JWSS - Journal of Water and Soil Science

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Solarization of field soil through mulching with transparent polyethylene sheets alone and in integration with farm yard manure (40 T/ha) and/or metalayl (25 kg/ha) during summer 1994-95 for five weeks in Isfahan resulted in an increase in temperature of 10 °C (± 1) and moisture preservation of up to 82%. Solarization effectively reduced foot-rot of cucumber (Phytophthora drechsleri) in the above treatments up to 88%, 96% and 95%, respectively.

Solarization, farm yard manure and their integration reduced the infection of Meloidogyne javanica up to 50%, 57% and 83% while plant parasitic nematodes were also reduced to 72%, 75% and 86%, respectively. The populations of free living nematodes increased only in farm yard manure and in integration with solarization by 30% and 53%, respectively.

Solarization effectively reduced the populations of almost all weeds by around 100%, except for Cyperus rotundus and Sonchus asper which were reduced by 59% and 44%, respectively.

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In an etiological study on stunted, nursery grown maple seedlings, a species of root-lesion nematode (Pratylenchus vulnus) and two species of fusarium (Fusarium oxysporum and F. solani) were isolated from the infected roots. Interactions between the nematode and the two fungi in a complete randomized design with six treatments and four replications were carried out under greenhouse conditions. Nematodes were surface sterilized and reared on sterilized carrot discs. Then the maple seedlings at two-leaf stage were inoculated with a population density of 40 nematodes/100 gr soil in pots. The results indicated singnificant differences between the treatments. Seven months after inoculation, the treatments with nematode alone showed a high mortality rate of about 75% along with a severe increase in population up to about 11.2 and 1266 nematodes/gr of soil and roots, respectively. In treatments with nematode and each of the two-fungus species, the deleterious effects of fungi on nematodes were observed through decrease of nematode population. It was also shown that F. oxysporum was more effective than F. solani in nematode control.

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OF2 and VAP genes, probably involved in signal transduction of sugarbeet nematode resistance, have already been cloned in bacterial vector by AFLP molecular marker and a two-hybrid system, respectively. To examine their capability to introduce resistance in sugarbeet, the genes were transferred to plant expression vectors. For this reason, OF2 gene after isolation was inserted within T-DNA of pAM194 binary vector, downstream of CaMV35S constitutive promoter and also inserted within T-DNA of modified pBin121 binary vector, downstream of HS1^pro-1 gene inducing promoter (responsible for nematode resistance). VAP gene after isolation was inserted within T-DNA of pAM194 plasmid, downstream of CaMV35S constitutive promoter. Thus, three new constructs were made in which genes of signal transduction pathway were expressed to give beet cyst nematode resistance. These plasmids were separately transferred to Agrobacterium rhizogenes, strain AR15834. In the next step, petiole explants of sugarbeet were inoculated with the bacterial cells. Transformation-derived hairy roots were analyzed by GUS staining and/or PCR and were then inoculated with nematode larvae. Primary results showed partial resistance against nematode larvae in some hairy roots. As a result, this resistance can be related to OF2 and VAP genes effect.

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In order to identify the plant-parasitic nematodes of important cereals (wheat, barley, and corn) in Isfahan Province, 120 soil and root samples were collected from the rhizosphere of these crops in various locations in the region during 1999 and 2000. Nematodes were extracted from soil by sieving and centrifugal sugar-flotation method. The specimens were killed and fixed in FGA (formaldehyde, glycerin and acetic acid 4:1:1) hot solution. The extracted nematodes were mounted in glycerin, permanent slides prepared, and studied by light microscope. The results revealed the presence of three species belonging to two genera of family criconematidae as follows: Criconemella antipolitana , C. xenoplax, Hemicycliophora poranga. H. poranga was extracted from rhizosphere of corn and reported for the first time from Iran. The most frequently occurring and dominant species was C. antipolitana that was isolated from soil around the roots of wheat and barley in several areas. C. xenoplax was extracted from a wheat field with the history of peach cultivation. These species are new records for Isfahan Province.

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Plant parasitic nematodes of Dorylaimida occur only in the Longidoridae. Longidorus is one of important genera in this family, which it,s species are parasite on many plants and act as a vector for some of plant viruses. From 1999 to 2001, more than 250 soil samples were collected from several localities of Iran. Nematodes were extracted, fixed (De Grisse, 1969) and mounted on glass slides in dehydrated glycerine. Using a light microscope equipped with a drawing tube, morphological and anatomical characteristics of nematodes were studied, body parts were drawn, measurements were carried out and the species of nematodes were determined. Species were compared with the original descriptions and variations reported from other regions of the world. Differences and similarities of the species with main description and closely related species were discussed. Three species, Longidorus africanus, L. iranicus and L. pisi were identified the latter species is first record for Iran. L. africanus were mainly occurred in southern regions while L.iranicus were mostly found in northern areas.

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The effect of Rathayibacter tritici on the movement of Anguina tritici larva and nematode function as vector of ear rot bacterium was conducted in the laboratory (Agarose plates) and greenhouse conditions. The results showed that the contact of nematode larva with high concentration of bacterium or long duration of nematode-bacteria contact can decrease the movement and the efficiency of nematode function as the vector of the disease, and in some cases it resulted in the mortality of the nematode. No differences were detected in the mobility of larva in the concentrations less than 10² CFU and less than 0.5 hour of nematode-bacteria contact times and their controls (exposed to water alone). Movement of the nematodes appeared to be random under these conditions. It can be concluded that Rathayibacter tritici did not act as an attractant to Anguina tritici larva. These results suggest that the attachment of a large number of bacteria to nematode (as an essential vector of the bacterium) would induce nematode weakness and mortality. So it is possible that ear rot bacterium can parasite ear cockle nematode, or the nematode is a host for this bacterium .

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  In this research, after optimization of sterilizing cyst and larvae of second stage of Heterodera schachtii, the possibility of using nematode on seedlings of sugar beet (Beta vulgaris L.) in in vitro condition was studied for developing larvae to cyst. For this purpose, non sterile cysts were extracted from infected soil and hatched into zinc chloride solution with concentration of 0.5gl^-1. Then, for preparation of sterile second stage larvae, several sterilizing treatments were used . Mean comparisons were performed between sterilized live larvae number by Duncan's method. Results showed that 70% ethanol for 1 minute followed by 5% sodium hypochlorite for 5 minutes and 0.1% sodium hypochlorite for 20 minutes were the best treatments for disinfecting cysts and larvae, respectively. In the next step, two nematode susceptible sugar beet varieties were applied to produce cyst from the larvae in in vitro culture. PG_oB medium containing different hormonal compositions was used to produce hairy roots and inoculation of seedling with sterilized larvae. After nematode inoculation tests, were the stained cysts were observed under stereomicroscope and counted 40 days later. Five to twelve cysts were formed on the roots of each seedling from two varieties. As a result, it seems that this technique can be used for sugar beet germplasm evaluation to screen nematode resistant genotypes in in vitro controlled condition.

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Polyphenol oxidase (PPO) activities were measured spectrophotometrically in tomato(Roma VF: resistant to Fusarium oxysporum f.sp. lycopersici and susceptible to Meloidogyne javanica) roots inoculated with nematode and fungi in four biological life stages of nematode. PPO specific activity showed a significant decrease in treatments (nematode + fungi) as compared with control plants. Results showed that nematode is able to change or suppress induction of this enzyme in spite of fungal induction. Nematode in all biological stages suppressed the induction of PPO activity in tomato roots but in young adult formation stage the suppression was higher than other life stages including giant cell formation, egg-laying and penetration. Native polyacrylamide gel revealed the presence of five isozymes of PPO with R_m values of 0.26, 0.31, 0.44, 0.50 and 0.63. Isoforms with Rm values of 0.26, 0.44, 0.50 and 0.53 showed significant changes in nematode + fungus inoculated tomato plants compared to plants inoculated with fungus alone. Based on the results obtained in this study, nematode infection resulted in suppression of tomato defenses against the fungi (as secondary pathogen) and intensified the fungal disease. Five PPO isoforms with R_m values of 0.26, 0.31, 0.44, 0.50 and 0.63 were shown in native polyacrylamide gel among which the isoform with R_m value of 0.31 had a thicker band in tomato plants inoculated with nematode or fungus alone. Two isoforms with R_m values of 0.44 and 0.53 in nematode-inoculated plants were similar to the corresponding bands in the healthy control. Isoforms with R_m values of 0.26, 0.44, 0.50 and 0.53 in nematode-fungus-inoculated tomato plants showed significant changes compared to plants inoculated with the fungus alone. Nematode infection of root tissue caused suppression of PPO specific activity and induced changes in PPO isoforms with R_m values of 0.26, 0.44, 0.50 and 0.63 more significantly than isoform 0.31.

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This research was conducted to study the effect of nematode (Meloidogyne javanica) on severity of tomato Fusarium wilt (Fusarium oxysporum f.sp. lycopersici race 1) and changes in Phenylalanine ammonia lyase (PAL) activity in split-root assays to show the possibility of systemic induced susceptibility caused by nematode to the fungus by local nematode infection. The results showed that the activity of nematode in one part of root confered susceptibility to Fusarium wilt in other part of the plant root and subsequently disease symptoms increased significantly in comparison to the control (without nematode inoculation). PAL specific activity showed a significant decrease in one half of split-root inoculated with fungus when the other half was inoculated with nematode as compared with the control (without nematode inoculation). Based on the results obtained in this study, nematode infection in one part of tomato plant root resulted in suppression of tomato defenses in all parts of the root, reduced tolerance or resistance to the fungus (as secondary pathogen) and intensified the fungal disease.

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Root knot nematodes (Meloidogyne spp.) cause yield loss in all countries, of which, M. javanica, is the most widespread species in Iran. In order to identify M. javanica, 100 infected root and soil samples of root knot nematode were collected from different regions of Kerman province. After purification of populations and identification of M. javanica based on morphological and morphometerical characters of females and second stage juveniles (J2), total DNA was extracted from eggs, J2 and female adults. Specific 670 and 1600 bp bands were amplified in all M. javanica populations using species-specific primer pairs including OPARjav / OPAFjav and Mjavf / Mjavr These specific bands could not be amplified in other species such as M. incognita and M. arenaria. It seems that, application of these species specific primers in comparison with morphological characters would be more applicable, leading to easier identification of M. javanica.

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In order to estimate the loss of rice caused by white tip nematode, (Aphelenchoides besseyi), to Alikazemi cv., two series of experiments were conducted in greenhouse and microplot conditions at Rice Research Institute, Rasht (Guilan province) during 2005-2006. The microplots sizes were 0.5×2 meter. Rice seedlings were inoculated by plastic tube method with 0, 100, 300, 500, 700 and 900 nematodes per plant. The experiments were arranged in greenhouse and microplot conditions as a completely randomized design and completely randomized block design with four replications, respectively. The factors under investigation consisted of symptoms (number of plants and Leaves infected), yield and population density of nematodes. The variance analysis of data showed significant differences among treatments in all parameters. The minimum infestation level leading to symptoms and yield loss in microplot condition (500 nematodes) was greater than greenhouse (300 nematodes). Regression analysis of data revealed that the yield reduction was mainly explained by population of nematodes in greenhouse and microplot (R2= 92.56, R2 = 91.55). Also, there was a positive correlation at 5% level of probability between disease incidence and nematode population (R2= 84.42, R2 = 75.27). The resulting equations from regression can be used to estimate crop loss from population of nematodes.

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Plant parasitic nematodes, especially root knot nematodes, cause damage to most of agricultural products, and many efforts have been done to control them. In recent years, application of industrial waste and wastewater sludge as organic fertilizers in agriculture has been increased. To investigate the effects of sewage sludge on root knot nematode pathogenicity in tomato, different weights of sewage sludge (0, 4, 8, 15 and 25% of sludge in the soil) in soil were added as a completely randomized design with six replications. Analysis variance and mean comparison of growth indices of plants showed significant effects of treatments. For example, means of stem length of plants increased in soil with more amount of sewage sludge. This relationship was also observed in other indices and nutrients elements. So using sewage increased nitrogen, phosphorous, calcium and magnesium in tomato shoots inoculated with nematodes while the change of potassium was very small. Application of sewage sludge decreased the number of galls, egg-masses and eggs in egg-mass of root knot nematode.