Response of seven populations of the two spotted spider mite (Tetranychus urticae Koch) for spiromesifen on cucumber under plastic houses in Jordan.
Two-spotted spider mite (TSSM), Tetranychus urticae Koch (Acari: Tetranychidae), is a polyphagous pest with over 200 host plant species. It is a major pest on field crops, and plastic-houses crops, horticultural crops. ornamentals and fruit trees [20,19,3]. The development of the resistance is also known to be accelerated under confined environmental conditions such as plastic-houses . Since the mite has a very short and prolific life cycle, its resistance to acaricides has more readily emerged than the rate of the resistance in other pests, In addition, the mites resistance to certain acaricides has been shown to have cross resistance to other acaricides., Thus, most commercial acaricides have been often proved to be ineffective to control the field mite populations [15,4,9].
From field screening of various acaricides, it was speculated that TSSM has developed resistance to the most conventional acaricides, but toxicological data are very scarce and poorly documented . It is quite possible that TSSM susceptibility to acaricides would differ from one location to another of cucumber cultivation in Jordan. Therefore, it was important to monitor the acaricide susceptibilities of T. urticae populations that were collected from cucumber cultivation in Jordan and to evaluate the efficacy of testing acaricides. This study showed the results of laboratory-based tests that determined the response of six field populations (Al-Ramtha, Baq'a, Zyzya, Krimeh, Deir-Alla, and Karamah) and one susceptible strain of T. urticae to spiromesiten. The total production of cucumber in 2009 was about 113,000 ton. Fifty percent of this production was exported to different countries .
The Jordanian farmers rely heavily on acaricides to control the two-spotted spider mite. Therefore, they have increased the rate of application, applied a mixture of acaricides and applied acaricides more frequently than they should. They have complained about unsatisfactory results in controlling T. urticae [13,14]. The total quantity of acaricides imported to Jordan in 2009 was 120,000 liters and/or kilograms. The imported acaricides belong to 18 active ingredients. From the most important imported active ingredient was spiromesifin .
In addition to imported acaricides, there were more than 15 local agrochemical factories that produce different formulations of active ingredients. The majority of these factories production is for export and the quantity of their products that goes to the local market is unknown. Although TSSM represents a real threat to cucumber plantation under plastic-houses in Jordan, few toxicological studies on this pest have been conducted. Therefore, this study aimed to evaluate the susceptibilities of TSSM collected from cucumber cultivation in Jordan to the studied spiromesifen acaricide. and to find the resistance factors to the tested among the (TSSM) populations.
Materials and Methods
Populations of the mite:
Seven T. urticae populations of different origin were used in this study. Six of these populations were collected from cucumber plants grown under plastic houses conditions in different regions of Jordan. These geographical regions include Al-Ramtha (100 Km North West of Amman), Baq'a (20 Km north west of Amman), Zyzya (30 Km South of Amman), Krimeh (South of Jordan Valley), Deir-Alla (Central of Jordan Valley) and Karamah (South of Jordan Valley). These regions are considered the main area for cucumber production in the country. Other T. urticae was strain was obtained from Lattakia Center for Rearing and Production of Biological Agents (LCRPBA) in Syria. This strain was reared there for 5 years and they did not spray it by acaricides at all.
Production of bean plants:
Bean (Phaseolus vulgaris L. cv. Bronco) was chosen for rearing and for the toxicological tests of the (TSSM) because it is one of the mite's favorite host, and for its ease of producing transplants. Seeds of bean (Bronco, Asgrow, USA) were directly sowed inside 10 cm. pots. Potting media used was Peat-moss and Perlite with 3:1 ratio. Plants were infested with SSS T. urticae when they reached the true leaf stage. These plants were irrigated and replaced as needed. No pesticides were applied on the plants except for later the tested acaricides. These plants were grown under greenhouse conditions at a temperature of 25-35C, relative humidity of 45% to 60% and a photoperiod of L16: D8.
For use in the toxicological tests, polystyrene trays of 84 cells were filled by Peat-moss and Perlite (3:1 ratio). Then, these trays were sown by 1-2 bean seeds for each cell, after complete germination the cotyledon leaves were used in the toxicological tests.
Rearing of the Syrian susceptible strain:
Syrian T.urticae strain (SSS) was reared and maintained on Phaseolus vulgaris at the Faculty of Agriculture, University of Jordan, at temperature between 27-+5C and 57-+8% and a photoperiod of L16:D8. plants were irrigated and replaced as needed.
P. vulgaris seeds were sowed in 84 polystyrene trays filled with Peat-moss and Perlite at ratio of 3:1. Each cell was sowed with one or two seeds. Prior to germination, cotyledon leaves of seedling were used for all experimental sets.
The Tested acaricide:
Spiromesifen 24SC (240W/V) (Oberon). Produced by Bayer Crop Science 2002,with higher recommended rate of 180 mg/L H2O. Its Molecular formula is C23H 30O4 .
Preparing of stock solution:
For each test, fresh stock solution was prepared by dissolving a calculated quantity of the acaricide enough to run the whole concentrations needed. The acaricide was used as its commercial formulation.
Bioassay of the acaricide toxicity:
Toxicological bioassays were conducted according to the procedures described by IRAC . Cotyledon leaves from untreated bean plants were placed, lower side up, in Petri-dishes lined with water-saturated cotton wool. 25 adult females of T. urticae were introduced into each cotyledon leaf by using a binocular microscope and a fine paintbrush. 24 hours after mite release, each Petri-dish was sprayed with a constant amount of the acaricide solutions for 2 seconds using a hand sprayer. The sprayed Petri-dishes were left to dry for 30 minutes, then they were placed under room temperature. Mites condition was assessed by gentle probing with a fine paintbrush. Mites were classed as dead when they didn't move or displayed some movement not exceeding their body length. Mites which were able to move were considered alive. This assessment of mites conditions was recorded 48 hours after acaricide application. The LC50, and LC90 values and their 95% confidence limits were calculated from probit regressions using the SPSS13 program [16,21].
The application of the acaricide was done with four replicates per each concentration, and with seven concentrations for the acaricide. These concentrations were chosen based on preliminary studies and they were different for each T. urticae population. Tap water was sprayed as control. These experiments were carried out in the Pesticide laboratory at temperature between 25,5C and 57,8% relative humidity.
Data were subject to probit analysis  which incorporated Abbott's correction for natural mortality . The SPSS (version 13 USA) computer program was used for data analysis to estimate LC50 and LC90 values, regression coefficient (slope) and its standard error, intercept and its standard error, Pearson goodness of fit chi-square, expected mortality, and 95% confidence limits (95%CL) for effective level of concentrations. This programmed used normal equivalent deviate (NED) instead of probit numbers. However, NED numbers can be readily adjusted to probit by adding 5 to each NED number . Y value for each line estimated by probit regression was equal to 0.0 and 1.28 when LC50 and LC90 (X) value was converted to log base 10, respectively.
LC50 and LC90 values were considered significant when (95% CL) did not overlap. To determine the resistance factor (RF) for each population, the LC50 of each acaricide of the field population was divided by the corresponding LC50 for the susceptible strain. The resistance factors were categorized according to Fukami  as follows: low. RF<10, moderate 10<RF [less than or equal to] 40, high 40-60 and very high resistance >60. LC90 values in ppm divided by the higher recommended field rate in ppm were calculated and tabulated for each TSSM strain (Ratio value). Goodness of line fitting was checked by Chi-square test X2. According to Finney , the value of X2 at 0.05 level of probability equals to 14.1 at 5 degree of freedom (df). Results obtained in this study revealed that (X2) Goodness of fit chi-square were less than that tabulated for each regression line indicating goodness of fit at 0.05 level of probability.
Susceptibility of seven populations of T.urticae collected from different locations of cucumber production is illustrated in Table (1).
The estimated LC50 values in ppm of spiromesifen against tested field populations were (131.63) for Baqa followed by (122.36) for karamah, (110.71) for krimeh, (89.49) for Zyzya, (87.49) for Deir-Alla and (56.33) for Al-Ramtha, (Table 1). The highest LC 90 was against Baqa population while the lowest LC50 was against Al-Ramtha population. The LC50 values against Karamah and Baqa were not significantly different, while they differed significantly from Al-Ramtha, Zyzya, krimeh, and Deir-Alla populations. According to the resistance factor figures, it was concluded that all the populations were resistant to spiromesifen action. Based on estimated [LC.sub.50], Baq'a population was the most resistant population (RF = 6.33) then Karamah (RF = 5.88), Krimeh (RF = 5.32), Zyzya (RF = 4.32), Deir-Alla (RF = 4.2), and Al-Ramtha (2.71).
The higher recommended field rate (180ppm) for spiromesifen was higher than the estimated [LC.sub.90] (145,05) for Al-Ramtha population, while it was less than that for other populations, which ranged from 191 to 321ppm.
Monitoring of local populations for susceptibility towards acaricides is the first step in resistance management of T. urticae. It is essential to carry out acaricide resistance tests regularly to avoid resistance development in target mites. In addition, control tactics must depend on the use of different acaricides to avoid or delay resistance. Toxicity of spiromesifen to the tested field populations that shown in Table (1) indicated that there was moderate resistance to spiromesifen. Baq'a population showed the highest RF to spiromesifen (RF = 6.33) while Al-Ramtha population showed the lowest RF (RF = 2.71). Depending on mode of action, Spiromesifen is classified as inhibitors of acetyl CoA carboxylase . In Serbia  explained that spiromesifen affected fecundity, fertility and population growth rates of the treated T. urticae females. They concluded that the females treated with 180 ppm of spiromesifen laid no eggs and most died within few days after treatment. The present obtained results do not agree with those obtained by Marcic et al.  in Serbia. These conflicting results might be due to the bioassay test used. In the present study, results were taken on mortality of treated adult females of TSSM while Marcic et al.  studied the toxicity of spiromesifen on fecundity and fertility and population growth rates of TSSM. In the present study, it was noticed that some spiromesifen treated females were unable to lay eggs at all, while others laid abnormal flaccid shape eggs . The most distinguished character of treated females were the unusual big size, accumulations of eggs in the body while the dead treated females mostly known by the adhering of egg to their abdomen. In agreement with this result, Lee et al  pointed out that the resistance factors to an other acaricide named abamectin, varied between 0.3 to 19.5 towards the same mite T.urticae eight populations collected from different locations in Korea. In conclusions all the tested field populations were susceptible to spiromesifen. The resistance factor ranged from 2.71 to 6.33. In contrast to toxicity effect spiromesifen was an effective acaricide since it had a good residual effect, lasting for nine days after application. However, in order to gain safe cucumber product and to manage resistance development by T.urticae to acaricides the following are recommended:
Regular monitoring should be carried out to detect the extent of resistance to the pesticides used. Restricting the use of acaricides to which the magnitude of resistance is high. Establishing of baseline LC50 to new acaricide before widespread use which allow better monitoring of changes in susceptibility over time. Applying acaricides that have different active ingredients and different mode of action. Training growers on alteration of acaricides based on mode of action to facilitate long term sustainable spider mite management for agriculture in Jordan. Studying the mechanisms of resistance to acaricides is very important to prevent cross resistance between closely related groups.
Thanks are extended to the Dean ship of Research at University of Jordan and Agriculture Materials company administration for finance support. Thanks are also to the staff at the Lattakia (enter for Rearing and production of biological Agents in Syria for providing us the sensitive strain of the spider mite.
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(2) Mohammad Raed Kame L. Al-Lala, (1) Tawfiq M. Al-Antary and (3) Marwan I. Abdel-Wali
(1) Prof.of Pesticides (Corresponding author), Plant Protection Dept, Faculty of Agriculture, University of Jordan, Amman, Jordan
(2) Ph.D student
(3) Plant Protection Dept., Head Center of Agriculture Research and Extension, Ministry of Agriculture, Amman, Jordan.
Tawfiq M. Al-Antary, Plant Protection Dept, Faculty of Agriculture, University of Jordan, Amman, Jordan
Table 1: Susceptibilities of field-collected populations of T. urticae adult females to spiromesifen (Higher recommended field rate = 180 mg/L). Population [LC.sub.50](mg/l) [LC.sub.90](mg/l) name 95% CL (1) 95% CL Ar-Ramtha 56.33 d (6) 50.33-61.85 145.06 c (6) 127.33-172.94 Baq'a 131.63 a 119.50-142.49 286.52 ab 255.56-335.74 Zyzya 89.84 c 80.44-98.43 228.38 ab 199.67-294.83 Krimeh 110.71 b 104.61-116.48 191.48 b 176.22-214.28 Deir-Alla 87.49 c 78.44-95.74 215.39 b 189.96-255.38 Karamah 122.36 a 121.40-147.54 321.01 a 278.46-390.95 SSS 20.8 e 18.37-23.09 64.92 d 55.49-80.30 Population L.E.P.R (2) Slope [+ or -] name Y = a + b(x) S.E (3). Ar-Ramtha Y = -5.46 + 3.12(x) 3.12 [+ or -] 0.29 Baq'a Y = -8.04 + 3.79(x) 3.79 [+ or -] 0.38 Zyzya Y = -6.18 + 3.16(x) 3.16 [+ or -] 0.31 Krimeh Y = -11.01 + 5.39(x) 5.39 [+ or -] 0.49 Deir-Alla Y = -6.36 + 3.28(x) 3.28 [+ or -] 0.31 Karamah Y = -6.39 + 3.06(x) 3.06 [+ or -] 0.30 SSS Y = -3.42 + 2.59(x) 2.59 [+ or -] 0.24 Population Ratio (4) RF (5) Chi Square name [LC.sub.90] [LC.sub.50] Calculated Ar-Ramtha 0.81 2.71 4.7 Baq'a 1.59 6.33 6.0 Zyzya 1.27 4.32 7.1 Krimeh 1.06 5.32 6.2 Deir-Alla 1.2 4.2 5.7 Karamah 1.78 5.88 6.9 SSS 0.36 -- 4.8 1. 95% Confidence limits for [LC.sub.50] or [LC.sub.90] in ppm. 2. L.E.P.R. = Line Estimated by Probit Regression. 3. S.E. = Standard error. 4. Ratio [LC.sub.90] = [LC.sub.90]/higher recommended field rate. 5. R.F. Resistance Factor = [LC.sub.50] of field population/ [LC.sub.50] of susceptible population (SSS). 6. [LC.sub.50] or [LC.sub.90] values having different letters are significantly different (95% CL did not overlap).
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|Title Annotation:||ORIGINAL ARTICLE|
|Author:||Lala, Mohammad Raed Kame L. Al-; Antary, Tawfiq M. Al-; Abdel-Wali, Marwan I.|
|Publication:||Advances in Environmental Biology|
|Date:||Sep 1, 2012|
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