Induction of host‐plant preference in Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae)

The coconut hispine beetle, Brontispa longissima, is a serious invasive pest of coconut palm (Cocos nucifera) in Southeast Asia and the Pacific region. Recently, however, it has been reported that on Ishigaki Island, Japan, where C. nucifera is rare, the endemic palm Satakentia liukiuensis serves as a regular host. To explore this phenomenon, we examined the preferences of the beetle for the two palm hosts by choice tests and measurements of feeding amount using four different groups: naïve adults obtained from a colony reared on C. nucifera (NC); naïve adults from a colony reared on S. liukiuensis (NS); wild adults collected on C. nucifera on Ishigaki Island (WC); and wild adults collected on S. liukiuensis on Ishigaki Island (WS). Both NC and NS consumed significantly more C. nucifera leaves than S. liukiuensis leaves and were observed more often on the leaves of C. nucifera. These results suggest that B. longissima has an innate preference for C. nucifera. In contrast, field‐collected adults (WC, WS) initially showed preferences for the species on which they were collected, but after exposure to the other host plant for 2 weeks, their preferences for that host increased. These results suggest that the feeding experience of B. longissima adults affected their subsequent host preferences. This flexible host preference may have facilitated the beetle’s successful establishment in novel Japanese habitats where the original host plant was rare.     

Life history traits and damage potential of an invasive pest Brontispa longissima (Coleoptera: Chrysomelidae) on Satakentia liukiuensis

The coconut hispine beetle Brontispa longissima has been causing serious damage to the coconut palm Cocos nucifera in the Pacific, and Southeast and East Asia. This beetle also attacks Satakentia liukiuensis, an endemic palm on Ishigaki and Iriomote Islands, Japan. To assess the potential impact of B. longissima on S. liukiuensis, we examined the development period, survival rate, egg production and body size of B. longissima on S. liukiuensis at 22–31°C and consumption of S. liukiuensis leaf at 28°C. We also examined these life history traits on C. nucifera at 28°C to compare with those on S. liukiuensis. Brontispa longissima completed their development and reproduced on S. liukiuensis at 22–30°C. Linear regression analysis was used to estimate the effective cumulative temperature (k) as 891.2 degree‐days, and the lower developmental threshold as 13.3°C. On the basis of these values and monthly average temperatures on Ishigaki Island, it was estimated that B. longissima has four generations per year on the island. Satakentia liukiuensis was less suitable for B. longissima than C. nucifera in terms of immature survival, development time, resultant adult size and reproduction. The low quality of S. liukiuensis as a host‐plant may prevent the outbreak of B. longissima, which has not occurred yet on Ishigaki and Iriomote Islands.     

Response of Brontispa longissima to coconut palm (Cocos nucifera) leaf volatiles

Brontispa longissima (Gestro) (Coleoptera: Hispidae) is a new invasive pest in China that has caused severe economic damage to palm trees (Arecaceae, Palmae). The response of this beetle to coconut palm (Cocos nucifera) leaf volatiles is investigated in laboratory bioassays. Both sexes are attracted to a mixture of β‐myrcene, (?)‐limonene and E‐2‐hexen‐1‐ol (1 : 6 : 1), which are key components of coconut palm leaf volatiles. A blend of β‐myrcene and (?)‐limonene (0.7 : 1–1 : 0.7) in low amounts (100 ng) elicits aggregation and oviposition in females. Chemical analyses of food‐deprived, gravid female B. longissima show high concentrations of β‐myrcene and (?)‐limonene in their accessory glands, suggesting that female beetles sequester both compounds and release them during oviposition.     

Experimental investigations into the feasibility ofex situ preservation of palm seeds; an alternative strategy for biological conservation of this economically important plant family

Given the widespread belief that the conservation of palms, especially the large-trunked species, is only accomplished throughin situ preservation or in plantations, this paper explores the feasibility of a third approach, i.e. cryogenic preservation of their seedsex situ. Seeds of the following palm species were subjected to air-drying to assess their tolerance of dessication:Washingtonia filifera (L. Linden) H. Wendl.;Sabal mexicana Mart. (syn. S. texana; Zong, 1990);Schippia concolor Burret;Orbignya cohune (Mart.) Dahlgren ex Standley;Acoelorraphe wrightii (Griseb. & H. Wendl.) H. Wendl. ex Becc.;Desmoncus orthacanthos Mart.;Attalea crassispatha (Mart.) Burret;Zambia antillarum (Desc.) L.H. Bailey;Pinanga malaiana (Mart.) Scheff.;Pinanga aff. polymorpha Becc.;Daemonorops verticillaris (Griff.) Mart. Of these, only two (W. filifera andS. mexicana) survived drying to moisture contents around 5% (fresh weight basis). Seeds of the remaining spp. would be difficult or impossible to conserveex situ in seedbanks or cryostores. Data are presented to show that the response ofO. cohune embryos to drying is similar to other recalcitrant (dessication intolerant) seeds, while seeds ofA. wrightii may belong to an intermediate seed storage category with limited tolerance of drying. The results are discussed in relation to the inadequacy of current knowledge as a basis for decisions on the broad scaleex situ conservation of palm germplasm.     

Parasitisation of Tetrastichus brontispae (Hymenoptera: Eulophidae), a biological control agent of the coconut hispine beetle Brontispa longissima (Coleoptera: Chrysomelidae)

The gregarious endoparasitoids Tetrastichus brontispae Ferrière is one of the important natural enemies of the coconut hispine beetle Brontispa longissima (Gestro), a serious invasive pest on coconut palm plants (Cocos nucifera L.) in Southeast Asia. Development at different temperatures, effect of host and female ages, effect of food and oviposition frequency and superparasitism were investigated in the laboratory. Females were allowed only one attack against one host in all experiments. The wasp developed in a host between 19 and 30°C, whilst no wasp completed its immature development at 16 and 31°C. Host and female ages affected parasitisation. Parasitoid emergence was high on day 0 and day 1 pupal hosts, and younger females produced more offspring than older females. The longevity of the female was affected not only by food supply, but also by oviposition frequency. The female survived longer when oviposition frequency was low. However, the total number of hosts parasitised by the female during her lifetime did not differ at different oviposition frequencies. In superparasitism, although the percentage of adult emergence and body size of offspring decreased with an increasing number of attacks per host, a host parasitised by up to four females could produce parasitoid offspring.     

Artificial diets for rearing the coconut hispine beetle Brontispa longissima and its parasitoid Asecodes hispinarum

We tested artificial diets for rearing the coconut hispine beetle, Brontispa longissima, a serious invasive pest of coconut (Cocos nucifera) in Southeast Asia. We examined three artificial diets that were identical except for their agar content. The survival rate from hatching to adult emergence was 26.0% when beetles were reared on a ‘soft diet’ (20 g/l agar), 16.0% on a ‘hard diet’ (40 g/l agar), and 41.0% on a ‘mixed diet’, in which the hard diet was used for the first instar and the soft diet for later instars. Females raised as larvae on the mixed diet and then as adults on the soft diet laid few eggs, which did not hatch. However, females reared on the mixed diet as larvae and then on fresh leaves as adults consistently laid eggs that hatched. We then examined the suitability of B. longissima larvae reared on the mixed diet as hosts for the larval parasitoid Asecodes hispinarum, a specialist parasitoid of this beetle. Of the oviposited hosts, 75.0% became mummified and 41.7% produced adult wasps. These results indicate that the mixed diet may be useful for rearing B. longissima larvae as hosts for the rearing of A. hispinarum.     

Comparative developmental and reproductive biology of geographical populations from two cryptic species in Brontispa longissima (Coleoptera: Chrysomelidae)

Brontispa longissima is a serious pest of the coconut palm Cocos nucifera, presumed to have originated in Papua New Guinea and Indonesia. It recently invaded Southeast and East Asia, where outbreaks have been reported. Mitochondrial DNA analysis reveals two cryptic species in B. longissima: one is distributed over a wide area including Asia and the Pacific region (the Asian clade) and the other in a limited area in the Pacific region (the Pacific clade). Recent invasions and outbreaks have been reported only from the area where the Asian clade has been found, suggesting that this clade has become a pest in Asia. To infer if the Asian clade has the ability to establish, spread and outbreak in novel habitats more effectively than the Pacific clade, we compared life‐history traits between the two populations of different clades. The net reproduction rate (R₀) was 130.0 and 94.0, the mean length of a generation (T) was 57.7 and 54.7 days, and the intrinsic rate of natural increase (r) was 0.084 and 0.083 per day for the population from Ishigaki Island, Japan (ISH) (the Asian clade) and for the population from Papua New Guinea (PNG) (the Pacific clade), respectively. Although the difference in r was little, the simulated population growth showed that the ISH population can be 1.6 times larger than that of the PNG after ten generations. The rapid population growth of the Asian clade would be partly responsible for its establishment, spread and frequent outbreaks in Asia.     

水椰八角铁甲和椰心叶甲对椰子和银海枣的寄主选择性

【目的】水椰八角铁甲和椰心叶甲均为棕榈科植物的重要入侵害虫,两者的外部形态、取食部位和危害特征相似。研究它们的寄主选择性有助于了解这2种害虫的扩散和成灾机制。【方法】在室内用椰子和银海枣2种寄主植物分别饲养水椰八角铁甲和椰心叶甲,研究在不同寄主植物上水椰八角铁甲和椰心叶甲的存活率、产卵率、发育历期等以及这2种害虫对不同寄主植物的选择性。【结果】水椰八角铁甲在2种寄主上的存活率差异显著,除了卵期和蛹期之外,幼虫期各虫态在银海枣上的存活率明显比在椰子上的存活率高;椰心叶甲在椰子上的存活率高于银海枣,各虫态平均存活率分别为95%和86%。取食银海枣的水椰八角铁甲达标准卵量概率为0.23,取食椰子不产卵,无法完成整个世代;取食椰子的椰心叶甲达标准卵量概率为0.86,取食银海枣不产卵,也无法完成整个世代;水椰八角铁甲取食银海枣完成世代的实验种群趋势指数为12.55,椰心叶甲取食椰子完成世代的实验种群趋势指数为66.55。【结论】水椰八角铁甲和椰心叶甲分别对银海枣和椰子这2种寄主植物具有明显的选择性。在海南椰子树的数量远远超过银海枣,该实验结果在一定程度上解释了椰心叶甲在海南岛广泛分布而水椰八角铁甲只是零星发生的原因。     

Cellular responses of leaf explants of Cocos nucifera L. in vitro

Leaf explants of Cocos nucifera L. (coconut palm) were studied in vitro in order to establish whether or not rapid cellular changes contribute to the well known recalcitrance of coconut cells in tissue culture. Segments from the base of immature leaves were cultured on modified Eeuwens' medium at 30°C in darkness. The mitotic index, nuclear DNA amounts, cell and nuclear size were measured both before and during culture (from 0 to 70 days). There was no basipetal gradient of cell division in immature coconut leaves; the mitotic index never exceeded 2% and showed neither a positional nor temporal relationship with leaf development. Moreover the vast majority of cells were in G1 of the cell cycle. This cell cycle pattern was maintained for most of the period in culture although at 70 days there was an increase in the proportion of cells in S- and G2-phases consistent with low rates of callus formation. The nuclear: cell size ratio was constant in cells within the immature leaf irrespective of developmental age. However upon transfer to culture media, cell size but not nuclear size increased. We suggest that this uncoupling of cell and nuclear size disrupts cell co-ordination and is a key contributor to recalcitrant cellular behaviour of this species in vitro.     

Are the palms Washingtonia filifera and Chamaerops humilis suitable hosts for the red palm weevil,Rhynchophorus ferrugineus (Col. Curculionidae)?

The red palm weevil, Rhynchophorus ferrugineus (Olivier) (Coleoptera, Curculionidae), is a widely distributed pest which has recently become the major pest of palms in the Mediterranean Basin. This weevil has been recorded on 19 different palm species, however, the host status of Washingtonia filifera and Chamaerops humilis remains unclear. The present study was carried out to check whether these species had either antixenotic or antibiotic mechanisms of resistance against this pest. Our results show that both W. filifera and C. humilis are resistant to R. ferrugineus. Resistance in W. filifera is based on antibiosis whereas in C. humilis resistance is based on antixenosis.     

Artificial diets for rearing the coconut hispine beetle,Brontispa longissima (Coleoptera: Chrysomelidae), and their suitability to two specialist parasitoids

The coconut hispine beetle, Brontispa longissima (Gestro), is a serious invasive pest that infests young unopened fronds of coconut palms (Cocos nucifera L.) in Southeast Asia. We previously developed the first artificial diet for rearing B. longissima larvae, which contained a leaf powder of young coconut fronds. Because the fronds are required for healthy growth of coconut palms, it is necessary to reduce their use for rearing the beetles. In this study, we tested two new artificial diets for the beetle larvae, which contained the leaf powders of mature coconut leaves or orchard grass (Dactylis glomerata L.). Brontispa longissima successfully developed from hatching to adulthood on both the mature coconut leaf diet and orchard grass diet. The beetles reared on the mature coconut leaf diet and orchard grass diet developed faster than those reared on the young coconut leaf diet. Fecundity and egg hatchability of beetles did not differ among the three diet treatments. We then examined the suitability of beetle larvae or pupae reared on each diet as hosts for two specialist endoparasitoids, Asecodes hispinarum Boucek and Tetrastichus brontispae Ferriere. The survival rate from oviposition to adult emergence for A. hispinarum was 43.8% in hosts reared on a young coconut leaf diet, 77.1% on a mature coconut leaf diet, and 85.7% on an orchard grass diet. For T. brontispae, the survival rate was 70.0% in hosts reared on the young coconut leaf diet, 38.1% on the mature coconut leaf diet, and 66.7% on the orchard grass diet. Our results indicate these artificial diets can be useful for rearing B. longissima and its two parasitoids, helping to reduce the costs of mass rearing these insects.     

Transient genetic transformation of embryogenic callus of <Emphasis Type="Italic">Cocos nucifera</Emphasis>

Coconut palm (Cocos nucifera) is a plant species recalcitrant to in vitro morphogenesis and no protocols for the genetic transformation of coconut tissues have been published. The present study aimed to develop a protocol for genetic transformation of this palm species; evaluating reporter genes, transformation methods, and conditions for the use of antibiotics to select transformed plant cells. The gene gusA was first used for Agrobacterium tumefaciens mediated transformation of coconut embryogenic calli. However, endogenous GUS-like activity was found in calli not co-cultured with bacteria. Then essays for Agrobacterium-mediated transformation were developed using green and red fluorescent genes. Both genes are suitable as reporter genes for coconut transformation. In order to establish a protocol for coconut genetic transformation, an approach was used that combined biobalistics to generate micro-wounds in explants, vacuum infiltration and co-culture with Agrobacterium tumefaciens (C58C1 + pER10W-35SRed containing the embryogenesis related gene WUSCHEL). Calli treated with the combined protocol showed red fluorescence with greater intensity and greater area than calli treated with either biobalistics or infiltration, followed by bacteria co-culture. PCR amplification of DNA extracts from transformed embryogenic callus produced a band with the expected size using WUSCHEL primers (862 bp). No band was obtained using the VirE2 primers. This is the first report of transient genetic transformation of C. nucifera and it is the first step toward a protocol that will be useful for the study of the role of genes of interest and for practical applications, such as the improvement of coconut micropropagation via somatic embryogenesis.     

Coconut malformation: An emerging disease caused by Fusarium proliferatum in southern Iran

Symptoms of vegetative malformation were observed on coconut palms (Cocos nucifera L.) in the Qeshm Island, Bandar Abbas and Minab, in Hormozgan province, southern Iran. The symptoms included misshapen and dwarfed leaves with shortened, thickened and tightened leaflets in wavy and zigzag form. The aim of this study was to identify the causal pathogen of coconut palm malformation and complete Koch's postulates for putative pathogen. Small pieces of surface‐disinfested malformed vegetative tissues of coconut palms were cultured on potato dextrose agar (PDA) medium. Fusarium isolates were permanently obtained from the symptomatic tissues. Sequence data from the internal transcribed spacer region (ITS1–5.8S‐ITS2) and translation elongation factor 1 alpha (TEF‐1α) gene were used for molecular identification of the isolates. BLAST search of the sequences showed 99%–100% identity to several Fusarium proliferatum strains in the GenBank, FUSARIUM‐ID and Fusarium MLST databases. A phylogeny inferred using individual sequence data from ITS region and TEF‐1α gene placed our isolates together with the other F. proliferatum sequences retrieved from the GenBank. Pathogenicity tests were carried out using one‐year‐old healthy coconut palm seedlings and conidial suspensions (10⁶ conidia/ml) of the F. proliferatum isolates. The first visible symptoms appeared on newly produced leaves of the inoculated seedlings during the 16th week after inoculation, wherease no disease symptoms were observed on the control plants until the end of the experiment. Reisolation from symptomatic tissues of the inoculated seedlings yielded isolates of F. proliferatum with morphological and molecular characteristics identical to those of the isolates used for inoculations. This is the first report of coconut palm malformation caused by F. proliferatum worldwide.     

Detection and differentiation of African coconut phytoplasmas: RFLP analysis of PCR-amplified 16S rDNA and DNA hybridisation

Phytoplasmas associated with lethal decline diseases of the coconut palm (Cocos nucifera) in west and east Africa were detected by the specific amplification of their 16S rRNA genes. The primers used were based on conserved mollicute-specific and coconut-phytoplasma 16S rRNA gene sequences. Phytoplasma 16S rDNA was amplified from all African decline affected palms, some periwinkle maintained phytoplasmas, but not from healthy palms, infected palms from Florida or from the Cocos spiroplasma and Acholeplasma sp. mollicutes. African phytoplasmas were also detected by DNA hybridisation using two probes from the palm lethal yellowing phytoplasma from Florida. Probes hybridised at moderate stringency to dot blots of lethal decline affected palms from Africa, indicating possible genetic relationships between different coconut phytoplasmas. RFLP analysis of rDNA fragments (length c. 1.45 kbp) detected polymorphisms, indicating that the pathogens found in west and east Africa are not identical. This provides a useful tool for further epidemiological studies of African coconut phytoplasmal diseases.     

A revision of the genus Sclerosperma (Arecaceae)

Summary  A taxonomic revision of the palm genus Sclerosperma (Arecaceae) is presented. Three species are recognised: S. mannii H. Wendl., which is relatively widespread from Liberia to the Democratic Republic of Congo; S. walkeri A. Chev., which is apparently confined to the interior of Gabon and a band along the Congo River; and S. profiziana, a new species previously considered conspecific to S. mannii that is found in southwest Ghana, Congo, the Democratic Republic of Congo and Angola. The taxonomic history, morphology, distribution and conservation status of the genus and each species are discussed.     

Two new species and one new combination of Agaricales from Japan

 Two new species and one new combination of Agaricales are described and illustrated from eastern Honshu, Japan, and Ishigaki Island, southwestern Japan: (1) Gymnopus oncospermatis comb. nov. (subsection Vestipedes of section Vestipedes), new to Japan, has pale brown to whitish, collybioid basidiomata with very close lamellae, and forms a distinct white extensive mycelial mat on the dead leaf bases of the palm Satakentia liukiuensis in Ishigaki Island; (2) Hydropus aurarius sp. nov. (subsection floccipedes of section floccipedes), forming clitocyboid basidiomata with a dark brown furfuraceous pileus and stipe and vivid yellow lamellae, found on fallen dead twigs in oak forests; and (3) Tylopilus castanoides sp. nov. (section Oxydabiles), forming medium to large, reddish-brown basidiomata with a minutely scurfy-punctate stipe, found in Carpinio quercion forests. Received: May 20, 2002 / Accepted: July 17, 2002 Acknowledgments I am grateful to Dr. Yousuke Degawa (KPM) for allowing the specimens cited to be kept in the Kanagawa Prefectural Museum of Natural History. Thanks are also owed to Mr. Sadayoshi Uehara, Mr. Wakao Ikeda, Mrs. Youko Ando, Dr. Yasushi Shibata, and Mr. Yasuhiko Goto for provision of specimens. Correspondence to:H. Takahashi     

Cold tolerance of the coconut hispine beetle, <Emphasis Type="Italic">Brontispa longissima</Emphasis> (Coleoptera: Chrysomelidae) in Japan

The coconut hispine beetle, Brontispa longissima (Gestro), supposedly originated from Papua New Guinea and Indonesia but has recently invaded Southeast and East Asian countries where it has been causing serious damage to Cocos nucifera L. This insect also occurs on the Southwest Islands off Kyushu Island in Japan. To evaluate the potential northward range expansion of this insect in Japan, we investigated its cold tolerance at 0, 5, and 10 °C (egg, larva, pupa, and adult), 13 °C (adult), and 15 °C (egg and hatched larva). At 15 °C, few eggs hatched, and the larvae that hatched died within a few days of hatching. At 13 °C, Ltime₉₅ was estimated to be 23 days for adults, with the most cold-tolerant developmental stage at 10 °C. At all developmental stages, Ltime₉₅ of B. longissima was estimated to be 19 days at 10 °C, 8 days at 5 °C, and 5 days at 0 °C, suggesting the cold tolerance of this beetle is very low. Considering average daily temperatures, it is unlikely that B. longissima can establish itself north of Amami-Oshima Island, located in the far south off the main island of Japan.     

椰心叶甲、椰子织蛾肠道菌群多样性分析

[目的] 椰心叶甲、椰子织蛾同属棕榈科食叶性入侵害虫,两者同时取食棕榈科植物却无明显竞争作用,本文通过分子生物学手段对两者的肠道菌群组成、肠道代谢通路进行比对,以此探讨两者食性之间相关性。[方法] 以16S rRNA菌群分析等预测手段对两者肠道微生物群落、代谢通路等进行差异分析。[结果] 椰心叶甲体内变形菌门群落丰度显著高于椰子织蛾,椰子织蛾体内蓝菌门、放线菌门、酸杆菌门、疣微菌门等菌群丰度显著高于椰心叶甲;椰心叶甲体内膦酸酯和亚膦酸盐代谢、N-聚糖合成等肠道代谢通路显著强于椰子织蛾,而椰子织蛾肠道中酮体的合成和降解、维生素B6代谢、核黄素代谢、黄酮和黄酮醇生物合成等代谢通路显著强于椰心叶甲。[结论] 椰子织蛾体内相对较高的酸杆菌门丰度可能导致了其偏好取食纤维素高的椰子老叶,而椰心叶甲则相反。同时,椰子织蛾肠道内显著较高的维生素B含量在其族群发展过程中起到了主要作用。两者细菌丰度以及代谢通路强度的差异可能影响了两者的食性。     

Phylogenetic relationships of coconut phytoplasmas and the development of specific oligonucleotide PCR primers

Using the polymerase chain reaction the 16S rRNA genes and the 16S-23S spacer regions of phytoplasmas associated with lethal decline diseases of coconut palm (Cocos nucifera), were amplified from infected plants from Florida and the Yucatan region in Mexico and from east and west Africa. Following sequencing of the rDNA products, phylogenetic analysis confirmed that these coconut phytoplasmas form a separate cluster within the phytoplasma clade and that the pathogen causing diseases in west Africa formed a new sub-clade within this cluster. Analysis of the 16S-23S intergenic spacer regions confirmed the sequence diversity of this region and enabled two primers to be designed which were specific for the diseases found in east and west Africa. None of these specific primers, when paired with a universal primer, produced PCR amplification products from healthy coconut DNA, infected coconut DNA from the Caribbean or DNA from a variety of periwinkle (Catharanthus roseus)-maintained phytoplasmas. These specific primers can serve as effective tools for identifying particular coconut phytoplasmas in field samples.     

Primulina lutescens sp. nov. (Gesneriaceae) from southern Guangxi,China

Primulina lutescens B. Pan & H. S. Ma, a new species of Gesneriaceae from Guangxi Zhuangzu Autonomous Region, China, is illustrated and described. The new species morphologically resembles Primulina carinata Y. G. Wei, F. Wen & H. Z. Lü, but it differs in leaf, floral characteristics and flowering time. The conservation status of P. lutescens is asessed as ‘Endangered’ (EN) according to the IUCN red list categories and criteria.