In this study,a high-throughput screening method was established through the 24-square deep-well microliter plate(MTP) fermentation and micro-plate detection for large-scale screening of the mutants.It was suitable fo...In this study,a high-throughput screening method was established through the 24-square deep-well microliter plate(MTP) fermentation and micro-plate detection for large-scale screening of the mutants.It was suitable for screening a large number of mutants and improving the breeding efficiency after heavy-ion beam irradiation.Seventeen strains showed higher cellulase activity compared with the initial strain after the screening of plate and MTP fermentation.The filter paper activity and β-glucosidase activity of Aspergillus niger H11201 had increased 38.74 and 63.23%separately compared with A.niger H11 by shaking flask fermentation,and it was genetically stable after being passaged to nine generations.The results indicate that the high-throughput screening method can be used for the quick breeding of A.niger with high cellulase activity.展开更多
Rice panicle traits serve as critical indicators of both yield potential and germplasm resource quality.However,traditional manual measurements of these traits,which typically involve threshing,are not only laborious ...Rice panicle traits serve as critical indicators of both yield potential and germplasm resource quality.However,traditional manual measurements of these traits,which typically involve threshing,are not only laborious and time-consuming but also prone to introducing measurement errors.This study introduces a high-throughput and nondestructive method,termed extraction of panicle traits(EOPT),along with the software Panicle Analyzer,which is designed to assess unshaped intact rice panicle traits,including the panicle grain number,grain length,grain width,and panicle length.To address the challenge of grain occlusion within an intact panicle,we define a panicle morphology index to quantify the occlusion levels among the rice grains within the panicle.By calibrating the grain number obtained directly from rice panicle images based on the panicle morphology index,we substantially improve the grain number detection accuracy.For measuring grain length and width,the EOPT selects rice grains using an intersection over union threshold of 0.8 and a confidence threshold of 0.7 during the grain detection process.The mean values of these grains were calculated to represent all the panicle grain lengths and widths.In addition,EOPT extracted the main path of the skeleton of the rice panicle using the Astar algorithm to determine panicle lengths.Validation on a dataset of 1,554 panicle images demonstrated the effectiveness of the proposed method,achieving 93.57%accuracy in panicle grain counting with a mean absolute percentage error of 6.62%.High accuracy rates were also recorded for grain length(96.83%)and panicle length(9713%).Moreover,the utility of EOPT was confirmed across different years and scenes,both indoors and outdoors.A genome-wide association study was conducted,leveraging the phenotypic traits obtained via EOPT and genotypic data.This study identified single-nucleotide polymorphisms associated with grain length,width,number per panicle,and panicle length,further emphasizing the utility and potential of this method in advancing rice breeding.展开更多
目的:使用高通量测序法辅助开发RAS相关区域家族1A(Ras-Association Domain Family 1A,RASSF1A)基因甲基化定性检测试剂盒(荧光PCR法)。方法:提取37例肺癌血浆样本DNA进行亚硫酸氢盐修饰转化,使用高通量测序方法检测样本的亚硫酸氢盐转...目的:使用高通量测序法辅助开发RAS相关区域家族1A(Ras-Association Domain Family 1A,RASSF1A)基因甲基化定性检测试剂盒(荧光PCR法)。方法:提取37例肺癌血浆样本DNA进行亚硫酸氢盐修饰转化,使用高通量测序方法检测样本的亚硫酸氢盐转化效率和RASSF1A基因CpG位点的甲基化水平,之后使用荧光PCR法检测样本RASSF1A基因的甲基化状态。结果:高通量测序显示样本DNA的平均转化效率为99.47%;根据CpG位点甲基化水平的双向聚类分析结果,筛选出第26~30号CpG位点作为荧光PCR法的检测位点,以甲基化水平5%作为阈值线,ΔCt Cut-off值为9,得出两种检测方法对37例样本的测定结果一致。结论:本研究借助高通量测序技术成功构建基于荧光PCR法的RASSF1A基因甲基化定性检测方法,操作简便、成本低、效率高,适用于临床RASSF1A基因甲基化状态常规化检测。展开更多
A new high throughput heat-treatment method with a continuous temperature gradient between 600 and 700 ？C was utilized on the Ti-5553 alloy（Ti-5 Al-5 Mo-5 V-3 Cr, mass fraction, %）. The temperature gradient was ind...A new high throughput heat-treatment method with a continuous temperature gradient between 600 and 700 ？C was utilized on the Ti-5553 alloy（Ti-5 Al-5 Mo-5 V-3 Cr, mass fraction, %）. The temperature gradient was induced by the variation of the axial section of sample, which was heated by the direct current. The variation of continuous cooling rates on the treated sample was realized by using the end quenching method. The microstructural evolution and mechanical properties under different heat treatment conditions were evaluated. The results show that the pseudo-spinodal decomposition of the alloy occurs at（617±1） ？C, and the size of the precipitated α phase is around 300 nm. Moreover, the highest microhardness is obtained after the heat treatment at the pseudo-spinodal decomposition temperature for 4 h. These indicate that the high throughput method is efficient and fast to determine the phase transformation temperature and corresponding microstructural evolution of alloys.展开更多
基金supported by the National Natural Science Foundation of China(No.11305225)the Science and Technology Service Network Initiative(KFJ-EW-STS-086)
文摘In this study,a high-throughput screening method was established through the 24-square deep-well microliter plate(MTP) fermentation and micro-plate detection for large-scale screening of the mutants.It was suitable for screening a large number of mutants and improving the breeding efficiency after heavy-ion beam irradiation.Seventeen strains showed higher cellulase activity compared with the initial strain after the screening of plate and MTP fermentation.The filter paper activity and β-glucosidase activity of Aspergillus niger H11201 had increased 38.74 and 63.23%separately compared with A.niger H11 by shaking flask fermentation,and it was genetically stable after being passaged to nine generations.The results indicate that the high-throughput screening method can be used for the quick breeding of A.niger with high cellulase activity.
基金supported by the National Key Research and Development Program of China(2021YFD1200504 and 2022YFD2002304)the Key Core Technology Project in Agri-culture of Hubei Province(HBNYHXGG2023-9)the Supporting Project for High-Quality Development of Seed Industry of Hubei Province(HBZY2023B001-06).
文摘Rice panicle traits serve as critical indicators of both yield potential and germplasm resource quality.However,traditional manual measurements of these traits,which typically involve threshing,are not only laborious and time-consuming but also prone to introducing measurement errors.This study introduces a high-throughput and nondestructive method,termed extraction of panicle traits(EOPT),along with the software Panicle Analyzer,which is designed to assess unshaped intact rice panicle traits,including the panicle grain number,grain length,grain width,and panicle length.To address the challenge of grain occlusion within an intact panicle,we define a panicle morphology index to quantify the occlusion levels among the rice grains within the panicle.By calibrating the grain number obtained directly from rice panicle images based on the panicle morphology index,we substantially improve the grain number detection accuracy.For measuring grain length and width,the EOPT selects rice grains using an intersection over union threshold of 0.8 and a confidence threshold of 0.7 during the grain detection process.The mean values of these grains were calculated to represent all the panicle grain lengths and widths.In addition,EOPT extracted the main path of the skeleton of the rice panicle using the Astar algorithm to determine panicle lengths.Validation on a dataset of 1,554 panicle images demonstrated the effectiveness of the proposed method,achieving 93.57%accuracy in panicle grain counting with a mean absolute percentage error of 6.62%.High accuracy rates were also recorded for grain length(96.83%)and panicle length(9713%).Moreover,the utility of EOPT was confirmed across different years and scenes,both indoors and outdoors.A genome-wide association study was conducted,leveraging the phenotypic traits obtained via EOPT and genotypic data.This study identified single-nucleotide polymorphisms associated with grain length,width,number per panicle,and panicle length,further emphasizing the utility and potential of this method in advancing rice breeding.
文摘目的:使用高通量测序法辅助开发RAS相关区域家族1A(Ras-Association Domain Family 1A,RASSF1A)基因甲基化定性检测试剂盒(荧光PCR法)。方法:提取37例肺癌血浆样本DNA进行亚硫酸氢盐修饰转化,使用高通量测序方法检测样本的亚硫酸氢盐转化效率和RASSF1A基因CpG位点的甲基化水平,之后使用荧光PCR法检测样本RASSF1A基因的甲基化状态。结果:高通量测序显示样本DNA的平均转化效率为99.47%;根据CpG位点甲基化水平的双向聚类分析结果,筛选出第26~30号CpG位点作为荧光PCR法的检测位点,以甲基化水平5%作为阈值线,ΔCt Cut-off值为9,得出两种检测方法对37例样本的测定结果一致。结论:本研究借助高通量测序技术成功构建基于荧光PCR法的RASSF1A基因甲基化定性检测方法,操作简便、成本低、效率高,适用于临床RASSF1A基因甲基化状态常规化检测。
基金Project(2014CB644002)supported by the National Basic Research and Development Project of ChinaProject(2015CX004)supported by the Innovation-driven Plan in Central South University,China
文摘A new high throughput heat-treatment method with a continuous temperature gradient between 600 and 700 ？C was utilized on the Ti-5553 alloy（Ti-5 Al-5 Mo-5 V-3 Cr, mass fraction, %）. The temperature gradient was induced by the variation of the axial section of sample, which was heated by the direct current. The variation of continuous cooling rates on the treated sample was realized by using the end quenching method. The microstructural evolution and mechanical properties under different heat treatment conditions were evaluated. The results show that the pseudo-spinodal decomposition of the alloy occurs at（617±1） ？C, and the size of the precipitated α phase is around 300 nm. Moreover, the highest microhardness is obtained after the heat treatment at the pseudo-spinodal decomposition temperature for 4 h. These indicate that the high throughput method is efficient and fast to determine the phase transformation temperature and corresponding microstructural evolution of alloys.
