رضا فرشباف پورآباد

Effects of cyflumetofen and clofentezine were evaluated on Tetranychus urticae Koch, under laboratory conditions. LC50 values of cyflumetofen and clofentezine against eggs were 0.8 μg a.i./mL and 0.42 μg a.i./mL, respectively. Sublethal effects of these compounds on the life table parameters of T. urticae were evaluated by exposing mites to LC50 and LC25 concentrations. The developmental time of male and female T. urticae were significantly affected by both acaricides. Moreover, exposure to LC25 and LC50 concentrations of cyflumetofen or clofentezine significantly reduced net reproductive rate (R0), finite rate of increase (λ), and intrinsic rate of increase (r). The LC25 and LC50 treatments showed strong effects on both survivorship, fecundity, and intrinsic rate of increase. Our findings indicated that lethal and sublethal exposure to cyflumetofen or clofentezine result in significant alteration of life-history characteristics of T. urticae. Our findings suggest that these acaricides are promising candidates for use in management programs of this major pest.

Potato tuber moth, Phthorimaea operculella is an important pest of potatoes which annually impose economic loss in potato production especially in stored places. Chemical control with spray in farms and fumigation in storages is a conventional method for depressing the population of this pest. In addition, with harmful effects of synthetic pesticides on environment and human health, developing resistant population of insects, compel researchers and plant protection institutions to utilizing new safe control methods. Delaying the resistance by keeping effectiveness of pesticides by using synergists and sublethal concentrations is a resistance management method. Bacillus thuringiensis is an effective microbial control agent against some insects. Its effectiveness decreased because of developing resistance in some population of insects. Combination of Bt with some botanical biopesticides increases its efficiency that reported in many studies. In this study combination of B. thuringiensis with hexane, ethyl acetate and methanol extracts of Celery, Apium graveolens were investigated on mortality and some biological parameters of PTM. Celery is a biennial plant in the apiaceae family, which is rich in some compounds such as phenolic and flavonoids. These compounds could affect biology and physiology of insects. Combination of sublethal concentrations of Bt with extracts of celery subjected for study of their effects on mortality, synergistic effects and some biological parameters on PTM larvae in this paper.

Potato tuber moth colony established by preparing larvae from a health colony in department of plant protection, Azarbaijan Shahid Madani university. Larva reared on potato tubers in plastic containers. Rearing carried out in controlled condition of 26±2ºC, 60% RH and a photoperiod of 16:8 (L: D) hrs.Foliage of celery after well washing with distilled water, dried in shadow in laboratory condition. Coarsely powdered dry plants used for extraction. Extraction carried out using the maceration method with three solvents, hexane, ethyl acetate and methanol respectively. Sub-lethal concentrations of both agents, including LC10, LC30 and LC50, individually and in combination, were assayed by the oral bioassay method. The same age and size of potato leaves were impregnated with sub-lethal concentrations of both experimental materials using the dipping method. The first larval instars of PTM were transferred on treated leaves, and mortality was recorded in 48, 96 and 144 hours’ intervals. Larval developmental period and adult’s emergence as an important biological parameterrecorded in all treatments. In addition, the inhibitory activity of extracts on α-amylase and protease enzyme of PTM larva was considered.Inhibitory activity of celery extracts in two concentrations 1 and 10%, on PTM larvae alpha-amylase activity was assayed by the dinitrosalicylic acid procedure, using 1% soluble starch as a substrate described by Bernfeld, (1955). And protease inhibitory activity assayed using azocasein as substrate, with comparing enzyme activity in controls and present of extracts in reactions.All experiments were replicated three times. On-way ANOVA analysis of variance in random design was used for statistical analysis. Duncan’s multiple range tests used for comparing means. For determining the synergistic, additive or antagonistic effects of combinations, chi-square test conducted between observed and expected results (df=1).

The results revealed that ethyl acetate extract of celery in combination with Bt synergistically increased the mortality of larvae. Compared to the control, the combination of methanolic extract with Bt didn’t affect larval mortality. However, the combination of sub-lethal concentrations significantly increased larval life span compared with the control and adult emergence decreased especially in LC50 combinations. Increasing the mortality with exposure time progressing, observed in all combinations and the most larval mortality recorded in 144 hours’ exposure time. Ethylacetate extract of celery alone and in combination with Bt more than other solvents affected biological parameters of PTM. Extracts in the concentration of 10 percent in reactions inhibited α-amylase and protease activity of PTM larvae by 27.97-37.77 and 11.48-25.51 percent, respectively.Numerous researchers have reported increased efficiency of Bt in combination with other plant extracts for controlling various insects. The observed synergistic effects can be attributed to factors such as disrupting midgut properties, inhibiting digestive enzyme activity, and disrupting the insect's immune system by plant extracts.Inhibitory activity of celery extracts on digestive enzymes activity of PTM larva, well documented in this research which is in alignment with other reports. Effects on larval life span that with prolonging it affects population dynamics of insect pests, reported in same studies. This biological effect has adverse effect on development and physiology of insects and asynchronies developmental stage with environmental variations which finally decrease the population of next generations or overwintering insect’s density. Adults and pupa emergence percentage decreased in combination of Bt with celery extracts that same results were reported in other researches.

This study aims to determine the effects of different extracts (prepared by different solvents) of celery in combination with Bt on potato tuber moth larvae. Effects of different solvents in separating effective compounds on PTM, was not same. In certain instances, the methanol extract of celery showed a minor effect in enhancing the potency of Bt. However, strong synergistic effects were observed with ethyl acetate and hexane extracts at various integrated concentrations. Time also played a crucial role in achieving the desired mortality results, with mortality and synergistic effects increasing over time in all extract combinations with Bt. Particularly, towards the end of the exposure period, a synergistic effect was notably observed in all integrations involving ethyl acetate extract of celery.In addition, with mortality, larval developmental period also affected by combinations. Increasing in life span observed in all treatments with different ratios. Adult emergence decreased strongly in different treatments and this is so important in decreasing the population of PTM. Because of importance of Bt as an effective and safe insecticide and increasing the resistance by different pests such as PTM, management of insects in integrated programs using plant extracts will improve the efficiency of Bt. Decreasing the sprayed concentrations in combination with other control methods are two important resistance management toll in IPM programs. Celery is a potent plant for this goal and additional studies are needed.

The greater wax moth, Galleria mellonella is one of the most important pests of beehives in Iran and the world. Which leads to the death of weak honey bee colonies. Also, this insect is an important laboratory host for the isolation and mass production of the entomopathogenic fungi and nematodes. In this research, the effect of pollens of different plants including poppy, wild rapeseed, rapeseed and apple in artificial diet, were evaluated on some physiological and biological parameters of this insect. The highest amount of the last instar larval gut protein was recorded in the diet containing poppy pollen (20.22 mg/ml) and no significant difference was observed between the other treatments. The highest relative activity of digestive alpha-amylase and pectinase in the last instar larval gut was related to the diets containing poppy pollen (100 %), and the lowest relative enzymes activity was recorded in the diet containing rapeseed pollen (51.12 and 76.35 %, respectively). Larvae fed with the diet containing poppy pollen and the resulting pupae had a higher weight than the other diets (325 and 175 mg, respectively). The highest percentage of larval survival and pupation was related to the diets containing poppy (77.5 and 77.5 %), apple (62.5 and 70 %), rapeseed (52.5 and 47.5 %) and wild rapeseed (47.5 and 30 %) pollens, respectively. However, there was no significant difference in the percentage of adult emergence in different diets. Consequently, the diets containing rapeseed and wild rapeseed pollens compared to poppy and apple, are not suitable diets for rearing G. mellonella and it is better to use poppy pollen in the preparation of artificial diet for the mass rearing of this insect.

The striped bug, Graphosoma lineatum L. is one of the pests of Apiaceae family such as parsley, carrot, celery and some medicinally important plants. The adults and nymphs of this insect feed on seeds of host plants. The eggs of the striped bug have an important role in mass rearing program of some parasitoids. Nutritional supplements undertake an essential function in mass rearing of insects and increase the efficiency of production. Vitamins such as B group have different functions in physiology of insects. The growth and development of insects hinge on acquiring essential nutrients through food materials. Apart from their primary food source, insects rely on essential amino acids and certain minerals for proper nutrition. These crucial components must be obtained from their food sources. Symptoms of nutritional deficiency, as reported in various research studies, include delayed growth, weight loss, prolonged immature stages, increased mortality, wing deformities, alterations in mating behaviors, and various physiological issues. In this study the effects of some vitamins and amino acids on anatomy of salivary glands, total protein of body, developmental time and fecundity of G. lineatum were evaluated in controlled conditions.

Insects reared on parsley seeds adhered on the inner side of big plastic containers in controlled condition. Supplements including vitamins B12, B6, B1 and Serine and Phenylalanine amino acids selected for evaluation. These supplements used for the treatmnet of G. lineatum in four different concentrations dissolved in water. For evaluating the effects of nutritional supplements on anatomy of salivary glands, both glands separate after dissection of adult’s thorax, and dimension of glands including width and length of posterior and anterior lobs, measured by micrometer apparatus located on stereomicroscope. Total body protein measured by Kjeldahl apparatus after well drying the total body of insects in oven followed by fine powder of dry materials. Total protein was determined using a standard method, calculated by multiplying the nitrogen amounts by 6.38, a constant value. In distinct experiments, the impact of nutritional supplements on the duration of each nymphal stage was investigated. One-day-old eggs were employed for this purpose. Following treatment justification, the time required for the development of each instar was recorded on a daily basis. All tests were replicated three times, with distilled water used for the control treatment. The data were analyzed by One-way ANOVA experimental randomized design and Duncan’s multiple range test using SAS software used for comparing the means.

The results disclosed a significant impact of all vitamins and amino acids on all studied parameters. Notably, in the case of salivary glands, the posterior lobe exhibited a more pronounced effect in insects fed with B1, B6, and Phenylalanine. Different concentrations of vitamin B6, B1 and Phenylalanine affected total protein content of male and female insect’s body. Vitamin B12 had non-significant effect on male and female total body protein. Nutritional supplements changed the rate of laid and hatched eggs in addition with nymph’s developmental time. Vitamin B12 in the concentration of 0.005 g/l, B6 in the concentration of 0.25 g/l, B1 in the concentration of 0.5 g/l, Serine and Phenylalanine both in the concentration of 0.5 g/l showed the highest rate of laid and hatched eggs. Vitamin B6 and B1 in the concentration of 1 and 1.5 g/l resulted in prolonging the nymph’s life span in comparing with controls. The negative effects of high concentrations of nutritional supplements on fecundity and normal developmental period of nymphs well documented in this study. Metabolism, cell division, hormone synthesis, enzymes activity and many physiological parameters affects by supplements especially vitamins. Any disorder in utilization, absorption and inhibition of their function leads to biological and behavioral problems in insects. In some cases, the function of symbionts for compensation essential supplements documented.   

The results of this study indicate important function of vitamins B12, B6 and B1 on G. lineatum nymphal developmental period. In high concentrations, rate of growth decreased in comparison with controls. Salivary glands anatomy showed some changes in dimensions especially in posterior lobs in dose-depending manner. Number of eggs laid and hatched decreased especially in high concentrations of nutritional supplements. The results of this study will help to justify better meridic diet for rearing striped bug. In semi-artificial diets for rearing this insect, corporation of little concentrations (optimum) will improve the fitness of insects.

The cotton bollworm, Helicoverpa armigera (Hübner), is one of the most important pests of crops. Like other insects, this insect has an immune system against external threats such as various diseases, parasitoids, and chemical compounds. Phenoloxidase is a key factor in the immunity of insects and other arthropods. The present study investigated the lethal effects of four insecticides, indoxacarb, profenofos, chlorfluazuron, and hexaflumuron, and their effects on phenolic oxidase activity in cotton bollworm. Among the studied insecticides, in two bioassays (mixture of insecticide with artificial food and immersion of sugar beet leaves), Chlorfluazuron had the highest toxicity (LC50 = 1.71 and 3.11 mg ai/liter, respectively). Also, the highest phenol-oxidase activity at 24 h was for larvae treated with chlorfluazuron and hexaflumuron. Also, when treated with chlorfluazuron and hexaflumuron, the larvae fed on an artificial diet had higher phenoloxidase activity than those fed on sugar beet cultivars.

The Indian meal moth, Plodia interpunctella (Lepidoptera: Pyralidae) (Hubner, 1813) is one of the most important and well-known stored-product pests in most parts of the world. The larvae of this insect are omnivorous and have the ability to grow and reproduce on a wide variety of grains, dried fruits, dried vegetables, nuts, oil seeds, chocolate, animal feed and various processed products. Therefore, it causes economic losses to various agricultural and stored products. On the other hand, this insect is used as a host in the rearing of various useful insect species. In general, the Indian moth eats carbohydrate-rich foods, and its growth and development is highly dependent on biochemical compounds and especially the quality and quantity of nutrients in its diet. The enzymes like amylase and pectinase in the digestive system of the lepidoperan larvae, which are secreted by the cells of the midgut, play an effective role in the digestion and absorption of food. Investigating some of the biological and physiological properties of this insect on various types of foods will help to optimize the rearing of this insect for management studies and also as a host for rearing of other pest insect’s predators and parasitoids.
In this study, we investigated the effects of different diets, namely raisin, fig, pistachio, peanut, almond, and walnut, which are known to be the main hosts of the Indian meal moth, on the protein content in the last instar larval gut. We compared their protein patterns and content using SDS-PAGE. Additionally, we measured the activities of the digestive enzymes alpha-amylase and pectinase using the DNS (Dinitrosalicylic acid) assay. Alpha-amylase activity was assessed using 1% starch as a substrate, while pectinase activity was measured using pectin as the substrate. To facilitate a better comparison of the relative enzymatic activities, the activities were calculated as a percentage of the highest enzymatic activity. Furthermore, we compared several biological parameters, including the incubation period, larval period, pupal period, adult longevity, life span, and adult emergence.
The results of the study revealed that the Indian meal moth larvae had the lowest amounts of gut protein when fed on figs and raisins, while the highest amounts were observed in larvae fed on pistachios, peanuts, almonds, and walnuts. The determination of protein levels using the Bradford method and SDS-PAGE gel electrophoresis yielded consistent results, clearly indicating the protein concentrations in the samples. The activity of the alpha-amylase enzyme in the larvae's gut was highest in those fed on pistachios and lowest in those fed on figs. Similarly, the pectinase enzyme showed the highest activity in larvae fed on pistachios and the lowest activity in those fed on raisins. The embryonic period of the Indian meal moth remained consistent across different diets. However, when comparing the duration of the larval period, pupal period, and overall growth period, the shortest duration was observed in larvae fed on pistachios, while the longest duration was seen in those fed on raisins. The length of the growth period tends to increase when the insect is fed on less desirable food sources. Regarding lifespan, the shortest duration was observed in the diets consisting of almonds and pistachios, while the longest lifespan was recorded in those fed on raisins. When examining adult emergence, the diets of almonds and figs showed the highest and lowest values, respectively. In conclusion, pistachios, almonds, and walnuts were found to be more favorable diets for the Indian meal moth larvae compared to peanuts, figs, and raisins.
This study aimed to determine the optimal diet for rearing the Indian meal moth. The investigated diets, which are major food sources for this insect, were found to have an impact on the amount and concentration of intestinal protein in the last instar larvae, as well as the activity of alpha-amylase and pectinase enzymes. Significant differences were observed in various physiological and biological components among the diets. The highest values for all these components were observed in the larvae reared on a pistachio diet, while the lowest values were found in those reared on fig and raisin diets. Based on the findings of this research, it can be concluded that pistachio, walnut, and almond diets are favorable and suitable for laboratory breeding of the Indian meal moth, compared to peanut, raisin, and fig diets. Among these diets, pistachio was identified as the most optimal choice.

Bacterial soft rot caused by Pectobacterium species is one of the important and common diseases in Potatoes and vegetables. Disinfection of tubers or seeds with chemicals is one of the methods of disease management. Copper nanoparticles, like silver and gold nanoparticles, have a strong inhibitory effect on bacterial cells, but they are much cheaper and more accessible than them. This research was conducted to determine the effect of copper nanoparticles alone and in combination with oxytetracycline and streptomycin antibiotics on potato, carrot and onion soft rot.

The pathogen was isolated from rotten tissues of potato, carrot and onion, purified and identified by studying the phenotypic and genetic characteristics of the Pel-gene region using Pectobacterium specific primers (Y1 and Y2). The effect of copper nanoparticles, oxytetracycline and streptomycin, and their combination with copper nanoparticles on the pathogen growth was investigated in a completely randomized design experiment with three replications for each treatment in vitro.

Pectobacterium odoriferum was identified as pathogen based on phenotypic characteristics and genetic affinity. Streptomycin compared to oxytetracycline inhibited the pathogen growth more and their effect in combination with copper nanoparticles increased by 12 and 19.5%, respectively.

Copper nanoparticles can inhibit the growth of P. odoriferum, and in combination with antibiotics increase their effect.

Unrestricted use of chemical insecticides, especially at high concentrations, has reduced their efficacy. Plant-based pesticides, due to their relatively high safety levels to humans and environment as well as effective control on insects, have become a suitable alternative to conventional pesticides. The aim of this study was to investigate the biological and physiological effects of the formulated essential oil of Trachyspermum ammi (L.) on the larvae of Helicoverpa armigera Hüb. The obtained essential oil was formulated into an emulsion using Tween 20 as emulsifier. Chemical analysis of the extracted essential oil indicated that Terpinen (14.4%), Cymene (18.5%), and Thymol (61.6%) were the major compounds. The essential oil of T. ammi have a significant repellency rate on third and sixth instar larvae of cotton bollworm at different observed time intervals, with an average repellency rate of 61-76%. Regarding fumigant toxicity, the highest concentration of the essential oil (500 µL/L) resulted in 100% larval mortality. The LC50 value for fumigant toxicity measured as 82.73 µL/L. The duration of larval development significantly increased in treatments compared to control. In EO concentrations of 2.5-1.25 percent an increase was observed respectively in the duration of the larval stage by 9 and 7 days. The pupae and adult’s emergence affected in dose dependent manner. At concentration of 50 µl/l, 89% inhibition of protease enzyme recorded. With regards to the biological and physiological effects observed in the formulated T. ammi essential oil, with complimentary researches, it could be used in integrated management programs of cotton bollworm.

The Sunn pest, Eurygaster integriceps (Puton) (Hem.: Scutelleridae), is an economic pest of wheat that reduces the quantitative and qualitative properties of wheat products. We investigated the effect of E. integriceps feeding on six different wheat cultivars (Ghermez (Red), Noodle, Antanius, Sabalan, Azar 2 (with two types of cultivation), and Sardari) on the protein content in the adult’s gut and fat body and their digestive enzymatic activity. All qualified values of the insect feeding on wheat cultivars differed significantly. The least amount of adult weight and protein content of gut and fat body were observed in the insects fed on Ghermez (Red) cultivar, and the highest amount belonged to the Sardari cultivar. The same results for protein content were obtained from SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Also, the lowest and highest gut α-amylase, pectinase, and protease activities were in Ghermez and Sardari cultivars, respectively. Therefore, it was concluded that the type of wheat cultivar affects the food preference of this insect and, thereby, physiological parameters of the insect gastrointestinal tract. Planting a wheat cultivar like “Ghermez,” which may be resistant, can be a suitable and cost-effective method to decrease the chemicals applied against this pest.

In current research, the effect of four artificial diets containing cowpea, red kidney bean, navy bean and corn seed flour on protein content of 4th instars whole body; 5th and 6th instars larval and pre-pupal gut and fat bodies; and pupal whole body of the cotton bollworm (Helicoverpa armigera Hübner), were investigated. Also, the effect of different diets on the digestive α-amylase and pectinase was studied. Protein patterns were also compared using SDS-PAGE. According to the results, the lowest amount of protein in 4 th instars whole body and 5 th instars gut and fat bodies was seen in corn as diet and highest one was in navy bean diet. Insects reared on the corn as diet did not developed to 6 th larval stage during the experiment. Except for the protein content of 6 th instars fat bodies, no significant differences were observed in the other treatments of the 6 th instars and pre-pupa. The lowest amount of pupal protein was found in red kidney bean diet and the highest one was in navy bean diet. Due to the decrease in protein content in different diets, insect weight and the activity of digestive α-amylase and pectinase were also affected and significant differences were observed. The least weight and activity of enzymes were recorded in corn diet. Diferences in the amount of protein was also detected in SDS-PAGE. Consequently, artificial diets of corn and red kidney bean are not suitable for this insect rearing and white bean and cowpea are reported as appropriate diets.

Anagasta kuehniella Zeller (Lepidoptera: Pyralidae) causes the quality loss of the bread by feeding on flour. Disruption in insect’s digestive enzymes through transgenic plants, expressing enzyme inhibitors, is a methods of pest control. So, the aim of the current study was to investigate the inhibitory effects of proteinaceous extracts of two varieties of wheat (Sivand, Sardari) and Triticale against activity of digestive α-amylase of fourth (L4) and fifth (L5) instars larvae of A.kuehniella. Ammonium sulfate (70%) was used to extract proteins from seeds. The optimal temperature and pH of α-amylase activity was found 40°C and 10, respectively. The results showed that the highest dose of proteinaceous extracts of Sivand and Sardari (13 µg ml-1 protein) and Triticale (16 µg ml-1 protein) reduced the enzyme activity, 73.08%, 71.07% and 65.88% of L5 and 58.16%, 42.14% and 50.25% of L4, respectively. The lowest dose of proteinaceous extracts of Sivand and Sardari (0.812 µg ml-1 protein) and Triticale (1 µg ml-1 protein) inhibited the enzyme activity, 19.93%, 17.8% and 25.26% of L5 and 11.78%, 12.81% and 10.09% of L4, respectively. In the gel electrophoresis of enzyme without usage of inhibitors, one band was observed. At the highest dose of extracts, in the L5, amylase band disappeared and in the L4, waned band clearly. Band resolution was increased by reducing inhibitor dose. It was concluded that Sivand, Sardari and Triticale proteinaceous extracts can be used for management of A.kuehniella.

Plants have a set of protein molecules that are able to interfere with the digestion of herbivore insects and can be used as an approach to pest control. In this study, digestive alpha-amylase inhibition of Colorado potato beetle, Leptinotarsa decemlineata (Say), by proteinaceous seed extract of rapeseed, Brassica napus L. cv. Karaj3 was investigated. The most inhibitory effect of proteinaceous extracts on the alpha-amylase activity was at pH 5. Four precipitated rapeseed proteinaceous fractions; 0-30, 30-50, 50-70, and 70-100% saturation by ammonium sulfate, caused 0, 52, 12 and 0% inhibition on the gut alpha-amylase activity of the fourth instar larvae, respectively. Fraction 30-50% resulted in 84, 86, 74, 52 and 37% reduction in the L1, L2, L3, L4 and adult’s gut alpha-amylase activity, respectively. The enzyme inhibition kinetic test was conducted by the fraction 30-50% on the alpha-amylase of adults and the type of inhibition was determined uncompetitive. In the zymogram and studying inhibition on the gel, the inhibitory effect was seen as a reduction in intensity of the bands. In bioassays, different cultivars of potato; Agria, Burren, Picasso and Marx were used. Potato leaves were treated with extract and fed by larvae. No significant difference was observed in the duration of molting from third to fourth-instar larvae on any cultivars. But, weight of fourth-instar larvae was declined on Picasso and Marx and the percentage of fourth instar larval evaluation was decreased on Agria, Picasso and Marx. The alphaamylase activity of fourth instar larvae declined on Agria, significantly compared to the control.

During evolution, various compounds against herbivors were developed in plant species. Disturbance in digestive system by plant enzyme inhibitors is one of the effective methods in integrated pest managment programs. In this study, the effects of total Goebelia alopecuroides and Vicia ervilia seed extracts and their fractions were assessed against proteolytic activity of cotton bollworm. Total extracts of G. alopecuroides was effectively inhibited digestive proteinase of cotton bollworm compared with that of V. ervilia. The inhibitory effect of G. alopecuroides on Bovine trypsin and chymotrypsin as well as insects’ trypsin and chymotrypsin was higher than that of V. ervilia. Total seed extracts of V. ervilia significantly decreased digestive chymotrypsin activity. Ammonium sulfate precipitation of seed extract also showed significant inhibitory effect among the fractions. The fifth fraction of G. alopecuroides showed the highest inhibititory efect agianst digestive proteolytic activity of cotton bollworm and bovine trypsin. The first fraction of V. ervilia significantly inhibited total proteolytic activity.The first fraction of both plants seed extracts inhibited digestive trypsin of cotton bollworm compared to the other fractions. The third fraction of V. ervilia and the forth fraction of G. alopecuroides showed higher inhibitory effect on digestive chymotrypsin of cotton bollworm comperd with the other fractions. Comparing crude extracts effects of both plant species on laboratory and wild populations of cotton bollworm showed that wild ones was susseptible compared with the laboratory population.

Cotton boll worm "Helicoverpa armigera" is one of the most important pests in cultivated crops. Physiological understanding is one of the basic steps to control pests by chemicals and pathogens. In this study, Lipase enzyme characteristics in mid gut of cotton boll worm were studied. Studying of enzyme activity and other treatments were done by Auto analyzer and chromogenic method. The highest and langest enzyme activity were observed in the 6th instar larvae and male adults, respectively, The highest enzyme activity was observed at 40°C. Optimum pH for lipase enzyme activity in the mid gut was 9.5. After eight week enzyme activity at -20°C was 17% of its activity in the first week. While enzyme activity at 4°C after five weeks, was 6% of its activity in the first week, then enzyme activity completely stopped. The results of the study of enzyme activity at different times of starvation showed that enzyme activity after four day starvation was reduced to approximately 9% of its activity when larvae had fed. Enzyme activity was decreased by ethanol 96%, and 3, 4 and 5mM of NaCl and 3, 4 and 5mM of CaCl2 and different concentrations of SDS. Enzyme activity was increased by 1 and 2mM of NaCl and 5mM of KH2PO4. Different concentrations of Tris, EDTA concentration up to 25% ethanol and 4mM of KH2PO4 also had no observed effect on enzyme activity.

Plodia interpunctella Hubner (Lepidoptera: Pyralidae) is a cosmopolitan species known to infest a wide range of dried plant materials, including legumes and cereals. In this study, after confirmation of the presence of alpha-amylase gene in samples using gene specific primers, total RNA was extracted from fifth instar larvae of P. interpunctella, grown on the five different nutrition regimes including pistachio, white mulatto, plum, wheat and artificial diet (as control). After synthesis of cDNA, change in gene expression was studied with Real-time PCR assay. PCR performance test indicates 99 and 98% amplification efficiency for SiAmy and SiAmyC primers, respectively. Different nutrition regimes caused differential change in the expression of the gene. In comparison with the control, pistachio regime caused 5% increase in the expression of the alpha-amylase gene, wherease 10%, 27% and 60% decreases were observed in the larva grown on plum, wheat and mulatto respectively. Maximum variation in expression of alpha-amylase gene was observed in the larvae fed on mulatto. The results showed that the inhibition of alpha-amylase gene expression varied in the different sources of glucose in larvae fed on different diets and there was a negative correlation between the amount of sugar in the diet and alpha-amylase gene expression.

In this study, larval midgut α-amylase activity of the Mediterranean flour moth reared on wheat flour under laboratory conditions was studied. Enzyme activity was evaluated by using a commercial kit and an autoanalayzer device. Specific enzyme activity was measured as international unit per mg protein (IU/mg Protein). In this study, some α-amylase enzyme properties such as optimum temperature and pH of enzyme activity, enzyme activity during different hours of the day and night, enzyme activity at different developmental stages of adult male and female, effect of starvation period, and the effect of storage conditions of enzyme at specified periods in two different temperatures were studied. The results revealed that incubation temperature significantly affected enzyme activity for both sexes and optimal enzyme activity was detected in 40˚C for both male and females. pH values also significantly affected enzyme activity and the optimal pHs for amylase activity in both sexes were 5. Effects of enzyme concentration on α-amylase activity were significant in both sexes. Increasing the concentration of enzyme in both sexes caused a sharp increase in enzyme activity. Storage period in -20˚C did not decreased enzyme activity significantly in both sexes, but storage period in 4˚C in males significantly decreased the activity in comparison with the control. The results of studying enzyme activity during 24 hr. periods showed significant differences during day and night and the most and the least enzyme activities in both sexes were recorded in 12 and 24 o’clock, respectively. Enzyme activity in different developmental stages showed significant differences and the most enzyme activities were detected in 5th female larvae, 5th male larvae, 4th male and females, female and male pre-pupa, and female and male adults, respectively. In male and female pupae, no detectable activity was observed.Enzyme activity in male and female fifth instar larvae showed Significant difference. Starvation periods significantly affected the digestive enzyme activity in both male and female larvae and the trends of enzyme activity during 4 days of starvation was the same in both sexes. The most and the least enzyme activity were detected 2 and 4 days after starvation, respectively.

The flies of the tribe Terellini (Tephritidae) with feeding on seeds of asteraceus plants cause decreasing the seed amount of them as well as their gradual and sustainable control. In order to identification of fruit flies of the tribe Terellini in Ajabshir region, a survey was conducted during 2009- 2011. In total, thirteen species belonging to four genera were identified which all are new recorded for the studied area and Chaetorellia jaceae Robineau-Desvoidy, 1830 is newly reported for the Iran insectfauna. Identification keys to genera and species are provided. All the identified species are as biocontrol agents of the asteraceus weeds.

In order to identify the fruit flies of the genus Tephritis Laterille, 1804 (Diptera: Tephritidae) in Ajabshir region- East Azarbaijan province, a faunistic study was conducted during 2009-2010. In total, seven species were identified which all of them are as new records for the studying area and one species marked with an asterisk is being newly reported for Iran insect fauna: Tephritis cometa Low, 1840; T. formosa Low, 1844; T. hurvitzi Freidberg, 1758; T hyoscyami L., 1981; *T. nigricauda Low, 1856; T. postica Low, 1844 and T. praecox Low, 1844. A key for identification of the species is provided.

Types of insects digestive enzymes are generally associated with their food regimes; and at least، the predominant presence of an enzyme indicates that the diets of the target insects contain high levels of its substrate. Insect digestive enzymes show considerable different activities in relation to their individual conditions and developmental stages. In this study، it was revealed that the type of food (several kinds of seeds of umbelliferous plants) had no significant effects on the enzyme activity of the stripped bug، Graphosoma lineatum (L.). The results showed that the enzyme activity in male and female adults will differ 4. 5 and 3. 5 times، respectively، if the insects’ biological and physiological conditions are not similar. In general، females’ enzyme activity was greater than that of the males. Environmental temperatures (4-30 ˚C) did not affect the α-amylase activity. In starved insects (0-48 hours)، the most enzyme activities were observed in 24-hour (12. 32 U/mg protein) and 36-hour (14. 13 U/mg protein) starved insects which had not significant difference to each other. Enzyme activities were similar between the two salivary glands of an individual bug which shows that these two glands respond to the related stimuli simultaneously. During the circadian cycle، enzyme activities were different. This was related to the different conditions of the used insects. α-amylase activities in insects obtained from 10th to 17th generations had not significant differences to each other. Despite from the similarities between some results of the present study with the results of other researchers، we confirm this theory that we are not able to establish a generalization about insect digestive enzymes properties and each species should be investigated separately.

In most Heteroptera, the salivary glands complex consists of two principal and two accessory glands. Principal glands are structurally bilobed and are divided into anterior and posterior lobes. They are located at the thorax on each side of the midgut. Accessory glands are of two types: tubular and vesicular. In the present study, we investigated the sizes of different parts of the salivary glands of second to fifth instar nymphs and newly emerged, 10-days old, and 30-days old adults of the stripped bug, Graphosoma lineatum (L.), including the length of the salivary gland, length of anterior and posterior lobes, width of anterior and posterior lobes, and width of accessory gland. All sizes were measured by using a calibrated micrometer and a stereomicroscope. Experiments were carried out as factorial in CRD. With each instar and stage, gland size grows larger, whereas the ratio of its length to the body length decreases. Both in nymphs and adults, the effect of salivary glands situation on the size of its different parts was not significant which shows that the shape and size of right and left salivary glands of nymphs and adults are completely similar to each other. In the case of nymphs, the effect of instar was significant at 1% level which shows that an increase in developmental stages significantly increases the length of their salivary glands. In the case of adults, the effects of sexuality and adults’ life span were significant at 1% level. These show that the sizes of different parts of the females’ salivary glands are greater than that of the same age males and an increase in the adults’ life span would increase their salivary glands length.

Digestive enzyme inhibitors are proteinacious or nonproteinacious compounds which reduce an enzyme activity through attaching to its active site and/or its substrate. Nowadays, plant enzyme inhibitors are of great importance because 1) these have considerable effects on insect digestive enzymes and as a result on their development; and 2) the transgenic plants expressing them are safe. In this study, the effects of NaCl, ethylenediamine tetraacetate disodium dehydrate (EDTA), Tris, sodium dodecyl sulfate (SDS) and an alpha-amylase inhibitor derived from wheat kernels (WAAI) on alpha-amylase activity of adults of the stripped bug Graphosoma lineatum (L.) during 60 minutes incubation was studied. Distilled water was considered as the control. The results showed that the effects of the inhibitor type on adult alpha-amylase activity was significant (P

Studying insect digestive enzymes is important for understanding the function of their digestive systems. Nowadays, many attentions have focused on plant enzyme inhibitors and their application for producing transgenic plants. Determining the properties of all digestive enzymes is the first step that should be executed. In the case of beneficial insects, digestive enzyme activators would be of great importance. In 2004, some properties of salivary α-amylase of the stripped bug, Graphosoma lineatum was investigated for the first time. The experiments were repeated four times. Enzyme was active at 25 to 50 °C and its considerable activities were observed at 25 to 40 °C. However, the highest activity occurred at 37 °C; so, its optimum temperature was 37 °C. Enzyme activity decrease at pH4, 4.5 and 10 in relation to the elongation of incubation period (30-110 minutes). At pH5 to 9.5, enzyme was active throughout the incubation period without any decreasing. At pH10.5, enzyme was active until 30 minutes after incubation; and at pH11, the enzyme became completely inactive. Optimum pH for enzyme activity was 6.5. There was a linear relationship between enzyme concentration (2-25 pairs of salivary glands per one ml of buffer solution) and its activity. Enzyme activities in phosphate buffer (41.53 U/mg protein), Ringer’s infusion (42.52 U/mg protein), 5% dextrose (43.10 U/mg protein) and distilled water (41.63 U/mg protein) solutions were statistically similar, but significant difference was observed between its activity in 96°C (40.06 U/mg protein) and 75°C (14.07) ethanol solutions. Preservation of the enzyme solution at 4°C for 40 days had no effects on its activity.