Characterization of Phytophthora nicotianae isolates in southeast Spain and their detection and quantification through a real-time TaqMan PCR.

BACKGROUND: The soil-borne pathogens Phytophthora nicotianae and P. capsici are the causal agents of root and stem rot of many plant species. Although P. capsici was considered the causal agent in one of the main pepper production areas of Spain to date, evidence of the presence of P. nicotianae was found. We aimed to survey the presence of P. nicotianae and study the variability in its populations in this area in order to improve the management of Tristeza disease. RESULTS: A new specific primer and a TaqMan probe were designed based on the internal transcribed spacer regions of ribosomal DNA to detect and quantify P. nicotianae. Both morphological and molecular analysis showed its presence and confirmed it to be the causal agent of the Phytophthora disease symptoms in the studied area. The genetic characterization among P. nicotianae populations showed a low variability of genetic diversity among the isolates. Only isolates of the A2 mating type were detected. CONCLUSIONS: Not only is a specific and early detection of P. nicotianae essential but also the study of genetic variability among isolates for the appropriate management of the disease, above all, in producing areas with favorable conditions for the advance of the disease.

Solarization and biosolarization using organic wastes for the bioremediation of soil polluted with terbuthylazine and linuron residues.

Strategies for remediation of polluted soils are needed to accelerate the degradation and natural attenuation of pesticides. This study was conducted to assess the effect of solarization (S) and biosolarization (BS) during the summer season using organic wastes (composted sheep manure and sugar beet vinasse) for the bioremediation of soil containing residues of terbuthylazine and linuron. The results showed that both S and BS enhanced herbicide dissipation rates compared with the non-disinfected control, an effect which was attributed to the increased soil temperature and organic matter. Linuron showed similar behavior under S and BS conditions. However, terbuthylazine was degraded to a greater extent in the biosolarization experiment using sugar beet vinasse than in the both the solarization and biosolarization experiments using composted sheep manure treatments. The main organic intermediates detected during the degradation of terbuthylazine and linuron were identified, enabling the main steps of degradation to be proposed. The results confirm that both S and BS techniques can be considered as a remediation tools for polluted soils containing these herbicides.

Impact of biofumigation with solarization on degradation of pesticides and heavy metal accumulation.

A greenhouse study was conducted to evaluate the effect of biofumigation (with sheep and chicken manure) combined with solarization on the dissipation of pesticides (pyrifenox, DDT and dieldrin), and on soil metals accumulation. The treatments consisted of a control, and soil disinfestations by biofumigation combined with solarization (B+S) for two, four, five, six consecutive years. B+S enhanced the dissipation of pyrifenox with regard to control treatment. Significant differences were not detected among plots disinfected with B+S for consecutive years. On the other hand, B+S had no effect on the dissipation of DDT and dieldrin, probably due to the resistance of these pesticides to microbial degradation and/or high temperatures. Biofumigation is considered to be an environmentally safe practice, since no accumulation of heavy metals such as Cd, Co, Cu, Cr, Pb, Ni, or Zn was observed after the subsequent application of organic matter through the treatments.

Chitosan Increases Tomato Root Colonization by Pochonia chlamydosporia and Their Combination Reduces Root-Knot Nematode Damage.

The use of biological control agents could be a non-chemical alternative for management of Meloidogyne spp. [root-knot nematodes (RKN)], the most damaging plant-parasitic nematodes for horticultural crops worldwide. Pochonia chlamydosporia is a fungal parasite of RKN eggs that can colonize endophytically roots of several cultivated plant species, but in field applications the fungus shows a low persistence and efficiency in RKN management. The combined use of P. chlamydosporia with an enhancer could help its ability to develop in soil and colonize roots, thereby increasing its efficiency against nematodes. Previous work has shown that chitosan enhances P. chlamydosporia sporulation and production of extracellular enzymes, as well as nematode egg parasitism in laboratory bioassays. This work shows that chitosan at low concentrations (up to 0.1 mg ml-1) do not affect the viability and germination of P. chlamydosporia chlamydospores and improves mycelial growth respect to treatments without chitosan. Tomato plants irrigated with chitosan (same dose limit) increased root weight and length after 30 days. Chitosan irrigation increased dry shoot and fresh root weight of tomato plants inoculated with Meloidogyne javanica, root length when they were inoculated with P. chlamydosporia, and dry shoot weight of plants inoculated with both P. chlamydosporia and M. javanica. Chitosan irrigation significantly enhanced root colonization by P. chlamydosporia, but neither nematode infection per plant nor fungal egg parasitism was affected. Tomato plants cultivated in a mid-suppressive (29.3 ± 4.7% RKN egg infection) non-sterilized clay loam soil and irrigated with chitosan had enhanced shoot growth, reduced RKN multiplication, and disease severity. Chitosan irrigation in a highly suppressive (73.7 ± 2.6% RKN egg infection) sterilized-sandy loam soil reduced RKN multiplication in tomato. However, chitosan did not affect disease severity or plant growth irrespective of soil sterilization. Chitosan, at an adequate dose, can be a potential tool for sustainable management of RKN.

Field and laboratory selection of Frankliniella occidentalis (Pergande) for resistance to insecticides.

Response of western flower thrips, Frankliniella occidentalis (Pergande), to selection for resistance to insecticides commonly used to control this pest in Murcia (south-east Spain) was studied under field and laboratory conditions. In the field, plots within sweet pepper crops in commercial and experimental greenhouses were treated under different selection strategies: insecticide rotation versus formetanate reiteration, formetanate reiteration versus acrinathrin reiteration, and formetanate reiteration versus methiocarb reiteration. Thrips populations were sampled monthly and bioassayed against methiocarb, methamidophos, acrinathrin, endosulfan, deltamethrin and formetanate. In the laboratory, F occidentalis strains were selected against each insecticide for several generations. To evaluate cross-resistance, each selected strain was bioassayed with the other insecticides. Frankliniella occidentalis populations showed a rapid development of acrinathrin resistance, reaching high levels in field and laboratory conditions. Formetanate and methiocarb resistance were also observed, although development was slower and at moderate levels. Cross-resistances between acrinathrin/deltamethrin and acrinathrin/formetanate were detected under field and laboratory conditions. Formetanate/methiocarb cross-resistance was suspected in laboratory selections, but not in field assays. Simultaneous moderate resistance levels to the three specific insecticides against thrips (formetanate, methiocarb and acrinathrin) were shown in laboratory selection strains, indicating a general mechanism of resistance, probably metabolic.

Insecticide resistance in field populations of Frankliniella occidentalis (Pergande) in Murcia (south-east Spain).

Thirty-nine field populations of Frankliniella occidentalis (Pergande) were collected from different crops (sweet pepper, tomato, lettuce, artichoke, melon, cucumber, carnation, broad bean, peach and plum) in Murcia (south-east Spain). All populations were reared separately in the laboratory to obtain enough individuals for bioassays. Female thrips were bioassayed, using a standard topical application method, against methiocarb, methamidophos, acrinathrin, endosulfan, deltamethrin and formetanate. Methiocarb was the only insecticide that showed a high efficacy against F occidentalis at field dose rates. Acrinathrin and methamidophos were moderately effective, while endosulfan and deltamethrin were ineffective. Only moderate levels of resistance (Resistance Ratios at LC50 of 10-30) were detected for the selective insecticides methiocarb, formetanate and acrinathrin used against F occidentalis in crops where these insecticides are used intensively. This generalized and low level of resistance to these insecticides, coupled with a lack of efficacy for the three broad-spectrum insecticides, was observed even in intensively managed vegetable crops. Implementation of IPM strategies in Murcia has contributed to more successful insecticide anti-resistance management.

Enhanced dissipation of oxyfluorfen, ethalfluralin, trifluralin, propyzamide, and pendimethalin in soil by solarization and biosolarization.

This study was conducted to assess the effects of solarization and biosolarization on the degradation of oxyfluorfen, ethalfluralin, trifluralin, propyzamide, and pendimethalin. The experimental design consisted of 17 L pots filled with clay-loam soil, which were contaminated with the studied herbicides. Then, soil disinfection treatments were applied during the summer season, including a control without disinfection (C), solarization (S), and biosolarization (BS). Soil from five pots per treatment was sampled periodically up to 90 days. Herbicide dissipation rates were higher in both S and BS treatments with regard to the control. Similar dissipation rates were observed under S and BS for most of the herbicides studied, except oxyfluorfen and pendimethalin, which were degraded to a greater extent in the BS than in the S treatment. The obtained results showed that both solarization and biosolarization can be considered, in addition to soil disinfection techniques, such as bioremediation tools for herbicide-polluted soils.