Variation in the Internal Transcribed Spacer Region of Phakopsora pachyrhizi and Implications for Molecular Diagnostic Assays.

Phakopsora pachyrhizi, the causal agent of soybean rust (SBR), is a global threat to soybean production. Since the discovery of SBR in the continental United States, quantitative polymerase chain reaction assays based on the internal transcribed spacer (ITS) ribosomal DNA locus were established for its rapid detection. However, insufficient data were initially available to test assays against factors that could give rise to misidentification. This study aimed to reevaluate current assays for (i) the potential for false-positive detection caused by nontarget Phakopsora species and (ii) the potential for false-negative detection caused by intraspecific variation within the ITS locus of P. pachyrhizi. A large amount of intraspecific and intragenomic variation in ITS was detected, including the presence of polymorphic ITS copies within single leaf samples and within single rust sori. The diagnostic assays were not affected by polymorphisms in the ITS region; however, current assays are at risk of false positives when screened against other species of Phakopsora. This study raises caveats to the use of multicopy genes (e.g., ITS) in single-gene detection assays and discusses the pitfalls of inferences concerning the aerobiological pathways of disease spread made in the absence of an evaluation of intragenomic ITS heterogeneity.

Coumestrol Confers Partial Resistance in Soybean Plants Against Cercospora Leaf Blight.

Although previous research showed that the purple symptom of Cercospora leaf blight (CLB) is associated with lower biomass of Cercospora cf. flagellaris and lower concentrations of cercosporin, a reactive oxygen species producer, as compared with blighted leaves, the hypothesis that the purple symptom is a plant reaction to the pathogen has never been tested. In this study, we demonstrated that high levels of coumestrol (COU) were associated with purple symptoms of CLB and that COU has strong antioxidant activity. Additionally, we found that COU is restricted to the pigmented areas of purple leaves, and the pigmentation is restricted to the adaxial surfaces, suggesting that COU may be acting as a sunscreen. Even though COU is associated with the purple leaf symptom, this coumestan is not the direct cause of discoloration in that COU is colorless. Quantification of chlorophyll a and b and total carotenoids suggested that blighted but not purple or asymptomatic leaves were undergoing photooxidative stress. Because the purple symptom is associated with high COU concentrations, lower biomass of C. cf. flagellaris, and lower cercosporin concentrations, we conclude that the purple symptom is a disease resistance reaction, mediated in part by COU, which provides a high level of antioxidant activity and, hence, partial resistance.

The Duffy antigen receptor for chemokines regulates asthma pathophysiology.

BACKGROUND: The Duffy antigen receptor for chemokines (DARC) is an atypical receptor that regulates pro-inflammatory cytokines. However, the role of DARC in asthma pathophysiology is unknown. OBJECTIVE: To determine the role of DARC in allergic airways disease in mice, and the association between DARC single nucleotide polymorphisms (SNPs) and clinical outcomes in patients with asthma. METHODS: Mice with targeted disruption of the Darc gene (Darc∆E2 ) or WT mice were challenged over 3 weeks with house dust mite (HDM) antigen. Allergic airways disease was assessed 24 hours and 7 days following the final challenge. Additionally, associations between DARC SNPs and clinical outcomes were analysed in a cohort of poorly controlled asthmatics. RESULTS: Total airway inflammation following HDM did not differ between Darc∆E2 and WT mice. At 24 hours, Darc∆E2 mice had increased airway hyperresponsiveness; however, at 7 days airway hyperresponsiveness had completely resolved in Darc∆E2 but persisted in WT mice. In poorly controlled asthmatics, DARC SNPs were associated with worse asthma control at randomization and subsequent increased risk of healthcare utilization (odds ratio 3.13(1.37-7.27), P=.0062). CONCLUSIONS AND CLINICAL RELEVANCE: Our animal model and human patient data suggest a novel role for DARC in the temporal regulation in asthma pathophysiology and symptoms.

Cercospora cf. flagellaris and Cercospora cf. sigesbeckiae Are Associated with Cercospora Leaf Blight and Purple Seed Stain on Soybean in North America.

Cercospora kikuchii has long been considered the causal agent of Cercospora leaf blight (CLB) and purple seed stain (PSS) on soybean, but a recent study found C. cf. flagellaris associated with CLB and PSS in Arkansas (United States) and Argentina. Here, we provide a broader perspective on the distribution of C. cf. flagellaris on soybean and alternate hosts within the United States (Arkansas, Louisiana, Mississippi, Missouri, and Kansas). We used a multilocus phylogenetic approach with data from actin, calmodulin, translation elongation factor 1-α, histone 3, the internal transcribed spacer region of rDNA and the mating-type locus to determine that two species, C. cf. flagellaris (200 of 205 isolates) and C. cf. sigesbeckiae (five of 205 isolates), are associated with CLB and PSS in the United States. In our phylogenetic analyses, species-level lineages were generally well-supported, though deeper-level evolutionary relationships remained unresolved, indicating that these genes do not possess sufficient phylogenetic signal to resolve the evolutionary history of Cercospora. We also investigated the potential for sexual reproduction in C. cf. flagellaris in Louisiana by determining the frequency of MAT1-1/MAT1-2 mating-type idiomorphs within the Louisiana population of C. cf. flagellaris. Though the MAT 1-2 idiomorph was significantly more common in our collection, the presence of both mating types suggests the potential for sexual reproduction exists.

Development of a Quantitative Polymerase Chain Reaction Detection Protocol for Cercospora kikuchii in Soybean Leaves and Its Use for Documenting Latent Infection as Affected by Fungicide Applications.

Cercospora leaf blight (CLB) of soybean, caused by Cercospora kikuchii, is a serious disease in the southern United States. A sensitive TaqMan probe-based real-time quantitative polymerase chain reaction (qPCR) assay was developed to specifically detect and quantify C. kikuchii in naturally infected soybean plants. The sensitivity was 1 pg of genomic DNA, which was equivalent to about 34 copies of genome of C. kikuchii. Using this qPCR assay, we documented a very long latent infection period for C. kikuchii in soybean leaves beginning at the V3 growth stage (as early as 22 days after planting). The levels of biomass of C. kikuchii remained low until R1, and a rapid increase was detected from the R2/R3 to R4/R5 growth stages shortly before the appearance of symptoms at R6. The efficacy of various fungicide regimens under field conditions also was evaluated over a 3-year period using this qPCR method. Our results showed that multiple fungicide applications beginning at R1 until late reproductive stages suppressed the development of C. kikuchii in leaves and delayed symptom expression. Different fungicide chemistries also had differential effects on the amount of latent infection and symptom expression during late reproductive growth stages.

Sudden Vegetation Dieback in Atlantic and Gulf Coast Salt Marshes.

Salt marshes rank as the most productive ecosystems on the planet. Biomass production can be greater than 3 kg dry matter/m2/year, which is 40% more biomass than tropical rainforests produce. Salt marshes provide multiple benefits to mankind. For example, coastal communities receive protection from storm surges and wave erosion. Salt marshes absorb excess nitrogen and phosphorus from sewage and fertilizer run-off into rivers, which, in turn, prevents algal blooms and hypoxia in coastal waters. In addition, these unique ecosystems provide habitat and shelter for many hundreds of species of shellfish, finfish, migratory and sedentary birds, and other marine animals. Despite the richness in animal species, the intertidal marshes of the salt marsh ecosystem are dominated by only a few plant species. Of these, the most prevalent plant species in a marsh are the tall and short forms of smooth cordgrass (Spartina alterniflora). The first recorded account of a dieback in a U.S. salt marsh was in the early 1990s in the Florida panhandle where patches of Sp. alterniflora as large as 1 ha died. This article explores possible causes of Sudden Vegetation Dieback.

Effects of Simplicillium lanosoniveum on Phakopsora pachyrhizi, the soybean rust pathogen, and its use as a biological control agent.

The fungus Simplicillium lanosoniveum was isolated from soybean leaves infected with Phakopsora pachyrhizi, the soybean rust pathogen, in Louisiana and Florida. The fungus did not grow or become established on leaf surfaces until uredinia erupted, but when soybean rust signs and symptoms were evident, S. lanosoniveum colonized leaves within 3 days and sporulated within 4 days. Development of new uredinia was suppressed by about fourfold when S. lanosoniveum colonized uredinia. In the presence of S. lanosoniveum, uredinia became increasingly red-brown, and urediniospores turned brown and germinated at very low rates. Assays using quantitative real time polymerase chain reaction revealed that the fungus colonized leaf surfaces when plants were infected with P. pachyrhizi, either in a latent stage of infection or when symptoms were present. However, when plants were inoculated before infection, there was no increase of DNA of S. lanosoniveum, suggesting that the pathogen must be present in order for the antagonist to become established on soybean leaf surfaces. We documented significantly lower amounts of DNA of P. pachyrhizi and lower disease severity when soybean leaves were colonized with S. lanosoniveum. These studies documented the mycophilic and disease-suppressive nature of S. lanosoniveum.

Reversible blunting of arousal from sleep in response to intermittent hypoxia in the developing rat.

Arousal is an important survival mechanism when infants are confronted with hypoxia during sleep. Many sudden infant death syndrome (SIDS) infants are exposed to repeated episodes of hypoxia before death and have impaired arousal mechanisms. We hypothesized that repeated exposures to hypoxia would cause a progressive blunting of arousal, and that a reversal of this process would occur if the hypoxia was terminated at the time of arousal. P5 (postnatal age of 5 days), P15, and P25 rat pups were exposed to either eight trials of hypoxia (3 min 5% O(2) alternating with room air) (group A), or three hypoxia trials as in group A, followed by five trials in which hypoxia was terminated at arousal (group B). In both groups A and B, latency increased over the first four trials of hypoxia, but reversed in group B animals during trials 5-8. Progressive arousal blunting was more pronounced in the older pups. The effects of intermittent hypoxia on heart rate also depended on age. In the older pups, heart rate increased with each hypoxia exposure. In the P5 pups, however, heart rate decreased during hypoxia and did not return to baseline between exposures, resulting in a progressive fall of baseline values over successive hypoxia exposures. In the group B animals, heart rate changes during trials 1-4 also reversed during trials 5-8. We conclude that exposure to repeated episodes of hypoxia can cause progressive blunting of arousal, which is reversible by altering the exposure times to hypoxia and the period of recovery between hypoxia exposures.

Population structure of Cercospora kikuchii, the causal agent of Cercospora leaf blight and purple seed stain in soybean.

Random amplified polymorphic DNA (RAPD) and microsatellite-primed polymerase chain reaction (MP-PCR) were used to characterize 164 isolates of Cercospora kikuchii, most of which were collected from Louisiana. Plant tissue (seeds versus leaves), but not host cultivar, had a significant impact on pathogen population differentiation. Cluster analysis showed that the Louisiana population was dominated by a primary lineage (group I) with only a few Louisiana isolates belonging to the minor lineage that also included the non-Louisiana isolates (group II). A previous study showed that isolates could be differentiated according to vegetative compatibility groups (VCGs). However, RAPD and MP-PCR data demonstrated that isolates of C. kikuchii were not generally clustered according to these VCGs. Furthermore, genetic relationships within and between VCGs were examined using sequences of the intergenic spacer region of rDNA. These analyses showed that VCG is not an indicator of evolutionary lineage in this fungus. Our results suggest the likely existence of a cryptically functioning sexual stage in some portion of the C. kikuchii population.

Viability of Phakopsora pachyrhizi Urediniospores Under Simulated Southern Louisiana Winter Temperature Conditions.

Soybean rust, caused by Phakopsora pachyrhizi, originally occurred in Asia. It has now spread to South America and the continental United States. This disease has the potential to cause severe economic losses to U.S. soybean growers, especially in the south, where the environmental conditions are more favorable to P. pachyrhizi survival during winter. In the present study, the effect of simulated southern Louisiana winter temperature conditions (12°C, 14-h days and 1°C, 10-h nights with 75% relative humidity) on soybean rust urediniospore viability was examined. It was found that urediniospore viability declined rapidly from 72 to 40% after 1 day and then decreased gradually to 17% after 7 days and 11% after 60 days. Spores stored under southern Louisiana winter conditions for 60 days still produced pustules on inoculated leaves. In comparison, the viability of spores stored at room temperature decreased gradually and reached 0% at the end of 60 days. Winter temperature treatment not only reduced spore viability but also decreased germ tube growth. In addition, soybean rust spores recovered from overwintered dry kudzu leaves were also found viable. This study indicates that soybean rust spores could survive southern Louisiana winter conditions and initiate a new cycle of infection in the next growing season.

First Report of Soybean Rust Caused by Phakopsora pachyrhizi in the Continental United States.

Asian soybean rust, caused by Phakopsora pachyrhizi Sydow, has been known to occur in the eastern hemisphere for nearly a century. More recently, it was reported from Hawaii in 1994, eastern and southern Africa from 1996-1998, Nigeria in 2001, and Brazil and Paraguay in 2002. Aerobiological models suggested that urediniospores of the pathogen would be disseminated on wind currents to the continental United States in association with tropical storms if the disease became established north of the equator during hurricane season (U.S. Soybean Rust Detection and Aerobiological Modeling online publication at www.aphis.usda.gov/ppq/ ep/soybean_rust/ ). Since soybean rust was observed at approximately 5°N latitude in South America before several hurricanes impacted the continental United States in September 2004, it seems likely that the introduction was associated with at least one of these tropical storms, especially hurricane Ivan. Symptoms of the disease were first observed on soybean (Glycine max (L.) Merr.) in the continental United States on November 6, 2004 in a field near Baton Rouge, LA. Typical pustules and urediniospores on infected leaves were readily apparent when viewed with a dissecting microscope. Urediniospores were obovoid to broadly ellipsoidal, hyaline to pale yellowish brown with a minutely echinulate thin wall, and measured 18 to 37 × 15 to 24 µm. Paraphyses were cylindric to clavate and slightly thickened at the apex, colorless to pale yellowish brown, and 25-50 × 6-14 µm in size. This morphology is typical of Phakopsora pachyrhizi and P. meibomiae, a less aggressive, western hemisphere species (2). DNA was extracted from leaves containing sori using the Qiagen DNeasy Plant Mini kit. P. pachyrhizi was detected using a real-time polymerase chain reaction (PCR) protocol (1) that differentiates between P. pachyrhizi and P. meibomiae performed in a Cepheid thermocycler with appropriate positive and negative controls. The PCR master mix was modified to include OmniMix beads (Cepheid). The field diagnosis of P. pachyrhizi was confirmed officially by the USDA/APHIS on November 10, 2004, and this was followed on November 11, 2004 by a wide-ranging survey of soybean and kudzu (Pueraria sp.) in soybean production areas in southern and central Louisiana. Collections from this survey also were assayed as described above, and six soybean specimens from five sites were confirmed positive. The disease was not found on kudzu samples. To our knowledge, this is the first report of P. pachyrhizi in the continental United States. Voucher specimens have been placed in the USDA National Fungus Collection. References: (1) R. D. Frederick et al. Phytopathology 92:217, 2002. (2) Y. Ono et al. Mycol. Res. 96:825, 1992.

Vegetative Compatibility Groups in Cercospora kikuchii, the Causal Agent of Cercospora Leaf Blight and Purple Seed Stain in Soybean.

ABSTRACT Nitrogen nonutilizing (Nit) mutants were used to assess vegetative compatibility of 58 isolates of Cercospora kikuchii, 55 of which were isolated from soybean plants in Louisiana. Two isolates were vegetatively self-incompatible. Of 56 self-compatible isolates, 16 were assigned to six multimember vegetative compatibility groups (VCGs), 01 to 06, with 2 or 3 isolates in each VCG. The other 40 isolates each belonged to a distinct VCG. All six multimember VCGs contained isolates from different soy bean cultivars, and three included isolates from different locations. Only one of six multimember VCGs included isolates both from soybean leaves and seed, while the other five included isolates from only leaves or seed. The likelihood of tissue specificity or preference was discussed. All isolates and tested Nit mutants produced cercosporin on potato dextrose agar under light. Significantly different amounts of cercosporin were produced among wild-type isolates, and two Nit mutants produced significantly more cercosporin than their wild-type counterparts. All isolates produced typical Cercospora leaf blight symptoms on soybean plants in greenhouse pathogenicity tests.

Factors Affecting Soybean Root Colonization by Calonectria ilicicola and Development of Red Crown Rot Following Delayed Planting.

Field studies were conducted in 1994, 1995, and 1996 to determine the effects of planting date, cultivar susceptibility, and soil pathogen population on soybean root colonization by Calonectria ilicicola and subsequent development of red crown rot. Early season colonization of roots was important for red crown rot symptom development. Symptom development in the more susceptible cultivar, Sharkey, was reduced following delayed planting and remained low in the less susceptible cultivar, Cajun, regardless of planting date. Taproot colonization was positively correlated with inoculum density during all three growing seasons but was strongest in 1994. Also, lateral root colonization correlated positively with inoculum density in 1994, the only year in which foliar symptoms were detected. A substantial decrease in inoculum density in 1995, along with reduced soybean root colonization, were attributed to high soil temperatures and probably low rainfall recorded during that summer. The effect of soybean plant age on root colonization was examined by exposing plants to the pathogen at different ages. Soybean plants were most susceptible to C. ilicicola during the first week after seedling emergence. By the second week, susceptibility was reduced by nearly half, and it remained near that level for the next several weeks.

Effects of Soil Temperature on Microsclerotia of Calonectria ilicicola and Soybean Root Colonization by this Fungus.

Field soil artificially infested with laboratory-produced microsclerotia of Calonectria ilicicola was incubated for 1, 2, 3, or 6 weeks at 20, 25, 30, 35, and 40°C. These temperatures approximate soil temperatures that were measured in soybean fields during the growing season in south Louisiana. Germinable microsclerotia were enumerated after incubation at different temperatures, and soybean seeds were planted in these soils. After 8 weeks, percent root colonization was determined as a measure of infectivity of microsclerotia. Results showed that soil temperature is a critical factor in survival of microsclerotia. The optimal soil temperature range for survival of microsclerotia was 20 to 30°C, and the maximum soil temperature limit was 35°C, above which microsclerotia did not survive. Effects of temperature on soybean root colonization were examined in growth chambers by growing soybean plants in soil infested with laboratory-grown microsclerotia for 4 weeks after seed germination. Maximum infection of young soybean roots by C. ilicicola occurred between 25 and 30°C but decreased with increasing temperatures and was negligible at 40°C. According to these results, soil temperature is a critical environmental factor controlling the development of red crown rot in soybeans in Louisiana. These findings suggest that, if red crown rot is a threat, soybean planting time should be based on soil temperature rather than calendar dates.

Origin of Race 3 of Fusarium oxysporum f. sp. lycopersici at a Single Site in California.

ABSTRACT Thirty-nine isolates of Fusarium oxysporum were collected from tomato plants displaying wilt symptoms in a field in California 2 years after F. oxysporum f. sp. lycopersici race 3 was first observed at that location. These and other isolates of F. oxysporum f. sp. lycopersici were characterized by pathogenicity, race, and vegetative compatibility group (VCG). Of the 39 California isolates, 22 were in VCG 0030, 11 in VCG 0031, and six in the newly described VCG 0035. Among the isolates in VCG 0030, 13 were race 3, and nine were race 2. Of the isolates in VCG 0031, seven were race 2, one was race 1, and three were nonpathogenic to tomato. All six isolates in VCG 0035 were race 2. Restriction fragment length polymorphisms (RFLPs) and sequencing of the intergenic spacer (IGS) region of rDNA identified five IGS RFLP haplotypes, which coincided with VCGs, among 60 isolates of F. oxysporum from tomato. Five race 3 isolates from California were of the same genomic DNA RFLP haplotype as a race 2 isolate from the same location, and all 13 race 3 isolates clustered together into a subgroup in the neighbor joining tree. Collective evidence suggests that race 3 in California originated from the local race 2 population.

Viability and stability of biological control agents on cotton and snap bean seeds.

Cotton and snap bean were selected for a multi-year, multi-state regional (south-eastern USA) research project to evaluate the efficacy of both commercial and experimental bacterial and fungal biological control agents for the management of damping-off diseases. The goal for this portion of the project was to determine the viability and stability of biological agents after application to seed. The biological seed treatments used included: (1) Bacillaceae bacteria, (2) non-Bacillaceae bacteria, (3) the fungus Trichoderma and (4) the fungus Beauveria bassiana. Seed assays were conducted to evaluate the following application factors: short-term (< or = 3 months) stability after seed treatment; quality (i.e. isolate purity); compatibility with chemical pesticides and other biocontrol agents; application uniformity between years and plant species. For the bacterial treatments, the Bacillaceae genera (Bacillus and Paenibacillus) maintained the greatest population of bacteria per seed, the best viability over time and the best application uniformity across years and seed type. The non-Bacillaceae genera Burkholderia and Pseudomonas had the least viability and uniformity. Although Beauveria bassiana was only evaluated one year, the seed fungal populations were high and uniform. The seed fungal populations and uniformity for the Trichoderma isolates were more variable, except for the commercial product T-22. However, this product was contaminated with a Streptomyces isolate in both the years that it was evaluated. The study demonstrated that Bacillaceae can be mixed with Trichoderma isolates or with numerous pesticides to provide an integrated pest control/growth enhancement package.

Host Specialization in the Charcoal Rot Fungus, Macrophomina phaseolina.

ABSTRACT To investigate host specialization in Macrophomina phaseolina, the fungus was isolated from soybean, corn, sorghum, and cotton root tissue and soil from fields cropped continuously to these species for 15 years in St. Joseph, LA. Chlorate phenotype of each isolate was determined after growing on a minimal medium containing 120 mM potassium chlorate. Consistent differences in chlorate sensitivity were detected among isolates from different hosts and from soil versus root. To further explore genetic differentiation among fungal isolates from each host, these isolates were examined by restriction fragment length polymorphism and random amplified polymorphic DNA (RAPD) analysis. No variations were observed among isolates in restriction patterns of DNA fragments amplified by polymerase chain reaction covering the internal transcribed spacer region, 5.8S rRNA and part of 25S rRNA, suggesting that M. phaseolina constitutes a single species. Ten random primers were used to amplify the total DNA of 45 isolates, and banding patterns resulting from RAPD analysis were compared with the neighbor-joining method. Isolates from a given host were genetically similar to each other but distinctly different from those from other hosts. Chlorate-sensitive isolates were distinct from chlorate-resistant isolates within a given host. In greenhouse tests, soybean, sorghum, corn, and cotton were grown separately in soil infested with individual isolates of M. phaseolina that were chosen based on their host of origin and chlorate phenotype. Root colonization and plant weight were measured after harvesting. More colonization of corn roots occurred when corn was grown in soil containing corn isolates compared with isolates from other hosts. However, there was no host specialization in isolates from soybean, sorghum, or cotton. More root colonization in soybean occurred with chlorate-sensitive than with chlorate-resistant isolates.

Antennal morphology as a physical filter of olfaction: temporal tuning of the antennae of the honeybee, Apis mellifera.

There are many different antennal morphologies for insects, yet they all have the same functional role in olfaction. Chemical signals are dispersed through two physical forces; diffusion and fluid flow. The interaction between antennal morphology and fluid flow generates a region of changing flow velocity called the boundary layer. The boundary layer determines signal dispersion dynamics and therefore influences the signal structure and information that arrives at the receptor cells. To investigate how the boundary layer changes the information in the signals arriving at receptor cells, we measured chemical dynamics within the boundary layer around the bee antennae using microelectrodes. We used two types of chemical signals: pulsed and continuous. The results showed that the boundary layer increased the decay time of the chemical signal for the pulsatile stimuli and increased the peak height for the continuous data. Spectral analysis of continuous signals showed that the temporal aspects of the chemical signal are changed by the boundary layer. Particularly the temporal dynamics of the signal are dampened at the slowest flow speed and amplified at the intermediate and fast flow speeds. By altering the structure of the chemical signal, the morphology will function as a sensory filter.

Sites of infection by pythium species in rice seedlings and effects of plant age and water depth on disease development.

ABSTRACT Seedling disease, caused primarily by several species of Pythium, is one of the major constraints to water-seeded rice production in Louisiana. The disease, also known as water-mold disease, seed rot, and seedling damping-off, causes stand reductions and growth abnormalities. In severe cases, fields must be replanted, which may result in delayed harvests and reduced yields. To develop more effective disease management tactics including biological control, this study was conducted primarily to determine sites of infection in seeds and seedlings; effect of plant age on susceptibility to P. arrhenomanes, P. myriotylum, and P. dissotocum; and minimum exposure times required for infection and seedling death. In addition, the effect of water depth on seedling disease was investigated. Infection rates of seed embryos were significantly higher than those of endosperms for all three Pythium spp. The development of roots from dry-seeded seedlings was significantly reduced by P. arrhenomanes and P. myriotylum at 5 days after planting compared with that of roots from noninoculated controls. Susceptibility of rice to all three species was sharply reduced within 2 to 6 days after planting, and seedlings were completely resistant at 8 days after planting. There was a steep reduction in emergence through the flood water, relative to the noninoculated control, following 2 to 3 days of exposure to inoculum of P. arrhenomanes and P. myriotylum. In contrast, P. dissotocum was much less virulent and required longer exposure times to cause irreversible seedling damage. Disease incidence was higher when seeds were planted into deeper water, implying that seedlings become resistant after they emerge through the flood water. These results suggest that disease control tactics including flood water management need to be employed for a very short period of time after planting. Also, given that the embryo is the primary site of infection and it is susceptible for only a few days, the disease should be amenable to biological control.

Sodium Hypochlorite: Effect of Solution pH on Rice Seed Disinfestation and Its Direct Effect on Seedling Growth.

Bacteria were completely eliminated from rice seeds following immersion in household bleach solutions (50% bleach and 2.6% NaOCl) adjusted to pH 7.0 in 0.5 M potassium phosphate, while fungi were eliminated at pH 5.0 and below. Mercuric chloride was used to disinfest non-dormant rice seeds in order to test the direct effect of sodium hypochlorite on rice seedling growth apart from an indirect effect related to elimination of microbial contaminants. Seeds were immersed for 2 h in each solution or water in each of the following treatments: (i) sterilewater followed by sterile water, (ii) HgCl2 (1,000 µg/ml) followed by sterile water, (iii) HgCl2 followed by KH2PO4 (0.3 M), (iv) HgCl2 followed by NaOCl, and (v) NaOCl followed by sterile water. Sodium hypochlorite solutions were prepared in 0.3 M potassium phosphate, which gave a final pH of 7.3. Seedling growth in treatments in which seeds were treated with HgCl2 followed by NaOCl, and NaOCl followed by sterile water, was significantly greater than those treatments in which seeds were treated with sterile water followed by sterile water, HgCl2 followed by sterile water, and HgCl2 followed by KH2PO4. Sodium hypochlorite stimulated rice seedling growth directly, as opposed to an indirect effect related to elimination of microbial contaminants or alleviation of dormancy.