Bioinspired inner microstructured tube controlled capillary rise.

Effective, long-range, and self-propelled water elevation and transport are important in industrial, medical, and agricultural applications. Although research has grown rapidly, existing methods for water film elevation are still limited. Scaling up for practical applications in an energy-efficient way remains a challenge. Inspired by the continuous water cross-boundary transport on the peristome surface of Nepenthes alata, here we demonstrate the use of peristome-mimetic structures for controlled water elevation by bending biomimetic plates into tubes. The fabricated structures have unique advantages beyond those of natural pitcher plants: bulk water diode transport behavior is achieved with a high-speed passing state (several centimeters per second on a milliliter scale) and a gating state as a result of the synergistic effect between peristome-mimetic structures and tube curvature without external energy input. Significantly, on further bending the peristome-mimetic tube into a "candy cane"-shaped pipe, a self-siphon with liquid diode behavior is achieved. Such a transport mechanism should inspire the design of next generation water transport devices.

Protein replenishment in pitcher fluids of Nepenthes × ventrata revealed by quantitative proteomics (SWATH-MS) informed by transcriptomics.

Carnivorous plants capture and digest insects for nutrients, allowing them to survive in soil deprived of nitrogenous nutrients. Plants from the genus Nepenthes produce unique pitchers containing secretory glands, which secrete enzymes into the digestive fluid. We performed RNA-seq analysis on the pitcher tissues and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis on the pitcher fluids of Nepenthes × ventrata to study protein expression in this carnivory organ during early days of pitcher opening. This transcriptome provides a sequence database for pitcher fluid protein identification. A total of 32 proteins of diverse functions were successfully identified in which 19 proteins can be quantified based on label-free quantitative proteomics (SWATH-MS) analysis while 16 proteins were not reported previously. Our findings show that certain proteins in the pitcher fluid were continuously secreted or replenished after pitcher opening, even without any prey or chitin induction. We also discovered a new aspartic proteinase, Nep6, secreted into pitcher fluid. This is the first SWATH-MS analysis of protein expression in Nepenthes pitcher fluid using a species-specific reference transcriptome. Taken together, our study using a gel-free shotgun proteomics informed by transcriptomics (PIT) approach showed the dynamics of endogenous protein secretion in the digestive organ of N. × ventrata and provides insights on protein regulation during early pitcher opening prior to prey capture.

Regulation of enzyme activities in carnivorous pitcher plants of the genus Nepenthes.

MAIN CONCLUSION: Nepenthes regulates enzyme activities by sensing stimuli from the insect prey. Protein is the best inductor mimicking the presence of an insect prey. Carnivorous plants of the genus Nepenthes have evolved passive pitcher traps for prey capture. In this study, we investigated the ability of chemical signals from a prey (chitin, protein, and ammonium) to induce transcription and synthesis of digestive enzymes in Nepenthes × Mixta. We used real-time PCR and specific antibodies generated against the aspartic proteases nepenthesins, and type III and type IV chitinases to investigate the induction of digestive enzyme synthesis in response to different chemical stimuli from the prey. Transcription of nepenthesins was strongly induced by ammonium, protein and live prey; chitin induced transcription only very slightly. This is in accordance with the amount of released enzyme and proteolytic activity in the digestive fluid. Although transcription of type III chitinase was induced by all investigated stimuli, a significant accumulation of the enzyme in the digestive fluid was found mainly after protein and live prey addition. Protein and live prey were also the best inducers for accumulation of type IV chitinase in the digestive fluid. Although ammonium strongly induced transcription of all investigated genes probably through membrane depolarization, strong acidification of the digestive fluid affected stability and abundance of both chitinases in the digestive fluid. The study showed that the proteins are universal inductors of enzyme activities in carnivorous pitcher plants best mimicking the presence of insect prey. This is not surprising, because proteins are a much valuable source of nitrogen, superior to chitin. Extensive vesicular activity was observed in prey-activated glands.

The effect of ramet mortality on clonal plant growth.

Clonal plants grow horizontally by producing multiple physiological individuals (ramets). We studied clonal growth in a homogeneous environment using a dynamic spatial model based on a stochastic cellular automaton. We investigated different growth forms from the aspect of ramet mortality. Non-steady-state and quasi-steady-state cases were defined, and we determined the number of steps suitable for making a reliable difference between these two types of cases. This given number of steps was used when testing for the proportion of quasi-steady-state cases in 1000 repetitions. We also tested the efficiency of occupation in these cases. Our expectation was that higher occupation would be associated with lower ramet mortality. The results only partially verified this hypothesis. Though with increasing ramet mortality, the average number of ramets tended to decrease, it was not the lowest ramet mortality that resulted in the highest occupation. Our results showed that very low ramet mortality was unfavourable for the plant, as the spreading front and the area behind this front were so packed that the plant was not able to return and recolonize the vacated sites in the central area. This resulted in a lower proportion of quasi-steady-state cases and lower occupation in these cases. Our results may contribute to a deeper understanding of clonal plant growth and its limiting factors.

Inter- and intra-variability of seed germination traits of Carpobrotus edulis N.E.Br. and its hybrid C. affine acinaciformis.

Invasions by alien Carpobrotus spp. have been recognised as one of the most severe threats to Mediterranean climate coastal ecosystems, and Carpobrotus is considered one of the most widespread invasive alien genera in the Mediterranean Basin. The aims of this study were to characterise seed germination of both C. edulis and its hybrid C. aff. acinaciformis, in terms of photoperiod, temperature and salinity. Inter- and intra-specific variability in the responses to photoperiod (12/12 h light and total darkness), constant temperatures (5, 10, 15, 20, 25 °C) and an alternating temperature regime (25/10 °C), salt stress (0, 125, 250, 500 mm NaCl) and the recovery of seed germination were evaluated for two seed lots of C. edulis and two of its hybrid C. aff. acinaciformis. All the tested seed lots achieved higher germination percentages in the light, with respect to total darkness. In relation to temperature, the two C. edulis seed lots did not show a preference, while the two C. aff. acinaciformis seed lots differed in their germination response, one germinating more at the lowest temperatures (5 and 10 °C) and one at the highest (20 and 25 °C). For all seed lots, highest germination occurred without NaCl (0 mm) and germination decreased with increasing salinity. Different germination requirements in a saline substrate were not detected for C. edulis, while they were observed for C. aff. acinaciformis. Marked differences were detected in recovery responses between the two taxa. C. edulis has the ability to germinate over a wide time window throughout the year. This study identified significant differences in seed production, seed mass, germination requirements (temperature) and salinity tolerance for both C. edulis and C. aff. acinaciformis. Our results indicate the extreme versatility of the hybrid forms to germinate in a wide range of natural conditions and habitats.

The role of organic acids on microbial deterioration in the Radix pseudostellariae rhizosphere under continuous monoculture regimes.

A three-year field monoculture trial of Radix pseudostellariae and complementary laboratory studies were conducted to further elucidate the underlying mechanism responsible for significant decreases in the biomass yield and quality of R. pseudostellariae under continuous monoculture regimes. HPLC analysis indicated that continuous monoculture soil was rich in organic acids, which had cumulative effects over time. Further analysis suggested that the application of a mixture of organic acids significantly promoted growth of pathogenic fungi, and increased the expression of chemotaxis-related gene (cheA) and biofilm formation of the specific pathogenic Kosakonia sacchari. However, opposite reactions were observed in the case of Bacillus megaterium and Bacillus pumilus. Concurrently, the present results revealed that the mixed organic acids stimulated the production of toxins, as well as H2O2 in the pathogenic fungi. Furthermore, the presence of organic acids reflecting environmental conditions under monocropping had negative effects on the expression of the biocontrol-related genes, which resulted in attenuated antagonistic activities of plant growth-promoting rhizobacteria (PGPR) to suppress mycelial growth of the pathogenic fungi. These results help to unveil the mechanisms associated with how accumulated organic acids differentially mediate deterioration of soil microbial composition and structure in monocropping system.