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Short basal internodes are important for lodging resistance of rice (Oryza sativa L.). Several canopy indices affect the elongation of basal internodes, but uncertainty as to the key factors determining elongation of basal internodes persists. The objectives of this study were (1) to identify key factors affecting the elongation of basal internodes and (2) to establish a quantitative relationship between basal internode length and canopy indices. An inbred rice cultivar, Yinjingruanzhan, was grown in two split-plot field experiments with three N rates (0, 75, and 150 kg N ha−1 in early season and 0, 90, and 180 kg N ha−1 in late season) as main plots, three seedling densities (16.7, 75.0, and 187.5 seedlings m−2) as subplots, and three replications in the 2015 early and late seasons in Guangzhou, China. Light intensity at base of canopy (Lb), light quality as determined from red/far-red light ratio (R/FR), light transmission ratio (LTR), leaf area index (LAI), leaf N concentration (NLV) and final length of second internode (counted from soil surface upward) (FIL) were recorded. Higher N rate and seedling density resulted in significantly longer FIL. FIL was negatively correlated with Lb, LTR, and R/FR (P 0.05). Stepwise linear regression analysis showed that FIL was strongly associated with Lb and LAI (R2 = 0.82). Heavy N application to pot-grown rice at the beginning of first internode elongation did not change FIL. We conclude that FIL is determined mainly by Lb and LAI at jointing stage. NLV has no direct effect on the elongation of basal internodes. N application indirectly affects FIL by changing LAI and light conditions in the rice canopy. Reducing LAI and improving canopy light transmission at jointing stage can shorten the basal internodes and increase the lodging resistance of rice. Keywords: Internode elongation, Leaf area index, Light intensity, Light quality, R/FR, Light transmission ratio, Leaf N concentration

Zinc homeostasis is crucial for the development and stress resistance of bacteria in the environment. Serial zinc sensing transcriptional regulators, zinc transporters and zinc binding proteins were found to maintain the zinc homeostasis in bacteria. Zur is a zinc uptake regulator that is widely distributed in species, and ZnuABC, as well as the Type VI Secretion System (T6SS4) function in zinc acquisition. Here, we report that the regulator Zur inhibits the expression of the ZnuABC which inhibition could be eliminated at low zinc level, and upregulates the T6SS4 operon in Yersinia pseudotuberculosis to facilitate Zn2+ uptake and oxidative stress resistance. Zur regulates the expression of ZnuABC and T6SS4 by directly binding to their promoter regions. Zur senses the Zn2+ concentration and represses ZnuABC in a Zn2+-containing environment. Zur works as an auxiliary regular activator of T6SS4, facilitating oxidative stress resistance. This study revealed the dual function of regulator Zur on ZnuABC and T6SS4, and enriched the knowledge of Zn2+ homeostasis maintenance in Y. pseudotuberculosis.

Bacterial type VI secretion systems (T6SSs) inject toxic effectors into adjacent eukaryotic and prokaryotic cells. It is generally thought that this process requires physical contact between the two cells. Here, we provide evidence of contact-independent killing by a T6SS-secreted effector. We show that the pathogen Yersinia pseudotuberculosis uses a T6SS (T6SS-3) to secrete a nuclease effector that kills other bacteria in vitro and facilitates gut colonization in mice. The effector (Tce1) is a small protein that acts as a Ca2+- and Mg2+-dependent DNase, and its toxicity is inhibited by a cognate immunity protein, Tci1. As expected, T6SS-3 mediates canonical, contact-dependent killing by directly injecting Tce1 into adjacent cells. In addition, T6SS-3 also mediates killing of neighboring cells in the absence of cell-to-cell contact, by secreting Tce1 into the extracellular milieu. Efficient contact-independent entry of Tce1 into target cells requires proteins OmpF and BtuB in the outer membrane of target cells. The discovery of a contact-independent, long-range T6SS toxin delivery provides a new perspective for understanding the physiological roles of T6SS in competition. However, the mechanisms mediating contact-independent uptake of Tce1 by target cells remain unclear., Bacteria can use type VI secretion systems (T6SSs) to inject toxic effector proteins into adjacent cells, in a contact-dependent manner. Here, the authors provide evidence of contact-independent killing by a T6SS effector that is secreted into the extracellular milieu and then taken up by other bacterial cells.

Potassium (K) fertilizer additions can result in high crop yields of good quality and low nitrogen (N) loss; however, the interaction between K and N fertilizer and its effect on N2O emissions and associated microbes remain unclear. We investigated this in a pot experiment with six fertilizer treatments involving K and two sources of N, using agricultural soil from the suburbs of Wuhan, central China. The aim was to determine the effects of the interaction between K and different forms of N on the N2O flux and the abundance of nitrifying and denitrifying microbial communities, using static chamber-gas chromatography and high-throughput sequencing methods. Compared with no fertilizer control (CK), the addition of nitrate fertilizer (NN) or ammonia fertilizer (AN) or K fertilizer significantly increased N2O emissions. However, the combined application (NNK) of K and NN significantly reduced the average N2O emissions by 28.3%, while the combined application (ANK) of K and AN increased N2O emissions by 22.7%. The abundance of nitrifying genes amoA in ammonia oxidizing archaea (AOA) and ammonia oxidizing bacteria (AOB) changed in response to N and/or K fertilization, but the denitrifying genes narG, nirK and norl were strongly correlated with N2O emissions. This suggests that N or K fertilizer and their interaction affect N2O emissions mainly by altering the abundance of functional genes of denitrifying microbes in the soil-plant system. The genera Paracoccus, Rubrivivax and Geobacter as well as Streptomyces and Hyphomicrobium play an important role in N2O emissions during denitrification with the combined application of N and K.

The transition between the planktonic state and the biofilm-associated state is a key developmental decision for pathogenic bacteria. Biofilm formation by Yersinia pestis is regulated by hmsHFRS genes (β-1, 6-N-acetyl-D-glucosamine synthesis operon) in its flea vector and in vitro. However, the mechanism of biofilm formation in Yersinia pseudotuberculosis remains elusive. In this study, we demonstrate that the LysR-type regulator RovM inversely regulates biofilm formation and motility in Y. pseudotuberculosis by acting as a transcriptional regulator of these two functions. RovM is strongly induced during growth in minimal media but strongly repressed in complex media. On one hand, RovM enhances bacterial motility by activating the expression of FlhDC, the master regulator of flagellar genes, via the recognition of an operator upstream of the flhDC promoter. On the other hand, RovM represses β-GlcNAc production under nutrition-limited conditions, negatively regulating hmsHFRS expression by directly binding to the −35 element of its promoter. Compared to wild-type bacteria, the rovM mutant established denser biofilms and caused more extensive mortality in mice and silkworm larvae. These results indicate that RovM acts as a molecular switch to coordinate the expression of genes involved in biofilm formation and motility in response to the availability of nutrients.

Mycothiol (MSH) plays a major role in protecting cells against oxidative stress and detoxification from a broad range of exogenous toxic agents. In the present study, we reveal that intracellular MSH contributes significantly to the adaptation to acidic conditions in the model organism Corynebacterium glutamicum. We present evidence that MSH confers C. glutamicum with the ability to adapt to acidic conditions by maintaining pHi homeostasis, scavenging reactive oxygen species (ROS), and protecting methionine synthesis by the S-mycothiolation modification of methionine synthase (MetE). The role of MSH in acid adaptation was further confirmed by improving the acid tolerance of C. glutamicum by overexpressing the key MSH synthesis gene mshA. Hence, our work provides insights into a previously unknown, but important, aspect of the C. glutamicum cellular response to acid stress. The results reported here may help to understand acid tolerance mechanisms in acid sensitive bacteria and may open a new avenue for improving acid resistance in industry strains for the production of bio-based chemicals from renewable biomass.

The type VI secretion system (T6SS) is a versatile molecular machinery widely distributed in Gram-negative bacteria. The activity of the T6SS is tightly regulated by various mechanisms, including quorum sensing (QS), iron concentration, and transcriptional regulators. Here we demonstrated that the stringent response regulator, RelA, contributes to bacterial resistance to multiple environmental stresses in Yersinia pseudotuberculosis. We also revealed that the stress resistance function of stringent response (SR) was partially mediated by the general stress response T6SS4 system. RelA positively regulates the expression of T6SS4 to combat various stresses in response to nutrition starvation collectively mediated by the RovM and RovA regulators. These findings revealed not only the important role of T6SS4 in SR induced stress resistance, but also a new pathway to regulate T6SS4 expression in response to starvation stress.

The type VI secretion system (T6SS) is a versatile secretion system widely distributed in Gram-negative bacteria that delivers multiple effector proteins into either prokaryotic or eukaryotic cells, or into the extracellular milieu. T6SS participates in various physiological processes including bacterial competition, host infection, and stress response. Three pathogenic Yersinia species, namely Yersinia pestis, Yersinia pseudotuberculosis, and Yersinia enterocolitica, possess different copies of T6SSs with distinct biological functions. This review summarizes the pathogenic, antibacterial, and stress-resistant roles of T6SS in Yersinia and the ion-transporting ability in Y. pseudotuberculosis. In addition, the T6SS-related effectors and regulators identified in Yersinia are discussed.

RpoS (σ(S)), the stationary phase/stress σ factor, controls the expression of a large number of genes involved in cellular responses to a variety of stresses. However, the role of RpoS appears to differ in different bacteria. While RpoS is an important regulator of flagellum biosynthesis, it is associated with biofilm development in Edwardsiella tarda. Biofilms are dense communities formed by bacteria and are important for microbe survival under unfavorable conditions. The type VI secretion system (T6SS) discovered recently is reportedly associated with several phenotypes, ranging from biofilm formation to stress sensing. For example, Vibrio anguillarum T6SS was proposed to serve as a sensor for extracytoplasmic signals and modulates RpoS expression and stress response. In this study, we investigated the physiological roles of RpoS in Yersinia pseudotuberculosis, including bacterial survival under stress conditions, flagella formation, biofilm development and T6SS expression. We found that RpoS is important in resistance to multiple stressors-including H2O2, acid, osmotic and heat shock-in Y. pseudotuberculosis. In addition, our study showed that RpoS not only modulates the expression of T6SS but also regulates flagellum formation by positively controlling the flagellar master regulatory gene flhDC, and affects the formation of biofilm on Caenorhabditis elegans by regulating the synthesis of exopolysaccharides. Taken together, these results show that RpoS plays a central role in cell fitness under several adverse conditions in Y. pseudotuberculosis.

Diatoms, with hierarchical micro/nanoscale porous silica structures, have promising application in micro-nanotechnology especially biochemical sensing. In order to explore their potential and prepare diatom based substrates for biochemical sensor application, a fabrication technology for bonding diatom frustules and Si substrates was developed. The bonding process was carried out at 75 °C and assisted by hydrofluoric acid (HF). The bonding mechanism was discussed and several bonding conditions were adjusted to keep the morphological integrity of diatom frustules after bonding. The bonding pressure was optimized from 2.0 × 104 Pa to 3.0 × 104 Pa and the HF concentration from 0.4% to 0.6%. And the optimal shear bonding strength achieved was 0.72 MPa. In addition, bonded diatom frustules were further used as masks to obtain nano gold pillar arrays for surface-enhanced Raman scattering (SERS) detection.

We proved the feasibility of using a microfluidic chip to culture diatom Bacillaria paradoxa, and analyzed the gliding characteristics of its self-organized colony in detail. The optimal cultivation parameters of B. paradoxa for the designed chip made with polydimethylsiloxane are as follows: the preferable cells injecting rate for keeping the cells alive is 0.2 mL/h, the initial cell density for fast reproduction is 5.5 × 10(4) cells/mL, and the optimal replacement period of culture medium is 4 days. B. paradoxa tends to form a colony during their growth, and the colony can glide with a steady period of 29 ± 3 s along its axial direction in a constant stream, the amplitude of the colony will not decay (e.g., 24 μm of two-cell colony at 1.1 mm/s flow rate), and the colony rapidly adjusts its direction of gliding to the direction of water flow. The successful culture of diatoms on a microfluidic platform may be used for biosensing chips and the creation of gasoline-producing diatom solar panels.

A filtration-aided acid cleaning method was used to collect biosilica structures from a diatom culture medium, natural seawater, or water bloom. Cell extraction, acid cleaning, and acid removal were all performed on a polytetrafluoroethylene (PTFE) filter cloth, significantly improving the treatment capacity and efficiency of the traditional acid wash method. Five typical diatoms were cultivated in the laboratory for acid cleaning. Different growth speeds were introduced, and different process parameters for acid cleaning were utilized. After the acid cleaning, biosilica structures were collected from the frustules of diatoms using different methods. Girdle bands and valves of Coscinodiscus sp. were separated by floating of the valves. Central spines of Ditylum brightwellii and valves of Skeletonema costatum were separately collected by settling or filtration. Rod-like frustules, such as those of Bacillaris paradoxa, are not suitable for large quantities of acid wash. The silica structures were observed and tested using an AFM-calibrated glass needle to determine their elasticity. Elasticity tests showed that ringent girdle bands are more flexible than complete ones (Coscinodiscus sp.) and that both long-chain clusters of Nitzschia palea and central spines of D. brightwellii have certain elasticities. The required pressure for deforming or breaking the biosilica structures of diatoms was also determined.

Based on the fact that SiO2 can dissolve in HF solution, three kinds of diatom frustules were treated with 1% HF solution at room temperature. Given the proper reaction times (0–2 h for the diatoms Coscinodiscus and Navicula, and 0–3 h for the diatom Melosira), the size of the pores on the frustules gradually increased and the structures of the frustules remained. While HF treatment does not affect the composition, chemical bonds, or photoluminescence signature of the diatom frustules, the treatment reduces their surface areas. This method may be beneficial to diatom studies, diatom nanotechnology, and diatom device applications that make use of diatom pores.

Diatom valves and girdle bands are useful micro–nano materials in nanotechnology and micro manufacturing. A settling method is used to extract high-purity diatom valves and girdle bands from Coscinodiscus diatomite. The average models of diatom valves and girdle bands are established. Stokes Law and hydrokinetic theories are used to analyze the settling velocity of valves and girdle bands. Based on the calculation results, settling experiments are carried out, by which clean diatom valves with purity of 80% and girdle bands with purity of 90% are obtained. This method can be applied to other particles separation problem to separate micro-particles with similar radius but different sectional area.

A photoassisted chemical bonding method was used to accurately position diatom frustules into complex microscale patterns on a PDMS surface. The micropores and fine features of the diatom frustules...

Weeding and fertilization are important farming practices. Integrated weed management should protect or improve the biodiversity of farmland weed communities for a better ecological environment with not only increased crop yield, but also reduced use of herbicides. This study hypothesized that appropriate fertilization would benefit both crop growth and the biodiversity of farmland weed communities. To study the effects of different fertilizing patterns on the biodiversity of a farmland weed community and their adaptive mechanisms, indices of species diversity and responses of weed species and wheat were investigated in a 17-year field trial with a winter wheat-soybean rotation. This long term field trial includes six fertilizing treatments with different N, P and K application rates. The results indicated that wheat and the four prevalent weed species (Galium aparine, Vicia sativa, Veronica persica and Geranium carolinianum) showed different responses to fertilizer treatment in terms of density, plant height, shoot biomass, and nutrient accumulations. Each individual weed population exhibited its own adaptive mechanisms, such as increased internode length for growth advantages and increased light interception. The PK treatment had higher density, shoot biomass, Shannon-Wiener and Pielou Indices of weed community than N plus P fertilizer treatments. The N1/2PK treatment showed the same weed species number as the PK treatment. It also showed higher Shannon-Wiener and Pielou Indices of the weed community, although it had a lower wheat yield than the NPK treatment. The negative effects of the N1/2PK treatment on wheat yield could be balanced by the simultaneous positive effects on weed communities, which are intermediate in terms of the effects on wheat and weeds.

Diatom, with delicate nanoscale porous SiO 2 structures, has a promising application in biosensing. We bond diatoms with SiO 2 substrate using spin-on glass (SOG) as interlayer to acquire a high performance diatom based fluorescence detecting biosensor. As diatoms can improve proteins density, diatom based biosensors have higher sensitivity compared with traditional protein sensors. This biosensor is fully made of SiO 2 which can be modified for biomass attachment. Nanoscale structures of bonded diatoms are better preserved than any related works. The highest bonding strength is 0.6 MPa. Human immunoglobulin G (IgG) is attached on traditional protein sensors and diatom based biosensors as antibody. After hybridized with Cy3-goat anti-human IgG, the diatom based biosensor provides a fluorescence detection signal 2.5 times higher than the traditional one. Given improved fluorescence detecting sensitivity, this new kind of diatom based biosensor can be widely used for various protein detections.

Diatoms have silica frustules with transparent and delicate micro/nano scale structures, two dimensional pore arrays, and large surface areas. Although, the diatom cells of Coscinodiscus sp. live underwater, we found that their valves can float on water and assemble together. Experiments show that the convex shape and the 40 nm sieve pores of the valves allow them to float on water, and that the buoyancy and the micro-range attractive forces cause the valves to assemble together at the highest point of water. As measured by AFM calibrated glass needles fixed in manipulator, the buoyancy force on a single floating valve may reach up to 10 μN in water. Turning the valves over, enlarging the sieve pores, reducing the surface tension of water, or vacuum pumping may cause the floating valves to sink. After the water has evaporated, the floating valves remained in their assembled state and formed a monolayer film. The bonded diatom monolayer may be valuable in studies on diatom based optical devices, biosensors, solar cells, and batteries, to better use the optical and adsorption properties of frustules. The floating assembly phenomenon can also be used as a self-assembly method for fabricating monolayer of circular plates.

Diatoms have delicate porous structures which are very beneficial in improving the absorbing ability in the bio-detection field. In this study, multi-layered hierarchical arrays were fabricated by packing Nitzschia soratensis (N. soratensis) frustules into Cosinodiscus argus (C. argus) frustules to achieve advanced sensitivity in bio-detection chips. Photolithographic patterning was used to obtain N. soratensis frustule arrays, and the floating behavior of C. argus frustules was employed to control their postures for packing N. soratensis frustule array spots. The morphology of the multi-layer C. argus–N. soratensis package array was investigated by scanning electron microscopy, demonstrating that the overall and sub-structures of the diatom frustules were retained. The signal enhancing effect of multi-layer C. argus–N. soratensis packages was demonstrated by fluorescent antibody test results. The mechanism of the enhancement was also analyzed, indicating that both complex hierarchical frustule structures and optimized posture of C. argus frustules were important for improving bio-detection sensitivities. The technique for fabricating multi-layer diatom frustules arrays is also useful for making multi-functional biochips and controllable drug delivery systems.

Diatom, with delicate three-dimensional porous structures and texture, has a promising application in micro-nanotechnology especially biosensing. In order to achieve a diatom-based compound substrate, a fabrication technique is developed for hydrofluoric acid (HF) bonding of diatom with SiO2-based substrate at a temperature as low as 80 °C. The bonding conditions are optimized with various HF concentrations and applied pressure. The optimized HF concentration is found to be in the range of 0.8% to 1.2% and applied pressure is from 0.4.0 MPa to 0.6.0 MPa. The morphological integrity and nano-microscale substructures of the diatoms after bonding are characterized. The bonding strength is approximately 0.435 MPa.

Diatom frustules, considered as novel bio-functional materials, display a diversity of patterns and unique micro- and nanostructures which may be useful in many areas of application. Existing devices directly use the original structure of the biosilica frustules, limiting their function and structural scale. Current research into the shapes, materials and structural properties of frustules are considered; a series of frustule processing methods including structure processing, material modification, bonding and assembly techniques are reviewed and discussed. The aim is to improve the function of diatom frustules allowing them to meet the design requirements of different types of micro devices. In addition, the importance of the comprehensive use of diatom processing methods in device research is discussed using biosensors and solar cells as examples, and the potential of bio-manufacturing technology based on diatom frustules is examined.