Photosynthetic and biochemical traits change in the green-tide-forming macroalga Ulva pertusa during sporulation1

The physiological and biochemical changes in the green macroalga Ulva pertusa during the progression of sporulation have been characterized. The transition from the vegetative to the sporulation stage was accompanied by an increase in chlorophyll a (Chl a), chlorophyll b (Chl b), and carotenoid content, as well as an increase in DPPH scavenging and responsiveness to diphenylamine. However, oxygen evolution and maximum electron transport rate decreased. The discrepancy between photosynthetic performance and pigment content might relate to the self-shading of spores within a sporangium. Spore-forming U. pertusa thalli were low-light-adapted, due to an increase in the number of photosynthetic units. Decreased electron transport during sporulation might trigger sporulation, as for some cyanobacteria and other Ulva spp., via oxidization of the plastoquinone pool and cyclic phosphorylation, thus producing ATP to generate carbon and nitrogen skeletons required for spores. It is thus concluded that carotenoids function both in spore initiation and/or maturation and in their photoprotection.     

Design and Optimization of Planting Process Parameters for 2ZYX-2 Type Green Onion Ditching and Transplanting Machine

Considering the current low level of mechanization for domestic green onion planting and the high labor intensity of artificial planting, a 2ZYX-2 green onion ditching and transplanting machine, which can complete ditching, ridging, transplanting, repression, soil covering and other operations, is designed in this study. The Central Composite test design method was carried out with the speed of the transplanting machine, the depth of the opener and the horizontal position of the opener as the experimental factors and with the qualification ratio of perpendicularity, the variation coefficient of the plant spacing and the qualification ratio of the planting depth as the test index. Through the analysis of the model interaction and response surface, the change laws that the influence the machine’s forward speed, the depth of the opener and the horizontal position of the opener were studied. The regression model was optimized by Design-Expert 8.0.6 software, and the accuracy of the predicted results was verified by experiments. The optimal working parameters showed that the forward speed of the machine was 0.06 m/s, the depth of the opener was 102 mm, and the horizontal position of the opener was 29 mm. Under conditions of optimal working parameters, the qualification rate of the verticality was 86.83%, the coefficient of variation for the plant spacing was 2.77, and the pass rate of planting depth was 88.26%. The research related to the thesis can provide a reference for the mechanized planting of green onion, which is of great significance to the cost-effectiveness of the green onion industry.     

Early detection of Agrilus planipennis: investigations into the attractive range of the sex pheromone (3Z)-lactone

The emerald ash borer (EAB), Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), is a highly destructive primary pest of ash (Fraxinus sp., Oleaceae) trees outside of its native range. Ash is an important component of many ecosystems and its loss would be detrimental to both the economy and the environment. The present study aimed to improve our understanding of the effectiveness of green sticky prism traps baited with host kairomone and insect pheromone lures for A. planipennis and to collect data for modelling the range of attraction of the pheromone (3Z)-dodecen-12-olide [(3Z)-lactone]. Traps were deployed over a single flight season in urban locations of Ontario, Canada, with low densities of EAB. Traps were placed in pairs of trees separated by not more than 25 m. All traps contained the host kairomone, (3Z)-hexenol, with the remaining half in each pairing additionally baited with (3Z)-lactone pheromone. Both lure types were highly effective in capturing EAB, with >90% detection rates overall. However, traps baited with the lactone pheromone and host volatile lures doubled trap captures of EAB over distances of at least 25 m from the nearest traps baited with only host volatiles. Although the baseline detection rate of traps containing (3Z)-hexenol alone is not significantly reduced compared with traps containing (3Z)-lactone, the overall trap effectiveness is significantly increased when (3Z)-lactone is present. The implications for the use of (3Z)-lactone at 3 mg per septum dose in an early warning trapping system are discussed. Trap layout methods and risk-based analysis models can now be further refined by including these data about the attractive range of lures and their behaviour in different plot environments.     

Current genetic engineering strategies for the production of antihypertensive ACEI peptides

Hypertension is a major and highly prevalent risk factor for various diseases. Among the most frequently prescribed antihypertensive first-line drugs are synthetic angiotensin I-converting enzyme inhibitors (ACEI). However, since  their use in hypertension therapy has been linked to various side effects, interest in the application of food-derived ACEI peptides (ACEIp) as antihypertensive agents is rapidly growing. Although promising, the industrial production of ACEIp through conventional methods such as chemical synthesis or enzymatic hydrolysis of food proteins has been proven troublesome. We here provide an overview of current antihypertensive therapeutics, focusing on ACEI, and illustrate how biotechnology and bioengineering can overcome the limitations of ACEIp large-scale production. Latest advances in ACEIp research and current genetic engineering-based strategies for heterologous production of ACEIp (and precursors) are also presented. Cloning approaches include tandem repeats of single ACEIp, ACEIp fusion to proteins/polypeptides, joining multivariate ACEIp into bioactive polypeptides, and producing ACEIp-containing modified plant storage proteins. Although bacteria have been privileged ACEIp heterologous hosts, particularly when testing for new genetic engineering strategies, plants and microalgae-based platforms are now emerging. Besides being generally safer, cost-effective and scalable, these “pharming” platforms can perform therelevant posttranslational modifications and produce (and eventually deliver) biologically active protein/peptide-based antihypertensive medicines.     

Honokiol: A non-adipogenic PPARγ agonist from nature

Background

Peroxisome proliferator-activated receptor gamma (PPARγ) agonists are clinically used to counteract hyperglycemia. However, so far experienced unwanted side effects, such as weight gain, promote the search for new PPARγ activators.

Methods

We used a combination of in silico, in vitro, cell-based and in vivo models to identify and validate natural products as promising leads for partial novel PPARγ agonists.

Results

The natural product honokiol from the traditional Chinese herbal drug Magnolia bark was in silico predicted to bind into the PPARγ ligand binding pocket as dimer. Honokiol indeed directly bound to purified PPARγ ligand-binding domain (LBD) and acted as partial agonist in a PPARγ-mediated luciferase reporter assay. Honokiol was then directly compared to the clinically used full agonist pioglitazone with regard to stimulation of glucose uptake in adipocytes as well as adipogenic differentiation in 3T3-L1 pre-adipocytes and mouse embryonic fibroblasts. While honokiol stimulated basal glucose uptake to a similar extent as pioglitazone, it did not induce adipogenesis in contrast to pioglitazone. In diabetic KKAy mice oral application of honokiol prevented hyperglycemia and suppressed weight gain.

Conclusion

We identified honokiol as a partial non-adipogenic PPARγ agonist in vitro which prevented hyperglycemia and weight gain in vivo.

General significance

This observed activity profile suggests honokiol as promising new pharmaceutical lead or dietary supplement to combat metabolic disease, and provides a molecular explanation for the use of Magnolia in traditional medicine.     

Root–root interactions: extending our perspective to be more inclusive of the range of theories in ecology and agriculture using in-vivo analyses

Background

There is a large body of literature on competitive interactions among plants, but many studies have only focused on above-ground interactions and little is known about root–root dynamics between interacting plants. The perspective on possible mechanisms that explain the outcome of root–root interactions has recently been extended to include non-resource-driven mechanisms (as well as resource-driven mechanisms) of root competition and positive interactions such as facilitation. These approaches have often suffered from being static, partly due to the lack of appropriate methodologies for in-situ non-destructive root characterization.

Scope

Recent studies show that interactive effects of plant neighbourhood interactions follow non-linear and non-additive paths that are hard to explain. Common outcomes such as accumulation of roots mainly in the topsoil cannot be explained solely by competition theory but require a more inclusive theoretical, as well as an improved methodological framework. This will include the question of whether we can apply the same conceptual framework to crop versus natural species.

Conclusions

The development of non-invasive methods to dynamically study root–root interactions in vivo will provide the necessary tools to study a more inclusive conceptual framework for root–root interactions. By following the dynamics of root–root interactions through time in a whole range of scenarios and systems, using a wide variety of non-invasive methods, (such as fluorescent protein which now allows us to separately identify the roots of several individuals within soil), we will be much better equipped to answer some of the key questions in root physiology, ecology and agronomy.     

Functional and evolutionary analysis of DXL1, a non-essential gene encoding a 1-deoxy-D-xylulose 5-phosphate synthase like protein in Arabidopsis thaliana

The synthesis of 1-deoxy-D-xylulose 5-phosphate (DXP), catalyzed by the enzyme DXP synthase (DXS), represents a key regulatory step of the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway for isoprenoid biosynthesis. In plants DXS is encoded by small multigene families that can be classified into, at least, three specialized subfamilies. Arabidopsis thaliana contains three genes encoding proteins with similarity to DXS, including the well-known DXS1/CLA1 gene, which clusters within subfamily I. The remaining proteins, initially named DXS2 and DXS3, have not yet been characterized. Here we report the expression and functional analysis of A. thaliana DXS2. Unexpectedly, the expression of DXS2 failed to rescue Escherichia coli and A. thaliana mutants defective in DXS activity. Coherently, we found that DXS activity was negligible in vitro, being renamed as DXL1 following recent nomenclature recommendation. DXL1 is targeted to plastids as DXS1, but shows a distinct expression pattern. The phenotypic analysis of a DXL1 defective mutant revealed that the function of the encoded protein is not essential for growth and development. Evolutionary analyses indicated that DXL1 emerged from DXS1 through a recent duplication apparently specific of the Brassicaceae lineage. Divergent selective constraints would have affected a significant fraction of sites after diversification of the paralogues. Furthermore, amino acids subjected to divergent selection and likely critical for functional divergence through the acquisition of a novel, although not yet known, biochemical function, were identified. Our results provide with the first evidences of functional specialization at both the regulatory and biochemical level within the plant DXS family.     

Subtractive phage display technology identifies zebrafish marcksb that is required for gastrulation

In the present study, we used a phage display technique to screen differentially expressed proteins from zebrafish post-gastrula embryos. With a subtractive screening approach, 6 types of single-chain Fv fragments (scFvs) were screened out from an scFv antibody phage display library by biopanning against zebrafish embryonic homogenate. Four scFv fragments (scFv1, scFv3, scFv4 and scFv6) showed significantly stronger binding to the tailbud embryos than to the 30%-epiboly embryos. A T7 phage display cDNA library was constructed from zebrafish tailbud embryos and used to identify the antigens potentially recognized by scFv1, which showed the highest frequency and strongest binding against the tailbud embryos. We acquired 4 candidate epitopes using scFv1 and the corresponding genes showed significantly higher expression levels at tailbud stage than at 30%-epiboly. The most potent epitope of scFv1 was the clone scFv1-2, which showed strong homology to zebrafish myristoylated alanine-rich C-kinase substrate b (Marcksb). Western blot analysis confirmed the high expression of marcksb in the post-gastrula embryos, and the endogenous expression of Marcksb was interfered by injection of scFv1. Zebrafish marcksb showed dynamic expression patterns during embryonic development. Knockdown of marcksb strongly affected gastrulation movements. Moreover, we revealed that zebrafish marcksb is required for cell membrane protrusion and F-actin alignment. Thus, our study uncovered 4 types of scFvs binding to zebrafish post-gastrula embryos, and the epitope of scFv1 was found to be required for normal gastrulation of zebrafish. To our knowledge, this was the first attempt to combine phage display technique with the embryonic and developmental study of vertebrates, and we were able to identify zebrafish marcksb that was required for gastrulation.     

Regulation of cell proliferation and migration in gallbladder cancer by zinc finger X-chromosomal protein

Gallbladder carcinoma (GBC) is one of the mostly aggressive and fatal malignancies. However, little is known about the oncogenic genes that contributed to the development of GBC. Zinc finger X-chromosomal protein (ZFX) was a novel member of the Krueppel C2H2-type zinc-finger protein family and its down-regulation led to impaired cell growth in human laryngeal squamous cell carcinoma. Here, we aim to investigate the function of ZFX in GBC cell proliferation and migration. Loss of function analysis was performed on GBC cell line (GBC-SD) using lentivirus-mediated siRNA against ZFX. The proliferation, in vitro tumorigenesis (colony-formation) ability as well as cell migration was significantly suppressed after GBC-SD cells which were infected with ZFX-siRNA-expressing lentivirus (Lv-shZFX). Our finding suggested that ZFX promoted the growth and migration of GBC cells and could present a potential molecular target for gene therapy of GBC.