Development of Two Early Forecasting Models for Predicting Incidence of Rice Panicle Blast in China.

Early forecasting of rice panicle blast is critical to the management of rice blast. To develop early forecasting models for rice panicle blast, the relationship between the seasonal maximum incidence of rice panicle blast (PBx) and the PBx in the preceding crop, weather conditions, location, and acreage of susceptible varieties was analyzed. Results revealed that PBx in the preceding crop, acreage of the susceptible varieties in class (SVC), altitude, weather conditions 120 to 180 days before the PBx date (dbPBx) and 30 to 90 dbPBx were significantly correlated with the PBx. Subsequently, a logistic model and a two-step hurdle model were developed to predict rice panicle blast. The logistic model was developed to predict whether the PBx was 0 or not based on the preceding PBx, altitude, acreage of susceptible varieties, the longest stretch of days with soil temperatures between 16 and 24°C for the period 120 to 150 dbPBx, and the longest stretch of rainy days in the period 120 to 180 dbPBx. The hurdle model predicted if the PBx was greater than 0 at the first step, and if the prediction was greater than 0, then a regression model was developed for predicting PBx based on the preceding PBx, SVC, altitude, and weather data 180 to 30 dbPBx. Validation with the test datasets showed that the logistic model could correctly predict whether PBx was 0 at a mean test accuracy of 78.39% and that the absolute prediction error of PBx by the two-step hurdle model was smaller than 6.16% for 90% of the records. The model developed in this study will be helpful in management decisions for rice growers and policy makers and offer a useful basis for further studies on the epidemiology and forecasting of rice panicle blast.

Superconducting ECR ion source: From 24-28 GHz SECRAL to 45 GHz fourth generation ECR.

The development of superconducting ECR source with higher magnetic fields and higher microwave frequency is the most straight forward path to achieve higher beam intensity and higher charge state performance. SECRAL, a superconducting third generation ECR ion source, is designed for 24-28 GHz microwave frequency operation with an innovative magnet configuration of sextupole coils located outside the three solenoids. SECRAL at 24 GHz has already produced a number of record beam intensities, such as 40Ar12+ 1.4 emA, 129Xe26+ 1.1 emA, 129Xe30+ 0.36 emA, and 209Bi31+ 0.68 emA. SECRAL-II, an upgraded version of SECRAL, was built successfully in less than 3 years and has recently been commissioned at full power of a 28 GHz gyrotron and three-frequency heating (28 + 45 + 18 GHz). New record beam intensities for highly charged ion production have been achieved, such as 620 eµA 40Ar16+, 15 eµA 40Ar18+, 146 eµA 86Kr28+, 0.5 eµA 86Kr33+, 53 eµA 129Xe38+, and 17 eµA 129Xe42+. Recent beam test results at SECRAL and SECRAL II have demonstrated that the production of more intense highly charged heavy ion beams needs higher microwave power and higher frequency, as the scaling law predicted. A 45 GHz superconducting ECR ion source FECR (a first fourth generation ECR ion source) is being built at IMP. FECR will be the world's first Nb3Sn superconducting-magnet-based ECR ion source with 6.5 T axial mirror field, 3.5 T sextupole field on the plasma chamber inner wall, and 20 kW at a 45 GHz microwave coupling system. This paper will focus on SECRAL performance studies at 24-28 GHz and technical design of 45 GHz FECR, which demonstrates a technical path for highly charged ion beam production from 24 to 28 GHz SECRAL to 45 GHz FECR.

Functional Interactions Between Major Rice Blast Resistance Genes, Pi-ta and Pi-b, and Minor Blast Resistance Quantitative Trait Loci.

Major blast resistance (R) genes confer resistance in a gene-for-gene manner. However, little information is available on interactions between R genes. In this study, interactions between two rice blast R genes, Pi-ta and Pi-b, and other minor blast resistance quantitative trait loci (QTLs) were investigated in a recombinant inbred line (RIL) population comprising 243 RILs from a Cybonnet (CYBT) × Saber (SB) cross. CYBT has the R gene Pi-ta and SB has Pi-b. Ten differential isolates of four Magnaporthe oryzae races (IB-1, IB-17, IB-49, and IE-1K) were used to evaluate disease reactions of the 243 RILs under greenhouse conditions. Five resistance QTLs were mapped on chromosomes 2, 3, 8, 9, and 12 with a linkage map of 179 single nucleotide polymorphism markers. Among them, qBR12 (Q1), was mapped at the Pi-ta locus and accounted for 45.41% of phenotypic variation while qBR2 (Q2) was located at the Pi-b locus and accounted for 24.81% of disease reactions. The additive-by-additive epistatic interaction between Q1 (Pi-ta) and Q2 (Pi-b) was detected; they can enhance the disease resistance by an additive 0.93 using the 0 to 9 standard phenotyping method. These results suggest that Pi-ta interacts synergistically with Pi-b.

Effects of Temperature and Duration of Preconditioning Cold Treatment on Sclerotial Germination of Claviceps purpurea.

Claviceps purpurea is an important ovary-infecting pathogen that replaces seed with sclerotia in Kentucky bluegrass grown for seed. Sclerotia overwinter in the soil and germinate in the spring to produce ascospores that infect grass seed ovaries. To better understand environmental conditions affecting ascospore production, the effects of preconditioning cold treatment and subsequent incubation temperature on germination of sclerotia were determined in growth chambers under controlled conditions. Preconditioning cold treatment was essential for germination only in treatments where the incubation temperature was high (at least higher than 20°C). At lower incubation temperatures (10 to 20°C), preconditioning also played a role in improving sclerotial germination. Preconditioning at 4°C (in darkness) for 4 to 8 weeks followed by incubation at 10 and 20°C (cycle of 12 h each of darkness and light), or constant 15°C (cycle of 12 h each of darkness and light), was optimal for ergot germination. When sclerotia were preconditioned for 4 weeks or longer, number of incubation days required for initiation of germination was not affected by temperature in the range from 10 to 25°C (cycle of 12 h each of darkness and light), although the duration of germination (or the progress speed of germination) was still affected by temperature. A simple model was developed based on laboratory results and validated with historic spore trap data collected from various Kentucky bluegrass fields in Oregon (Willamette Valley, central Oregon, and Grande Ronde Valley). The prediction model could predict ascospore onset well and explained 55% of variation in the data.

A Comparison of Different Operating Systems for Femtosecond Lasers in Cataract Surgery.

The introduction of femtosecond lasers is potentially a major shift in the way we approach cataract surgery. The development of increasingly sophisticated intraocular lenses (IOLs), coupled with heightened patient expectation of high quality postsurgical visual outcomes, has generated the need for a more precise, highly reproducible and standardized method to carry out cataract operations. As femtosecond laser-assisted cataract surgery (FLACS) becomes more commonplace in surgical centers, further evaluation of the potential risks and benefits needs to be established, particularly in the medium/long term effects. Healthcare administrators will also have to weigh and balance out the financial costs of these lasers relative to the advantages they put forth. In this review, we provide an operational overview of three of five femtosecond laser platforms that are currently commercially available: the Catalys (USA), the Victus (USA), and the LDV Z8 (Switzerland).

Multiple displacement amplification of whole genomic DNA from urediospores of Puccinia striiformis f. sp. tritici.

Biotrophic fungi, such as Puccinia striiformis f. sp. tritici, because they cannot be cultured on nutrient media, to obtain adequate quantity of DNA for molecular genetic analysis, are usually propagated on living hosts, wheat plants in case of P. striiformis f. sp. tritici. The propagation process is time-, space- and labor-consuming and has been a bottleneck to molecular genetic analysis of this pathogen. In this study we evaluated multiple displacement amplification (MDA) of pathogen genomic DNA from urediospores as an alternative approach to traditional propagation of urediospores followed by DNA extraction. The quantities of pathogen genomic DNA in the products were further determined via real-time PCR with a pair of primers specific for the ß-tubulin gene of P. striiformis f. sp. tritici. The amplified fragment length polymorphism (AFLP) fingerprints were also compared between the DNA products. The results demonstrated that adequate genomic DNA at fragment size larger than 23 Kb could be amplified from 20 to 30 urediospores via MDA method. The real-time PCR results suggested that although fresh urediospores collected from diseased leaves were the best, spores picked from diseased leaves stored for a prolonged period could also be used for amplification. AFLP fingerprints exhibited no significant differences between amplified DNA and DNA extracted with CTAB method, suggesting amplified DNA can represent the pathogen's genomic DNA very well. Therefore, MDA could be used to obtain genomic DNA from small precious samples (dozens of spores) for molecular genetic analysis of wheat stripe rust pathogen, and other fungi that are difficult to propagate.

A model for multiseasonal spread of verticillium wilt of lettuce.

Verticillium wilt, caused by Verticillium dahliae, is a destructive disease in lettuce, and the pathogen is seedborne. Even though maximum seed infestation rates of <5% have been detected in commercial lettuce seed lots, it is necessary to establish acceptable contamination thresholds to prevent introduction and establishment of the pathogen in lettuce production fields. However, introduction of inoculum into lettuce fields for experimental purposes to determine its long term effects is undesirable. Therefore, we constructed a simulation model to study the spread of Verticillium wilt following pathogen introduction from seed. The model consists of four components: the first for simulating infection of host plants, the second for simulating reproduction of microsclerotia on diseased plants, the third for simulating the survival of microsclerotia, and the fourth for simulating the dispersal of microsclerotia. The simulation results demonstrated that the inoculum density-disease incidence curve parameters and the dispersal gradients affect disease spread in the field. Although a steep dispersal gradient facilitated the establishment of the disease in a new field with a low inoculum density, a long-tail gradient allowed microsclerotia to be dispersed over greater distances, promoting the disease spread in fields with high inoculum density. The simulation results also revealed the importance of avoiding successive lettuce crops in the same field, reducing survival rate of microsclerotia between crops, and the need for breeding resistance against V. dahliae in lettuce cultivars to lower the number of microsclerotia formed on each diseased plant. The simulation results, however, suggested that, even with a low seed infestation rate, the pathogen would eventually become established if susceptible lettuce cultivars were grown consecutively in the same field for many years. A threshold for seed infestation can be established only when two of the three drivers of the disease-(i) low microsclerotia production per diseased plant, (ii) long-tail dispersal gradient, and (iii) low microsclerotia survival between lettuce crops-are present.

First Report of Fusarium solani on Utah Sweetvetch in the United States.

Utah sweetvetch (Hedysarum boreale Nutt.) is a native American perennial nitrogen fixing legume used mainly in rangeland reclamation, soil rejuvenation, and erosion control. In June 2011, a field of Utah sweetvetch grown for seeds in central Oregon had approximately 15% of the plants exhibiting chlorosis, defoliation, stunting, wilting, and/or death. Dissection of the crown of symptomatic plants revealed discolored pinkish brown vascular tissue. Symptomatic tissues from six random plants were surface sterilized, placed on acidified potato dextrose agar (PDA) medium, and cultured for 7 days at room temperature, which allowed six fungal isolates (SS1 through SS6) to be collected. On PDA, all six isolates had rapid, creamy white colored growth. Based on observations of 1-week-old isolates, microconidia were oval to kidney shaped, single celled, 8 to 10 × 2.5 to 4 µm, and formed at the tips of long unbranched monophialides. Macroconidia were three to four septate, cylindrical to slightly curved, with characteristic foot shaped basal cell and blunt apical cell, 37 to 49 × 4.4 to 5.3 µm. Chlaymydospores observed were 8.5 to 11 × 7.6 to 9 µm. Based on fungal references (1,2,3), the isolates were identified as Fusarium solani (Mart.) Sacc. Identification of the isolates at the molecular level was determined by amplification of the internal transcribed spacer (ITS) region using PCR and amplicon sequencing. Botrytis cinerea and F. graminearum cultures were used as controls for the extraction, amplification, and sequencing steps. Genomic DNA was extracted from mycelia using protocols of the MOBIO Ultraclean Soil DNA Isolation Kit (MO-BIO Laboratories Inc, Carlsbad, CA, USA). PCR was performed using ITS1/ITS4 primers and resulted in 563- to 573-bp amplicons, which were sequenced. Analysis of the ITS sequences (GenBank Accession Nos. JX524018 to JX524023) for the six fungal isolates using BLASTn revealed a 99% sequence identity with F. solani strains (AB470903, AB513851, AJ608989, EF152426, EU029589, and HM214456). Pathogenicity was confirmed on Utah sweetvetch plants in the greenhouse. Seeds of Utah sweetvetch were first plated on acidified PDA for germination; healthy seedlings were then selected and transplanted into pots with sterilized soil after 2 weeks of growth. The plants were kept in a greenhouse at Central Oregon Agricultural Research Center, Madras, Oregon. Ten 40-day-old healthy vetch plants were inoculated by drenching with a mixed conidial suspension (107 conidia/ml) of the six F. solani isolates. Ten plants drenched with sterile distilled water were included as controls. Symptoms of chlorosis and stunting similar to those in the commercial field were observed within 30 days of inoculation on 8 of 10 inoculated plants, while control plants were symptomless. Fungal isolates identical to F. solani were reisolated from the symptomatic plants. To our knowledge, this is the first report of F. solani on Utah sweetvetch plants. References: (1) C. Booth. The Genus Fusarium. CMI, Kew, Surrey, UK, 1971. (2) P. E. Nelson et al. Fusarium species: An illustrated manual for identification. The Pennsylvania State University Press, USA, 1983. (3) H. I. Nirenberg. A simplified method for identifying Fusarium spp. occurring on wheat. Can. J. Bot. 59:1599, 1980.

Interactions between Coniothyrium minitans and Sclerotinia minor affect biocontrol efficacy of C. minitans.

Coniothyrium minitans, marketed as Contans, has become a standard management tool against Sclerotinia sclerotiorum in a variety of crops, including winter lettuce. However, it has been ineffective against lettuce drop caused by S. minor. The interactions between C. minitans and S minor were investigated to determine the most susceptible stage in culture to attack by C. minitans, and to determine its consistency on S minor isolates belonging to four major mycelial compatibility groups (MCGs). Four isolates of S. minor MCG 1 and 5 each from MCGs 2 and 3 and one from MCG 4 were treated in culture at purely mycelial, a few immature sclerotial, and fully mature sclerotial phases with a conidial suspension of C. minitans. Sclerotia from all treatments were harvested after 4 weeks, air dried, weighed, and plated on potato dextrose agar for recovery of C. minitans. S. minor formed the fewest sclerotia in plates that received C. minitans at the mycelial stage; C. minitans was recovered from nearly all sclerotia from this treatment and sclerotial mortality was total. However, the response of MCGs was inconsistent and variable. Field experiments to determine the efficacy of C. minitans relative to the registered fungicide, Endura, on lettuce drop incidence and soil inoculum dynamics were conducted from 2006 to 2009. All Contans treatments had significantly lower numbers of sclerotia than Endura and unsprayed control treatments, and drop incidence was as low as in Endura-treated plots (P > 0.05). Although the lower levels of lettuce drop in Contans treatments were correlated with significantly lower levels of sclerotia, the lower levels of lettuce drop, despite the presence of higher inoculum in the Endura treatment, was attributable to the prevention of infection by S. minor. A useful approach to sustained lettuce drop management is to employ Contans to lower the number of sclerotia in soil and to apply Endura to prevent S. minor infection within a cropping season.

Impact of consumer-driven changes to crop production practices on lettuce drop caused by Sclerotinia sclerotiorum and S. minor.

Increasing demands for value-added salad products have triggered revolutionary changes in the production practices of vegetable salad crops in recent years. One of the pivotal changes is the adaptation of 2-m-wide beds for increased vegetable biomass per unit area. The move away from the traditional 1-m-wide raised beds in cool-season vegetable production and the associated irrigation practices potentially can have a major influence on diseases affecting cool-season vegetables. To assess the potential impacts of this shift on lettuce drop caused by Sclerotinia minor and S. sclerotiorum, the two bed widths and different irrigation frequencies within each were compared in two separate field experiments over four lettuce crops in 2 years. Treatments included 1- and 2-m bed widths with twice-weekly, weekly and biweekly drip irrigation serving as subplot treatments that were begun immediately following thinning. Incidence of lettuce drop was evaluated weekly thereafter until maturity. For S. sclerotiorum, 36 half-liter soil samples were also collected once each season and assayed for the number of sclerotia, and apothecia were counted weekly in a 10-m(2) area for each plot. Regardless of the species, the effects of bed width and irrigation frequency were both significant. Twice-weekly irrigation and 2-m bed width resulted in higher lettuce drop incidence than other treatments. For S. sclerotiorum, twice-weekly irrigation and 2-m bed width also significantly increased the number of apothecia per unit area and the accumulation of soilborne sclerotia over multiple cropping seasons. Results demonstrated that the 2-m bed width combined with the practiced frequency of irrigations can result in higher lettuce drop caused by S. minor and increased incidence of airborne infection by S. sclerotiorum in the Salinas Valley where, historically, it has not been a serious threat. Increased incidence of S. sclerotiorum infection in commercial lettuce fields in the Salinas Valley between 2001 and 2006 validates these experimental results. These relatively new crop production practices can alter the balance of the two Sclerotinia spp. that has long existed in California.

Effects of soil temperature, moisture, and burial depths on carpogenic germination of Sclerotinia sclerotiorum and S. minor.

Extensive studies have been conducted on the carpogenic germination of Sclerotinia sclerotiorum, but carpogenic germination in S. minor has not been studied adequately. It remains unclear why apothecia of this pathogen have seldom been observed in nature. In this study, a new method was developed to produce apothecia in the absence of soil or sand, and carpogenic germination without preconditioning was recorded for 95 of the 96 S. sclerotiorum isolates tested. Carpogenic germination of the two species was compared under a variety of temperature, soil moisture, burial depths, and short periods of high temperature and low soil moisture. The optimal temperatures for rapid germination and for maximum germination rates were both lower for S. minor than for S. sclerotiorum. The temperature range for carpogenic germination was also narrower for S. minor than for S. sclerotiorum. A 5-day period at 30 degrees C, either starting on the 10th or 20th day of incubation, did not significantly affect carpogenic germination of S. sclerotiorum. For both S. minor and S. sclerotiorum, the percentage of carpogenically germinated sclerotia increased as soil water potential increased from -0.3 to -0.01 MPa. In the greenhouse, a 10- or 20-day dry period completely arrested carpogenic germination of S. sclerotiorum, and new apothecia appeared after an interval of 35 days following rewetting, similar to the initial carpogenic germination regardless of when the dry period was imposed. In naturally infested fields, the number of sclerotia in 100 cc of soil decreased as depth increased from 0 to 10 cm before tillage, but became uniform between 0 and 10 cm after conventional tillage for both species. Most apothecia of S. minor were, however, produced from sclerotia located at a depth shallower than 0.5 cm while some apothecia of S. sclerotiorum were produced from sclerotia located as deep as 4 to 5 cm. These results provide the much needed information to assess the epidemiological roles of inoculum from sexual reproduction in diseases caused by the two Sclerotinia species in different geographical regions. However, more studies on effects of shorter and incompletely dry periods are still needed to predict production of apothecia of S. sclerotiorum in commercial fields under fluctuating soil temperature and moisture.

Comparative survival of Sclerotia of Sclerotinia minor and S. sclerotiorum.

Survival of sclerotia of Sclerotinia minor and S. sclerotiorum was compared in irrigated fields during the summer in two major lettuce production areas in California. More than 50% sclerotia of S. sclerotiorum compared with 4 and 35% of S. minor remained viable after 24 weeks of burial at 15 and 5 cm depths, respectively, in the San Joaquin Valley while >80% of sclerotia survived in the Salinas Valley for both species. The results explain in part the lower infections from S. minor in the San Joaquin Valley. To identify factors that contribute to the rapid decline in the viability of sclerotia, the effects of soil moisture, temperature, and oxygen levels were studied in laboratory. More than 90% of sclerotia of both species survived for at least 3 months in sterilized dry soils at temperatures between 15 and 40 degrees C. Soil moisture did not affect survival at 15 and 25 degrees C. At 35 degrees C, however, survival rates were significantly lower at high (-0.3 to -0.01 MPa) water potential than at low (<-1.0 MPa) water potential. Incubation under ultralow oxygen concentration (0.01%) significantly reduced survival of sclerotia in nonautoclaved moist soils at 25 degrees C, with less than 2% sclerotia surviving over 4 weeks compared with about 45% sclerotia surviving at the ambient oxygen level (21%). The combination of high temperature, high soil moisture, and reduced oxygen in irrigated fields contribute to the lower survival of both Sclerotinia species and the responses of the two species to these conditions shape their relative geographical distribution.

Nonlinear colony extension of Sclerotinia minor and S. sclerotiorum.

Fungal colonies initially extend exponentially and reach a constant linear extension rate determined solely by their growth in the peripheral zone. However the radial extension rates of Sclerotinia sclerotiorum and S. minor accelerate over time on PDA. Experiments were conducted to analyze the variable extension rates of the two Sclerotinia species and compare them with those of Verticillium dahliae and Cladosporium sp. In addition, the effects of starter disk size, disk position in the parent colony, the age of the parent colony, the concentration of potato dextrose broth and of incubation temperature also were determined. While the growth of Cladosporium sp. and V. dahliae followed established linear trends, the radial extension of S. sclerotiorum and S. minor colonies continuously accelerated over time until they reached the edge of the (150 mm diam) Petri dish. A polynomial model fitted the radial extension of colonies of Sclerotinia spp. Furthermore the accelerating colony extension rate was partly due to increasing colony radius. The rates of extension from mycelial disks transferred from the parental colony were positively correlated with the radius of the mycelial disks transferred. The rates of extension also were dependent on where the transferred disks were taken from parent colonies and the age and radius of the parent colony. On potato dextrose agar medium the extension rates of colonies of S. sclerotiorum and S. minor also were affected by broth concentration and temperature. With increasing nutrient concentration colony extension rates increased and were highest at 25 C. This study revealed a novel pattern of radial growth for Sclerotinia spp. that diverged from the established growth patterns of fungal colonies. Knowledge of the differences in growth behavior may be exploited in the laboratory studies on fungal competition and hyperparasitism and potentially in disease control strategies.

Incubation of excised apothecia enhances ascus maturation of Sclerotinia sclerotiorum.

Synchronized maturation of ascospores of Sclerotinia sclerotiorum is desirable for establishing a transformation system, conducting genetic analyses of the pathogen, defining the precise epidemiological roles of ascospores and screening plant germplasm for resistance. In general, fresh apothecia collected from germinated sclerotia contained primarily immature or discharged asci. This study was undertaken to investigate whether maturation of asci and ascospores could be enhanced by incubation of excised apothecia and to determine the effects of factors such as temperature, excision time, light and ventilation on maturation of asci and ascospores in excised apothecia. Maturation of asci was compared between intact and excised apothecia that were incubated under similar conditions. Results demonstrated that temperature was an important factor affecting ascus maturation of S. sclerotiorum during incubation of excised apothecia, and the optimum temperature was around 21 C. After incubation at 21 C for 30 h, the percentage of undischarged mature asci in excised apothecia increased up to 70-80%. This increase was accompanied by a significant increase in ascospore production of up to 5 x 10(5) ascospores per apothecium. Detailed time course studies indicated that mature asci peaked at 30-36 h of postexcision incubation. Mature asci and the number of ascospores were higher in open incubation than in closed incubation, suggesting that accumulation of volatile substances was not required for ascus/ascospore maturation during postexcision incubation and ventilation could enhance the maturation process. Light also did not affect the maturation of asci during the incubation of excised apothecia. Germination rates for ascospores from excised apothecia under various treatments were similar to those from untreated apothecia but declined slightly with time postexcision. The incubation of excised apothecia promoted ascus maturation compared with intact apothecia.

Surface modification and initial adhesion events for intestinal epithelial cells.

Rapid resealing of the mucosal epithelia is imperative following injuries to the small intestine because the mucosa is responsible for the adsorption of nutrients as well as providing a barrier to noxious agents present in the lumen. Tissue engineering may provide a possible solution for treating intestinal erosions, ulcerations, inflammatory bowel disease, and infection. Cell-biomaterial interaction is a critical component in tissue engineering that can determine the success of the tissue construct. Cell-biomaterial interactions can be enhanced by various types of surface modification, which promote integrin ligation leading to increased cell function. In order to relate the effect of surface adhesion molecules to signaling events and macroscopic cell response, an intestinal epithelial cell line, IEC-6, was plated on fibronectin (receptor-mediated) and poly-L-lysine (non-specific) surfaces. Focal adhesion kinase (FAK) phosphorylation, cell spreading, and cell adhesion strength were measured. Results showed increases in FAK phosphorylation generally corresponded to increases in cell spreading and adhesion strength for IEC-6 cells. Therefore, in a simplified system, initial adhesion and signaling mechanisms appeared to correspond to subsequent physical responses in IEC-6 cells relevant to tissue engineering applications.

The effects of short-term stimulation on fibroblast spreading in an in vitro 3D system.

Although the short-term effects of mechanical and biochemical stimulation on cell protein and DNA production have recently begun to be evaluated using 3D models, the effects that such stimulation have on cell morphology and adaptation remains unclear. Using a custom-built bioreactor, we were able to create a systematic model to examine the short-term effects of stimulation on cell morphology in a 3D model, specifically by evaluating cell spreading as the short-term indicator of cell adaptation. Fibroblasts were seeded on a porous poly(L-lactic acid) scaffold and cultured in a computerized bioreactor for 24 h under various uniaxial strains (0, 0.6, 3, 6%) and frequencies (0.0125, 0.125 Hz). Also, the effects of transforming growth factor (TGF-beta1) (1, 10, 100 ng/mL) were examined on static, nonstimulated cells-scaffold constructs after 24 h. Fibroblasts that had been subjected to mechanical stretching were found to exhibit significantly more spreading than the static control group. Conversely, TGF-beta1 between 1 and 100 ng/mL did not produce any significant difference in fibroblast spreading from the control groups after 24 h. Collectively, the findings suggest that cell morphology and adaptation may be affected by short-term mechanical stimulation, as seen by increased cell spreading by the fibroblasts under these experimental conditions.

Analyses of Lettuce Drop Incidence and Population Structure of Sclerotinia sclerotiorum and S. minor.

ABSTRACT To understand the geographical distribution of lettuce drop incidence and the structure of Sclerotinia minor and S. sclerotiorum populations, commercial lettuce fields were surveyed in the Salinas, San Joaquin, and Santa Maria Valleys in California. Lettuce drop incidence, pathogen species, and mycelial compatibility groups (MCGs) were determined and analyzed using geostatistic and geographical information system tools. Lettuce drop incidence was lowest in the San Joaquin Valley, and not significantly different between the other two valleys. Semivariogram analysis revealed that lettuce drop incidence was not spatially correlated between different fields in the Salinas Valley, suggesting negligible field-to-field spread or influence of inoculum in one field on other fields. Lettuce drop incidence was significantly lower in fields with a surface drip system than in fields with furrow or sprinkler irrigation systems, suggesting that the surface drip system can be a potential management measure for reducing lettuce drop. In the San Joaquin Valley, S. sclerotiorum was the prevalent species, causing drop in 63.5% of the fields, whereas S. minor also was identified in 25.4% of the fields. In contrast, in the Salinas Valley, S. minor was the dominant species (76.1%) whereas S sclerotiorum only observed in only 13.6% fields, in which only a few plants were infected by S. sclerotiorum. In the Santa Maria Valley, both species frequently were identified, with S. minor being slightly more common. Although many MCGs were identified in S. minor, most of them consisted of only one or two isolates. In all, approximately 91.4% of the isolates belonged to four MCGs. Among them, MCG-1 was the most prevalent group in all three valleys, accounting for 49.8% of total isolates. It was distributed all over the surveyed areas, whereas other MCGs were distributed more or less locally. Populations of S. sclerotiorum exhibited greater diversity, with 89 isolates collected from the Salinas and San Joaquin Valleys belonging to 37 different MCGs. Among them, the most recurrent MCG-A contained 16 isolates, and 30 MCGs contained only 1 isolate each. Many MCGs occurred within only one or a part of the two valleys. Potential reasons for this abundant diversity are discussed.

Analyses of the Relationships Between Lettuce Downy Mildew and Weather Variables Using Geographic Information System Techniques.

Previous studies in coastal California suggested that morning leaf wetness duration and temperature immediately after the prolonged leaf wetness period affect infection of lettuce by the downy mildew pathogen, Bremia lactucae. In this study, spatial analysis tools in a geographic information system were used to interpolate disease assessment data and then relate them to weather variables measured in 1995 and 1997 at weather stations in the Salinas Valley. Among the variables monitored at these weather stations, midday temperature (10:00 A.M. to 2:00 P.M.) was related most strongly to the interpolated downy mildew incidence in a circular area (radius = 5 km) around each station (r = 0.52, P < 0.0001); the higher the midday temperature, the lower the disease incidence. High humidity and prolonged morning leaf wetness duration also were associated with high downy mildew incidence. Cluster analysis resulted in distinct regions with different midday temperatures, which overlapped well (92.2% of the total area) with regions distinguished in previous cluster analyses of downy mildew incidence. Clusters of morning relative humidity showed similar patterns, although they overlapped less well with clusters of disease incidence. These results confirmed that midday temperature is an important determining factor for lettuce downy mildew, and its effects should be incorporated into a disease warning system for coastal California. Cluster analyses based on the effects of temperature have great potential for use in regional downy mildew risk assessment.

Quantitative analyses of Streptococcus mutans biofilms with quartz crystal microbalance, microjet impingement and confocal microscopy.

Microbial biofilm formation can be influenced by many physiological and genetic factors. The conventional microtiter plate assay provides useful but limited information about biofilm formation. With the fast expansion of the biofilm research field, there are urgent needs for more informative techniques to quantify the major parameters of a biofilm, such as adhesive strength and total biomass. It would be even more ideal if these measurements could be conducted in a real-time, non-invasive manner. In this study, we used quartz crystal microbalance (QCM) and microjet impingement (MJI) to measure total biomass and adhesive strength, respectively, of S. mutans biofilms formed under different sucrose concentrations. In conjunction with confocal laser scanning microscopy (CLSM) and the COMSTAT software, we show that sucrose concentration affects the biofilm strength, total biomass, and architecture in both qualitative and quantitative manners. Our data correlate well with previous observations about the effect of sucrose on the adherence of S. mutans to the tooth surface, and demonstrate that QCM is a useful tool for studying the kinetics of biofilm formation in real time and that MJI is a sensitive, easy-to-use device to measure the adhesive strength of a biofilm.