Optimization of sampling and monitoring of vegetative flushing in citrus orchards.

Citrus trees produce flushes throughout the year, but there are no criteria established for a precise shoot monitoring in orchards under tropical climate. Methods for quantification of flush dynamics would be useful for horticultural and pest management studies because different insect vectors feed and reproduce on flushes. We estimated the minimum number and distribution of trees for sampling and determined the flushing pattern over time in 'Valencia Late' orange trees grafted onto 'Swingle' citrumelo rootstock. Shoots within a square frame (0.25 m2) on two sides of the canopy were counted and classified by their phenological stage. The minimum number of samples was estimated using the mean number of shoots and area under the flush shoot dynamics (AUFSD). The temporal and spatial distribution analysis was performed by Taylor's power law and by multiple correspondence analysis (MCA). Additionally, a shoot maturity index (SMI) based on visual qualitative assessment of flushes is proposed. Considering the mean number of shoots, it was necessary to sample two sides of 16 trees to reach a relative sampling error (Er) of 25%, whereas by the AUFSD, only five trees were necessary to reach an Er of 10%. Flushes were predominantly randomly distributed over time and space. Testing eight transects, sampled trees should be distributed throughout the block, avoiding sampling concentration in a certain area. MCA showed that the west side and the upper sampling positions of trees were more likely to be associated with younger shoots. AUFSD and the evaluation of both sides of the canopy yielded a smaller number of trees to be assessed. The SMI was a reliable metric to estimate the shoot phenology of orange trees, and correlated well (R2 > 70%) with the mean number of shoots within the square frame. Therefore, SMI has the potential to make shoot monitoring in the field more practical.

Long-term, sustained feeding by Asian citrus psyllid disrupts salicylic acid homeostasis in sweet orange.

BACKGROUND: Phloem-feeding insects are known to modulate the salicylic acid (SA) signaling pathway in various plant-insect interaction models. Diaphorina citri is a phloem feeding vector of the deadly phytopathogens, Candidatus Liberibacter americanus and Candidatus Liberibacter asiaticus, and the interactions of D. citri with its host that may modulate plant defenses are not well understood. The objectives of this study were to investigate the molecular mechanisms involved in transcriptional regulation of SA modification and activation of defense-associated responses in sweet orange (Citrus sinensis) exposed to various durations (7-, 14- and 150- days) of continuous feeding by D. citri. RESULTS: We quantified expression of genes involved in SA pathway activation and subsequent modification, as well as, associated SA metabolites (SA methyl ester, 2,3-DHBA, and SA 2-O-ß-D-glucoside). NPR1 and PR-1 expression was upregulated in plants exposed to continuous feeding by D. citri for 14 days. Expression of BSMT-like, MES1-like and DMR6-like oxygenase, as well as, accumulation of their respective SA metabolites (SA methyl ester, 2,3-DHBA) was significantly higher in plants exposed to continuous feeding by D. citri for 150 days than in those without D. citri infestation. Concomitantly, expression of UGT74F2-like was significantly downregulated and its metabolite, SA 2-ß-D-glucoside, was highly accumulated in trees exposed to 150 d of feeding compared to control trees without D. citri. CONCLUSIONS: D. citri herbivory differentially regulated transcription and SA-metabolite accumulation in citrus leaves, depending on duration of insect feeding. Our results suggest that prolonged and uninterrupted exposure (150 d) of citrus to D. citri feeding suppressed plant immunity and inhibited growth, which may highlight the importance of vector suppression as part of huanglongbing (HLB) management in citrus.

Fruit fly identification, population dynamics and fruit damage during fruiting seasons of sweet oranges in Rusitu Valley, Zimbabwe.

In 2003, the pest species Bactrocera dorsalis (Hendel) was reported for the first time in Kenya, Africa, and subsequently on many other African countries. In this work, 20 locations along the Rusitu Valley (Zimbabwe) were sampled in 2014 during the sweet oranges fruiting seasons, to verify the fruit fly taxonomy, invasion source, population dynamics, and fruit damage. The trapped fruit flies were identified using morphological traits and molecular techniques, as B. dorsalis. The haplotype network analysis revealed that Zimbabwe COI sequences were identical to other African B. dorsalis sequences. Fruit fly trappings per day varied during the year, although it remained always high. The same applies to fruit damage, most likely due to the permanent availability of cultivated and wild fruit varieties during the year. Rusitu Valley was invaded by B. dorsalis, most likely from neighbouring countries. Ten years after the first report in Kenya, the complete or near complete invasion of Africa has been achieved by B. dorsalis. In northern Africa the distribution is clearly limited by the Sahara desert. The large population size, the polyphagous nature of the species, and the continuous availability of suitable host fruit species during the year complicates the eradication of this species.

Orange jasmine as a trap crop to control Diaphorina citri.

Novel, suitable and sustainable alternative control tactics that have the potential to reduce migration of Diaphorina citri into commercial citrus orchards are essential to improve management of huanglongbing (HLB). In this study, the effect of orange jasmine (Murraya paniculata) as a border trap crop on psyllid settlement and dispersal was assessed in citrus orchards. Furthermore, volatile emission profiles and relative attractiveness of both orange jasmine and sweet orange (Citrus × aurantium L., syn. Citrus sinensis (L.) Osbeck) nursery flushes to D. citri were investigated. In newly established citrus orchards, the trap crop reduced the capture of psyllids in yellow sticky traps and the number of psyllids that settled on citrus trees compared to fallow mowed grass fields by 40% and 83%, respectively. Psyllids were attracted and killed by thiamethoxam-treated orange jasmine suggesting that the trap crop could act as a 'sink' for D. citri. Additionally, the presence of the trap crop reduced HLB incidence by 43%. Olfactometer experiments showed that orange jasmine plays an attractive role on psyllid behavior and that this attractiveness may be associated with differences in the volatile profiles emitted by orange jasmine in comparison with sweet orange. Results indicated that insecticide-treated M. paniculata may act as a trap crop to attract and kill D. citri before they settled on the edges of citrus orchards, which significantly contributes to the reduction of HLB primary spread.

Temperature-dependent development of Asian citrus psyllid on various hosts, and mortality by two strains of Isaria.

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is a devastating pest of Citrus spp. The aim of present study was to investigate the development and mortality of ACP on citrus (Citrus sinensis) (healthy and Huánglóngbìng- (HLB) diseased) and jasmine (Murraya paniculata) plants at various temperatures. Two new Isaria strains were collected from citrus orchards of Fuzhou (China), and HLB-diseased plants were verified by running PCR for 16S gene of Candidatus Liberibacter asiaticus (CLas). Development observations were recorded for egg, nymph and adult stages on all plants and three different temperatures (20, 25 and 30 °C) whereas mortality observations were recorded for the nymph (fifth instar) and adults on all plants at 25 °C. Field collected Isaria strains were belonged to previously reported Chinese strains under Maximum Parsimony (MP) and Maximum Likelihood methods, as well as, CLas isolates were belonged to previously reported Chinese isolates under MP and Neighbor-Joining methods. The fastest development and mortality was observed on HLB-diseased plants whereas longest time was taken by development and mortality completion on jasmine plants at all temperatures. The fastest developmental times of egg, nymph (first to fourth and fifth instar) and adult stages were ranged from 3.02 to 3.72 d, to 7.63-9.3 d, 5.35-5.65 d and 24.46-28.47 d on HLB-diseased plants at 30-20 °C, respectively. On the other hand, I. javanica caused the fastest mortality of nymphs and adults (32.21 ± 4.47% and 19.33 ± 4.51%) on HLB-diseased plants with the concentration of 1 × 108 conidia.mL-1 after 3 d and 7 d, respectively. It is concluded that there is a need for extensive molecular work to understand the extra-development and mortality of ACP on diseased plants, because, CLas bacterium can be supportive to uptake more sap from plant phloem.

Spiroplasma affects host aphid proteomics feeding on two nutritional resources.

Bacterial symbionts are broadly distributed among insects, influencing their bioecology to different degrees. Aphids carry a number of secondary symbionts that can influence aphid physiology and fitness attributes. Spiroplasma is seldom reported as an aphid symbiont, but a high level of infection has been observed in one population of the tropical aphid Aphis citricidus. We used sister isolines of Spiroplasma-infected (Ac-BS) and Spiroplasma-free (Ac-B) aphids reared on sweet orange (optimum host) and orange jasmine (suboptimum host) to demonstrate the effects of Spiroplasma infection in the aphid proteome profile. A higher number of proteins were differently abundant in aphids feeding on orange jasmine, indicating an impact of host plant quality. In both host plants, the majority of proteins affected by Spiroplasma infection were heat shock proteins, proteins linked to cell function and structure, and energy metabolism. Spiroplasma also induced changes in proteins involved in antimicrobial activity, carbohydrate processing and metabolism, amino acid synthesis and metabolism in aphids feeding on orange jasmine. We discuss on how the aphid host proteome is differentially affected by Spiroplasma infection when the host is exploiting host plants with different nutritional values.

Effects of transgenic expression of Brevibacterium linens methionine gamma lyase (MGL) on accumulation of Tylenchulus semipenetrans and key aminoacid contents in Carrizo citrange.

KEY MESSAGE: Carrizo transgenic plants overexpressing methionine-gamma-lyase produced dimethyl sulfide. The transgenic plants displayed more resistance to nematode attacks (Tylenculus semipenetrans) and may represent an innovative strategy for nematode control. Tylenchulus semipenetrans is a nematode pest of many citrus varieties that causes extensive damage to commercial crops worldwide. Carrizo citrange vr. (Citrus sinensis L. Usb × Poncirus trifoliate L. Raf) plants overexpressing Brevibacterium linens methionine-gamma-lyase (BlMGL) produced the sulfur volatile compound dimethyl sulfide (DMS). The aim of this work was to determine if transgenic citrus plants expressing BlMGL showed increased tolerance to T. semipenetrans infestation and to determine the effect on the content of key amino acids. While transgenic lines emitted dimethyl sulfide from leaves and roots, no sulfur-containing volatiles were detectable in wild-type Carrizo in the same tissues. Significant changes detected some key amino acids from leaves of transgenic plants such as aspartate, lysine, glycine, leucine and threonine with no changes in the amounts of methionine and α-ketobutyrate. In roots only glycine showed significant changes across all transgenic lines in comparison to wild-type plants. Transgenic plants expressing BlMGL and emitting DMS had less T. semipenetrans aggregation and more biomass than infected WT control plants, indicating that they may represent an innovative management alternative to pesticide/nematicide-based remedies.

Utilização de óleo essencial de duas variedades de laranja na eclosão de Meloidogyne javanica / Use of essential oil from two varieties of orange in the hatching of Meloidogyne javanica / Utilización de aceite esencial de dos variedades de naranja en la eclosión de Meloidogyne javanica

O Brasil é o maior produtor de laranja do mundo. Cultura rica em óleos essenciais de ampla aplicação, como no controle de pragas e patógenos, dentre eles os fitonematoides, causadores de prejuízos relevantes à agricultura. Assim, o objetivo do trabalho foi avaliar a porcentagem de eclosão de juvenis de M. javanica submetidos à diferentes concentrações de óleos essenciais de duas variedades de laranja, Pêra Rio e Folha Murcha, em condições in vitro. Os óleos essenciais foram extraídos das variedades de laranja (Citrus sinensis) Pêra Rio e Folha Murcha, pela técnica de hidrodestilação. O experimento foi instalado em arranjo fatorial 2X8, DIC, sendo oito concentrações (0, 25, 50, 100, 200, 300, 400 e 500 µg mL-1) e dois tipos de OE, avaliados em três períodos (4 e 16 dias). Em cada unidade experimental, tubos de ensaio 10x75 mm, adicionou-se 1 mL de suspensão de nematoides contendo 50 ovos e 1 mL de emulsão para cada concentração, avaliando-se a porcentagem de juvenis eclodidos. As concentrações dos óleos essenciais das variedades influenciaram (p≤0,05) na porcentagem de eclosão de juvenis de M. javanica, em todos os períodos avaliados. Para a maioria das concentrações, aos quatro e 16 dias, o óleo essencial da var. Pêra Rio foi mais eficiente na redução da eclosão, quando comparado a var. Folha Murcha.(AU)

Brazil is the largest producer of oranges in the world. This crop is rich in essential oils which are used in a wide range of applications, such as controlling pests and pathogens. Phytonematodes are among such pests, and are responsible for causing harmful damages to agriculture. Thus, the purpose of this study was to evaluate the percentage of hatching of M. javanica juveniles submitted to different concentrations of orange essential oil under in-vitro conditions. Oils were extracted from two varieties of orange (Citrus sinensis) popularly known in Brazil as Pera Rio and Folha Murcha, using the hydro-distillation technique. The experiment was carried out in a 2X8 factorial arrangement, at eight concentrations (0, 25, 50, 100, 200, 300, 400 and 500 µg mL-1) and two types of essential oils, evaluated in two periods (4 and 16 days). In each experimental unit, 10x75-mm test tubes received 1-mL nematode suspension containing 50 eggs and 1-mL of emulsion added for each concentration. Then, the percentage of hatched juveniles was evaluated. The concentration of essential oils from the varieties influenced (p≤0.05) the percentage of hatching of M. javanica juveniles in all evaluated periods. For most concentrations, at 4 and 16 days, the Pera Rio variety was more efficient at reducing hatching when compared to the Folha Murcha variety.(AU)

Brasil es el mayor productor de naranja del mundo. Cultura rica en aceites esenciales de amplia aplicación, como en el control de plagas y patógenos, entre ellos los fitonematodos, causantes de perjuicios relevantes a la agricultura. Así, el objetivo del trabajo fue evaluar el porcentaje de eclosión de juveniles de M. javanica sometidos a diferentes concentraciones de aceites esenciales de dos variedades de naranja, Pera Río y Hoja Mustia, en condiciones in vitro. Los aceites esenciales fueron extraídos de las variedades de naranja (Citrus sinensis) Pera Río y Hoja Mustia, por la técnica de hidrodestilación. El experimento fue instalado en un arreglo factorial 2x8, DIC, siendo ocho concentraciones (0, 25, 50, 100, 200, 300, 400 y 500 µg mL-1) y dos tipos de OE, evaluados en tres períodos (4 y 16 días). En cada unidad experimental, tubos de ensayo 10x75 mm, se añadieron 1 mL de suspensión de nematodos conteniendo 50 huevos y 1 mL de emulsión para cada concentración, evaluando el porcentaje de juveniles eclosionados. Las concentraciones de los aceites esenciales de las variedades influenciaron (p≤0,05) en el porcentaje de eclosión de juveniles de M. javanica, en todos los períodos evaluados. Para la mayoría de las concentraciones, a los cuatro y 16 días, el aceite esencial de la var. Pera Río fue más eficiente en la reducción de la eclosión, cuando comparado a var. Hoja Mustia.(AU)

Global Potential Distribution of Bactrocera carambolae and the Risks for Fruit Production in Brazil.

The carambola fruit fly, Bactrocera carambolae, is a tephritid native to Asia that has invaded South America through small-scale trade of fruits from Indonesia. The economic losses associated with biological invasions of other fruit flies around the world and the polyphagous behaviour of B. carambolae have prompted much concern among government agencies and farmers with the potential spread of this pest. Here, ecological niche models were employed to identify suitable environments available to B. carambolae in a global scale and assess the extent of the fruit acreage that may be at risk of attack in Brazil. Overall, 30 MaxEnt models built with different combinations of environmental predictors and settings were evaluated for predicting the potential distribution of the carambola fruit fly. The best model was selected based on threshold-independent and threshold-dependent metrics. Climatically suitable areas were identified in tropical and subtropical regions of Central and South America, Sub-Saharan Africa, west and east coast of India and northern Australia. The suitability map of B. carambola was intersected against maps of fruit acreage in Brazil. The acreage under potential risk of attack varied widely among fruit species, which is expected because the production areas are concentrated in different regions of the country. The production of cashew is the one that is at higher risk, with almost 90% of its acreage within the suitable range of B. carambolae, followed by papaya (78%), tangerine (51%), guava (38%), lemon (30%), orange (29%), mango (24%) and avocado (20%). This study provides an important contribution to the knowledge of the ecology of B. carambolae, and the information generated here can be used by government agencies as a decision-making tool to prevent the carambola fruit fly spread across the world.

Nucleotides, micro- and macro-nutrients, limonoids, flavonoids, and hydroxycinnamates composition in the phloem sap of sweet orange.

Currently, the global citrus production is declining due to the spread of Huanglongbing (HLB). HLB, otherwise known as citrus greening, is caused by Candidatus Liberibacter asiaticus (CLas) and is transmitted by the Asian citrus psyllids (ACP), Diaphorina citri Kuwayama. ACP transmits CLas bacterium while feeding on the citrus phloem sap. Multiplication of CLas in the phloem of citrus indicates that the sap contains all the essential nutrients needed for CLas. In this study, we investigated the micro- and macro-nutrients, nucleotides, and others secondary metabolites of phloem sap from pineapple sweet orange. The micro- and macro-nutrients were analyzed using inductively coupled plasma-mass spectroscopy (ICP-MS) and inductively coupled plasma-optical emission spectroscopy (ICP-OES). Nucleotides and other secondary metabolites analysis was accomplished by reversed phase HPLC coupled with UV, fluorescence detection, or negative mode electrospray ionization mass spectrometry (ESI-MS). Calcium (89 mM) was the highest element followed by potassium (38.8 mM) and phosphorous (24 mM). Magnesium and sulfur were also abundant and their concentrations were 15 and 9 mM, respectively. The rest of the elements were found in low amounts (< 2mM). The concentrations of ATP, ADP, and AMP were 16, 31, and 3 µ mole/Kg fwt, respectively. GTP, GMP. NAD, FMN, FAD, and riboflavin were found at concentrations below (3 µ mole/Kg fwt). The phloem was rich in nomilin 124 mM and limonin 176 µ mole/Kg fwt. Hesperidin, vicenin-2, sinensetin, and nobiletin were the most predominant flavonoids. In addition, several hydroxycinnamates were detected. The results of this study will increase our knowledge about the nature and the chemical composition of citrus phloem sap.

Differences in antennal sensillae of male and female peach fruit flies in relation to hosts.

Antennal sensillae of male and female peach fruit flies, Bactrocera zonata (Saunders) (Diptera: Tephritidae), obtained from three different host fruit species (guava, Psidium guajava L. (Myrtales: Myrtaceae); peach, Prunus persica (L.) Stokes (Rosales: Rosaceae); and orange, Citrus sinensis (L.) Osbeck (Sapindales: Rutaceae)), were studied with scanning electron microscopy. This study was carried out to describe the different types of sensillae present on the three antennal segments (scape, pedicel, and flagellum or funiculus) of both sexes of B. zonata on different host fruit. The antennal segments of females tended to be larger than those of males feeding on peach and guava fruit. On orange, both sexes were similar (no significant differences were found). The first two antennal segments, scape and pedicel, are reinforced by some bristles and have different types of sensillae, including trichoid I, II, S; basiconic II; and sensilla chaetica in different numbers on different host fruit species. Numerous microtrichia, as well as trichoid (I, II), basiconic (I), clavate, and coeloconic (I, II) sensillae were observed on the funiculus with a great variation in number and length. As a result of feeding on different hosts, differences were found between sexes and some plasticity in size, number, distribution, and position of some sensillae, including trichoid, basiconic, chaetica, and clavate on the antennae of the female B. zonata. These sensillae were significantly larger in females. Also, some morphological and morphemetric differences have been found according to their feeding on different host fruit.

Why the aphid Aphis spiraecola is more abundant on clementine tree than Aphis gossypii?

Aphis spiraecola and Aphis gossypii cause harmful damages on clementine tree orchards. Weekly surveys measured the abundance of aphids (larvae, winged and wingless adults) as well as of auxiliary insects and parameters of energy metabolism. Correlatively, soluble carbohydrates, total free amino acids, free proline and condensed tannins were quantified in control and infested leaves. Both aphid species showed parallel temporal variations, but A. spiraecola was consistently more abundant regardless of the stage. Amino acids had a positive effect on both aphid species abundance, but neither condensed tannins nor auxiliary insects seemed to modulate aphid populations. Interestingly, the leaf carbohydrate content was positively correlated with the abundance of A. spiraecola, but not with that of A. gossypii. Moreover, A. gossypii's abundance was significantly down-regulated by high proline concentrations. Thus, the higher abundance of A. spiraecola could be explained by a better tolerance to high proline contents and a better conversion of foliar energy metabolites.

[Spider diversity in cultures of Citrus sinensis (Rutaceae) in Corrientes province, Argentina]. / Diversidad de Araneae en cultivos de Citrus sinensis (Rutaceae) de la Provincia de Corrientes, Argentina.

Spiders are predators that reduce insect pest populations in agroecosystems. In spite that the presence of spider assemblages has been described in different crop plants, no reports have been done for Citrus species in Argentina. We studied the spider community associated with cultures of Citrus sinensis in the province of Corrientes, Argentina, in two plots (AM1 irrigated and AM2 unirrigated) of 0.82 hectares each. Spiders were collected monthly by using pitfall traps, hand capture, agitation of foliage and sieving of soil litter. A total of 7174 specimens were collected (33 families, 44 genera and 200 species/morphospecies). The AM1 recorded 3811 individuals (33 families, 179 species/morphospecies) and AM2 3363 individuals (31 families, 174 species/morphospecies). November, December and January showed the highest abundance in both plots. A total of 70% of collected spiders were juveniles. Eight families were the most abundant and represented 75% of spiders collected in both plots, the largest being Lycosidae. We identified eight guilds in both plots, for which the ground runners resulted the most abundant (AM1 n = 1341, s=39, n=999 AM2, s = 33), followed by orb weavers (AM1 n = 637, s = 36; AM2 n = 552, s = 33), the stalkers (AM1 n = 471, s = 43, AM2 n = 453, s = 47) and the space web-builders (AM1 n = 446, s = 23; AM2 n = 342, s = 25). The analysis of alpha diversity in both plots (AM1H' = 4.161, J' = 0.802; AM2 H' = 4.184, J' = 0.811) showed no significant differences (t = 1.083 p = 0.279). The linear dependences model was the one with the best fit results, where the species observed were more than estimated. Clench model estimated 90.9% of the spiders observed in the unit with irrigation and 90.6% in the unit without irrigation.

Diversidad de Araneae en cultivos de Citrus sinensis (Rutaceae) de la Provincia de Corrientes, Argentina / Spider diversity in cultures of Citrus sinensis (Rutaceae) in Corrientes province, Argentina

Spiders are predators that reduce insect pest populations in agroecosystems. In spite that the presence of spider assemblages has been described in different crop plants, no reports have been done for Citrus species in Argentina. We studied the spider community associated with cultures of Citrus sinensis in the province of Corrientes, Argentina, in two plots (AM1 irrigated and AM2 unirrigated) of 0.82 hectares each. Spiders were collected monthly by using pitfall traps, hand capture, agitation of foliage and sieving of soil litter. A total of 7 174 specimens were collected (33 families, 44 genera and 200 species/morphospecies). The AM1 recorded 3 811 individuals (33 families, 179 species/morphospecies) and AM2 3 363 individuals (31 families, 174 spe- cies/morphospecies). November, December and January showed the highest abundance in both plots. A total of 70% of collected spiders were juveniles. Eight families were the most abundant and represented 75% of spiders collected in both plots, the largest being Lycosidae. We identified eight guilds in both plots, for which the ground runners resulted the most abundant (AM1 n=1341, s=39, n=999 AM2, s=33), followed by orb weavers (AM1 n=637, s=36; AM2 n=552, s=33), the stalkers (AM1 n=471, s=43, AM2 n=453, s=47) and the space web- builders (AM1 n=446, s=23; AM2 n=342, s=25). The analysis of alpha diversity in both plots (AM1H’=4.161, J’=0.802; AM2 H’=4.184, J’=0.811) showed no significant differences (t=1.083 p=0.279). The linear dependences model was the one with the best fit results, where the species observed were more than estimated. Clench model estimated 90.9% of the spiders observed in the unit with irrigation and 90.6% in the unit without irrigation.

Las arañas son ideales como indicadores de cambios ecológicos por su diversidad y abundancia. En la provincia de Corrientes, Argentina, se estudiaron arañas de Citrus sinensis en parcelas de 0.82 hectáreas, con riego (AM1) y sin riego (AM2). Mediante pitfall, agitación de follaje, captura manual y tamizado de hojarasca se recolectaron 7 174 ejemplares (33 familias, 44 géneros y 200 especies/morfoespecies), el 70% fueron juveniles. AM1 registró 3 811 individuos (33 familias, 179 especies/morfoespecies) y AM2 3 363 (31 familias, 174 especies/morfoespecies). Durante el verano se observó la mayor abundancia. En ambas parcelas, ocho familias representaron el 75% del total, Lycosidae fue la más numerosa y se identificaron ocho gremios; las vagabundas de suelo fueron las más abundantes (AM1 n=1341, s=39; AM2 n=999, s=33) seguida por las constructoras de telas orbiculares (AM1 n=637, s=36; AM2 n=552, s=33), las cazadoras al acecho (AM1 n=471, s=43; AM2 n=453, s=47) y las de telas espaciales (AM1 n=446, s=23; AM2 n=342, s=25. La diversidad alfa en ambas parcelas (AM1 H’=4.161, J’=0.802; AM2 H’=4.184, J’=0.811) no mostró diferencias significativas (t=1.083, p=0.279). El modelo de dependencia lineal fue el que mejor ajustó los resultados. El modelo de Clench estimó el 90.9% de las observadas en AM1 y el 90.6% en la AM2.

Mistletoes and mutant albino shoots on woody plants as mineral nutrient traps.

BACKGROUND AND AIMS: Potassium, sulphur and zinc contents of mistletoe leaves are generally higher than in their hosts. This is attributed to the fact that chemical elements which are cycled between xylem and phloem in the process of phloem loading of sugars are trapped in the mistletoe, because these parasites do not feed their hosts. Here it is hypothesized that mutant albino shoots on otherwise green plants should behave similarly, because they lack photosynthesis and thus cannot recycle elements involved in sugar loading. METHODS: The mineral nutrition of the mistletoe Scurrula elata was compared with that of albino shoots on Citrus sinensis and Nerium oleander. The potential for selective nutrient uptake by the mistletoe was studied by comparing element contents of host leaves on infected and uninfected branches and by manipulation of the haustorium-shoot ratio in mistletoes. Phloem anatomy of albino leaves was compared with that of green leaves. KEY RESULTS: Both mistletoes and albino leaves had higher contents of potassium, sulphur and zinc than hosts or green leaves, respectively. Hypothetical discrimination of nutrient elements during the uptake by the haustorium is not supported by our data. Anatomical studies of albino leaves showed characteristics of release phloem. CONCLUSIONS: Both albino shoots and mistletoes are traps for elements normally recycled between xylem and phloem, because retranslocation of phloem mobile elements into the mother plant or the host is low or absent. It can be assumed that the lack of photosynthetic activity in albino shoots and thus of sugars needed in phloem loading is responsible for the accumulation of elements. The absence of phloem loading is reflected in phloem anatomy of these abnormal shoots. In mistletoes the evolution of a parasitic lifestyle has obviously eliminated substantial feeding of the host with photosynthates produced by the mistletoe.

Cold treatment of Ceratitis capitata (Diptera: Tephritidae) in oranges using a larval endpoint.

South Africa currently exports fresh citrus (Citrus spp.) fruit to Japan using an in-transit cold treatment protocol of 14 d or 12 d at temperatures <0 degrees C for treatment of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) in 'Clementine' mandarins (Citrus reticulata Blanco) and other citrus types, respectively. To reduce the risk of chilling injury with this treatment, research was conducted with temperatures >0 degrees C. Earlier South African research had shown that young (6-d-old) larvae were slightly more tolerant of cold treatment and that there were no significant differences between cold tolerance of these larvae in different citrus types [oranges, Citrus sinensis (L.) Osbeck; grapefruits, Citrus paradisi Macfad.; lemons, Citrus limon (L.) Burm.f.; and mandarins). Due to their ready availability, 'Valencia' oranges were used in this study. When 62,492 larvae in total were treated in three replicates at a mean temperature of 1.5 degrees C for 16 d, there were three larval survivors. The trial was therefore repeated with oranges using a 16-d period at a mean temperature of 1.0 degrees C and a mean of 1.4 degrees C for the hourly maximum probe readings. Three replicates were again conducted and the resultant mean mortality in the control was 8.1% of 21,801 larvae, whereas the cold treatment mortality was 100% of 71,756 larvae. This treatment at a mean temperature of 1 degree C exceeded the Japanese confidence level requirement and also exceeded the Probit-9 mortality level, but not at a confidence level of 95%. These data support the establishment of a treatment protocol of 16 d at temperatures <1.4 degrees C, commencing once all fruit pulp probes reach a temperature of 1 degree C or lower.

Cold susceptibility and disinfestation of Bactrocera invadens (Diptera: Tephritidae) in oranges.

To develop a cold disinfestation treatment for the fruit fly Bactrocera invadens Drew, Tsuruta & White (Diptera: Tephritidae) that is rapidly spreading across Africa, research was conducted in Nairobi, Kenya, using flies from a laboratory culture and 'Valencia' orange (Citrus sinensis L. Osbeck) as the host. The developmental rate of B. invadens in Valencia oranges was determined at 28 degrees C, and the third instar was found to be the least susceptible of the egg and larval life stages to cold treatment at 1.1 degrees C in oranges. When 22,449 B. invadens third instars were exposed in oranges to a cold treatment with an approximate midpoint of 1.1 +/- 0.5 degrees C, the results suggested that a period of 16 d would be worthwhile verifying on a larger scale in oranges. Results from the first replicate of 16,617 larvae showed no survivors, but the second replicate of 23,536 larvae had three survivors. Because a longer cold treatment based on a mean temperature of 1.1 degrees C would create logistical difficulties for some export markets, further replicates were conducted at an approximate midpoint of 0.5 degrees C and at mean hourly maximum of 0.9 +/- 0.5 degrees C, for 16 d. After three replicates, in which 65,752 B. invadens third instars in total were treated with no survivors, the Japanese requirement of 99.99% mortality at the 95% confidence level was surpassed. The following treatment protocol for B. invadens larvae in oranges can therefore be recommended: fruit pulp to be maintained at temperatures of 0.9 degrees C or lower for 16 consecutive days.

Host status of grapefruit and Valencia oranges for Anastrepha serpentina and Anastrepha ludens (Diptera: Tephritidae).

Anastrepha serpentina (Wiedemann) (Diptera: Tephritidae) is sporadically captured in the Rio Grande Valley of Texas. Although its preferred hosts are in the Sapotaceae family, several varieties of Citrus, including grapefruit and oranges are listed as alternate hosts. Although Mexican fruit fly, Anastrepha ludens (Loew), is known to be a major pest of Citrus, doubt exists as to the status of Citrus as a breeding host for A. serpentina. To evaluate the host status of commercial Citrus for A. serpentina we compared oviposition and development with that of A. ludens under laboratory conditions with 'Rio Red' grapefruit (Citrus paradisi MacFayden) and 'Valencia' oranges [Citrus sinensis (L.) Osbeck] in different stages of maturity. Both fly species oviposited in early season fruit in which the eggs and larvae died in the fruit albedo. Survival of either species to the adult stage occurred in later season grapefruit. In oranges, no A. serpentina larvae survived compared with 150 A. ludens surviving to adults. Survival on both Citrus species was much lower for A. serpentina, only approximately 5% of eggs eclosed into larvae in grapefruit compared with approximatley 50% for A. ludens. In oranges approximately 16% of A. serpentina eggs eclosed compared with approximately 76% for A. ludens. In grapefruit, only one fourth as many A. serpentina larvae survived to the adult stage compared with A. ludens. Additional experiments were performed in a greenhouse on small, caged trees of la coma (Sideroxylon celastrinum H.B.K.), a Texas species of Sapotaceae. The A. serpentina females readily oviposited into these berries and normal adults emerged. The present low incidence of the adults, coupled with the high mortality during development of the larvae, suggests that Texas citrus is unlikely to support a breeding population of A. serpentina.

Terpene down-regulation in orange reveals the role of fruit aromas in mediating interactions with insect herbivores and pathogens.

Plants use volatile terpene compounds as odor cues for communicating with the environment. Fleshy fruits are particularly rich in volatiles that deter herbivores and attract seed dispersal agents. We have investigated how terpenes in citrus fruit peels affect the interaction between the plant, insects, and microorganisms. Because limonene represents up to 97% of the total volatiles in orange (Citrus sinensis) fruit peel, we chose to down-regulate the expression of a limonene synthase gene in orange plants by introducing an antisense construct of this gene. Transgenic fruits showed reduced accumulation of limonene in the peel. When these fruits were challenged with either the fungus Penicillium digitatum or with the bacterium Xanthomonas citri subsp. citri, they showed marked resistance against these pathogens that were unable to infect the peel tissues. Moreover, males of the citrus pest medfly (Ceratitis capitata) were less attracted to low limonene-expressing fruits than to control fruits. These results indicate that limonene accumulation in the peel of citrus fruit appears to be involved in the successful trophic interaction between fruits, insects, and microorganisms. Terpene down-regulation might be a strategy to generate broad-spectrum resistance against pests and pathogens in fleshy fruits from economically important crops. In addition, terpene engineering may be important for studying the basic ecological interactions between fruits, herbivores, and pathogens.

An attempt of postharvest orange fruit rot control using essential oils from Mediterranean plants.

Twelve essential oils from Mediterranean aromatic plants were tested at different doses against four fungi known as causal agents of post-harvest orange fruit rot: Botrytis cinerea, Penicillium italicum, Phytophthora citrophthora, and Rhizopus stolonifer. Essential oils were obtained from Hyssopus officinalis, Lavandula angustifolia, Majorana hortensis, Melissa officinalis, Ocimum basilicum, Origanum vulgare, Salvia officinalis, and Thymus vulgaris (Family Lamiaceae), Verbena officinalis (Family Verbenaceae), and Pimpinella anisum, Foeniculum vulgare, and Carum carvi (Family Apiaceae). Because preliminary in vitro experiments showed that only the oils from V. officinalis, T. vulgaris, and O. vulgare exhibited some fungistatic activity against the above-named fungi, these three essential oils were used in successive in vivo tests carried out to protect healthy "Washington navel" orange fruits from artificial infection by the same micromycetes. The essential oil of T. vulgaris, at a 2,000 ppm dose, controlled fruit rot by B. cinerea, P. citrophthora, and R. stolonifer but was ineffective against P. italicum. Essential oils of V. officinalis and O. vulgare inhibited infection by the first two fungi and only by P. citrophthora, respectively. This finding represents an important result, with the goal of using the essential oils as natural preservatives for food products, due to their positive effect on their safety and shelf life.