Liquid chromatography-mass spectrometry （LC-MS）-based metabolomics has been facilitated by the con- struction of MSz spectral tag （MS2T） library from the total scan ESI MS/MS data, and the development of widely ta...Liquid chromatography-mass spectrometry （LC-MS）-based metabolomics has been facilitated by the con- struction of MSz spectral tag （MS2T） library from the total scan ESI MS/MS data, and the development of widely targeted metabolomics method using MS/MS data gathered from authentic standards. In this report, a novel strategy called step- wise multiple ion monitoring-enhanced product ions （stepwise MIM-EPI） was developed to construct the MS2T library, in which stepwise MIM was used as survey scans to trigger the acquisition of EPI. A total number of 698 （almost） non- redundant metabolites with MS2 spectra were obtained, of which 135 metabolites were identified/annotated. Integrating the data gathered from our MS2T library and other available multiple reaction monitoring （MRM） information, a widely targeted metabolomics method was developed to quantify 277 metabolites, including some phytohormones. Evaluation of the dehydration responses and natural variations of these metabolites in rice leaf not only suggested the coordinated regulation of abscisic acid （ABA） with metabolites such as serotonin derivative（s）, polyamine conjugates under drought stress, but also revealed some C-glycosylated flavones as the potential markers for the discrimination of indica and japonica rice subspecies. The new MS2T library construction and widely targeted metabolomics strategy could be used as a tool for rice functional genomics.展开更多
Volatile organic compounds play essential roles in plant environment interactions as well as determining the fragrance of plants.Although gas chromatography-mass spectrometry-based untargeted metabolo-mics is commonly...Volatile organic compounds play essential roles in plant environment interactions as well as determining the fragrance of plants.Although gas chromatography-mass spectrometry-based untargeted metabolo-mics is commonly used to assess plant volatiles,it suffers from high spectral convolution,low detection sensitivity,a limited number of annotated metabolites,and relatively poor reproducibility.Here,we report a widely targeted volatilomics(WTV)method that involves using a“targeted spectra extraction”algorithm to address spectral convolution,constructing a high-coverage MS2 spectral tag library to expand volatile annotation,adapting a multiple reaction monitoring mode to improve sensitivity,and using regression models to adjust for signal drift.The newly developed method was used to profile the volatilome of rice grains.Compared with the untargeted method,the newly developed WTV method shows higher sensitivity(for example,the signal-to-noise ratio of guaicol increased from 4.1 to 18.8),high annotation coverage(the number of annotated volatiles increased from 43 to 132),and better reproducibility(the number of volatiles in quality control samples with relative standard deviation value below 30.0%increased from 14 to 92 after normalization).Using the WTV method,we studied the metabolic responses of tomato to environmental stimuli and profiled the volatilomes of different rice accessions.The results identified benzothiazole as a potential airborne signal priming tomato plants for enhanced defense and 2-nonanone and 2-heptanone as novel aromatic compounds contributing to rice fragrance.These case studies suggest that the widely targeted volatilomics method is more efficient than those currently used and may considerably promote plant volatilomics studies.展开更多
Ms2 is an important dominant male-sterile gene in wheat,but the biochemical function of Ms2 and the mechanism by which it causes male sterility remain elusive.Here,we report the molecular basis underlying Ms2-induced ...Ms2 is an important dominant male-sterile gene in wheat,but the biochemical function of Ms2 and the mechanism by which it causes male sterility remain elusive.Here,we report the molecular basis underlying Ms2-induced male sterility in wheat.We found that activated Ms2 specifically reduces the reactive oxygen species(ROS)signals in anthers and thereby induces termination of wheat anther development at an early stage.Furthermore,our results indicate that Ms2 is localized in mitochondria,where it physically interacts with a wheat homolog of ROS modulator 1(TaRomo1).Romo1 positively regulates the ROS levels in humans but has never been studied in plants.We found that single amino acid substitutions in the Ms2 protein that rescue the ms2 male-sterile phenotype abolish the interaction between Ms2 and TaRomo1.Significantly,Ms2 promotes the transition of TaRomo1 proteins from active monomers to inactive oligomers.Taken together,our findings unravel the molecular basis of Ms2-induced male sterility and reveal a regulatory mechanism in which ROS act as essential signals guiding the anther development program in wheat.展开更多
The male-sterile lines with Ms2 gene were highly evaluated in recurrent selection in wheat （Triticum aestivum L.）. Three populations C6 （population after six cycles of selection）, C7 （population after seven cycle...The male-sterile lines with Ms2 gene were highly evaluated in recurrent selection in wheat （Triticum aestivum L.）. Three populations C6 （population after six cycles of selection）, C7 （population after seven cycles of selection）, and C8 （population after eight cycles of selection） were constructed through recurrent selection with 12 parental materials （P）. Acid polyacrymide gel electrophoresis （A-PAGE） analysis was used to identify gliadin patterns and evaluate the genetic diversity in 12 parents and three populations. A total of 63 bands were identified, of which 17 polymorphic bands and 7 unique bands were present in populations and seven polymorphic bands and four unique bands were present in parents. The number of polymorphic and unique bands decreased gradually from C6 to C8, especially for to- and y-gliadins. The genetic distances in C6, C7, and C8 were calculated. The distributions of genetic distance were different in three recurrent selection populations. From C6 to C8, the genetic distance was 0.2687, 0.2652 and 0.1987, respectively. Statistically significant differences were detected between C7 and C8 with the T value of 37.9718. The result of cluster analysis based on genetic similarity matrix of three populations fitted well to those of principle coordinates analysis （PCoA）. Compared with 12 parents, almost all individuals of three populations are new genotypes. Most of the individuals from C6 and C7 could be divided into two groups, while most individuals of C8 were in one cluster. In conclusion, the results indicated that the genetic diversity was decreased severely according to the information revealed by A-PAGE, although some variations could be created in the recurrent selection. It was necessary to introduce diverse germplasm based on the genetic database of recurrent population to maintain and improve the breeding efficiency in the further program.展开更多
This paper describes the use of log-linear energy phase-space projections to analyze microwave-assisted inactivation of bacteria and viruses under different fomite conditions within multimode microwave ovens. The oven...This paper describes the use of log-linear energy phase-space projections to analyze microwave-assisted inactivation of bacteria and viruses under different fomite conditions within multimode microwave ovens. The ovens are operated at a cavity-magnetron frequency of 2.45 ± 01 GHz. Porous fomites (moist face towels, cotton swabs, kitchen sponges, and scrubbing pads, cigarette filters and N95-like respirators);along with non-porous hard surface syringe fomites are studied. The fomites are classed as dielectric;and absorb microwave energy to varying degrees depending on their complex dielectric permittivity. Microorganism resilience to microwave stress (defined as ≥4 log<sub>10</sub> reduction in inactivation) when mapped using iso-volume trend-lines in energy phase-space reveals the persistence imparted by the fomite, and can be mapped between different microwave ovens. Microorganism resilience to thermal microwave-assisted treatment increases from vegetative Gram-negative to vegetative Gram-positive and on to Gram-positive spores. Bacteriophage MS2 and influenza viruses have an intermediate resilience dependency. It is shown that linear-scaled fomite temperature against process time graphs can differentiate between non-thermal and thermal micro-wave-assisted treatment of microorganisms.展开更多
基金the National High Technology R&D Program of China (863 Program)，the Major State Basic Research Development Program of China (973 Program)，by the National Natural Science Foundation of China，by the Program for New Century Excellent Talents in University of Ministry of Education in China (NCET-09-0401).A patent for this method has been approved by the State Intellectual Property Office of China
文摘Liquid chromatography-mass spectrometry （LC-MS）-based metabolomics has been facilitated by the con- struction of MSz spectral tag （MS2T） library from the total scan ESI MS/MS data, and the development of widely targeted metabolomics method using MS/MS data gathered from authentic standards. In this report, a novel strategy called step- wise multiple ion monitoring-enhanced product ions （stepwise MIM-EPI） was developed to construct the MS2T library, in which stepwise MIM was used as survey scans to trigger the acquisition of EPI. A total number of 698 （almost） non- redundant metabolites with MS2 spectra were obtained, of which 135 metabolites were identified/annotated. Integrating the data gathered from our MS2T library and other available multiple reaction monitoring （MRM） information, a widely targeted metabolomics method was developed to quantify 277 metabolites, including some phytohormones. Evaluation of the dehydration responses and natural variations of these metabolites in rice leaf not only suggested the coordinated regulation of abscisic acid （ABA） with metabolites such as serotonin derivative（s）, polyamine conjugates under drought stress, but also revealed some C-glycosylated flavones as the potential markers for the discrimination of indica and japonica rice subspecies. The new MS2T library construction and widely targeted metabolomics strategy could be used as a tool for rice functional genomics.
基金This work was supported by the Hainan Province Major Research Project(modern agriculture)ZDYF2020066the Hainan Provincial Natural Science Foundation of China(320MS011)the Hainan Major Science and Technology Project(Nno.ZDKJ202002).
文摘Volatile organic compounds play essential roles in plant environment interactions as well as determining the fragrance of plants.Although gas chromatography-mass spectrometry-based untargeted metabolo-mics is commonly used to assess plant volatiles,it suffers from high spectral convolution,low detection sensitivity,a limited number of annotated metabolites,and relatively poor reproducibility.Here,we report a widely targeted volatilomics(WTV)method that involves using a“targeted spectra extraction”algorithm to address spectral convolution,constructing a high-coverage MS2 spectral tag library to expand volatile annotation,adapting a multiple reaction monitoring mode to improve sensitivity,and using regression models to adjust for signal drift.The newly developed method was used to profile the volatilome of rice grains.Compared with the untargeted method,the newly developed WTV method shows higher sensitivity(for example,the signal-to-noise ratio of guaicol increased from 4.1 to 18.8),high annotation coverage(the number of annotated volatiles increased from 43 to 132),and better reproducibility(the number of volatiles in quality control samples with relative standard deviation value below 30.0%increased from 14 to 92 after normalization).Using the WTV method,we studied the metabolic responses of tomato to environmental stimuli and profiled the volatilomes of different rice accessions.The results identified benzothiazole as a potential airborne signal priming tomato plants for enhanced defense and 2-nonanone and 2-heptanone as novel aromatic compounds contributing to rice fragrance.These case studies suggest that the widely targeted volatilomics method is more efficient than those currently used and may considerably promote plant volatilomics studies.
基金supported by grants from the National Natural Science FoundationofChina(31871623,31991213)the National Key Research andDevelopment Program of China(2016YFD0100302)+1 种基金the Central Public-interest Scientific Institution Basal Research Fund(S2022D02)the Talent Program and Agricultural Science and Technology Innovation Program of CAAS.WethankLingli Zheng and Yuhong Liuforper forming the wheat transformation and planting,and Drs.Guan-Zhu Han and Yun Zhou forthe experimental assistance.
文摘Ms2 is an important dominant male-sterile gene in wheat,but the biochemical function of Ms2 and the mechanism by which it causes male sterility remain elusive.Here,we report the molecular basis underlying Ms2-induced male sterility in wheat.We found that activated Ms2 specifically reduces the reactive oxygen species(ROS)signals in anthers and thereby induces termination of wheat anther development at an early stage.Furthermore,our results indicate that Ms2 is localized in mitochondria,where it physically interacts with a wheat homolog of ROS modulator 1(TaRomo1).Romo1 positively regulates the ROS levels in humans but has never been studied in plants.We found that single amino acid substitutions in the Ms2 protein that rescue the ms2 male-sterile phenotype abolish the interaction between Ms2 and TaRomo1.Significantly,Ms2 promotes the transition of TaRomo1 proteins from active monomers to inactive oligomers.Taken together,our findings unravel the molecular basis of Ms2-induced male sterility and reveal a regulatory mechanism in which ROS act as essential signals guiding the anther development program in wheat.
基金funded by the National Basic Research Program of China (973 Program of China,2009CB118301)the National 863 Program of China(2006AA100102)
文摘The male-sterile lines with Ms2 gene were highly evaluated in recurrent selection in wheat （Triticum aestivum L.）. Three populations C6 （population after six cycles of selection）, C7 （population after seven cycles of selection）, and C8 （population after eight cycles of selection） were constructed through recurrent selection with 12 parental materials （P）. Acid polyacrymide gel electrophoresis （A-PAGE） analysis was used to identify gliadin patterns and evaluate the genetic diversity in 12 parents and three populations. A total of 63 bands were identified, of which 17 polymorphic bands and 7 unique bands were present in populations and seven polymorphic bands and four unique bands were present in parents. The number of polymorphic and unique bands decreased gradually from C6 to C8, especially for to- and y-gliadins. The genetic distances in C6, C7, and C8 were calculated. The distributions of genetic distance were different in three recurrent selection populations. From C6 to C8, the genetic distance was 0.2687, 0.2652 and 0.1987, respectively. Statistically significant differences were detected between C7 and C8 with the T value of 37.9718. The result of cluster analysis based on genetic similarity matrix of three populations fitted well to those of principle coordinates analysis （PCoA）. Compared with 12 parents, almost all individuals of three populations are new genotypes. Most of the individuals from C6 and C7 could be divided into two groups, while most individuals of C8 were in one cluster. In conclusion, the results indicated that the genetic diversity was decreased severely according to the information revealed by A-PAGE, although some variations could be created in the recurrent selection. It was necessary to introduce diverse germplasm based on the genetic database of recurrent population to maintain and improve the breeding efficiency in the further program.
文摘This paper describes the use of log-linear energy phase-space projections to analyze microwave-assisted inactivation of bacteria and viruses under different fomite conditions within multimode microwave ovens. The ovens are operated at a cavity-magnetron frequency of 2.45 ± 01 GHz. Porous fomites (moist face towels, cotton swabs, kitchen sponges, and scrubbing pads, cigarette filters and N95-like respirators);along with non-porous hard surface syringe fomites are studied. The fomites are classed as dielectric;and absorb microwave energy to varying degrees depending on their complex dielectric permittivity. Microorganism resilience to microwave stress (defined as ≥4 log<sub>10</sub> reduction in inactivation) when mapped using iso-volume trend-lines in energy phase-space reveals the persistence imparted by the fomite, and can be mapped between different microwave ovens. Microorganism resilience to thermal microwave-assisted treatment increases from vegetative Gram-negative to vegetative Gram-positive and on to Gram-positive spores. Bacteriophage MS2 and influenza viruses have an intermediate resilience dependency. It is shown that linear-scaled fomite temperature against process time graphs can differentiate between non-thermal and thermal micro-wave-assisted treatment of microorganisms.
