetin. Callus was transferred to media containing various combinations of 2,4-D and kinetin for improvement of somatic embryogenesis. Somatic embryos were produced on media with several growth regulator combinations. The somatic embryos developed from proembryos, which developed long suspensors. A dicotyledonary embryo formed at the distal end of the suspensor. The embryos turned green in light. When transferred to a medium containing 1 mg l^–1 ABA the somatic embryos matured. The suspensors desiccated and these embryos rooted when transferred to a medium without phytohormones.Abbreviations ABA abscisic acid - 2,4-D 2,4-dichlorophenoxyacetic acid