The vacuole is one of the most conspicuous organelles in plant cells, participating in a series of physiological processes, such as storage of ions and compartmentalization of heavy metals. Isolation ...of intact vacuoles and elemental analysis provides a powerful method to investigate the functions and regulatory mechanisms of tonoplast transporters. Here, we present a protocol to isolate intact vacuoles from
root protoplasts and analyze their elemental content by inductively coupled plasma mass spectrometry (ICP-MS). In this protocol, we summarize how to prepare the protoplast, extract the vacuole, and analyze element concentration. This protocol has been applied to explore the function and regulatory mechanisms of tonoplast manganese (Mn) transporter MTP8, which is antagonistically regulated by CPK4/5/6/11 and CBL2/3-CIPK3/9/26. This protocol is not only suitable for exploring the functions and regulatory mechanisms of tonoplast transporters, but also for researching other tonoplast proteins. Graphical abstract.
Measurements of photosynthetically active radiation (PAR), leaf photosynthesis, canopy leaf area index (LAI) and crop yield were carried from flowering to maturity to study the effects of tree ...shading on crop yield in a Paulownia-wheat intercropping system in China. We found that the tree shading reduced the amount of incoming PAR within the intercropping system by 22%, 44% and 56% during flowering, grain-filling and maturing, respectively. The amount of PAR intercepted by the wheat crop inside the intercropping system was 34%, 55% and 68% less than the crop outside the intercropping system during flowering, grain-filling and maturing. Estimates of PAR-saturated leaf photosynthetic rate were not affected by tree shading, and the differences between the wheat yield inside the intercropping system and outside the intercropping system can be explained by the difference in the amount of PAR intercepted. Total grain numbers and grain dry weight per 1000 grains were linearly correlated with the amount of PAR intercepted during 7 days prior to anthesis and during anthesis and grain-filling, respectively. As compared with the wheat crop outside the intercropping system, grain numbers and grain dry weight were reduced by 36% and 25%, respectively. As a result wheat yield inside the intercropping system was 51% lower than that outside the intercropping system. Our results suggest a much higher reduction in crop yield and much smaller economic gain of the Paulownia-wheat intercropping system than previous studies on the similar Paulownia-wheat intercropping systems in China.
The genus
Armeniaca
Scop. is well known for its popular cultivated edible fruit trees such as
Armeniaca vulgaris
Lam. and ornamental flowers such as
A. mume
Sieb. Another species,
A. cathayana
Fu et ...al., one of six important dry fruits (kernel-using apricot), is cultivated for its edible seeds in North China. In the present study, DNA from 70 individuals of
Armeniaca
, including 38 of
A. cathayana
, 18 of
A. vulgaris
, 12 of
A. sibirica
, 1 of
A. dasycarpa
and 1 of
A. mume
, was extracted and analyzed using microsatellites and capillary electrophoresis. For 20 polymorphic loci selected, 339 alleles and 140.7 effective alleles were detected. The number of alleles per locus ranged from 8 to 28, with an average of 16.95 alleles per locus. The observed heterozygosity (Ho) and the expected heterozygosity (He) ranged from 0.427 to 0.971 and from 0.737 to 0.912, respectively. The polymorphism information content varied from 0.708 to 0.905, with an average of 0.827. Based on the genetic similarity among 70 individuals, a UPGMA was used to establish the phylogenetic relationships. The taxonomic positions among five species were clearly revealed, and
A. cathayana
was more closely related to
A. vulgaris
than to
A. sibirica
. The results will provide a scientific basis for research on the taxonomy, germplasm resources and breeding of
Armeniaca
, especially for
A. cathayana
.
Bio‐oils, produced by biomass pyrolysis, have become promising candidates for feedstocks of high value‐added chemicals and alternative sources for transportation fuels. Bio‐oil is such a complicated ...mixture that contains nonpolar hydrocarbons and polar components which cover almost all kinds of organic oxygenated compounds such as carboxylic acids, alcohols, aldehydes, ketones, esters, furfurals, phenolic compounds, sugar‐like material, and lignin‐derived compounds. Comprehensive characterization of bio‐oil and its subfractions could provide insight into the conversion process of biomass processing, as well as its further utilization as transportation fuels or chemical raw materials. This review focuses on advanced analytical strategies on in‐depth characterization of bio‐oil, which is concerned with gas chromatography, high‐resolution mass spectrometry, FTIR spectroscopy and NMR spectroscopy, offering complementary information for previous reviews.
Mass transfer from liquid to gas phase is a crucial process in chemical engineering. Due to the heterogeneity and dynamic nature of the phase transition process, monitoring the structural changes of ...compounds during this process is challenging. In this work, a combining of the mass spectrometry‐based method and quantum chemistry calculations was used to study the structural changes during liquid‐to‐gas phase transitions. Electrospray is used to induce the transition of phenolic acid molecules from the liquid phase to the gas phase. The deprotonated structures were captured and separated using ion mobility mass spectrometry. Deprotonated structures helped to identify the most acidic functional group within the molecule. These deprotonated structures were confirmed by collision‐induced dissociation and molecular collision cross‐section measurements. Quantum chemistry calculations revealed the mechanisms about the variation of intramolecular acid site and “kinetic trapping.” Three liquid‐to‐gas phase transformation models were proposed, demonstrating that the variation in free energy among different deprotonated structures governs the thermodynamic/kinetic process of the liquid‐to‐gas phase conversion.
This study proposes a new DNA band display technology of microsatellite DNA called Fluorescent Imaging Technology. In comparison with Silver Stain Technology, this technology is worth popularizing in ...the laboratory because of its high resolution, efficiency, simplicity and clear background.
Multifunctional compounds may form different prototropic isomers under different conditions, which are known as protomers/deprotomers. In biological systems, these protomer/deprotomer isomers affect ...the interaction modes and conformational landscape between compounds and enzymes and thus present different biological activities. Study on protomers/deprotomers is essentially the study on the acidity/basicity of each intramolecular functional group and its effect on molecular structure. In recent years, the combination of mass spectrometry (MS) and computational chemistry has been proven to be a powerful and effective means to study prototropic isomers. MS-based technologies are developed to discriminate and characterize protomers/deprotomers to provide structural information and monitor transformations, showing great superiority than other experimental methods. Computational chemistry is used to predict the thermodynamic stability of protomers/deprotomers, provide the simulated MS/MS spectra, infrared spectra, and calculate collision cross-section values. By comparing the theoretical data with the corresponding experimental results, the researchers can not only determine the protomer/deprotomer structure, but also investigate the structure–activity relationship in a given system. This review covers various MS methods and theoretical calculations and their devotion to isomer discrimination, structure identification, conformational transformation, and phase transition investigation of protomers/deprotomers.
Summary
Manganese (Mn) is an essential micronutrient in plants. However, excessive Mn absorption in acidic soils can cause Mn toxicity, which adversely affects plant growth and crop yields. At ...present, acidic soils cover c. 30% of the Earth's surface. However, the mechanism underpinning Mn uptake remains largely unknown.
We identified cbl1/9 and cipk23 mutants exhibiting high‐Mn‐sensitive phenotype through the reverse genetics method. Furthermore, we identified the CIPK23 phosphorylated NRAMP1 through a variety of protein interaction techniques and protein kinase assays.
Here, we demonstrated that two calcineurin B‐like proteins, CBL1/9, and their interacting kinase CIPK23 positively regulated the tolerance of Mn toxicity in Arabidopsis. The cbl1 cbl9 double mutant and cipk23 mutants exhibited high‐Mn‐sensitive phenotypes, which manifested as decreased primary root length, biomass, and chlorophyll concentration, and higher accumulation of Mn. In addition, CIPK23 interacted with and phosphorylated the Mn transporter NRAMP1 primarily at Ser20/22 in vitro and in vivo, and thereby induced clathrin‐mediated endocytosis of NRAMP1 to reduce its distribution on the plasma membrane and enhance plant tolerance to Mn toxicity.
In summary, we found that the CBL1/9–CIPK23–NRAMP1 module regulates the tolerance to high‐Mn toxicity and provide insight into a mechanism of the tolerance of plants to Mn toxicity.
Vitiligo is an acquired chronic depigmentary disorder affecting approximately 0.5% to 1% of individuals worldwide. The compound glycyrrhizin (CG), a complementary medicine, has been reported for ...treatment of vitiligo, but the evidence has not been systematically evaluated. We systematically assessed the efficacy and safety of CG in combination with conventional therapy for the treatment of vitiligo.
We searched Embase, Web of Science, PubMed, The Cochrane Library, Chinese BioMedical Literature Database (CBM), Wanfang Data, China National Knowledge Infrastructure (CNKI), and VIP information from inception to July 2022. Randomized controlled trials comparing CG combined with conventional therapy with conventional therapy alone for vitiligo were included in our analysis. The primary outcome was treatment response, which defined as >50% repigmentation rate of vitiligo after treatment. The secondary outcome was incidence of adverse events. Meta-analysis was performed using the Review Manager 5.4 software. Statistical heterogeneity was evaluated with chi-square and I2 statistics, dichotomous data were expressed as risk ratios (RR) with 95% confidence intervals using the Mantel-Haenszal method.
Thirty-nine studies enrolling with 3994 participants were subjected to this review. The results of our meta-analysis indicated that addition of CG had superior effectiveness on repigmentation rate than phototherapy (RR = 1.28; P < .001), immunosuppressant (RR = 1.76; P < .001), traditional Chinese medicine (RR = 1.38; P < .001), combination of phototherapy and immunosuppressant (RR = 1.42; P < .001), and combination of phototherapy and traditional Chinese medicine (RR = 1.37; P < .001). In addition, CG did not increase the incidence of adverse events for vitiligo (RR = 0.79; P = .05).
CG as a complementary medicine has a potential benefit in treatment of vitiligo. However, since the methodological flaws in the studies we included, more high-quality randomized controlled trials are warranted.
