Screening of biological activities of fungi associated with pistachio bark beetle, Chaetoptelius vestitus (Coleoptera, Curculionidae), infesting pistachio cultivations in Tunisia.

AIMS: In this study, we aimed to explore the enzymatic diversity, the entomopathogenic and the antimicrobial potentialities of fungi associated with the pistachio bark beetle, Chaetoptelius vestitus. METHODS AND RESULTS: A total of 40 isolates were screened for enzymatic diversity. Most of them, 92·5%, were able to produce at least two of the screened enzymes. Pathogenic assays performed on C. vestitus showed a high entomopathogenic activity of the isolates Ata_io_1 (A. tamarii), Fve_io_1 (F. verticillioides), Tpi_io_1 (T. pinophilus), Pal_io_1 (P. album), Pbi_io_2 (Penicillium bilaiae) and Pch_io_1 (P. chrysogenum), as based on mean mortality of C. vestitus. A screening of antimicrobial activity using well diffusion method showed that the isolates Tro_io_1 (T. pinophilus), Tat_io_1 (T. atroroseus) and Pch_io_1 (P. chrysogenum) presented the highest antibacterial activity. Furthermore, Mgu_io_1 (M. guilliermondii), Asc_io_1 (A. sclerotiorum), Ata_io_1 (A. tamarii), G. lavendula (Gla_io_1), Pva_io_1 (P. variotii), Pul_io_1 (P. ulaiense), Tat_io_1 (T. atroroseus) and Tro_io_1 (T. roseum) were active against at least two of the three tested fungal phytopathogens. CONCLUSIONS: Fungal isolates representing entomopathogenic activity and a broad spectrum of antimicrobial activities can be considered as promising resources for biological pistachio trees protection. SIGNIFICANCE AND IMPACT OF THE STUDY: Fungi associated with C. vestitus were investigated for detecting their potential biotechnological applications.

Endozoochory of frugivorous birds as potential dispersal of seed parasites from Pistacia chinesis: experimental evidence.

The plant-disperser-fruit pest triads involve 3 interacting animals or groups (plants, vertebrates and seed parasites), and the dispersal of both seeds and seed parasites, which can both benefit from endozoochory via defecation or regurgitation by frugivorous vertebrates. However, we have very limited knowledge about the ecological and evolutionary consequences of these plant-disperser-fruit pest triads. Across central Northern China, several seed wasps (mainly Eurytoma plotnikov attack Pistacia chinensis fruits, and seed wasp larvae can develop, diapause and finally emerge as adults inside a seed during the following 1-3 years. In this study, we experimentally investigated whether frugivorous birds discriminated P. chinesis fruits with or without seed parasites, and whether bird endozoochory (by defecation or regurgitation) affected larval survival of seed parasites. The infestation rate by seed parasites was 37% of the P. chinesis fruit crop but with up to 48% of aborted fruits. We found that all 5 bird species can discriminate and then reject all unhealthy fruits (including aborted and insect-infested). However, 4 of the 5 bird species, in particular bulbul species, consumed 15-41% of aborted and insect-infested fruits as complementary food only when these unhealthy fruits were provided. Moreover, all larva of seed parasites remained alive after bird defecation or regurgitation. In conclusion, our study demonstrates that endozoochory by frugivorous birds could lead to potential dispersal of seed parasites of P. chinensis but with a very low probability.

Quantitative differences detected in the histology of galls induced by the same aphid species in different varieties of the same host.

Plant galls are abnormal growths caused by an inducer that determines their morphology and anatomy. We qualitatively and quantitatively compared the histological anatomy of five aphid species (Paracletus cimiciformis, Forda marginata, Forda formicaria, Baizongia pistaciae and Geoica wertheimae) that induce galls in Pistacia terebinthus shrubs growing in Israel. We also quantitatively compared these galls to those that the aphids create on the same host in Spain. Histological study was conducted following methods described previously by the authors. Quantitative differences among the galls were found in five of 12 common anatomical traits: gall thickness, stomatal number in the epidermis-air, size of vascular bundles, distance of phloem ducts from the lumen and number of intraphloematic schizogenous ducts. Other structures were particular to one or some species: number of cracks in the epidermis-lumen, a sclereid layer, trichomes and microcrystal inclusions. Fisher's tests of combined probabilities showed that the galls induced in Israel were statistically different from those in Spain. In particular, the number of intraphloematic schizogenous ducts was higher in the galls induced in P. terebinthus in Israel. Such differences were also found in other traits related to defence of the gall inhabitant. In conclusion, while the gall shape and size are determined mainly by the cecidogenic insect, it seems that the host plant also plays an important role in determining the number/size of quantitative traits, in this case mainly protective structures.

Differences in Monoterpene Biosynthesis and Accumulation in Pistacia palaestina Leaves and Aphid-Induced Galls.

Certain insect species can induce gall formation on numerous plants species. Although the mechanism of gall development is largely unknown, it is clear that insects manipulate their hosts' anatomy, physiology, and chemistry for their own benefit. It is well known that insect-induced galls often contain vast amounts of plant defensive compounds as compared to non-colonized tissues, but it is not clear if defensive compounds can be produced in situ in the galled tissues. To answer this question, we analyzed terpene accumulation patterns and possible independent biosynthetic potential of galls induced by the aphid Baizongia pistaciae L. on the terminal buds of Pistacia palaestina Boiss. We compared monoterpene levels and monoterpene synthase enzyme activity in galls and healthy leaves from individual trees growing in a natural setting. At all developmental stages, monoterpene content and monoterpene synthase activity were consistently (up to 10 fold on a fresh weight basis) higher in galls than in intact non-colonized leaves. A remarkable tree to tree variation in the products produced in vitro from the substrate geranyl diphosphate by soluble protein extracts derived from individual trees was observed. Furthermore, galls and leaves from the same trees displayed enhanced and often distinct biosynthetic capabilities. Our results clearly indicate that galls possess independent metabolic capacities to produce and accumulate monoterpenes as compared to leaves. Our study indicates that galling aphids manipulate the enzymatic machinery of their host plant, intensifying their own defenses against natural enemies.

Molecular and morphological identification of pistachio armored scale insects (Hemiptera: Diaspididae), with description of a new species.

Members of the family Diaspididae (Hemiptera: Coccomorpha) can be devastating pests that suck parenchyma cell contents from crops and cause severe damage to pistachio trees (Pistacia vera L.). The current research collected and characterized diaspidid species from pistachio orchards in Kerman province, Iran, according to their morphological and molecular features. Lepidosaphes pistaciae Archangelskaya, Suturaspis davatchi (Balachowsky & Kaussari) and Melanaspis inopinata (Leonardi) are redescribed and a new species, Melanaspis pistaciae Hosseininaveh & Kaydan sp. n., is described. Phylogenetic trees based on molecular analysis of COI and 28S rDNA fragments placed all the species in separated clades and confirmed M. pistaciae as a new taxon which is concluded by morphological differences. Molecular analysis suggests non-monophyly of the populations of each species. Melanaspis pistaciae sp. n. has spread to most cultivated pistachio areas in Iran and has probably been misidentified as M. inopinata in the past. Further investigation of the biology of this species may lead to development of more effective approaches for controlling this pest.

Ooencyrtus (Hymenoptera: Encyrtidae), egg parasitoids of the pistachio green stink bugs (Hemiptera: Pentatomidae) in Iran.

This paper deals with three species of Ooencyrtus Ashmead (Hymenoptera: Encyrtidae) obtained from eggs of the green stink bug, Brachynema germarii (Kolenati) (Hemiptera: Pentatomidae) infesting pistachio plants in Iran. Two new species, Ooencyrtus iranicus Hayat & Mehrnejad, sp. nov. and O. pistaciae Hayat & Mehrnejad, sp. nov., are described, and O. telenomicida (Vassiliev) is newly recorded from Iran, redescribed and illustrated. A key to some species considered similar to the species treated here is also given.

Contribution of gall microscopic structure to taxonomy of gallicolous aphids on Pistacia.

Aphids inducing galls on Pistacia plants belong to the tribe Fordini. According to the Heie & Wegierek classification, the genera are grouped into three subtribes. Previous microscopic studies showed that this taxonomy is not consistent with the histological characteristics of the galls. In this paper, galls induced by Aplonerura lentisci, Asiphonella cynodonti, Forda riccobonii, Slavun wertheimae and Smynthurodes betae were analyzed for the first time, as well as nine other galls previously described. Based on histological features three groups of galls can be establish: the first group comprises closed galls, induced by Baizongia pistaciae, Geoica utricularia, Rectinasus buxtoni and Slavun wertheimae; the second group includes two species of Geopemphigus (G. blackmani and G. torsus), and the third one is divided into two subgroups, the first comprises Aplonerura lentisci, Asiphonella cynodonti and Geopemphigus morral, and the second that includes Forda formicaria, F. marginata, F. riccobonii, Paracletus cimiciformis and Smynthurodes betae. An identification key of species based on microscopic features of galls is presented.

Antioxidant activity of insect gall extracts of Pistacia integerrima.

BACKGROUND: Pistacia integerrima (P. integerrina) insect galls are widely used in ayurveda and siddha system of medicine as karkatasringi. The use of leaf galls as a rejuvenator may be attributed to antioxidant property, however there is less scientific evidence. Therefore, the aim of this study was to evaluate the chemical composition and the antioxidant potential of leaf gall extracts (aqueous and ethanol) of P. integerrina, which is extensively used in the preparation of traditional medications. METHODS: The antioxidant activities of aqueous and ethanolic leaf gall extracts were examined using diphenylpicrylhydrazyl (DPPH), hydroxyl scavenging and ferric reducing power (FRAP) methods. RESULTS: The presences of phenolics, tannins, phytosterols, triterpenoids, saponins, flavonoids and reducing sugars were identified in both the extracts. In comparison to the aqueous extract, the ethanolic extract had the highest total phenolic and flavonoid content at 234 ±2.4 mg of GAE/g d.w. and 95.5 ±3.2 mg of QUE/g d.w., respectively. This higher content of total phenolics and flavonoids found in the ethanolic extract was directly associated with higher antioxidant activity. CONCLUSIONS: This study demonstrates the poetnet antioxidant activities of P. integerrima leaf gall extracts. Further, there was a strong association between the higher antioxidant activities with that of higher total phenolic and flavonoid content in the ethanolic leaf gall extracts of P. integerrima. The results encourage the use of P. integerrima leaf gall extracts for medicinal health, functional food and nutraceuticals applications, due to their antioxidant properties. Future work will be interesting to learn the chemical composition and better understand the mechanism of action of the antioxidants present in the extract for development as a drug for therapeutic application.

The histo structure of galls induced by aphids as a useful taxonomic character: the case of Rectinasus (Hemiptera, Aphididae, Eriosomatinae).

Morphological differentiation of gall tissues induced on plants may play a role to characterize the real taxonomic position of the gall inducer. We verified this hypothesis with galls induced by Rectinasus buxtoni on Pistacia palaestina. There is controversy about the taxonomic localization of genus Rectinasus: in one classification it is situated with the genera Forda and Paracletus while in another it is linked to the genera Geoica and Baizongia. Histological examination of the walls of the galls reveals the presence of two opposed vascular bundles and an inner surface of the gall with cavities. These features place Rectinasus in the same group as Geoica and Baizongia, and not with Paracletus and Forda, whose galls have a different histological structure, as generally admitted.

Evaluation of monitoring traps with novel bait for navel orangeworm (Lepidoptera: Pyralidae) in California almond and pistachio orchards.

Experiments conducted in three almond, Prunus dulcis (Rosales: Rosaceae), orchards and three pistachio, Pistacia vera (Sapindales: Anicardiaceae), orchards in 2009 and 2010, and determined that sticky bottom wing traps baited with ground pistachio mummies, or a combination of ground pistachio plus ground almond mummies, trapped more adult female navel orangeworm, Amyelois transitella (Walker) (Lepidoptera: Pyralidae), than did traps baited with ground almond mummies alone. During both years of this study, 2.9 and 1.8 more moths were caught in traps baited with pistachio mummies compared with traps baited with almond mummies in almond orchards and pistachio orchards, respectively. Also, traps located in pistachio orchards caught 5.9 and 8.3 times more navel orangeworm than were trapped from almond orchards in 2009 and 2010, respectively. Implications for use of this novel baited trap in almond and pistachio orchard integrated pest management programs are discussed.

Study on the physiology of diapause, cold hardiness and supercooling point of overwintering pupae of the pistachio fruit hull borer, Arimania comaroffi.

The pistachio fruit hull borer, Arimania comaroffi (Ragonot) (Lepidoptera: Pyralidae), is a key pest of pistachio orchards in Iran. This pest passes the winter as diapausing pupae. In this study, some physiological changes in relation to environmental temperature were investigated in field collected pupae. The relationship between supercooling point, cold hardiness and physiological changes of a wild population of this pest was also investigated. The glycogen content decreased with decrease in environmental temperature. Decrease in glycogen content was proportional to increase in total body sugar, trehalose, myo-inositol and sorbitol contents. In January with mean ambient temperature of 5.4°C, glycogen (5 mg/g fresh body weight) content was at the lowest level whereas total body sugar (10.3 mg/g fresh body weight), trehalose (8.6 mg/g fresh body weight), myo-inositol (5.3 mg/g fresh body weight) and sorbitol (2.6 mg/g fresh body weight) were at the highest levels. Total body sugar, trehalose, myo-inositol and sorbitol contents increased as mean temperature decreased from 22.7°C in October to 5.4°C in January. Total body lipid decreased during overwintering and reached to the lowest level at the end of March. Supercooling points were decreased from October to January and reached to the lowest level (-16°C) in January with minimum ambient temperature of -10°C. Survival at low temperature after 24 h was also greatest in January with 72% survival at -10°C, 39% survival at -15°C and 0% survival at -20°C. Increase in temperature from February onward, was proportional with increase in supercooling points and decrease in survival rate. Regardless of sampling date, all pupae died after 24 h at -20°C, whereas none pupae died after 24 h at -5°C. This indicates that this insect is freeze-intolerant.

Energy allocation changes in overwintering adults of the common pistachio Psylla, Agonoscena pistaciae Burckhardt & Lauterer (Hemiptera: Psyllidae).

The common pistachio psylla, Agonoscena pistaciae Burckhardt & Lauterer (Hemiptera: Psyllidae), is known as the key pest of pistachio orchards in Iran. This pest passes the winter as adults. In this study, energy allocation changes in relation to ambient temperature were investigated in field-collected adults by measuring total body sugar, trehalose, glucose, sorbitol, myoinositol, glycogen, lipid, and protein contents. Glycogen content decreased with decrease in ambient temperature. The decrease in glycogen content was proportional to the increase in total body sugar, trehalose, myoinositol, and sorbitol contents. In January, with mean ambient temperature of 5.4°C, glycogen content was at the lowest level, whereas total body sugar, trehalose, glucose, and sorbitol were at the highest level. Total body sugar, trehalose, myoinositol, and sorbitol contents increased as temperature decreased from 22.7°C in October to 5.4°C in January. In conclusion, low molecular weight carbohydrates and polyols may play a role in winter survival and adaptation to cold of the common pistachio psylla by providing the required cryoprotection. Also, overwintering adults of the common pistachio psylla may store energy in the form of lipid for later utilization during the overwintering.

Variable developmental rate and survival of navel orangeworm (Lepidoptera: Pyralidae) on pistachio.

A series of laboratory and field studies were conducted using two lines of navel orangeworm, reared on different stages of new crop and mummy pistachios, Pistacia vera L. This study demonstrated the potential importance of malformed pistachios (pea splits) to the population dynamics of navel orangeworm, because these nuts, which are available as early as two months before mature nuts, supported navel orangeworm development and survival. Overall, the developmental rate on new crop pistachios is fastest on mature nuts, 422.3 +/- 123 degree-days (DD, degrees C), but other factors such as exposure to insecticide residue also sped development, although survival decreased. Development took the longest on unharvested nuts (mummies) dried at 90 degrees C for 24 h, 2664.7 +/- 131.4 DD. In most trials development was variable and two generations could develop at the fastest rate before the slowest individual completed development, which in turn calls into question the concept of discrete generations. Generally, survival was highest on mature pistachios and other stages of new crop nut and lowest on mummies collected in May. Survival was also higher on the new varieties 'Lost Hills' and 'Golden Hills' (24.7 and 32.0%, respectively) than on the most extensively planted variety 'Kerman' (13.3%). In our trials, both the rate of development and survival were dependent on nut stage, age, variety, and quality, indicating that pistachios, like almonds, Prunus dulcis (Mill.) D. A. Webb, are a dynamic rather than a static nutrient source for navel orangeworm.

Effect of bait formulation and number of traps on detection of navel orangeworm (Lepidoptera: Pyralidae) oviposition using egg traps.

Egg traps are the primary tool for monitoring egg deposition of the navel orangeworm, Amyelois transitella (Walker) (Lepidoptera: Pyralidae), and for timing treatments for this pest in almonds, Prunus amygdalus Batsch, and pistachios, Pistacia vera L. We compared, in almond and pistachio orchards, the number of eggs per trap in traps baited with almond meal, pistachio meal, or the current standard commercial bait. When considering cumulative eggs captured over an extended period, traps baited with pistachio meal prepared from previous-crop nuts generally captured a similar number of eggs compared with the commercial bait, and more eggs than those baited with almond meal prepared from previous-crop nuts. However, differences in eggs per trap between bait formulations were not as evident when examining individual weeks, particularly in weeks with few eggs per trap, as is typical when treatment decisions are made. The variance in eggs per trap was generally greater than the mean and increased with the mean and, when mean eggs per trap was low, most traps did not have eggs. We discuss implications of these findings for the relative importance of bait type and trap numbers for monitoring, and for experiments comparing egg trap performance.

Protracted emergence of overwintering Amyelois transitella (Lepidoptera: Pyralidae) from pistachios and almonds in California.

The navel orangeworm, Amyelois transitella (Walker), is the primary insect pest of pistachios and almonds in California. Four years of research (2002-2006) were conducted in Madera and Kern Counties to elucidate the pattern of adult emergence of the overwintering navel orangeworm population. Springtime emergence from unharvested (mummy) nuts was protracted (600 degree-days or more from 1 January of each year) and in 2004 and 2006 extended to mid-July. The population structure, sex ratio, and timing of emergence differed between pistachio and almond mummies. Pistachio populations had a significantly greater proportion of late stage individuals compared with almond mummies, 85.7 versus 34.1%. The sex ratio of adults emerging from pistachio mummies was significantly skewed with a ratio 57:43 male:female compared with 50:50 in almond mummies. Emergence from mummies held outdoors (variable temperature) began in early March and continued through early June in both pistachio mummies and almond mummies. The adult emergence pattern from pistachio mummies contained a single emergence peak, whereas emergence from almond mummies occurred in multiple peaks. These same patterns occurred when mummies were held at constant temperature, and the emergence peak from pistachio mummies occurred sooner. The impact of these findings on understanding navel orangeworm population dynamics and current control recommendations is discussed.

Economic injury level of the psyllid, Agonoscena pistaciae, on Pistachio, Pistacia vera cv. Ohadi.

The pistachio psylla, Agonoscena pistaciae Burckhardt and Lauterer (Hemiptera: Psyllidae) is a major pest of pistachio trees, Pistacia vera L. (Sapindalis: Anacardiaceae) throughout pistachio-producing regions in Iran. Different density levels of A. pistaciae nymphs were maintained on pistachio trees by different insecticide dosages to evaluate the relationship between nymph density and yield loss (weight of 1000 nuts). Psylla nymph densities were monitored weekly by counting nymphs on pistachio terminal leaflets. There was a significant reduction in weight of 1000 nuts as seasonal averages of nymphs increased. Regression analysis was used to determine the relationship between nymph density and weight of 1000 nuts. The economic injury levels varied as a function of market values, management costs, insecticide efficiency and yield loss rate and ranged from 7.7 to 30.7 nymphal days per terminal leaflet, based on weight of 1000 nuts.

Effects of mating disruption treatments on navel orangeworm (Lepidoptera: Pyralidae) sexual communication and damage in almonds and pistachios.

Two experiments in 2003 examined the effects of different ways of dispensing the principal sex pheromone component on sexual communication among and crop damage by the navel orangeworm, Amyelois transitella (Walker) (Lepidoptera: Pyralidae) in Nonpareil almonds and pistachios. A third experiment in 2004 compared the effect on navel orangeworm damage to several almond varieties using one of these dispensing systems by itself or with phosmet, phosmet alone, and an untreated control. Additional data are presented estimating release rates from timed aerosol release devices (PuffersNOW, Suterra LLC, Bend, OR) and hand-applied membrane dispensers. In 2003, puffers placed peripherally around 16-ha blocks, evenly spaced Puffers, and hand-applied dispensers reduced males captured in virgin-baited traps by > or = 95% and mating in sentinel females by > or = 69%, with evenly placed Puffers showing greater reduction of males captured and females mated compared with the other dispensing systems. Mating disruption with gridded Puffers or hand-applied devices in almonds resulted in an approximately 37% reduction of navel orangeworm damage (not significant), whereas peripheral Puffers resulted in a 16% reduction of navel orangeworm damage to almonds. In pistachios neither peripheral nor gridded Puffers reduced navel orangeworm damage, whereas insecticide reduced damage by 56%. In 2004, Puffers alone, insecticide alone, and both in combination significantly reduced navel orangeworm damage in Nonpareil almonds. In other, later harvested varieties, the insecticide treatments reduced damage, whereas the mating disruption treatment alone did not. We discuss application of these findings to management of navel orangeworm in these two crops.

Assessment of geographic and host-associated population variations of the carob moth, Ectomyelois ceratoniae, on pomegranate, fig, pistachio and walnut, using AFLP markers.

The carob moth, Ectomyelois ceratoniae (Zeller 1839) (Lepidoptera: Pyralidae) is the most important pest of pomegranate, Punica granatum L. (Myrtales: Ponicaceae), in Iran. In this study, 6 amplified fragment length polymorphism primer combinations were used to survey the genetic structure of the geographic and putative host-associated populations of this pest in Iran. An AMOVA was performed on test populations. Pairwise differences, Mantel test, multidimensional analysis, cluster analysis and migration rate were calculated for 5 geographic populations of E. ceratoniae sharing the same host, pomegranate. In another part of the study, 3 comparisons were performed on pairwise populations that were collected on different hosts (pomegranate, fig, pistachio and walnut) in same geographic regions. The results showed high within population variation (85.51% of total variation), however geographic populations differed significantly. The Mantel test did not show correlations between genetic and geographic distances. The probable factors that affect genetic distances are discussed. Multidimensional scaling analysis, migration rate and cluster analysis on geographic populations showed that the Arsanjan population was the most different from the others while the Saveh population was more similar to the Sabzevar population. The comparisons didn't show any host fidelity in test populations. It seems that the ability of E. ceratoniae to broaden its host range with no fidelity to hosts can decrease the efficiency of common control methods that are used on pomegranate. The results of this study suggest that in spite of the effects of geographic barriers, high within-population genetic variation, migration rate and gene flow can provide the opportunity for emerging new phenotypes or behaviors in pest populations, such as broadening host range, changing egg lying places, or changing over-wintering sites to adapt to difficult conditions such as those caused by intensive control methods.

Demographic parameters of wrinkling aphid of pistachio leaf, Forda hirsuta Mordv. (Hem.:Pemphigidae) insitu of Rafsanjan's orchards.

Wrinkling aphid of pistachio leaf, Forda hirsuta Mordv. (Hem.:Pemphigidae) is one of the pests of pistachio trees. This aphid caused the shrinkage, thickening, and changing the color of the pistachio leaves. Since insect feeds from leave edges, the thick and rolled upward wrinkles were formed, which its green color turned into red. Therefore, its economical damages are out of direct feeding from plant extraction, twisting pistachio leaves, and the decrease of photosynthesis. In this research two orchards and 10 trees that each of them were selected in Rafsanjan region and 58 fundatrix galls, 120 nymphal galls were marked and the demographic parameters for apterus parthenogenesis female of this aphid were calculated via daily observations. The results indicated that intrinsic rates of increase (r) for 1, 2 and 3 generations were 0.01, 0.0638 and 0.0575 femal/femal/days respectively, Doubling time (DT) were 69.31, 10.52 and 12.04 days, respectively, net fecundity rates were 1.71, 11.5 and 7.37 femal/femal/days, respectively, Net fertility rates were 1.11, 8.87 and 5.01 days, respectively, and mean generation times (Tc) were calculated to be 31.5, 32.2 and 31.87 days, respectively. Other reproductive parameters such as gross hatch rate, gross fecundity rate, gross fertility rate, mean age gross fecundity and fertility, mean age net fecundity and fertility, mean age hatch, finite rate of increase (lambda), intrinsic birth rate (b) intrinsic death rate (d) and daily reproductive rate were also calculated. The results revealed that population parameters especially r in the first and second generations were lowest and highest, respectively and mean longevity of fundatrix was 30 days and the born nymphs made separate galls in the edge of pistachio leaves.

Bioecology of pistachio twig borer moth Kermania pistaciella Amsel, in Rafsanjan and Isfahan-Iran.

Pistachio woodborer, Kermania pistaciella Amsel. (Lep.: Oinophylidae) is one of the most important pests of pistachio in Iran. Some of morphological characters and its biology were studied in two orchards in Rafsanjan. For this study, 80 pistachio branches were selected and visited once a week. The larval emergence was recorded and cocoon formation was located. Seasonal occurrence of adults were recorded based on marked cocoons on branches, caged cocoons and the number of captured males in cylindrical pheromone traps bated with virgin females. To determine sex ratio, six hundred cocoons were caged and emerged moths (males or females) were recorded. For determination of the pest oviposition period, totally 10 trees and from each tree two branches and two fruit clusters were randomly selected. Number of hatched and unhatched eggs was recorded in two days interval. By visual observation, larval tunneling behavior and locomotion were recorded on 50 branches and 50 fruit clusters. By visiting 15 pistachio producing regions in Isfahan, distribution areas, and percent of pest damage was determined. For this study, 110 pistachio branches were selected and visited twice a week. Percent of infestation of the pest in Burkhart, Ardestan, Najafabad, Kashan, Natanz, Naiin and Shahreza were 88.9+/-17, 15.25, 41, 0.0, 23.46+/-32 and 0, respectively. Overwintering was identified as 4th larval instars. This study disclosed 7th and 15th of March and 7th of April as larval emergence initiation, its peak, and termination, respectively and this takes usually 32 days whereas the pupae period was identified one month. Cocoons were formed underside of branches. Larval emergence time was not affected by location of branches and their vertical and horizontal position. By using marked cocoons, adults' emergence initiation, its peak and termination was recognized 18th and 20th of April and 2nd of May, respectively. Start, peak and end of male moth capture in pheromone traps was 18th and 26th of April and 2nd of May, respectively. The number of captured males in two last days at traps with two and four day's old females was not significantly different. Start, peak, and end of oviposition were 24th of April and 4-10th of May, respectively. The eggs hatched almost after one week and larvae tunneled in the pedicel of fruit cluster and branches and began to feed. This study revealed that, there are four larval instars in pistachio woodborer life cycle and different larval instars could be separated by measuring the width of head-capsule and pronotum. Sex ratio was also determined as 1 per 2.79. Also by collecting larvae and pupae from five different geographical areas and placing them in cages the emerged natural enemies were collected. Three parasitoids and two predators were collected. The parasitoids were identified as: Chelonus sp (Hym.: Braconidae), pteromalus sp (Hym.: Pteromalidae) and one unidentified species from Ichneumonidae. An ant as a pupal predator and a spider feeding on the larvae were collected.