上海地区红棕象甲的耐寒性研究

红棕象甲Rhynchophorus ferrugineus (Olivier)是我国危害棕榈科植物的重要入侵害虫, 为探明其越冬抗寒性, 利用过冷却点测定仪测定了红棕象甲上海种群不同虫态的过冷却点, 在低温箱内测定了该虫不同虫态的耐寒性, 然后结合田间越冬模拟试验、气象资料和寄主分布情况, 初步确定了红棕象甲在我国的越冬北界。结果表明： 红棕象甲过冷却点随虫态的发育程度的升高而下降, 卵、1龄幼虫、5龄幼虫、9龄幼虫和成虫的平均过冷却点分别为-5.92, -6.42, -7.19, -7.43和-11.84℃, 过冷却点由高到低的顺序依次为: 卵>幼虫>成虫。在6, 24, 48和72 h 4个时间处理下, 各虫态在低温与存活率之间呈显著或极显著的logistic回归, 半致死温度（Ltemp₅₀）均随处理时间的延长而上升, 不同虫态在处理72 h 后, 卵、1龄幼虫、5龄幼虫、9龄幼虫和成虫之间的Ltemp₅₀分别为1.61, -1.67, -2.39, -2.40和-0.40℃, 各虫态耐寒性由弱到强的顺序依次为: 卵<成虫<幼虫。红棕象甲不同发育阶段的过冷却点与其耐寒性并不完全相关, 幼虫和成虫均可能是该虫的越冬虫态。连续两年的田间模拟越冬试验表明, 在上海地区, 红棕象甲的幼虫和成虫的越冬存活率均在60％以上, 说明红棕象甲在上海地区是可以越冬的。根据这些结果, 结合寄主分布情况, 初步将红棕象甲在我国的越冬北界定于北纬35°附近, 即1月份0℃等温线左右。     

Potential role of microbial pathogens in control of red palm weevil (Rhynchophorus ferrugineus) ‐ A Review

The invasive Red Palm Weevil (RPW), Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae), is one of the most destructive pests of ornamental and economically important palms globally. It has been found in 50 % of date‐growing and 15 % of coconut‐producing countries in the world. Synthetic organic insecticides have been the default method to combat this pest, but they are clearly inefficient due to the secretive nature of the insect and there is concern about non‐target effects from blanket spraying. For this reason, there is increasing interest in biological control methods such as the possible use of microbial entomopathogens, which might be incorporated into IPM approaches. In this review we summarize research work on microbial control agents, their effectiveness against RPW and their integration with other control measures.     

Potential of certain natural extracts for the control of the red palm weevil,Rhynchophorus ferrugineus (Oliver)

Abstract

Laboratory bioassays were conducted to evaluate the potential of 13 plant extracts for the control of the red palm weevil, Rhynchophorus ferrugineus. Results indicated that the ethanol extracts of sea ambrosia (Ambrosia maritima), French cotton (Calotropis procera) and curcuma (Curcuma longa) have superior toxicity as compared to the extracts of other tested plants, estimated LC 50 were 0.723, 0.978 and 1.092%, respectively. In field experiments, the percentage of recovered date palm trees reached 90.91% within 15 days when A. maritima extract was applied. The injection of C. procera and C. longa extracts into the tunnels of infested trees resulted in 86.36% and 85.00% of tree recovery, respectively.     

A large-scale gene discovery for the red palm weevil Rhynchophorus ferrugineus （Coleoptera： Curculionidae）

The red palm weevil （RPW; Rhynchophorusferrugineus） is a devastating pest of palms, prevalent in the Middle East as well as many other regions of the world. Here, we report a large-scale de novo complementary DNA （cDNA） sequencing effort that acquired ~5 million reads and assembled them into 26 765 contigs from 12 libraries made from samples of different RPW developmental stages based on the Roche/454 GS FLX platform. We annotated these contigs based on the publically available known insect genes and the Tribolium castaneum genome assembly. We find that over 80% of coding sequences （CDS） from the RPW contigs have high-identity homologs to known proteins with complete CDS. Gene expression analysis shows that the pupa and larval stages have the highest and lowest expression levels, respectively. In addition, we also identified more than 60 000 single nucleotide polymorphisms and 1 200 simple sequence repeat markers. This study provides the first large-scale eDNA dataset for RPW, a much-needed resource for future molecular studies.     

Sex ratio and the role of mild relative humidity in mating behaviour of red date palm weevil Rhynchophorus ferrugineus Oliv. (Coleoptera: Curculionidae) gamma‐irradiated adults

The influence of gamma radiation on the mating behaviour of the red date palm weevil (RDPW), Rhynchophorus ferrugineus (Olivier), and the efficacy of sterile insect technique (SIT) under different levels of relative humidity (RH) were studied. No adverse effects of gamma radiation were observed on the mating behaviour parameters of the RDPW, such as mate recognition time, mating duration, mating frequency within a 30‐min period and duration between consecutive matings. However, the weevils were sexually stimulated during aggregation. RH significantly affected egg laying as well as egg hatching. Significantly lower egg laying and hatching were recorded at 25% RH than at higher humidity levels, suggesting that low humidity conditions are better for successful SIT manipulations in the field. Lifespan of irradiated RDPW males was significantly shorter than that of un‐irradiated controls, irrespective of RH level.     

警惕外来危险害虫红棕象甲的扩散

红棕象甲Rhynchophorus ferrugineus(Olivier)是棕榈科植物外来危险害虫,该虫原产印度,20世纪80年代扩散到中东,目前在我国主要分布在海南、广西、广东、台湾、云南、西藏(墨脱)等地区,存在进一步扩张的趋势。文章对该虫的地理起源及分布、生物学特性、寄主及防治等方面作了描述。     

Entomopathogenic fungi-mediated biological control of the red palm weevil Rhynchophorus ferrugineus

The red palm weevil (RPW), Rhynchophorus ferrugineus, is an important pest of palms, and difficult to control by conventional methods. Therefore, microbial control is an alternative strategy for controlling RPW. Herein, a total of 15 entomopathogenic fungi (EPFs) were subjected to primary pathogenicity screening against last stage of RPW larvae. The preliminary data showed that four Beauveria bassiana isolates (JEF-484, 158, 462 and 507) and one Isaria fumosorosea isolate (JEF-014) resulted in 100 % mortality within 5–10 days post inoculation (d.p.i.), respectively. According to the time required for RPW mortality, JEF-484, 158, 462 and 014 were further subjected to bioassays using 10⁷ conidia/ml suspensions by spraying method. Based on the results, JEF-484 showed the highest mortality and shortest LT₅₀ on the last stage of RPW larvae, followed by JEF-158. The two isolates also showed good conidial production and high thermal stability compared to the other isolates. Therefore, JEF-484 and JEF-158 were selected for bioassays against RPW egg and the last larval stage with different concentrations of 10⁵, 10⁶ and 10⁷ conidia/ml conidial suspensions by spraying method. For the bioassay at the egg stage, JEF-158 showed a significantly higher ovicidal effect than JEF-484. In the larval bioassay, both EPF isolates showed a dosage-dependent effect on the RPW larvae. JEF-484 caused higher mortality in RPW larvae than JEF-158. In summary, the combination of the 2 promising EPF isolates might provide an opportunity for the practical microbial control of RPW at different life stages in palm tree fields.     

Nano essential oils against the red palm weevil,Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae)

The red palm weevil, Rhynchophorus ferrugineus, is the biggest pest of various palm species. The aim of this study was to evaluate the impact of the essential oils of purslane, mustard and castor (bulk and nano) on R. ferrugineus and to evaluate the oviposition deterrent effect to prevent new infestations. Prepared concentrations of essential oils (bulk and nano) were tested for their impact on R. ferrugineus larvae, pupae and adults. The most effective oil was purslane oil (bulk and nano), followed by mustard and the least effective was castor. The percent mortality of larvae was 75.2, 45.3 and 17.9% (bulk phase) and 92.5, 84.4 and 65.5% (nano phase) in purslane, mustard and castor, respectively, when treated with the high concentration. The percent mortality of adults was 67.2, 40.0 and 15.6% (bulk phase) and 83.5, 72.9 and 60.0% (nano phase) in purslane, mustard and castor, respectively, when treated with the high concentration. The number of eggs laid decreased with increasing concentration of the tested oils. Purslane essential oil showed good impact on R. ferrugineus larvae and adults followed by mustard and castor oils as bulk and as nano.     

Isolation and identification of bacterial strains with insecticidal activities from Rhynchophorus ferrugineus Oliver (Coleoptera: Curculionidae)

Several pathogenic bacteria strains were isolated from dead larvae of the weevil, Rhynchophorus ferrugineus Oliver, collected on palm trees in Taiwan, China. Strains of pure cultures were temporarily named TW‐1, TW‐2, TW‐3, TW‐4, TW‐5, TW‐6, TW‐7, TW‐8, TW‐9, TW‐10, TW‐11, TW‐12, TW‐13 and TW‐14. These strains were identified by synthetically using colony morphological characters, physiological and biochemical tests and molecular biological analysis. Our results showed that these 14 isolated strains belonged to four genera and five species. The expected approximate 1400‐bp bands were obtained through 16S rDNA universal primer amplification. The sequencing results obtained from a commercial sequencing company were initially compared with a corresponding database from NCBI using BLAST, and a phylogenetic tree was constructed using mega 5.1 software. The phylogenetic results indicated that these isolated strains had more than 99% homology with type strains. To further confirm these strains, numerous physiological and biochemical indexes were confirmed. TW‐1, TW‐2, TW‐3, TW‐9, TW‐12, TW‐13 and TW‐14 were identified as Serratia marcescens; TW‐4, TW‐5, TW‐8 and TW‐10 as Staphylococcus sciuri; TW‐6 as Klebsiella pneumonia subsp. pneumonia; TW‐7 as Proteus vulgaris and TW‐11 as Proteus mirabilis. The preliminary single‐dose bioassay on fourth instar larvae showed that S. marcescens had a higher virulence at a density of 1.89 OD₆₀₀ (56.37% mortality) than the other strains. The corrected mortalities of the other strains were all less than 50%. The lowest toxicities were found for P. vulgaris and P. mirabilis where the corrected mortalities were only 28.18% and 25.57%, respectively. LC₅₀ of S. marcescens was 1.2 × 10⁷ CFU/ml inoculums. Our results indicated that S. marcescens from facultative cultivable bacterial flora isolated from R. ferrugineus could potentially be used as a microbial control agent for this widespread pest.     

Pseudomonas aeruginosa,a facultative pathogen of red palm weevil,Rhynchophorus ferrugineus

Pseudomonas aeruginosa was identified as a facultative pathogen of red palm weevil. Intra-haemocoelic injection of the pathogen within larvae and pre-pupae was more effective at killing the insects [with a median lethal dose (LD₅₀) of 9×10² to 2×10³ bacteria/insect] than inoculation by force feeding (LD₅₀ of 10⁵ to 4×10⁵ bacteria/insect) or by wading the insects in a suspension of the pathogen (LD₅₀ of 10⁵ to 2×10⁵ bacteria/insect). Injection of 3×10³ bacteria/insect killed 69% of larvae; small larvae were more susceptible (LD₅₀ of 9×10⁵ bacteria/larva) than either larger larvae (LD₅₀ of 10³ bacteria/larva) or pre-pupa. The median time to death of the small larvae following injection of P. aeruginosa was about 6 days but that following force feeding or wading was about 8 days. A secondary invader, Serratia marcescens, had no effect on the pathogenicity of P. aeruginosa but hastened death of larvae by about 3 days.A. Banerjee and T.K. Dangar were with the Central Plantation Crops Research Institute, Regional Station, Kayangulam 690 533, Kerala, India. They are now with the Central Rice Research Institute. Cuttack 753 006, Orissa, IndiaCPCRI research paper no. 870.     

Assessing Spirulina platensis as a dietary supplement and for toxicity to Rhynchophorus ferrugineus (Coleoptera: Dryopthoridae)

Insects are important for humanity; play role in crop pollination, and biocontrol of harmful pests. The red palm weevil, Rhynchophorus ferrugineus, is a major pest of date palms and has become a serious threat. Scientists needs ample numbers of insects for bioassays to explore control options. The alga Spirulina platensis, is enriched by protein, natural vitamins, minerals, and amino acids, stimulate the development of organisms that feed on it. I assessed the value of Spirulina as a nutritional supplement for red palm weevil larvae by adding its various percentages to the artificial diet. Once a week, the larvae were removed from the containers, washed with distilled water, dried, weighed using an electronic scale, returned to a new container, and supplied with Spirulina mixed fresh diet. Larvae fed with lower concentrations showed vigorous growth and significant weight gain. Particularly, larvae fed 0.5%, 1%, and 2% Spirulina powder supplementation to their diet were healthier and gained more weight than larvae reared with >5% concentration. Overall 40% mortality was recorded in larvae fed with 10% concentration. Higher concentrations were lethal, and all larvae died within two weeks when fed 20% Spirulina. The present research findings indicate that Spirulina used in concentrations from 0.5% to below 5% had a beneficial effect on red palm weevil larval growth but a detrimental effect and even mortality was recorded when used ≥5%.     

Integration of entomopathogenic fungi and eco-friendly insecticides for management of red palm weevil,Rhynchophorus ferrugineus (Olivier)

Red palm weevil (Rhynchophorus ferrugineus) is a voracious pest of date palm worldwide. Pakistan ranks sixth in date palm production globally. Losses to date palm plantations in Pakistan sometimes surpass 10%-20%. Most of the traditional management strategies used by farmers have been found insignificant to combat this voracious pest. The entomopathogenic fungi, Beauveria bassiana [QA-3(L) and QA-3(H)] and insecticides, Nitenpyram (Active 10% SL) [NIT (L) and NIT (H)] were applied to larval (2nd, 4th, and 6th), pupal and adult stages of R. ferrugienus. Integration or alone application of fungi with insecticides at different concentration under laboratory conditions. Combined application was depicted additive and synergistic interactions. Contrarily, highest cumulative mortality (100%) was recorded in 2nd instar larvae as compared to later instar larvae at combined application. The maximum pupal and adult mortality remained 89% and 66% respectively after treatment with [QA-3 (H) + NIT (L)]. The combination of B. bassiana at higher concentration whereas Nitenpyram at lower dose was found more lethal to larvae, pupae and adults of R. ferrugineus. This signifies the need of combining B. bassiana and bio-rational insecticides that can reduce the cost of management with least harm to environment and natural enemies.     

Biochemical and ultrastructural changes in the ovaries of red palm weevil,Rhynchophorus ferrugineus (Coleoptera: Curculionidae) following acute imidacloprid poisoning

This study aims to inspect the effects of Imidacloprid (IMI) acute toxicity in the ovaries of the red palm weevil (Rhynchophorus ferrugineus). The LC₅₀ of IMI was determined to be 15.7 ppm. We assessed the biochemical changes resulting from treatment with four IMI concentrations (10, 15, 20 and 30 ppm). IMI reduced acetylcholine esterase activity in a dose-dependent manner. Glutathione (GSH) content decreased with increasing concentration of IMI; however, GSH was not affected and accompanied by an increase in catalase activity with the highest concentration of IMI. Lipid peroxidation increased steadily as IMI concentration increased. Based on the ultrastructural analyses of the follicle cells and the ooplasm of developing oocytes, acute IMI toxicity produced damaged yolk bodies, pale nuclei, and shrinking microvilli. Fractionation of rough endoplasmic reticulum, deformed mitochondria, and autophagic bodies were also observed. In conclusion, these ovarian pathobiochemical changes could be used as biomarkers of acute IMI exposure.     

Digestive proteases during development of larvae of red palm weevil, Rhynchophorus ferrugineus (Olivier, 1790) (Coleoptera: Curculionidae).

The evolution of digestive proteases during larval development of Rhynchophorus ferrugineus (Olivier, 1790) has been studied. A progressive increase of protease activity has been found. The optimum pH for proteolytic activity against azocasein was determined. Caseinograms revealed an active complex of alkaline proteases from the early stages of the development. From the apparent molecular masses, three groups of proteases have been found - high molecular-mass proteases, medium molecular-mass proteases, and low molecular-mass proteases. Studies using specific protease inhibitors showed the major presence of serine proteases in gut extracts. The results obtained from larvae reared on different substrates have made possible a comparative assessment of the influence of diet on the development of the digestive enzymatic system. Larvae fed on an artificial diet showed a complete pattern of digestive proteases. Data suggest that this diet seems to be suitable for future research with this insect pest.     

Immune function differences between two color morphs of the red palm weevil Rhynchophorus ferrugineus (Coleoptera: Curculionidae) at different life stages

Several studies demonstrated that in insects cuticle melanism is interrelated with pathogen resistance, as melanin‐based coloration and innate immunity possess similar physiological pathways. For some insects, higher pathogen resistance was observed in darker individuals than in individuals with lighter cuticular coloration. Here, we investigated the difference in immune response between two color morphs (black and red) and between the life stages (pupa and adult) of the red palm weevil Rhynchophorus ferrugineus (Coleoptera: Curculionidae). Here in this study, cuticle thickness, microbial test (antimicrobial activity, phenoloxidase activity, and hemocyte density), and immune‐related gene expression were evaluated at different stages of RPW. Study results revealed that cuticle thickness of black phenotype was thicker than red phenotype at old‐pupa stage, while no significant difference found at adult stage. These results may relate to the development processes of epidermis in different stages of RPW. The results of antimicrobial activity, phenoloxidase (PO) activity, and hemocyte density analyses showed that adults with a red phenotype had stronger pathogen resistance than those with a black phenotype. In addition to antimicrobial activity and PO activity, we tested relative gene expression in the fat body of old pupae. The results of hemolymph antimicrobial analysis showed that old pupae with a red phenotype were significantly different from those with a black phenotype at 12 hr after Staphylococcus aureus injection, suggesting that red phenotype pupae were more sensitive to S. aureus. Examination of gene expression in the fat body also revealed that the red phenotype had a higher immune response than the black phenotype. Our results were inconsistent with the previous conclusion that dark insects had increased immune function, suggesting that the relationship between cuticle pigmentation and immune function in insects was not a direct link. Additional possible factors that are associated with the immune response, such as life‐history, developmental, physiological factors also need to be considered.     

Identification and enzymatic characterisation of digestive glucosidases from gut of red palm weevil,Rhynchophorus ferrugineus (Olivier, 1790) (Coleoptera: Curculionidae)

Red palm weevil, Rhynchophorus ferrugineus (Olivier, 1790) (Coleoptera: Curculionidae) is a serious pest of date worldwide. Thus, damage to palms (almost exclusively to Phoenix canariensis Hort) has been recorded in various places. Thus, the aim of the current study was to investigate two major digestive enzymes of this insect, α-D-glucosidase and β-D-glucosidase. The results showed that α-D-glucosidase and β-D-glucosidase are present in the insect gut mainly in the midgut and hindgut but trace amounts of the both enzymes were found in the foregut. Optimum temperature for α- and β-glucosidases was found to be 50 and 40?°C, respectively, and pH values were 4.0. The activity of glucosidases were increased by NaCl and KCl and inhibited by some compounds such as MgCl₂ and CaCl₂. Kinetic parameters showed that K _m of α and β-D-glucosidases was 3.15 and 4.11?mM, respectively. Therefore, it is concluded that in this insect species, both α-glucosidase and β-glucosidase are active but with different amounts. Understanding of the digestive physiology and glucosidase activity of red palm weevil is important when new management strategies based on interfering in the gut physiology of the insects are devised.     

A review of the biology and control of the vine weevil, Otiorhynchus sulcatus (Coleoptera: Curculionidae)

At the turn of the century, damage by Otiorhynchus sulcatus was sporadic and limited to small areas. Increasing horticultural intensification and the adoption of husbandry techniques favourable to the weevil, such as the use of polythene mulches, increased its pest status. The development of the early inorganic pesticides reduced the number of serious outbreaks of this pest and weevil control was further improved by the development of the persistent organochlorine insecticides in the 1940's. The banning of a number of the more persistent insecticides over recent years has now left the horticultural industry in a very vulnerable position. O. sulcatus is now a pest on a range of horticultural crops throughout the temperate regions of the world. Infestations are most common in Europe (where it originated) and the USA, and nearly 150 plants species have been identified as potential hosts to O. sulcatus. Damage is most frequently caused by the root feeding larval stage. Populations as low as one larva plant can kill sensitive species such as Cyclamen. Severe damage by the leaf feeding adults is less common, although low levels of damage or contamination by adults may be unacceptable in certain situations. There is one generation a year. Oviposition by the flightless parthenogenetic females occurs over the summer months with oviposition rates of c. 500 and 1200 eggs adult^-1for outdoor and laboratory populations, respectively. O. sulcatus mainly overwinters as larvae, although significant numbers of adults may survive in areas where winter temperatures are not too severe. A number of natural enemies, such as hedgehogs, frogs and predatory beetles, help to maintain O. sulcatus populations at a low level in natural environments, but they are less successful in intensive horticultural systems where persistent chemicals have been heavily relied on to maintain the population below the economic threshold level. Increasing environmental concern is now forcing growers to consider new pest control strategies. Controlled release formulations of non-persistent products, such as fonofos and chlorpyrifos, have shown potential as control agents for O. sulcatus larvae. Biological control agents, such as insect parasitic nematodes, have been developed commercially and new microbial control agents are in the process of development. Most of the new control products are directed towards control of O. sulcatus larvae. Adult vine weevils are nocturnal and a much more difficult target for the new control agents. It is likely that an integrated approach to pest control will be required to maintain O. sulcatus populations below their economic threshold level.     

Lower temperature thresholds for oviposition and egg hatching of the red palm weevil, Rhynchophorus ferrugineus (Coleoptera: Curculionidae), in a Mediterranean climate

The red palm weevil, Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae), is an economically important pest of palms worldwide. To better understand the phenology of R. ferrugineus populations in the Mediterranean basin, the objective of the present work has been to estimate the effect of temperatures within the range 10-25°C on the reproductive parameters of R. ferrugineus and to determine its lower temperature thresholds for oviposition and egg hatching. Our results confirm that oviposition in R. ferrugineus is strongly affected by temperature. Lower thresholds for oviposition and egg hatching (15.45° and 13.95°C, respectively) are below the mean monthly temperatures registered in winter in most of the northern shore of the Mediterranean basin. Under these circumstances, new palm infestations would be difficult during most of the winter. These results should be taken into account when planning some palm management practices, such as pruning or pesticide treatments, in areas under a Mediterranean climate.     

The culturable bacterial community of frass produced by larvae of Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae) in the Canary island date palm

Aims: Larvae of the red palm weevil (RPW) Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae) feed inside palm stem tissues, making galleries and producing a wet fermenting frass. We characterized the culturable bacteria associated with frass produced by tunnelling larvae inside the Canary island date palms and investigated the role of frass and gut bacteria in plant polymers breakdown. Methods and Results: A culture‐dependent method was used to isolate bacteria from frass and noninfested palm tissues. Bacterial isolates were grouped into operational taxonomic units based on polymorphisms in the ITS‐PCR profiles, and representative isolates were identified by partial sequencing of the 16S rRNA gene. Frass bacteria were dominated by 2,3‐butanediol fermenter Enterobacteriaceae. None of the bacterial isolates was able to degrade cellulose; however, cellulolytic and hemicellulolytic bacteria were isolated from the larval gut enrichment cultures. Conclusions: Frass bacteria are specifically associated with the RPW larvae and might play beneficial roles for RPW, other than nutritional, that deserve further investigations. Breakdown of plant polymers probably occurs inside the larvae digestive system. Significance and Impact of the Study: Frass and gut micro‐organisms of R. ferrugineus should be included in studies of the interactions between RPW, its plant hosts, and its enemies.