南瓜类胡萝卜素高通量提取检测体系建立及应用

为了建立南瓜类胡萝卜素高通量提取检测体系,以南瓜果肉为试材,通过单因素试验和L27(313)正交试验,确定和优化南瓜类胡萝卜素提取的最佳提取液、提取温度和料液比。结果表明,乙醇是提取南瓜类胡萝卜素的最佳提取液,在55℃条件下,以料液比1:28,浸提90 min,类胡萝卜素的得率显著高[(7.56±0.14) mg/100 g]。在南瓜类胡萝卜素的提取条件中,提取温度对提取效果影响最大,料液比×提取时间和提取时间×温度的互作次之。利用高通量提取检测体系比较分析23份不同类型南瓜材料类胡萝卜素含量,发现印度南瓜‘贝栗4号’和中国南瓜×印度南瓜远缘杂种自交系20F8的类胡萝卜素含量分别为(31.40±0.35) mg/100g和(24.72±0.72) mg/100 g,显著高于其他供试南瓜材料,具有显著的营养利用价值。试验获得的南瓜类葫芦卜素高效检测体系,省事省力,结果准确,可提高南瓜营养品质育种效率。

稻米直链淀粉含量的96微孔高通量快速测定方法

直链淀粉含量的快速高通量测定方法对国际标准法(简称"国际法",BS EN ISO6647-2:2015《稻米直链淀粉含量的测定第2部分常规方法》)进行了改良,可以极大地提高水稻育种材料的筛选效率、适应当前水稻稻米品质育种的新需求。结果表明,高通量法测得的吸光度与直链淀粉含量有良好的相关性,相关系数达0.9911,该方法直链淀粉含量的测定值与国际法的测定值无显著差异;同时,高通量法还具有简单、易操作、样品量少、批量(96个样本)测定等优点,能用于水稻诱变群体后代或品种选育过程中的海量株系大规模初筛,可极大地提高稻米品质研究的效率。

From Phenotypes to Molecules: Revolutionizing Gut Microbiota Identification Methods

The gut microbiota is a complex ecosystem composed of many bacteria and their metabolites.It plays an irreplaceable role in human digestion,nutrient absorption,energy supply,fat metabolism,immune regulation,and many other aspects.Exploring the structure and function of the gut microbiota,as well as their key genes and metabolites,will enable the early diagnosis and auxiliary diagnosis of diseases,new treatment methods,better effects of drug treatments,and better guidance in the use of antibiotics.The identification of gut microbiota plays an important role in clinical diagnosis and treatment,as well as in drug research and development.Therefore,it is necessary to conduct a comprehensive review of this rapidly evolving topic.Traditional identification methods cannot comprehensively capture the diversity of gut microbiota.Currently,with the rapid development of molecular biology,the classification and identification methods for gut microbiota have evolved from the initial phenotypic and chemical identification to identification at the molecular level.This review integrates the main methods of gut microbiota identification and evaluates their application.We pay special attention to the research progress on molecular biological methods and focus on the application of high-throughput sequencing technology in the identification of gut microbiota.This revolutionary method for intestinal flora identification heralds a new chapter in our understanding of the microbial world.

农业反刍动物重要乳性状表型测定方法及研究进展

乳性状作为农业反刍动物的重要经济性状,其表型测定指标内容丰富,测定方法繁多复杂.为进一步凝练重要乳性状表型测定内容,以更好地培育改良中国特色的乳用动物种质资源、创制产量品质优异的新基因资源,本文从乳性状表型测定的常规乳性状表型与特色乳营养成分性状表型测定指标与测定方法、中红外光谱检测法,以及特色乳用畜种简介等四个方面,对我国主要农业反刍动物的重要乳性状表型的详细测定方法及其研究进展进行综述.

Understanding human diseases with high-throughput quantitative measurement and analysis of molecular signatures

Microarray and deep sequencing technologies have provided unprecedented opportunities for mapping genome mutations,RNA transcripts,transcription factor binding,and histone modifications at high resolution at the genome-wide level.This has revolutionized the way in which transcriptomes,regulatory networks and epigenetic regulations have been studied and large amounts of heterogeneous data have been generated.Although efforts are being made to integrate these datasets unbiasedly and efficiently,how best to do this still remains a challenge.Here we review major impacts of high-throughput genome-wide data generation,their relevance to human diseases,and various bioinformatics approaches for data integration.Finally,we provide a case study on inflammatory diseases.

High-throughput sequencing-based genome-wide identification of micro RNAs expressed in developing cotton seeds

Micro RNAs(mi RNAs) have been shown to play critical regulatory roles in gene expression in cotton. Although a large number of mi RNAs have been identified in cotton fibers, the functions of mi RNAs in seed development remain unexplored. In this study, a small RNA library was constructed from cotton seeds sampled at 15 days post-anthesis(DPA) and was subjected to high-throughput sequencing. A total of 95 known mi RNAs were detected to be expressed in cotton seeds. The expression pattern of these identified mi RNAs was profiled and 48 known mi RNAs were differentially expressed between cotton seeds and fibers at 15 DPA. In addition, 23 novel mi RNA candidates were identified in 15-DPA seeds. Putative targets for 21 novel and 87 known mi RNAs were successfully predicted and 900 expressed sequence tag(EST) sequences were proposed to be candidate target genes, which are involved in various metabolic and biological processes, suggesting a complex regulatory network in developing cotton seeds. Furthermore, mi RNA-mediated cleavage of three important transcripts in vivo was validated by RLM-5′ RACE. This study is the first to show the regulatory network of mi RNAs that are involved in developing cotton seeds and provides a foundation for future studies on the specific functions of these mi RNAs in seed development.

High throughput monoclonal antibody generation by immunizing multiple antigens

Recognizing proteins via the production of highly specific monoclonal antibodies(mAbs)is crucial to identifying proteins for proteomic research.However,traditional mAb generation is time-consuming with low efficiency.In this study,we assessed the high throughput method of producing mAbs by immunizing mice with multiple antigens in order to obtain hybridomas against these multiple antigens in one cell fusion.We selected eight proteins that play important roles in human physiological or pathological processes.These proteins were mixed and simultaneously administered to one mouse.We observed the immunizing period for 10 d,and determined the effect of liquid medium and semi-solid medium in hybridoma generation.As a result,all eight immunogens induced antibodies in the immunized mouse in one cell fusion,we obtained hybridomas specific to all eight proteins by enzyme-linked immuno sorbent assay(ELISA)screening,hybridomas against five out of eight showed specific positive in Western-blotting assays.This indicates that we generated mAbs specific to eight proteins in one cell fusion,greatly increasing the efficiency of mAb generation.Furthermore,we observed that hybridomas selected from the liquid medium and semi-solid medium showed different reactivity to antigens.Our study established high-throughput and time-saving methods for production of mAbs.These results provide alternative approaches for increasing the efficacy of mAb generation.

Development of a colorimetric assay for rapid quantitative measurement of clavulanic acid in microbial samples

We developed a colorimetric assay to quantify clavulanic acid （CA） in culture broth of Streptomyces clavuligerus, to facilitate screening of a large number of S. clavuligerus mutants. The assay is based on a β-1actamase-catalyzed reaction, in which the yellow substrate nitrocefin （λmax=390 nm） is converted to a red product （λmax=486 nm）. Since CA can irreversibly inhibit β-1actamase activity, the level of CA in a sample can be measured as a function of the A390]A486 ratio in the assay mixture. The sensitivity and detection window of the assay were determined to be 50 μg L -1 and 50 μg L to 10 mg L-1, respectively. The reliability of the assay was confirmed by comparing assay results with those obtained by HPLC. The assay was used to screen a pool of 65 S. clavuligerus mutants and was reliable for identifying CA over-producing mutants. Therefore, the assay saves time and labor in large-scale mutant screening and evaluation tasks. The detection window and the reliability of this assay are markedly better than those of previously reported CA assays. This assay method is suitable for high throughput screening of microbial samples and allows direct visual observation of CA levels on agar plates.