Chlorella triggers stomatal closure mediated by NADPH oxidase and improves instantaneous water use efficiency in Vicia faba.

Stomatal closure induced by live microbes and microbe-associated molecular patterns (MAMPs) has been associated with early defense responses of plants. Chlorella is a unicellular autotrophic microorganism that can synthesize many bioactive substances with positive effects on humans, animals and plants. However, its effects on stomatal movement and instantaneous intrinsic water use efficiency (WUEi) in plants have been not explored yet. Our present work showed that application of Chlorella to isolated epidermal peels of Vicia faba induced stomatal closure in a dose-and time-dependent manner. Pharmacological study revealed that the Chlorella-triggered stomatal closure was mainly mediated by reactive oxygen species (ROS) production via NADPH oxidase. Additionally, exogenous application of optimal concentrations of Chlorella suspension caused an obvious reduction in leaf transpiration rate (E) without a parallel reduction in net photosynthetic rate (Pn), favoring the improvement of WUEi in Vicia faba. The chlorophyll fluorescence and content analysis further indicated that Chlorella had no effects on plant photosynthetic reactions center after short-term foliar application.

Chlorella induces stomatal closure via NADPH oxidase-dependent ROS production and its effects on instantaneous water use efficiency in Vicia faba.

Reactive oxygen species (ROS) have been established to participate in stomatal closure induced by live microbes and microbe-associated molecular patterns (MAMPs). Chlorella as a beneficial microorganism can be expected to trigger stomatal closure via ROS production. Here, we reported that Chlorella induced stomatal closure in a dose-and time-dependent manner in epidermal peels of Vicia faba. Using pharmacological methods in this work, we found that the Chlorella-induced stomatal closure was almost completely abolished by a hydrogen peroxide (H2O2) scavenger, catalase (CAT), significantly suppressed by an NADPH oxidase inhibitor, diphenylene iodonium chloride (DPI), and slightly affected by a peroxidase inhibitor, salicylhydroxamic acid (SHAM), suggesting that ROS production involved in Chlorella-induced stomatal closure is mainly mediated by DPI-sensitive NADPH oxidase. Additionally, Exogenous application of optimal concentrations of Chlorella suspension improved instantaneous water use efficiency (WUEi) in Vicia faba via a reduction in leaf transpiration rate (E) without a parallel reduction in net photosynthetic rate (Pn) assessed by gas-exchange measurements. The chlorophyll fluorescence and content analysis further demonstrated that short-term use of Chlorella did not influence plant photosynthetic reactions center. These results preliminarily reveal that Chlorella can trigger stomatal closure via NADPH oxidase-dependent ROS production in epidermal strips and improve WUEi in leave levels.

Saccharomyces cerevisiae-induced stomatal closure mainly mediated by salicylhydroxamic acid-sensitive peroxidases in Vicia faba.

Saccharomyces cerevisiae induced stomatal closure in a dose-dependent manner on Vicia faba L. (cv. Daqingpi). Using pharmacological inhibitors in this study, we found that stomatal closure was completely inhibited by salicylhydroxamic acid (SHAM) and reduced glutathione (GSH), whereas slightly inhibited by diphenyleneiodonium chloride (DPI), suggesting that H2O2 was mostly produced by cell wall peroxidases. The specific NO scavenger (cPTIO), NO synthase (NOS) inhibitor NG-nitro-l-arginine methyl ester (l-NAME) and sodium azide (NaN3; inhibitor of nitrate reductase) prevented yeast-induced stomatal closure, suggesting that NO in guard cells of V. faba is derived from both NOS-like enzyme and nitrate reductase. Results of HgCl2 and ß-mercaptoethanol (ME) treatment (as a functional inhibitor of water channels and its reversing agent, respectively) suggest that water channels are involved in yeast-induced stomatal movements. CoCl2 (the blocker of calcium channel), LaCl3 (Ca(2+) antagonist) and EGTA (Ca(2+) chelator) also impaired yeast-induced stomatal closure. Thus, it is concluded that H2O2, NO, water channels and Ca(2+) are involved in yeast-induced stomatal closure.

Scaling relationship between tree respiration rates and biomass.

The WBE theory proposed by West, Brown and Enquist predicts that larger plant respiration rate, R, scales to the three-quarters power of body size, M. However, studies on the R versus M relationship for larger plants (i.e. trees larger than saplings) have not been reported. Published respiration rates of field-grown trees (saplings and larger trees) were examined to test this relationship. Our results showed that for larger trees, aboveground respiration rates RA scaled as the 0.82-power of aboveground biomass MA, and that total respiration rates RT scaled as the 0.85-power of total biomass MT, both of which significantly deviated from the three-quarters scaling law predicted by the WBE theory, and which agreed with 0.81-0.84-power scaling of biomass to respiration across the full range of measured tree sizes for an independent dataset reported by Reich et al. (Reich et al. 2006 Nature 439, 457-461). By contrast, R scaled nearly isometrically with M in saplings. We contend that the scaling exponent of plant metabolism is close to unity for saplings and decreases (but is significantly larger than three-quarters) as trees grow, implying that there is no universal metabolic scaling in plants.

Reproductive allocation patterns in different density populations of spring wheat.

The effects of increased intraspecific competition on size hierarchies (size inequality) and reproductive allocation were investigated in populations of the annual plant, spring wheat (Triticum aestivum). A series of densities (100, 300, 1000, 3000 and 10,000 plants/m(2)) along a gradient of competition intensity were designed in this experiment. The results showed that average shoot biomass decreased with increased density. Reproductive allocation was negatively correlated to Gini coefficient (R(2) = 0.927), which suggested that reproductive allocation is inclined to decrease as size inequality increases. These results suggest that both vegetative and reproductive structures were significantly affected by intensive competition. However, results also indicated that there were different relationships between plant size and reproductive allocation pattern in different densities. In the lowest density population, lacking competition (100 plants/m(2)), individual reproductive allocation was size independent but, in high density populations (300, 1000, 3000 and 10,000 plants/m(2)), where competition occurred, individual reproductive allocation was size dependent: the small proportion of larger individuals were winners in competition and got higher reproductive allocation (lower marginal reproductive allocation; MRA), and the larger proportion of smaller individuals were suppressed and got lower reproductive allocation (higher MRA). In conclusion, our results support the prediction that elevated intraspecific competition would result in higher levels of size inequality and decreased reproductive allocation (with a negative relationship between them). However, deeper analysis indicated that these frequency- and size-dependent reproductive strategies were not evolutionarily stable strategies.

Trade-offs between the metabolic rate and population density of plants.

The energetic equivalence rule, which is based on a combination of metabolic theory and the self-thinning rule, is one of the fundamental laws of nature. However, there is a progressively increasing body of evidence that scaling relationships of metabolic rate vs. body mass and population density vs. body mass are variable and deviate from their respective theoretical values of 3/4 and -3/4 or -2/3. These findings questioned the previous hypotheses of energetic equivalence rule in plants. Here we examined the allometric relationships between photosynthetic mass (M(p)) or leaf mass (M(L)) vs. body mass (beta); population density vs. body mass (delta); and leaf mass vs. population density, for desert shrubs, trees, and herbaceous plants, respectively. As expected, the allometric relationships for both photosynthetic mass (i.e. metabolic rate) and population density varied with the environmental conditions. However, the ratio between the two exponents was -1 (i.e. beta/delta = -1) and followed the trade-off principle when local resources were limited. Our results demonstrate for the first time that the energetic equivalence rule of plants is based on trade-offs between the variable metabolic rate and population density rather than their constant allometric exponents.

Network regulation of calcium signal in stomatal development.

AIM: Each cell is the production of multiple signal transduction programs involving the expression of thousands of genes. This study aims to gain insights into the gene regulation mechanisms of stomatal development and will investigate the relationships among some signaling transduction pathways. METHODS: Nail enamel printing was conducted to observe the stomatal indices of wild type and 10 mutants (plant hormone mutants, Pi-starvation induced CaM mutants and Pi-starvation-response mutant) in Arabidopsis, and their stomatal indices were analyzed by ANOVA. We analyzed the stomatal indices of 10 Arabidopsis mutants were analyzed by a model PRGE (potential relative effect of genes) to research relations among these genes. RESULTS: In wild type and 10 mutants, the stomatal index did not differ with respect to location on the lower epidermis. Compared with wild type, the stomatal indices of 10 mutants all decreased significantly. Moreover, significant changes and interactions might exist between some mutant genes. CONCLUSION: It was the stomatal intensity in Arabidopsis might be highly sensitive to most mutations in genome. While the effect of many gene mutations on the stomatal index might be negative, we also could assume the stomatal development was regulated by a signal network in which one signal transduction change might influence the stomatal development more or less, and the architecture might be reticulate. Furthermore, we could speculate that calcium was a hub in stomatal development signal regulation network, and other signal transduction pathways regulated stomatal development by influencing or being influenced by calcium signal transduction pathways.

[Effects of exogenous nitric oxide, salicylic acid and hydrogen peroxide on free amino acid and soluble protein contents in tobacco leaves].

The work focused on the effects of signal molecules of nitric oxide (NO), salicylic acid (SA) and hydrogen peroxide (H(2)O(2)) on compounds of nitric metabolites of free amino acid, soluble protein and proline in tobacco. The results indicated that NO, SA and H(2)O(2) were able to regulate the proline, free amino acid and soluble protein content in tobacco. Lower concentration of NO and H(2)O(2) raised the level of proline; while higher concentration of NO and H(2)O(2) lowered the proline, free amino acid content in tobacco. The above three signal molecules also showed similar effects on proline, free amino acid and soluble protein content in tobacco.

Impact of elevated CO2 concentration under three soil water levels on growth of Cinnamomum camphora.

Forest plays very important roles in global system with about 35% land area producing about 70% of total land net production. It is important to consider both elevated CO(2) concentrations and different soil moisture when the possible effects of elevated CO(2) concentration on trees are assessed. In this study, we grew Cinnamomum camphora seedlings under two CO(2) concentrations (350 micromol/mol and 500 micromol/mol) and three soil moisture levels [80%, 60% and 40% FWC (field water capacity)] to focus on the effects of exposure of trees to elevated CO(2) on underground and aboveground plant growth, and its dependence on soil moisture. The results indicated that high CO(2) concentration has no significant effects on shoot height but significantly impacts shoot weight and ratio of shoot weight to height under three soil moisture levels. The response of root growth to CO(2) enrichment is just reversed, there are obvious effects on root length growth, but no effects on root weight growth and ratio of root weight to length. The CO(2) enrichment decreased 20.42%, 32.78%, 20.59% of weight ratio of root to shoot under 40%, 60% and 80% FWC soil water conditions, respectively. And elevated CO(2) concentration significantly increased the water content in aboveground and underground parts. Then we concluded that high CO(2) concentration favours more tree aboveground biomass growth than underground biomass growth under favorable soil water conditions. And CO(2) enrichment enhanced lateral growth of shoot and vertical growth of root. The responses of plants to elevated CO(2) depend on soil water availability, and plants may benefit more from CO(2) enrichment with sufficient water supply.