Concepts in plant disease resistance

Resistance to nearly all pathogens occurs abundantly in our crops. Much of the resistance exploited by breeders is of the major gene type. Polygenic resistance, although used much less, is even more abundantly available. Many types of resistance are highly elusive, the pathogen apparently adapting very easily them. Other types of resistance, the so-called durable resistance, remain effective much longer. The elusive resistance is invariably of the monogenic type and usually of the hypersensitive type directed against specialised pathogens. Race-specificity is not the cause of elusive resistance but the consequence of it. Understanding acquired resistance may open interesting approaches to control pathogens. This is even truer for molecular techniques, which already represent an enourmously wide range of possibilities. Resistance obtained through transformation is often of the quantitative type and may be durable in most cases.

Effect of extracts from citric biomass, rusted coffee leaves and coffee berry husks on Phoma costarricencis of coffee plants

Phoma leaf spot, caused by Phoma costarricensis poses a serious threat to coffee (Coffea arabica) production, especially in the highlands of the state of Minas Gerais, Brazil. Extracts of citric biomass, coffee berry husks and coffee leaves severely affected by rust caused by Hemileia vastatrix, were evaluated against P. costarricensis. In an in vitro assay, aqueous extracts of rusted leaves and berry husks plus the commercial extracts based on citric biomass named Ecolife® and Agromil® were tested at various dilutions on the mycelial growth inhibition of P. costarricensis. In vivo, coffee seedlings maintained in glasshouse, were sprayed with these extracts seven days before inoculation of P. costarricensis. Only extracts from citric biomass had inhibitory effects on the fungus. In vivo, Ecolife® (5 ml/l), Agromil® (5 g/l) and the aqueous extract of rusted coffee leaves (dilution 1:6) reduced Phoma leaf spot. Both, Ecolife® and the extract of rusted coffee leaves were significantly more effective in reducing the area under the lesion progress curve when applied at lower doses, indicating a possible effect on the induction of resistance.

Identification of Races of Selected Isolates of Uromyces appendiculatus from Minas Gerais (Brazil) Based on the new international classification system

Twelve single-pustule isolates of Uromyces appendiculatus, the etiological agent of common bean rust, were collected in the state of Minas Gerais, Brazil, and classified according to the new international differential series and the binary nomenclature system proposed during the 3rd Bean Rust Workshop. These isolates have been used to select rust-resistant genotypes in a bean breeding program conducted by our group. The twelve isolates were classified into seven different physiological races: 21-3, 29-3, 53-3, 53-19, 61-3, 63-3 and 63-19. Races 61-3 and 63-3 were the most frequent in the area. They were represented by five and two isolates, respectively. The other races were represented by just one isolate. This is the first time the new international classification procedure has been used for U. appendiculatus physiological races in Brazil. The general adoption of this system will facilitate information exchange, allowing the cooperative use of the results obtained by different research groups throughout the world. The differential cultivars Mexico 309, Mexico 235 and PI 181996 showed resistance to all of the isolates that were characterized. It is suggested that these cultivars should be preferentially used as sources for resistance to rust in breeding programs targeting development lines adapted to the state of Minas Gerais.

Damage quantification and reaction of bean genotypes (Phaseolus vulgaris L.) to Meloidogyne incognita race 3 and M. javanica

The damage and the resistance levels of cultivars and accessions of common beans rescued in the South and mountain regions of Espírito Santo State, Brazil, to M. incognita race 3 and M. javanica parasitism were evaluated under a greenhouse. Four rescued bean genotypes ("FORT-10", "FORT-13", "FORT-16" and "FORT-19") and 2 commercial cultivars: "Pérola", and "Aporé", were tested. The cultivar "Rico-23" was included as standard of susceptibility to nematodes and non-inoculated plants constituted the control. Thus, the experiment was carried out in a completely randomized design in 3 (treatments considering nematodes) x 7 (genotypes and bean cultivars) factorial arrangement, with 7 replicates. Data were measured at 50 days after plant inoculation. For damage quantification, the following variables were evaluated: plant height (PHE), number of nodes (NNO), number of trifoliate leaves (NRT), fresh matter weight (FWE) and dry matter weight (DWE) of shoots, root weight (RWE), number of root nodules (NRO) and final population (FPO) of nematodes per root system. There were no significant differences between the effects caused by M. incognita and M. javanica, but both species showed inferior values of PHE, NNO, NRT, RWE, FWE and DWE compared to controls. Concerning the levels of resistance of bean plants to M. incognita, the genotypes "FORT-10", "FORT-13", "Aporé" and "FORT-16" behaved as moderately resistant, the cultivars "Rico 23" and "Pérola" low resistant, and the genotype "FORT-19" as highly susceptible. When parasitized by M. javanica, the beans "FORT-19", "Rico-23", "FORT-16" and "FORT-13" were low resistant, "Pérola" and "Aporé" susceptible and "FORT-10" highly susceptible.

Fetal heart rate responses during maternal resistance exercise: a pilot study

PURPOSE: To determine fetal heart rate (FHR) responses to maternal resistance exercise for the upper and lower body at two different volumes, and after 25 minutes post-exercise.METHODS: Ten pregnant women (22-24 weeks gestation, 25.2±4.4 years of age, 69.8±9.5 kg, 161.6±5.2 cm tall) performed, at 22-24, 28-32 and 34-36 weeks, the following experimental sessions: Session 1 was a familiarization with the equipment and the determination of one estimated maximum repetition. For sessions 2, 3, 4 and 5,FHR was determined during the execution of resistance exercise on bilateral leg extension and pec-deck fly machines, with 1 and 3 sets of 15 repetitions; 50% of the weight load and an estimated repetition maximum. FHR was assessed with a portable digital cardiotocograph. Results were analyzed using Student's ttest, ANOVA with repeated measures and Bonferroni (α=0.05; SPSS 17.0).RESULTS: FHR showed no significant differences between the exercises at 22-24 weeks (bilateral leg extension=143.8±9.4 bpm, pec-deck fly=140.2±10.2 bpm, p=0.34), 28-30 weeks (bilateral leg extension=138.4±12.2 bpm, pec-deck fly=137.6±14.0 bpm, p=0.75) and 34-36 weeks (bilateral leg extension=135.7±5.8 bpm, pec-deck fly=139.7±13.3 bpm, p=0.38), between the volumes(bilateral leg extension at 22-24 weeks: p=0.36, at 28-30 weeks: p=0.19 and at 34-36 weeks: p=0.87; pec-deck fly at 22-24 weeks: p=0.43, at 28-30 weeks: p=0.61 and at 34-36 weeks: p=0.49) and after 25 minutes post-exercise.CONCLUSION: Results of this pilot study would suggest that maternal resistance exercise is safe for the fetus.

Antimicrobial resistance and R-plasmid in Salmonella spp from swine and abattoir environments

Salmonella serovars isolated from swine are of particular interest not only because of the pathogenic potential for this animal species, but also due to its relevance with regard to public health. On basis of the profile of resistance to antimicrobials, 13 Salmonella strains were selected which belonged to the serovars Muenster (7), Derby (4), Typhimurium (1), and Braenderup (1). They were isolated from healthy swine as well as from the abattoir environment in the state of Rio de Janeiro. All strains of Salmonella were subjected to bacterial conjugation, and the E. coli K12 Nal r Lac+ F standard strain was used as receptor, with the purpose to verify the ability to transfer the resistance marks. Gene transfer phenomenon was detected in seven strains, and except SalmonellaTyphimurium which transconjugated to Sm, Tc and Su, the remaining ones were characterized by transferring mark Su only. By plasmidial analysis of strains used and their respective transconjugants, 63 Kb plasmid was found, which was probably related to S. Typhimurium resistance.

Resistance to β-lactam and tetracycline in Campylobacter spp.isolated from broiler slaughterhouses in southern Brazil

Abstract The study was carried out to screen and analyze the genetic characteristics of antimicrobial resistance in Campylobacter spp. from poultry sources. A total of 141 strains of Campylobacter isolated from samples of broilers of slaughterhouses in southern Brazil was identified by phenotypic and genotypic methods. Campylobacter isolates were evaluated for its antimicrobial susceptibility and the presence of resistance genes. The strains were investigated for antimicrobial susceptibility against two agents (ampicillin and tetracycline) by disk diffusion method. PCR assay was used to confirm the specie and the presence of ampicillin (blaOXA-61), tetracycline tet(O), and the energy-dependent multi-drug efflux pump (cmeB) genes. Campylobacter jejuni was the most ubiquitous; its presence was determined in 140 samples out of 141 (99.3%), whereas Campylobacter coli was found only in one of the contaminated samples (0.70%). The results obtained showed 65% and 35.5% of Campylobacter isolates resistant to β-lactams and tetracyclines, respectively. The cmeB gene responsible for multidrug resistance was detected in 26 isolates out 141 strains (18.5%). Moreover, 36 out of 141 Campylobacter strains (25.6%) were found to be resistant to at least two different antimicrobia resistance markers (β-lactams and tetracyclines).

A field survey of crop management practices and distribution of ALS resistant Euphorbia heterophylla in two states in southern Brazil

Herbicide resistance was reported in Brazil almost ten years ago. One of the main weeds with herbicide resistance is wild poinsettia (Euphorbia heterophylla). This work evaluates the distribution of ALS-resistant E. heterophylla in two states in southern Brazil and determines the major contributing management causes for weed resistance selection in the area. E. heterophylla seeds from 148 sites located in Paraná and Rio Grande do Sul were sampled during 2001 and 2002. Farmers provided specific site data for weed control, tillage system, crop rotation and harvesting operations during previous years. ALS resistant E. heterophylla biotypes were found widely distributed in the survey area. Data analysis suggests seed dissemination is unlikely to explain the widespread distribution of resistance. The most probable factor for the selection of the resistant E. heterophylla is the persistent high use of ALS-inhibiting herbicides over time. Indirect evidence is presented demonstrating the need to educate legislators and farmers about the importance of herbicide mixtures as a strategy to prevent herbicide resistance.

Selectivity of glyphosate in tank mixtures for RR soybean in sequential applications with mixtures only in the first or second application

Tank mixtures among herbicides of different action mechanisms might increase weed control spectrum and may be an important strategy for preventing the development of resistance in RR soybean. However, little is known about the effects of these herbicide combinations on soybean plants. Hence, two experiments were carried out aiming at evaluating the selectivity of glyphosate mixtures with other active ingredients applied in postemergence to RR soybean. The first application was carried out at V1 to V2 soybean stage and the second at V3 to V4 (15 days after the first one). For experiment I, treatments (rates in g ha-1) evaluated were composed by two sequential applications: the first one with glyphosate (720) in tank mixtures with cloransulam (30.24), fomesafen (125), lactofen (72), chlorimuron (12.5), flumiclorac (30), bentazon (480) and imazethapyr (80); the second application consisted of isolated glyphosate (480). In experiment II, treatments also consisted of two sequential applications, but tank mixtures as described above were applied as the second application. The first one in this experiment consisted of isolated glyphosate (720). For both experiments, sequential applications of glyphosate alone at 720/480, 960/480, 1200/480 and 960/720 (Expt. I) or 720/480, 720/720, 720/960 and 720/1200 (Expt. II) were used as control treatments. Applications of glyphosate tank mixtures with other herbicides are more selective to RR soybean when applied at younger stages whereas applications at later stages might cause yield losses, especially when glyphosate is mixed with lactofen and bentazon.

Limited occurrence of resistant radish (Raphanus sativus) to AHAS-inhibiting herbicides in Argentina

Radish has developed feral and weedy biotypes, which is a concern for agriculture around the world. In Argentina, it is one of the most widespread and troublesome crop weeds. In Brazil, this species has developed herbicide-resistance to acetohydroxyacid synthase (AHAS) inhibiting herbicides. The objective of this study was to record the presence of herbicide-resistant weedy radish plants in Argentina. In spring 2008, we found a small population of radish at the end of the flowering stage in an imidazolinone-tolerant canola field treated with imazethapyr. Screening and dose-response tests were conducted to two successive generations. They proved the biotype resistant status, and showed extensive survival (between 50 and 80% of control) to the application of a double dose of four AHAS‑inhibiting herbicides from two different chemical families (imidazolinones and sulfonylureas). Dose-response assays exhibited very high resistance for imazethapyr (LD50 = 2452.5 g a.i. ha-1, GR50 = 2926.9 g a.i. ha-1) and intermediate for metsulfuron (LD50 = 3.0 g a.i. ha-1, GR50 = 43.2 g a.i. ha-1). The acquisition of cross-resistance to different herbicide families would confer an adaptive and invasive advantage in agricultural environments to this biotype. Due to the herbicide rotation conducted in the field, the dispersion of this biotype was restricted. This is the first report of resistance in weedy radish in Argentina.

Reduction of seed production of red rice escapes in clearfield rice

Roguing is a practice used to reduce the seed source of red rice escapes to control in Clearfield-rice areas. However, there is great difficulty in performing it in large and heavily infested rice fields. This objective of this work was to evaluate the effects of the use of imazamox herbicide, applied in different rates and times, on plants of Clearfield-rice and red rice. Four experiments were conducted during the 2007/08 and 2008/09 growing seasons, in completely randomized block design and treatments arranged in factorial design, using three replications per treatment. The treatments had increasing rates of imazamox, application times and rice cultivars. The rice cultivars tested were IRGA 417, IRGA 422 CL, Avaxi CL and Puitá INTA CL. The variables evaluated were the number of panicles m-2, number of grains panicle-1, spikelet sterility in rice and red rice; and, rice grain yield and its components. The imazamox reduced the seed production of red rice escapes in a simulated situation of commercial Clearfield-rice area. The greater percentage reductions were obtained when this herbicide was applied at final formation of the panicle or panicle exertion of the red rice plant escapes to control. The Puitá INTA CL cultivar has high level of resistance to imazamox, independent of rate and application times tested, becoming the only alternative to the use of this practice.

Glyphosate influence on the physiological parameters of Conyza bonariensis biotypes

The objective of this study was to determine changes in gas exchange and inhibition of EPSPs, based on the accumulation of shikimic acid in horseweed biotypes resistant and sensitive to glyphosate. Two experiments were conducted in a factorial model. The first one evaluated horseweed biotypes (one resistant and one susceptible to glyphosate), and herbicide rates (0 and 1,080 g a.e. ha ¹) applied on the weed. In the second experiment, the horseweed biotypes (susceptible and resistant to glyphosate) were evaluated in five periods as following: 0, 3, 7, 10, and 14 days after herbicide application (DAH). The photosynthetic rate, transpiration, carboxylation efficiency, and water efficiency were determined using an infrared gas analyzer (IRGA), and shikimic acid concentration by HPLC. The application of glyphosate damaged the photosynthetic parameters of the susceptible biotype, causing complete inhibition of the photosynthetic rate, transpiration rate, carboxylation efficiency and water use efficiency, starting from the 7 DAH. On the other hand, total inhibition of the photosynthetic parameters was not observed for the resistant biotype. Shikimic acid accumulation occurred in both biotypes after glyphosate application but the susceptible biotype had the highest concentrations, indicating greater sensitivity of the enzyme EPSPs. The accumulation of shikimic acid in the resistant biotype indicates that the mechanism of resistance is not related to the total insensitivity of the enzyme EPSPs to glyphosate and/or that other resistance mechanisms may be involved.

Estimation of weed dry biomass and grain yield as a function of growth and yield traits under allelopathic weed management in maize

Growing concerns about toxicity and development of resistance against synthetic herbicides have demanded looking for alternative weed management approaches. Allelopathy has gained sufficient support and potential for sustainable weed management. Aqueous extracts of six plant species (sunflower, rice, mulberry, maize, brassica and sorghum) in different combinations alone or in mixture with 75% reduced dose of herbicides were evaluated for two consecutive years under field conditions. A weedy check and S-metolachlor with atrazine (pre emergence) and atrazine alone (post emergence) at recommended rates was included for comparison. Weed dynamics, maize growth indices and yield estimation were done by following standard procedures. All aqueous plant extract combinations suppressed weed growth and biomass. Moreover, the suppressive effect was more pronounced when aqueous plant extracts were supplemented with reduced doses of herbicides. Brassica-sunflower-sorghum combination suppressed weeds by 74-80, 78-70, 65-68% during both years of study that was similar with S-metolachlor along half dose of atrazine and full dose of atrazine alone. Crop growth rate and dry matter accumulation attained peak values of 32.68 and 1,502 g m-2 d-1 for brassica-sunflower-sorghum combination at 60 and 75 days after sowing. Curve fitting regression for growth and yield traits predicted strong positive correlation to grain yield and negative correlation to weed dry biomass under allelopathic weed management in maize crop.

Molecular Basis for Resistance to Fluazifop-P-Butyl in Itchgrass (Rottboellia cochinchinensis) from Costa Rica

Rottboellia cochinchinensis is an annual grass weed species known as itchgrass, or "caminadora" in America´s Spanish speaking countries, and has become a major and troublesome weed in several crops. The application of fluazifop-P-butyl at recommended rates (125 g a.i. ha-1) was observed to be failing to control itchgrass in a field in San José, Upala county, Alajuela province, Costa Rica. Plants from the putative resistant R. cochinchinensis population survived fluazifop-P-butyl when treated with 250 g a.i. ha-1 (2X label rate) at the three- to four-leaf stage under greenhouse conditions. PCR amplification and sequencing of partial carboxyl transferase domain (CT) of the acetyl-CoA carboxylase (ACCase) gene were used to determine the molecular mechanism of resistance. A single non-synonymous point mutation from TGG (susceptible plants) to TGC (putative resistant plants) that leads to a Trp-2027-Cys substitution was found. This Trp-2027-Cys mutation is known to confer resistance to all aryloxyphenoxyproprionate (APP) herbicides to which fluazifop-P-butyl belongs. To the best of our knowledge, this is the first report of fluazifop-P-butyl resistance and a mutation at position 2027 for a Costa Rican R. cochinchinensis population.

Gastroduodenal resistance and neural mechanisms involved in saline flow decrease elicited by acute blood volume expansion in anesthetized rats

We have previously demonstrated that blood volume (BV) expansion decreases saline flow through the gastroduodenal (GD) segment in anesthetized rats (Xavier-Neto J, dos Santos AA & Rola FH (1990) Gut, 31: 1006-1010). The present study attempts to identify the site(s) of resistance and neural mechanisms involved in this phenomenon. Male Wistar rats (N = 97, 200-300 g) were surgically manipulated to create four gut circuits: GD, gastric, pyloric and duodenal. These circuits were perfused under barostatically controlled pressure (4 cmH2O). Steady-state changes in flow were taken to reflect modifications in circuit resistances during three periods of time: normovolemic control (20 min), expansion (10-15 min), and expanded (30 min). Perfusion flow rates did not change in normovolemic control animals over a period of 60 min. BV expansion (Ringer bicarbonate, 1 ml/min up to 5% body weight) significantly (P<0.05) reduced perfusion flow in the GD (10.3 ± 0.5 to 7.6 ± 0.6 ml/min), pyloric (9.0 ± 0.6 to 5.6 ± 1.2 ml/min) and duodenal (10.8 ± 0.4 to 9.0 ± 0.6 ml/min) circuits, but not in the gastric circuit (11.9 ± 0.4 to 10.4 ± 0.6 ml/min). Prazosin (1 mg/kg) and yohimbine (3 mg/kg) prevented the expansion effect on the duodenal but not on the pyloric circuit. Bilateral cervical vagotomy prevented the expansion effect on the pylorus during the expansion but not during the expanded period and had no effect on the duodenum. Atropine (0.5 mg/kg), hexamethonium (10 mg/kg) and propranolol (2 mg/kg) were ineffective on both circuits. These results indicate that 1) BV expansion increases the GD resistance to liquid flow, 2) pylorus and duodenum are important sites of resistance, and 3) yohimbine and prazosin prevented the increase in duodenal resistance and vagotomy prevented it partially in the pylorus