Fungal Phyllosphere Communities are Altered by Indirect Interactions Among Trophic Levels

Trophic interactions involving predators, herbivores, and plants have been described in terrestrial systems. However, there is almost no information on the effect of trophic interactions on microbial phyllosphere community abundance, diversity, or structure. In this study, the interaction between a parasitoid, an insect herbivore, and the fungal phyllosphere community is examined. Parasitoid wasps have an indirect negative impact on fungal community diversity. On the citrus phyllosphere, the exotic wasp species, Amitus hesperidum and Encarsia opulenta, may parasitize the citrus blackfly (Aleurocanthus woglumi). If parasitism levels are low, the blackfly may produce significant amounts of honeydew secretions on the surface of the leaf. Honeydew deposition provides a carbon-rich substrate for the development of fungal growth persisting as sooty mold on the leaves. Leaves from sooty mold-infested grapefruit (Citrus paradisi) trees were collected from multiple orchards in south Texas. The effect of different levels of exotic parasite activity, citrus blackfly, and sooty mold infestation on phyllosphere mycobiota community structure and diversity was examined. Our results suggest the presence of the parasitoid may lead to a top–down trophic cascade affecting phyllosphere fungal community diversity and structure. Additionally, persistent sooty mold deposits that have classically been referred to as Capnodium citri (and related asexual morphological forms) actually comprise a myriad of fungal species including many saprophytes and potential fruit and foliar pathogens of citrus.     

Molecular characterization of a novel pathogen-responsive receptor kinase-like in <Emphasis Type="Italic">Citrus limon</Emphasis>

Infection by pathogenic microorganisms is a source of biological stress in plants. Understanding the interaction between plants and microbial infection at molecular level might contribute to our understanding for the effective control of the disease. Here, we isolated a novel putative receptor kinase-like protein (Citrus limon P5) that is the first Lec-receptor kinase-like protein isolated in lemon during a pathogen infection. C. limon P5 cDNA fragment was detected by differential display assay in C. limon during Capnodium citri (sooty mold) infection. The deduced amino acid sequence of P5 full-length cDNA revealed a 83% sequence homology with a receptor kinase-like protein from Arabidopsis thaliana characterized by a N-terminal lectin domain and C-terminal serine/threonine conserved domain. The inhibition of the pathogen-responsive P5 by the protein kinase inhibitor staurosporine and by a mitogen-activated protein kinase inhibitor (PD 98059) indicated a defense mechanism in C. limon against pathogens mediated via signal transduction pathway. To our knowledge, this is the first evidence that C. limon uses this putative defense mechanism against pathogens. The role of this protein is discussed as a starting point to understanding the molecular mechanisms in the C. limon in response to C. citri infection.     

Genetic diversity in the orange subfamily Aurantioideae. II. Genetic relationships among genera and species

Genetic relationships were studied by means of ten isoenzymatic systems, at the genus and species level, using two distances and four methods of aggregation in a germplasm collection of 198 cultivars and accessions of 54 species belonging to Citrus and 13 related genera. The most consistent results were obtained by the chord distance and the neighbor-joining clustering method. Citrus species were distributed in two main groups: the orange-mandarin group and the lime lemon-citron-pummelo group. The species C. halimii and C. tachibana are not included in these groups. Mandarin species fall into three main subgroups: one includes C. sinensis; the second, C. aurantium, the third, small-fruit species. The citron, the pummelo and the ancient lemon subgroups form a cluster to which the species belonging to subgenus Papeda and the cultivated limes, lemons and bergamots are related. Microcitrus spp, to which Severinia buxifolia and Atalantia ceylanica seem to be related, cluster with the lime lemon-citron-pummelo group while Fortunella is close to the orange-mandarin group. Poncirus trifoliata, the most important species for citrus rootstock improvement is located far from Citrus but connected to it through Fortunella spp. A broad distribution of species has been found that should be taken into account to sample new genotypes in the search of desired characters in order to fully and efficiently use genetic resources for citrus improvement.     

Acclimatation sur la côte d'azur et en corse deSerangium parcesetosum [Col.: Coccinellidae] prédateur de l'aleurode des citrus,Dialeurodes citri [Hom.: Aleyrodidae]

Résumé La CoccinelleSerangium parcesetosum Sicard originaire de l'Inde a été importée en France en 1985 de Géorgie soviétique où elle avait été introduite pour lutter contre l'Aleurode des CitrusDialeurodes citri Ashmead. Les premiers lachers ont été effectués sur la C?te d'Azur et en Corse où elle semble désormais bien acclimatée.      

利用虫生真菌生物防治柑橘木虱的研究进展

柑橘木虱是柑橘黄龙病的重要传播虫媒。目前对柑橘黄龙病的防治尚缺有效的药剂和抗病品种,加强对柑橘木虱的防治,对控制柑橘黄龙病的蔓延具有重要意义。目前防治柑橘木虱多采用化学防治,杀虫剂的频繁使用造成了农药残留、环境污染、生物多样性被破坏和害虫产生抗药性等诸多问题,生物防治以其高效、低毒、低残留、不易产生抗药性等优点逐渐受到重视。昆虫病原真菌能侵入昆虫寄主体内,导致昆虫发病死亡,具有良好的病害流行潜力及生产应用便利性,利用昆虫病原真菌防治柑橘木虱具有广阔的发展空间。本文总结了用于柑橘木虱生物防治的虫生真菌种类,重点介绍了国内外利用球孢白僵菌、玫烟色棒束孢、淡紫紫孢菌、宛氏拟青霉、蜡蚧菌等虫生真菌在防治柑橘木虱中的应用,并对虫生真菌防治柑橘木虱的发展前景进行了展望,以期为柑橘黄龙病的防控提供借鉴。     

Host location and discrimination mediated through olfactory stimuli in two species of Encyrtidae

The role of olfactory stimuli in host detection and evaluation was studied in two encyrtid Hymenoptera. The first, Epidinocarsis lopezi De Santis, is a monophagous parasitoid of the cassava mealybug Phenacoccus manihoti Matile-Ferrero, itself feeding exclusively on cassava, Manihot esculenta Crantz. The second, Leptomastix dactylopii Howard, is a monophagous parasitoid of the Citrus mealybug, Planococcus citri Risso, but this latter is highly polyphagous. The behaviour of females of both parasitoids (attaction and locomotion) was compared in a tubular olfactometer for the odours of their respective hosts on cassava and poinsettia. Tests were made using: 1) healthy host-plant alone; 2) host-plant infested with unparasitized mealybugs; 3) unparasitized mealybugs only; 4) host-plant infested with parasitized mealybugs and 5) parasitized mealybugs only. Only E. lopezi was attracted by the odour of the host-plant alone, but both species were attracted by the odour of an infested host-plant and that of unparasitized mealybugs. The odour of parasitized mealybugs, alone or on host-plant, induced an undirected activity. The attraction of E. lopezi to the odour of the host-plant alone could be linked to the monophagous diet of its host, whereas the attraction of the two species of parasitoids to the odours of infested host-plants and unparasitized mealybugs could be due to the fact that both parasitoids are specialists. The behavioural response of both species to the odour of parasitized mealybugs revealed a new aspect in host discrimination: the identification of parasitized hosts could be partly mediated through olfactory stimuli, and not only through gustatory stimuli.     

The spiders of a citrus grove in israel and their role as biocontrol agents ofCeroplastes floridensis [Homoptera: Coccidae]

A 12 month survey was carried out of the spider population in a commercial citrus grove at Kibbutz Afeq in Northern Israel. The spiders collected from grapefruit foliage and ground cover were identified; young stages were reared to maturity and then identified.Chiracanthium mildei L. Koch represented 52% of all spiders captured during the year andTheridion sp. accounted for 34%. In the undercover,Gnaphosidae spiders represented 43% of the total spiders captured in pitfall traps andLycosidae 35%. A field experiment was carried out to evaluate the effectivness of spiders in biological control of the scaleCeroplastes floridensis Mask. in the citrus ecosystem. Wherever spiders were undisturbed on tree branches, populations ofC. floridensis were not able to develop to a level sufficient to cause economic damage. The increase in the number of scales was minimal: from 47 initially to 56. There was no damage to leaves nor was honeydew or sooty mold observed. During the same time period, on the 3 branches from which spiders had been eliminated, the number of scales increased seven-fold from 44 to 309. Leaves were heavily infested with sooty mold fungi that developed on the honeydew produced by the scales. Contribution from the Agricultural Research Organization, Neve Ya'ar, Regional Experiment Station. P.O. Haifa 31999, Israel. No. 1314-E 1984, series.     

利用DNA片段测序方法探究枳属和富民枳的分类地位

富民枳(Poncirus polyandra)属于芸香科(Rutaceae)枳属(Poncirus Raf.).自发表以来,分类地位一直备受争议,其中在Flora of China中认为富民枳为柑橘杂交种(Poncirus polyandra),把枳属归并于柑橘属(Citrus).该研究选取枳属的富民枳、枳(Ponci...     

Notes on natural enemies attackingLepidosaphes species [Homoptera: Diaspididae] associated withCitrus in Costa Rica

Aphytis lepidosaphes Compere (Hymenoptera: Aphelinidae) parasitize two species ofLepidosaphes onCitrus in Costa Rica.Zagloba beaumonti andPentilia discors (Col.: Coccinellidae) also feed on both species.      

Survey of natural enemies of spider mites (Acari: Tetranychidae) in citrus orchards in eastern Spain

Field surveys were conducted from 2004 to 2007 to determine the species composition and relative abundance of natural enemies associated with colonies of either the citrus red mite, Panonychus citri, or the two spotted spider mite, Tetranychus urticae, in Valencian citrus orchards (eastern Spain). Fourteen species were recorded, six phytoseiid mites and eight insect predators. Two of them are reported for the first time on citrus in Spain and two more are first reports as predators associated with T. urticae. The community of predators associated with T. urticae and P. citri was almost identical, and the Morisita–Horn index of similarity between both natural enemy complexes was close to one, suggesting that predators forage on both pest species. Quantifying the presence of many known spider mites predators in Valencian citrus orchards is an important first step towards spider mite control. A challenge for future studies will be to establish conservation and/or augmentation management strategies for these predators, especially to improve T. urticae biological control.     

Feeding behavior ofEuseius stipulatus andTyphlodromus phialatus on the citrus red mitePanonychus citri [Acari: Phytoseiidae,Tetranychidae]

Observations were made on the feeding behavior of the two main phytoseiid species in Spanish Citrus orchards,Euseius stipulatus (Athias-Henriot) andTyphlodromus phialatus Athias-henriot. The experiences were carried out by rearing the predatory mites on excised orange leaves, and always with an excess of the prey the Citrus Red Mite (=CRM)Panonychus citri (McGregor). In experiments with all stages of CRM, the number of prey killed per hour was 5.12 and 2.00, the percentage of successful attacks, 58% and 21%, and the mean time spent feeding on each prey was 5.1 and 12.2 minutes for starving females ofE. stipulatus andT. phialatus respectively.E. stipulatus feeds on all stages of the prey except eggs, andT. philatus, on all stages, except males. Both species attack much less successfully females ofP. citri rather than immatures. In experiments with adult females and eggs ofP. citri as prey, the mean number of prey killed daily was 4.51 females forE. stipulatus, and 2.01 females and 2.12 eggs forT. philatus. Considering this killing rate and the number of eggs laid by the predators in the same period, it can be concluded thatE. stipulatus consumes only 30% of the content of the preys killed, whereasT. philatus consumes a percentage of prey variable between individuals and ranging from 40% to 100%. These differences in feeding behavior between the two species could partly explain differences in their efficiency as biocontrol agents ofP. citri observed in the field.      

Biological control of citrus mealybug,Planococcus citri with an introduced parasite,Leptomastix dactylopii in India

Planococcus citri (Risso) is one of the major pests of citrus orchards in India. For the control ofP. citri, an encyrtid parasite,Leptomastix dactylopii How. was introduced from West Indies in 1983. The parasite was mass bred and inoculative releases were made in 2 selected citrus orchards where infestation of mealybug on fruits (sweet orange, seedless lime and acid lime) ranged from 38 to 65 per cent. Establishment of the parasite in the 2 release orchards resulted in complete control of the mealybug within 3 to 4 months. No insecticidal sprays were required subsequently for the control ofP. citri in the following seasons. Contribution No. 152/85 of the IIHR, Bangalore — 560089.     

The plastome of Citrus. Physical map,variation among Citrus cultivars and species and comparison with related genera

Summary A physical plastome map was constructed for Citrus aurantium, and the plastomes of species and cultivars of Citrus and of two Citrus relatives were analysed by Southern blot-hybridisation of labelled total tobacco cpDNA to digests of total Citrus DNA. A resemblance was found between the plastomes of cultivars of C. limon (lemon), C. sinensis (orange), C. aurantium (sour orange), C. paradisii (grapefruit) and C. grandis (pomello). The plastomes of other Citrus types such as mandarin (C. reticulata) and citron (C. medico) differed from each other as well as from the plastomes of the aforementioned group. The plastomes of Poncirus trifoliata and Microcitrus sp. are distinct from each other as well as from the Citrus types.     

Ethylene-binding characteristics in Phaseolus,Citrus, and Ligustrum plants

A number of plants were tested for their ability to bind ethylene and the number of binding sites present in each was calculated. Primary leaves of laboratory-grown beans (Phaseolus vulgaris) bound 140 dpm/g fwt (1794 dpm/g dry wt) when exposed to 1.0 Ci/1 of [¹⁴C]ethylene (110 ci/mol). Phytotron-grown leaves were less succulent but only bound 90 dpm/g fwt (1046 dpm/g dry wt). Bean roots bound 30 dpm/g fwt. Citrus and Ligustrum bound 207 and 240 dpm/g fwt, respectively. The time required to achieve equilibrium of leaves with the gas phase was 15 min for bean, 30 min for Citrus, and 30–60 min for Ligustrum. The time for 1/2 of the bound ethylene to diffuse out of the leaves was 20 min for bean, 10 min for Citrus, and 30 min for Ligustrum. The amount of ethylene needed to occupy 1/2 of the binding sites was obtained from Scatchard plots. This value (K_d) was 0.2 l/1 for bean, 0.15 for Citrus, and 0.31 for Ligustrum. The quantity of binding sites in the tissues was 2.0×10^-9 mol of binding sites/kg tissue for bean leaves, 5.7×10^-9 for Citrus leaves, and 6.8×10^-9 for Ligustrum. Pretreatment with indoleacetic acid (IAA), ehtylene, and cycloheximide (1 mg/1) had little effect on the level of ethylene-binding sites in Citrus.Contribution from the Department of Biochemistry, School of Agriculture and Life Sciences and School of Physical and Mathematical Sciences, North Carolina State University. Paper No. 8445 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina 27695-7601.North Carolina-Israel exchange Scholar for 1981 at the Department of Biochemistry, North Carolina State University Raleigh, North Carolina, USA     

Refinement of the Citrus Tristeza Virus Resistance Gene (Ctv) Positional Map in Poncirus trifoliata and Generation of Transgenic Grapefruit (Citrus paradisi) Plant Lines with Candidate Resistance Genes in this Region

Citrus tristeza virus (CTV) is a major pathogen of Citrus. A single dominant gene Ctv present in the trifoliate relative of Citrus, Poncirus trifoliata confers broad spectrum resistance against CTV. Refinement of genetic maps has delimited this gene to a 121 kb region, comprising of ten candidate Ctv resistance genes. The ten candidate genes were individually cloned in Agrobacterium based binary vector and transformed into three CTV susceptible grapefruit varieties. Two of the candidate R-genes, R-2 and R-3 are exclusively expressed in transgenic plants and in Poncirus trifoliata, while five other genes are also expressed in non-transformed Citrus controls. Northern blotting with a CTV derived probe for assessment of infection in virus inoculated plants over a span of three growth periods, each comprising of six to eight weeks, indicates either an absence of initiation of infection or it’s slow spread in R-2 plant lines or an initial appearance of infection and it’s subsequent obliteration in some R-1 and R-4 plant lines. Limited genome walk up- and downstream form R-1 gene, based on it’s 100% sequence identity between Poncirus and Citrus, indicates promoter identity of 92% between the two varieties. Further upstream and downstream sequencing indicates the presence of an O-methyl transferase and a Copia like gene respectively in Citrus instead of the amino acid transporter like gene upstream and a sugar transporter like gene downstream in Poncirus. The possibility of recombinations in the resistance locus of Citrus and the need for consistent monitoring for virus infection and gene expression in the transgenic Citrus trees is discussed. Electronic supplementary material Electronic supplementary material is available for this article at and accessible for authorised users.     

On the nomenclature ofPan paniscus

In 1929 a subspecies of chimpanzee was classified asPan satyrus paniscus, a subspecies of theeastern chimpanzee, and elevated to species level,Pan paniscus, in 1933. Review of the literature indicates that this ape type was known since 1881 from several locations throughout the Congo Basin and first scientifically described in 1887 asTroglodytes niger var.marungensis, a subspecies of thewestern chimpanzee. The evidence presented here demonstrates that the synonymmarungensis was known and used as an earlier name. According to the International Code of Zoological Nomenclature, the earliest name awarded to a species is the one which has to be recognized in use. However, maintaining the emphasis on stability and assuming that to change the name would cause considerable confusion in the literature, it is this author's recommendation that the earlier namemarungensis be suppressed and the later synonympaniscus be conserved while an application is being considered for ruling by the International Commission on Zoological Nomenclature.     

Life-history traits of the six Panonychus species from Japan (Acari: Tetranychidae)

Six species of the genus Panonychus are known from Japan. Life-history parameters of all six species were investigated at 25 °C, and for three species two strains of different geographical origin were included. The intrinsic rate of natural increase (r _m ) ranged from 0.172/day for the P. osmanthi albino strain to 0.209/day for P. citri, while the net reproductive rate (R ₀) varied from 23.98 in the thelytokous species P. thelytokus to 46.61 in P. citri. Both values were higher in the polyphagous species (P. ulmi, P. mori and P. citri), which are considered crop pests, than those in the oligophagous species (P. thelytokus and P. osmanthi), considered non-pests. The only exception was P. bambusicola, an oligophagous non-pest species with R ₀- and r _m -values closely resembling those of the three polyphagous species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Suitability of Asian citrus psyllid, Diaphorina citri, as prey for ladybeetles

Given the apparent importance ofladybeetles as biological control agents of theinvasive Asian citrus psyllid, Diaphorinacitri Kuwayama (Homoptera: Psyllidae), aseries of experiments were undertaken to assessthe nutritional suitability of this pest as aprey item. Five species of Coccinellidae wereshown to develop successfully on a diet ofpsyllid nymphs and four species produced viableeggs. Eggs of the flour moth, Ephestiakuehniella Zeller (Lepidoptera: Pyralidae)were used as a reference diet. Larvae of Curinus coeruleus Mulsant, Cyclonedasanguinea L., Exochomus childreniMulsant, Harmonia axyridis Pallas, andOlla v-nigrum Mulsant all had survival ona diet of D. citri nymphs not differentfrom 100%, although developmental times wereextended and adult dry weights were reducedrelative to the Ephestia egg diet. Species were ranked (highest to lowest) forlarval performance on the D. citri dietrelative to the Ephestia egg diet as:E. childreni, 0.85;O. v-nigrum,0.82; C. coeruleus, 0.80;H.axyridis, 0.71;C. sanguinea, 0.48. Most females of C. sanguinea ceasedoviposition on the second day followingtransferal to the D. citri diet, but thefecundity and fertility of females of the otherspecies were not different from those feedingon Ephestia eggs. Generation times onthe Ephestia egg diet at 24°C(time to egg hatch + larval developmental time+ pupation time + adult prereproductive period)were (mean ± SEM): C. coeruleus, 56.7± 1.32 d;C. sanguinea, 28.0 ±0.88 d;E. childreni, 60.8 ± 1.96 d;H. axyridis, 32.2 ± 1.47 d;O.v-nigrum, 25.8 ± 1.12 d. Adult females ofC. coeruleus, H. axyridis and O.v-nigrum consumed the most psyllids in oneh, C. sanguinea was intermediate, andE. childreni consumed the least.     

佛冈县不同类型橘园柑橘木虱的种群动态

[目的] 明确佛冈县不同类型橘园柑橘木虱的种群动态,为当地柑橘木虱及柑橘黄龙病的防治工作提供科学依据。[方法] 通过2015-2016年的系统调查,对广东省佛冈县砂糖橘园的柑橘木虱种群消长动态进行研究,对比分析了不同处理措施对柑橘木虱种群数量的影响。[结果] 1-3月柑橘木虱发生危害较轻,随着嫩梢增多,其种群数量开始上升,6-9月为种群发生高峰期,期间出现多个发生高峰,10月之后种群数量逐渐进入消退期,种群数量维持在较低水平;2015、2016年佛冈县柑橘木虱的季节性消长动态基本一致,但种群发生量存在明显差别;3种不同类型橘园内柑橘木虱种群发生数量也存在显著差异,常年失管橘园柑橘木虱种群发生数量最高,常规水肥管理橘园次之,二者均显著高于常规水肥管理及化学防治橘园。[结论] 化学防治可在一定程度上控制柑橘木虱的种群数量,及时清理失管橘园对控制柑橘木虱和柑橘黄龙病传播至关重要。     

A simple chromosomal marker can reliably distinguishes <Emphasis Type="Italic">Poncirus</Emphasis> from <Emphasis Type="Italic">Citrus</Emphasis> species

Several chromosome types have been recognized in Citrus and related genera by chromomycin A₃ (CMA) banding patterns and fluorescent in situ hybridization (FISH). They can be used to characterize cultivars and species or as markers in hybridization and backcrossing experiments. In the present work, characterization of six cultivars of P. trifoliata (“Barnes”, “Fawcett”, “Flying Dragon”, “Pomeroy”, “Rubidoux”, “USDA”) and one P. trifoliata × C. limonia hybrid was performed by sequential analyses of CMA banding and FISH using 5S and 45S rDNA as probes. All six cultivars showed a similar CMA⁺ banding pattern with the karyotype formula 4B + 8D + 6F. The capital letters indicate chromosomal types: B, a chromosome with one telomeric and one proximal band; D, with only one telomeric band; F, without bands. In situ hybridization labeling was also similar among cultivars. Three chromosome pairs displayed a closely linked set of 5S and 45S rDNA sites, two of them co-located with the proximal band of the B type chromosomes (B/5S-45S) and the third one co-located with the terminal band of a D pair (D/5S-45S). The B/5S-45S chromosome has never been found in any citrus accessions investigated so far. Therefore, this B chromosome can be used as a marker to recognize the intergeneric Poncirus × Citrus hybrids. The intergeneric hybrid analyzed here displayed the karyotype formula 4B + 8D + 6F, with two chromosome types B/5S-45S and two D/5S-45S. The karyotype formula and the presence of two B/5S-45S chromosomes clearly indicate that the plant investigated is a symmetric hybrid. It also demonstrates the suitability of karyotype analyses to differentiate zygotic embryos or somatic cell fusions involving trifoliate orange germplasm. During the submission of this paper, we analyzed 25 other citrus cultivars with the same methodology and we found that the chromosome marker reported here can indeed distinguish Poncirus trifoliata from grapefruits, pummelos, and one variegated access of Citrus, besides the previously reported access of limes, limons, citrons, and sweet-oranges. However, among 14 mandarin cultivars, two of them displayed a single B/5S-45S chromosome, whereas in Citrus hystrix D.C., a far related species belonging to the Papeda subgenus, this chromosome type was found in homozygosis. Since these two mandarin cultivars are probably of hybrid origin, we assume that for almost all commercial cultivars and species of the subgenus Citrus this B type chromosome is a useful genetic marker.