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The subtilisin-like serine protease SDD1 mediates cell-to-cell signaling during Arabidopsis stomatal development.

Wild-type stomata are distributed nonrandomly, and their density is controlled by endogenous and exogenous factors. In the Arabidopsis mutant stomatal density and distribution1-1 (sdd1-1), the establishment of the stomatal pattern is disrupted, resulting in stomata clustering and twofold to fourfold increases in stomatal density. The SDD1 gene that encodes a subtilisin-like Ser protease is expressed strongly in stomatal precursor cells (meristemoids and guard mother cells), and the SDD1 promoter is controlled negatively by a feedback mechanism. The encoded protein is exported to the apoplast and probably is associated with the plasma membrane. SDD1 overexpression in the wild type leads to a phenotype opposite to that caused by the sdd1-1 mutation, with a twofold to threefold decrease in stomatal density and the formation of arrested stomata. While SDD1 overexpression was effective in the flp mutant, the tmm mutation acted epistatically. Thus, we propose that SDD1 generates an extracellular signal by meristemoids/guard mother cells and demonstrate that the function of SDD1 is dependent on TMM activity.

Arabidopsis↗

RPT2 is a signal transducer involved in phototropic response and stomatal opening by association with phototropin 1 in Arabidopsis thaliana.

Phototropin 1 (phot1) and phot2, which are blue light receptor kinases, function in blue light-induced hypocotyl phototropism, chloroplast relocation, and stomatal opening in Arabidopsis (Arabidopsis thaliana). Previous studies have shown that the proteins RPT2 (for ROOT PHOTOTROPISM2) and NPH3 (for NONPHOTOTROPIC HYPOCOTYL3) transduce signals downstream of phototropins to induce the phototropic response. However, the involvement of RPT2 and NPH3 in stomatal opening and in chloroplast relocation mediated by phot1 and phot2 was unknown. Genetic analysis of the rpt2 mutant and of a series of double mutants indicates that RPT2 is involved in the phot1-induced phototropic response and stomatal opening but not in chloroplast relocation or phot2-induced movements. Biochemical analyses indicate that RPT2 is purified in the crude microsomal fraction, as well as phot1 and NPH3, and that RPT2 makes a complex with phot1 in vivo. On the other hand, NPH3 is not necessary for stomatal opening or chloroplast relocation. Thus, these results suggest that phot1 and phot2 choose different signal transducers to induce three responses: phototropic response of hypocotyl, stomatal opening, and chloroplast relocation.

Arabidopsis↗

Dynamics of stomatal water relations following leaf excision.

We examined the stomatal response to leaf excision in an evergreen woody shrub, Photinia x fraseri, using a novel combination of gas exchange, traditional water relations and modelling. Plants were kept outdoors in mild winter conditions (average daily temperature range: -1 to 12 degrees C) before being transferred to a glasshouse (temperature range: 20-30 degrees C) and allowed to acclimate for different periods before experiments. 'Glasshouse plants' were acclimated for at least 9 d, and 'outdoor plants' were acclimated for fewer than 3 d before laboratory gas exchange experiments. The transient stomatal opening response to leaf excision was roughly twice as long in outdoor plants as in glasshouse plants. To elucidate the reason for this difference, we inferred variables of stomatal water relations (epidermal and guard cell turgor pressures and guard cell osmotic pressure: Pe, Pg and pi g, respectively) from stomatal conductance (gs) and bulk leaf water potential (psi l), using a hydromechanical model of gs. psi l was calculated from cumulative post-excision transpirational water loss using empirical relationships between psi l and relative water content obtained on similar leaves. Inferred Pg and Pe both declined immediately after leaf excision. Inferred pi g also declined after a lag period. The kinetics of pi g adjustment after the lag were similar in outdoors and glasshouse plants, but the lag period was much longer in outdoor plants. This suggests that the longer transient opening response in outdoor plants resulted from slower induction, not slower execution, of guard cell osmoregulation. We discuss the implications of our results for the mechanism of short-term stomatal responses to hydraulic perturbations, for dynamic modelling of gs and for leaf water status regulation.

Plant Leaves↗

Long-term growth of soybean at elevated [CO2] does not cause acclimation of stomatal conductance under fully open-air conditions.

Accurately predicting plant function and global biogeochemical cycles later in this century will be complicated if stomatal conductance (g(s)) acclimates to growth at elevated [CO(2)], in the sense of a long-term alteration of the response of g(s) to [CO(2)], humidity (h) and/or photosynthetic rate (A). If so, photosynthetic and stomatal models will require parameterization at each growth [CO(2)] of interest. Photosynthetic acclimation to long-term growth at elevated [CO(2)] occurs frequently. Acclimation of g(s) has rarely been examined, even though stomatal density commonly changes with growth [CO(2)]. Soybean was grown under field conditions at ambient [CO(2)] (378 micromol mol(-1)) and elevated [CO(2)] (552 micromol mol(-1)) using free-air [CO(2)] enrichment (FACE). This study tested for stomatal acclimation by parameterizing and validating the widely used Ball et al. model (1987, Progress in Photosynthesis Research, vol IV, 221-224) with measurements of leaf gas exchange. The dependence of g(s) on A, h and [CO(2)] at the leaf surface was unaltered by long-term growth at elevated [CO(2)]. This suggests that the commonly observed decrease in g(s) under elevated [CO(2)] is due entirely to the direct instantaneous effect of [CO(2)] on g(s) and that there is no longer-term acclimation of g(s) independent of photosynthetic acclimation. The model accurately predicted g(s) for soybean growing under ambient and elevated [CO(2)] in the field. Model parameters under ambient and elevated [CO(2)] were indistinguishable, demonstrating that stomatal function under ambient and elevated [CO(2)] could be modelled without the need for parameterization at each growth [CO(2)].

Acclimatization↗

Stomatal patterning and differentiation by synergistic interactions of receptor kinases.

Coordinated spacing and patterning of stomata allow efficient gas exchange between plants and the atmosphere. Here we report that three ERECTA (ER)-family leucine-rich repeat-receptor-like kinases (LRR-RLKs) together control stomatal patterning, with specific family members regulating the specification of stomatal stem cell fate and the differentiation of guard cells. Loss-of-function mutations in all three ER-family genes cause stomatal clustering. Genetic interactions with a known stomatal patterning mutant too many mouths (tmm) revealed stoichiometric epistasis and combination-specific neomorphism. Our findings suggest that the negative regulation of ER-family RLKs by TMM, which is an LRR receptor-like protein, is critical for proper stomatal differentiation.

Arabidopsis↗

Alteration of stimulus-specific guard cell calcium oscillations and stomatal closing in Arabidopsis det3 mutant.

Cytosolic calcium oscillations control signaling in animal cells, whereas in plants their importance remains largely unknown. In wild-type Arabidopsis guard cells abscisic acid, oxidative stress, cold, and external calcium elicited cytosolic calcium oscillations of differing amplitudes and frequencies and induced stomatal closure. In guard cells of the V-ATPase mutant det3, external calcium and oxidative stress elicited prolonged calcium increases, which did not oscillate, and stomatal closure was abolished. Conversely, cold and abscisic acid elicited calcium oscillations in det3, and stomatal closure occurred normally. Moreover, in det3 guard cells, experimentally imposing external calcium-induced oscillations rescued stomatal closure. These data provide genetic evidence that stimulus-specific calcium oscillations are necessary for stomatal closure.

Abscisic Acid↗

Dominant negative guard cell K+ channel mutants reduce inward-rectifying K+ currents and light-induced stomatal opening in arabidopsis.

Inward-rectifying potassium (K+(in)) channels in guard cells have been suggested to provide a pathway for K+ uptake into guard cells during stomatal opening. To test the proposed role of guard cell K+(in) channels in light-induced stomatal opening, transgenic Arabidopsis plants were generated that expressed dominant negative point mutations in the K+(in) channel subunit KAT1. Patch-clamp analyses with transgenic guard cells from independent lines showed that K+(in) current magnitudes were reduced by approximately 75% compared with vector-transformed controls at -180 mV, which resulted in reduction in light-induced stomatal opening by 38% to 45% compared with vector-transformed controls. Analyses of intracellular K+ content using both sodium hexanitrocobaltate (III) and elemental x-ray microanalyses showed that light-induced K+ uptake was also significantly reduced in guard cells of K+(in) channel depressor lines. These findings support the model that K+(in) channels contribute to K+ uptake during light-induced stomatal opening. Furthermore, transpirational water loss from leaves was reduced in the K+(in) channel depressor lines. Comparisons of guard cell K+(in) current magnitudes among four different transgenic lines with different K+(in) current magnitudes show the range of activities of K+(in) channels required for guard cell K+ uptake during light-induced stomatal opening.

Arabidopsis↗

[Calcium involved in the signaling pathway of jasmonic acid induced stomatal closure of Vicia faba L].

Ca2+, an ubiquitous second messenger in the signal transudation pathway, is required for various physiological and developmental processes in plant. Jasmonic acid (JA) has been known to induce the stomatal closure. By monitoring the changes of [Ca2+]cyt with fluorescent probe Fluo-3 AM under the confocal microscopy, we observed that exogenous JA increased [Ca2+]cyt in guard cells of Vicia faba L. while the control and linolenic acid (LA), which is a precursor of JA, could hardly affect the change of [Ca2+]cyt. EGTA, a chelator of Ca2+ completely blocked JA-induced stomatal closure. After epidermis pretreated with EGTA, JA failed to result in [Ca2+]cyt increasing. Ruthenium red that blocked Ca2+ released from intracellular Ca2+ store could not significantly change JA-induced stomatal closure, while JA still increased [Ca2+]cyt. Furthermore, Ca2+ channel inhibitor of nifedipine (NIF) reduced the effectiveness of JA-induced stomatal closure and JA-induced increasing fluorescent intensity in guard cells. The results demonstrated that Ca2+ is involved in the signal transduction of JA induced stomatal closure, and the source of [Ca2+]cyt increasing in guard cells induced by JA might derive mainly from the external stores.

Calcium↗

Immune responses of cattle and mice to the G glycoprotein of vesicular stomatitis virus.

A subunit vaccine for vesicular stomatitis was developed from a purified vesicular stomatitis virus preparation by selectively removing the immunogenic G glycoprotein of the virus with the dialyzable, nonionic detergent, beta-D-octylglucoside. Cattle immunized intramuscularly with a single dose of 112 micrograms of G glycoprotein preparation in complete Freund's adjuvant did not develop vesicular disease following challenge by intralingual inoculation of 400 times the infectious dose of the virus. Similarly, mice vaccinated subcutaneously with a single dose of 10 micrograms of G glycoprotein preparation, with or without complete Freund's adjuvant, were protected from lethal encephalitis caused by vesicular stomatitis virus. A subunit vaccine for vesicular stomatitis of cattle, horses, and swine avoids the hazards associated with attenuated and inactivated vaccines, such as vaccine breaks, reversion to virulence, or introduction of virus into potential wild reservoirs or arthropod hosts. Further, it is possible to distinguish serologically animals vaccinated with the subunit preparation from those that have had the clinical disease or that have been vaccinated with whole virus. This is an essential consideration both for epidemiological studies and for disease control or establishment of quarantine programs.

Animals↗

The role of C. albicans in denture stomatitis.

Denture stomatitis is an erythematous disorder of the denture-bearing tissues, especially the maxillary hard palate. Frequently it mimics many oral lesions. Early diagnosis of the lesion is essential to assure rational therapy. A diagnostic procedure based upon the enumeration of Candida albicans appeared to be feasible. A control group of twenty fully dentate subjects with no clinical evidence of stomatitis was sampled to establish normal concentrations of C. albicans in the oral flora. Samples were taken from a triangular area of the hard palate, serially diluted in broth, and all dilutions were incubated aerobically. In the control population, C. albicans never exceeded 100 organisms per square centimeter per subject sample. A population of edentulous patients with denture stomatitis, prior to any therapy, was similarly sampled. In many of these patients the concentration of C. albicans always exceeded 10,000 organisms per square centimeter per patient. The patients received denture replacements and antimycotic antibiotic therapy. Follow-up cultures were taken 4 to 6 weeks after completion of therapy. All patients were found to be free of denture stomatitis.

Antifungal Agents↗

Oral hygiene, dental visits and age of denture for prevalence of denture stomatitis.

A representative sample of Finnish denture wearers (n = 3875) aged 30 or over were examined clinically and interviewed about their oral hygiene habits, use of dental services and possible prosthetic treatment during last visit to a dentist. Of these denture wearers, over 80% brushed their dentures at least once a day, and 16% visited a dentist at least once in 2 yr. For denture stomatitis the age-standardized risk ratio decreased with brushing frequency in women and increased in men. For frequency of dental visits the prevalence of denture stomatitis increased in both men and women when visits were made over 2-yr intervals. The prevalence of denture stomatitis was higher in the group with dentures over 1 yr old. It seems that the oral hygiene measures as they are generally carried out and regular dental visits are not effective enough methods in preventing denture stomatitis.

Adult↗

Serological monitoring of vesicular stomatitis New Jersey virus in enzootic regions of Costa Rica.

The activity of vesicular stomatitis viruses was monitored on 3 dairy farms in Costa Rica. Antibody levels were measured and clinical disease monitored in 165 cattle during a 20 month period (1986-1988). Vesicular stomatitis New Jersey (VS NJ) virus was shown to be enzootic on these farms by a 94.2% prevalence of neutralizing antibody; this did not vary significantly between herds. The mean prevalence of antibody to vesicular stomatitis Indiana (VS IN) virus was 15.2%, but was significantly higher in 1 herd. A total of 25 cases (annual incidence rate of 9%) of clinical vesicular stomatitis (VS) was reported. VS NJ virus was identified as the causal agent by detection of VS NJ virus antigens by the complement fixation test. VS NJ virus was isolated in 11 cases. All episodes of disease occurred between November and January, the beginning of the dry season. Most animals maintained stable neutralizing antibody titers throughout the study, and all diseased animals were previously seropositive to VS NJ virus. A total of 31 animals with neutralizing antibodies to VS NJ virus had a VS NJ virus-specific IgM response, and 6 animals had IgM responses that persisted for as long as 6 months. There was no relation between IgM responses and clinical disease occurrence. VS NJ virus persisted predominantly as a subclinical infection in cattle throughout the year in enzootic areas of Costa Rica. The humoral response did not prevent reinfection with VS NJ virus.

Animals↗

[Denture cleansing and denture stomatitis. Clinical investigation].

Three hundred patients with complete dentures were examined, to find out if there exists a relationship between denture cleansing and denture stomatitis prevalence. The study revealed that: 1. Denture stomatitis prevalence was 46%. 2. Mechanical means for denture cleansing were used by 84.67% of all patients, chemical means for denture cleansing were used by 5.33% and a combination of mechanical and chemical means by 10%. 3. Dentures with satisfactory cleanliness were used by 50.33% of the whole sample of patients and dentures with unsatisfactory cleanliness by 49.67% of all patients. 4. There exists a strong significant relationship (p less than 0.001) between dentures with unsatisfactory cleanliness and denture stomatitis prevalence. 5. There exists no significant relationship (p less than 0.05) between methods for denture cleansing and denture stomatitis prevalence.

Dentifrices↗

Oral cryotherapy for the prevention of high-dose melphalan-induced stomatitis in allogeneic hematopoietic stem cell transplant recipients.

GOALS: The purpose of this study was to evaluate the efficacy of oral cryotherapy to prevent high-dose melphalan-induced stomatitis. PATIENTS AND METHODS: Eighteen consecutive recipients of allogeneic hematopoietic stem cell transplant conditioned with high-dose melphalan (140 mg/m2) in combination with fludarabine alone or with fludarabine and additional chemotherapy or radiation were enrolled. The severity of stomatitis was graded according to the National Cancer Institute Common Toxicity Criteria. Patients were kept on oral cryotherapy using ice chips and ice-cold water shortly before, during, and for additional 90 min after completion of melphalan administration. RESULTS: Only two of 18 patients (11.1%) developed grade 2 or 3 stomatitis while six of seven patients in the historical control developed it (85.7%; P=0.001). CONCLUSION: These results suggested that oral cryotherapy could effectively prevent stomatitis caused by high-dose melphalan, and we recommend that it should be incorporated into the conditioning regimen with high-dose melphalan.

Antineoplastic Agents, Alkylating↗

Brief oral cryotherapy for the prevention of high-dose melphalan-induced stomatitis in allogeneic hematopoietic stem cell transplant recipients.

We previously reported the efficacy of oral cryotherapy for the prevention of high-dose melphalan-induced stomatitis. The purpose of this study was to evaluate whether the further shortening of the duration of oral cryotherapy could minimize its side effects while sparing its efficacy. Seventeen consecutive recipients of allogeneic hematopoieic stem cell transplant conditioned with high-dose melphalan in combination with fludarabine alone or with fludarabine and additional radiation were enrolled in the study. The severity of stomatitis was graded according to the National Cancer Institute-Common Toxicity Criteria. Patients were kept on oral cryotherapy shortly before, during, and for additional 30 min after the completion of melphalan administration (60-min oral cryotherapy). Patients who were also enrolled in our previous study received the same type of oral cryotherapy but for additional 90 min after the completion of melphalan administration (120-min oral cryotherapy), and they served as controls. Only 2 (11.8%) of 17 patients receiving 60-min oral cryotherapy and 2 (11.1%) of 18 patients receiving 120-min oral cryotherapy developed grade 2 or 3 stomatitis, respectively. The difference between groups was not statistically significant (P = 0.677). The incidence of unpleasant symptoms such as chills and nausea during oral cryotherapy decreased significantly with 60-min oral cryotherapy, as compared with that associated with 120-min oral cryotherapy (P < 0.01). These results suggest that 60-min oral cryotherapy is as effective as 120-min oral cryotherapy at preventing high-dose melphalan-induced stomatitis, and shorter treatment might have contributed to relieve patient discomfort during oral cryotherapy.

Administration, Oral↗

Distribution and phospholipase activity of Candida species in different denture stomatitis types.

The aim of this study was to evaluate the correlation between frequency and phospholipase activity of Candida species and denture stomatitis according to Newton's classification. Seventy-five complete denture wearers were evaluated for the presence of yeasts on the palatal mucosa by culture method. In addition, the number of yeast isolates producing phospholipase and amount of this enzyme were determined using egg yolk agar plate method. According to Newton's classification, 25 denture wearers were with healthy palatal mucosa while 50 were with any types of denture stomatitis. The frequency of yeasts was linked to whether subjects had Type II or Type III, but not Type I denture stomatitis. Candida albicans was the most frequently isolated species in denture wearers with and without clinical signs of denture stomatitis and it was the only species produced phospholipase. Although the amount of phospholipase produced by the C. albicans isolates from denture wearers in control and Type II and III DS groups was not significantly different, there was statistically significant difference in the number of C. albicans isolates producing phospholipase between patients with and without clinical signs of DS.

Adult↗

Hematologic status in recurrent aphthous stomatitis compared with other oral disease.

The hematologic status of 144 consecutive patients in the United Kingdom with oral disease (69 with recurrent aphthous stomatitis; 75 with other disorders) was examined in an attempt to resolve the controversy as to whether levels of hemoglobin, blood cell numbers and indices, and blood film are adequate in the screening of patients with aphthae. Though hemoglobin levels and red blood cell indices were normal in patients with recurrent aphthous stomatitis, there was still a small minority of patients with deficiencies of iron (low serum ferritin), folate (low red blood cell levels), or vitamin B12 (low serum levels) that would have remained undetected. Full hematologic screening of patients with recurrent aphthous stomatitis thus reveals latent deficiency states, at least in some parts of the world, and there are good reasons for elucidating the underlying cause and correcting these deficiencies, particularly vitamin B12 deficiency. Therapeutic studies are now required to establish the frequency with which deficiencies actually predispose to recurrent aphthous stomatitis.

Adult↗

Denture stomatitis: a role for Candida biofilms.

OBJECTIVE: To assess the contribution of Candida biofilms to the etiology of denture stomatitis. STUDY DESIGN: Samples of denture acrylic were retrieved from patients with denture stomatitis and subjected to scanning electron microscopy (SEM) analysis. Oral swab and swish samples were taken from the same group of patients and representative C albicans isolates recovered were used to investigate the kinetics of biofilm development in vitro. RESULTS: Candida biofilms could be visualized by SEM directly from denture samples from patients with denture stomatitis. These biofilms showed a propensity to adhere along cracks and imperfections of the denture acrylic. C albicans clinical isolates were able to form biofilms in vitro, although differences in the extent of biofilm formation were observed for different isolates recovered from the same patient. Susceptibility testing indicated that the resulting biofilms showed increased resistance to antifungal treatment. Presence of serum and saliva conditioning films increased the initial adherence of selected isolates but had little effect in overall biofilm formation. CONCLUSIONS: Candida biofilms play a role in denture stomatitis.

Acrylic Resins↗