Current protocols in plant biology最新文献_第5页

Morphogenic Regulator-Mediated Transformation of Maize Inbred B73 形态发生调控因子介导的玉米自交系B73的转化

Current protocols in plant biology Pub Date : 2018-10-22 DOI: 10.1002/cppb.20075

Muruganantham Mookkan, Kimberly Nelson-Vasilchik, Joel Hague, Albert Kausch, Zhanyuan J. Zhang

{"title":"Morphogenic Regulator-Mediated Transformation of Maize Inbred B73","authors":"Muruganantham Mookkan, Kimberly Nelson-Vasilchik, Joel Hague, Albert Kausch, Zhanyuan J. Zhang","doi":"10.1002/cppb.20075","DOIUrl":"10.1002/cppb.20075","url":null,"abstract":"Maize B73 is a reference genome and has long been a major resource for genetics and molecular biology research. We have developed an efficient B73 transformation protocol by enabling somatic embryogenesis through differential co-expression of maize morphogenic regulators BBM and WUS2. We describe a successful protocol that utilizes Agrobacterium tumefaciens strain AGL1 harboring binary vector PHP78891 that comprises a BBM and WUS2 expression cassette as well as a green fluorescent protein (GFP) reporter cassette. The PHP78891 vector also contains, within the T-DNA region, a CRE/lox recombination system flanking the CRE/BBM/WUS2 co-expression cassette driven by the desiccation inducible RAB17 promoter that allows removal of the BBM/WUS2 cassette. Introduction and co-expression of BBM and WUS2 induced direct somatic embryogenesis (SE) in non-regenerable maize B73 from immature embryo explants. Removal of the CRE/BBM/WUS2 cassette is essential to allow regeneration to fertile plants. The GFP expression cassette outside the lox excision sites is retained in the transgenic plant genome, allowing subsequent phenotypic analysis of calli and regenerated transgenic events. This transformation system enables a selectable marker-free transformation process by taking advantage of BBM/WUS2-induced SE as a developmental selection system. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":10932,"journal":{"name":"Current protocols in plant biology","volume":"3 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cppb.20075","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36623999","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 8

Transformation of Recalcitrant Sorghum Varieties Facilitated by Baby Boom and Wuschel2 “生育高峰”和“乌舍尔”促进了高粱品种的转化

Current protocols in plant biology Pub Date : 2018-10-17 DOI: 10.1002/cppb.20076

Kimberly Nelson-Vasilchik, Joel Hague, Muruganantham Mookkan, Zhanyuan J. Zhang, Albert Kausch

引用次数: 27

Rapid and Efficient Genetic Transformation of Sorghum via Agrobacterium-Mediated Method 农杆菌介导的高粱快速高效遗传转化研究

Current protocols in plant biology Pub Date : 2018-10-12 DOI: 10.1002/cppb.20077

Phat Tien Do, Hyeyoung Lee, Kimberly Nelson-Vasilchik, Albert Kausch, Zhanyuan J. Zhang

引用次数: 11

Protocol for Chromatin Immunoprecipitation of Meiotic-Stage-Specific Tomato Anthers 减数分裂期特异性番茄花药染色质免疫沉淀方法

Current protocols in plant biology Pub Date : 2018-09-12 DOI: 10.1002/cppb.20074

Jihed Chouaref, Esther de Boer, Paul Fransz, Maike Stam

{"title":"Protocol for Chromatin Immunoprecipitation of Meiotic-Stage-Specific Tomato Anthers","authors":"Jihed Chouaref, Esther de Boer, Paul Fransz, Maike Stam","doi":"10.1002/cppb.20074","DOIUrl":"10.1002/cppb.20074","url":null,"abstract":"Interactions occurring between DNA and proteins across the nuclear genome regulate numerous processes, including meiosis. Meiosis ensures genetic variation and balanced segregation of homologous chromosomes. It involves complex DNA-protein interactions across the entire genome to regulate a broad range of processes, including formation and repair of double-strand DNA breaks (DSBs), chromosome compaction, homolog pairing, synapsis, and homologous recombination. The latter meiotic event, meiotic recombination, often occurs at discrete locations in a genome, within a tight time window. The identification of genomic binding sites of meiotic proteins is a major step toward understanding the molecular mechanisms underlying meiotic recombination and provides important information for plant breeding. Collecting meiotic cells from plants is challenging, tedious, and time consuming, since the meiocyte-producing organs, the anthers, are generally small and limited to certain developmental stages of plants. Here we provide a protocol to isolate meiotic-stage-specific anthers and perform ChIP on this material. We have developed a ChIP protocol specifically suited to (1) small amounts of input material and (2) proteins that bind transiently to chromatin and at very low frequency. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":10932,"journal":{"name":"Current protocols in plant biology","volume":"3 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cppb.20074","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36480957","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Labeling Maize (Zea mays L.) Leaves with 15NH4+ and Monitoring Nitrogen Incorporation into Amino Acids by GC/MS Analysis 标签玉米(Zea mays L.)15NH4+处理叶片及氨基酸中氮的GC/MS分析

Current protocols in plant biology Pub Date : 2018-09-10 DOI: 10.1002/cppb.20073

Caroline Cukier, Peter J. Lea, Rafael Cañas, Anne Marmagne, Anis M. Limami, Bertrand Hirel

{"title":"Labeling Maize (Zea mays L.) Leaves with 15NH4+ and Monitoring Nitrogen Incorporation into Amino Acids by GC/MS Analysis","authors":"Caroline Cukier, Peter J. Lea, Rafael Cañas, Anne Marmagne, Anis M. Limami, Bertrand Hirel","doi":"10.1002/cppb.20073","DOIUrl":"10.1002/cppb.20073","url":null,"abstract":"The human body contains approximately 3.2% nitrogen (N), mainly present as protein and amino acids. Although N exists at a high concentration (78%) in the air, it is not readily available to animals and most plants. Plants are however able to take up both nitrate (NO3−) and ammonium (NH4+) ions from the soil and convert them to amino acids and proteins, which are excellent sources for all animals. Most N is available as the stable isotope 14N, but a second form, 15N, is present in very low concentrations. 15N can be detected in extracts of plants by gas chromatography followed by mass spectrometry (GC/MS). In this protocol, the methods are described for tracing the pathway by which plants are able to take up 15N-labeled nitrate and ammonium and convert them into amino acids and proteins. A protocol for extracting and quantifying amino acids and 15N enrichment in maize (Zea mays L.) leaves labeled with 15NH4+ is described. Following amino acid extraction, purification, and separation by GC/MS, a calculation of the 15N enrichment of each amino acid is carried out on a relative basis to identify any differences in the dynamics of amino acid accumulation. This will allow a study of the impact of genetic modifications or mutations on key reactions involved in primary nitrogen and carbon metabolism. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":10932,"journal":{"name":"Current protocols in plant biology","volume":"3 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cppb.20073","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36474712","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7

An Efficient Cell Sorting Protocol for Maize Protoplasts 玉米原生质体的高效细胞分选方法

Current protocols in plant biology Pub Date : 2018-08-23 DOI: 10.1002/cppb.20072

Carlos Ortiz-Ramírez, Edgar Demesa Arevalo, Xiaosa Xu, David P. Jackson, Kenneth D. Birnbaum

{"title":"An Efficient Cell Sorting Protocol for Maize Protoplasts","authors":"Carlos Ortiz-Ramírez, Edgar Demesa Arevalo, Xiaosa Xu, David P. Jackson, Kenneth D. Birnbaum","doi":"10.1002/cppb.20072","DOIUrl":"10.1002/cppb.20072","url":null,"abstract":"Maize is one the most widely cultivated crops worldwide and an important model system for the study of genetics and cytogenetics. Although the availability of a genome sequence has enabled new quantitative genomic studies, developing methods to isolate specific types of cells will enable useful approaches for transcriptomic analysis in the crop plant. Fluorescence-activated cell sorting (FACS) is a powerful technique for cell isolation and the study of transcriptional profiles from specific cell populations. The use of FACS on plant cells requires the generation of protoplasts by tissue digestion and cell wall removal. Although some protocols are available, they mainly focus on dicot species and obtaining sufficient protoplasts from inner tissue layers has been challenging in both monocots and dicots. Here, we report a new protocol that dramatically increases protoplast yield from maize for subsequent cell isolation by FACS. This protocol is efficient in generating protoplasts from root and shoot inner layers and can also be applied successfully to Arabidopsis thaliana. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":10932,"journal":{"name":"Current protocols in plant biology","volume":"3 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cppb.20072","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36419683","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 20

Profiling Interactome Networks with the HaloTag-NAPPA In Situ Protein Array 利用HaloTag-NAPPA原位蛋白阵列分析相互作用组网络

Current protocols in plant biology Pub Date : 2018-08-14 DOI: 10.1002/cppb.20071

Junshi Yazaki, Mary Galli, Alice Y. Kim, Joseph R. Ecker

引用次数: 5

High-Quality Yeast-2-Hybrid Interaction Network Mapping 高质量酵母-2-混合相互作用网络映射

Current protocols in plant biology Pub Date : 2018-06-26 DOI: 10.1002/cppb.20067

Melina Altmann, Stefan Altmann, Claudia Falter, Pascal Falter-Braun

引用次数: 13

Lotus japonicus Genetic, Mutant, and Germplasm Resources 日本莲遗传、突变体及种质资源

Current protocols in plant biology Pub Date : 2018-06-19 DOI: 10.1002/cppb.20070

Masatsugu Hashiguchi, Hidenori Tanaka, Melody Muguerza, Ryo Akashi, Niels Nørgaard Sandal, Stig Uggerhøj Andersen, Shusei Sato

{"title":"Lotus japonicus Genetic, Mutant, and Germplasm Resources","authors":"Masatsugu Hashiguchi, Hidenori Tanaka, Melody Muguerza, Ryo Akashi, Niels Nørgaard Sandal, Stig Uggerhøj Andersen, Shusei Sato","doi":"10.1002/cppb.20070","DOIUrl":"10.1002/cppb.20070","url":null,"abstract":"A quarter of a century has passed since Lotus japonicus was proposed as a model legume because of its suitability for molecular genetic studies. Since then, a comprehensive set of genetic resources and tools has been developed, including recombinant inbred lines, a collection of wild accessions, published mutant lines, a large collection of mutant lines tagged with LORE1 insertions, cDNA clones with expressed sequence tag (EST) information, genomic clones with end-sequence information, and a reference genome sequence. Resource centers in Japan and Denmark ensure easy access to data and materials, and the resources have greatly facilitated L. japonicus research, thereby contributing to the molecular understanding of characteristic legume features such as endosymbiosis. Here, we provide detailed instructions for L. japonicus cultivation and describe how to order materials and access data using the resource center websites. The comprehensive overview presented here will make L. japonicus more easily accessible as a model system, especially for research groups new to L. japonicus research. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":10932,"journal":{"name":"Current protocols in plant biology","volume":"3 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cppb.20070","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36243857","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

High Enrichment [13C]-Labeling of Plants Grown Hydroponically from Seed to Seed in a Controlled 13C-Carbon Dioxide Atmosphere Enclosure 高富集[13C]-在可控13C-二氧化碳环境下水培植物从种子到种子的标记

Current protocols in plant biology Pub Date : 2018-06-19 DOI: 10.1002/cppb.20069

Calvin P. Peters, Erin M. Evans, Jerry D. Cohen, Adrian D. Hegeman

{"title":"High Enrichment [13C]-Labeling of Plants Grown Hydroponically from Seed to Seed in a Controlled 13C-Carbon Dioxide Atmosphere Enclosure","authors":"Calvin P. Peters, Erin M. Evans, Jerry D. Cohen, Adrian D. Hegeman","doi":"10.1002/cppb.20069","DOIUrl":"10.1002/cppb.20069","url":null,"abstract":"In vivo isotopic labeling empowers proteomic and metabolomic analyses to resolve relationships between the molecular composition, environment, and phenotype of an organism. Carbon-13 is particularly useful for plant labeling as it can be introduced via 13CO2 gas and readily assimilated into plant metabolic systems through natural carbon fixation. While short-term labeling experiments can be performed within a simple sealed enclosure, long-term growth in an isolated environment raises many challenges beyond nutrient availability and buildup of metabolic waste. Viable growth conditions must be maintained by means that do not compromise the integrity of the carbon-13 enrichment. To address these issues, an automated growth chamber equipped with countermeasures to neutralize stresses and ensure high isotopic enrichment throughout the life cycle of the plant has been developed. The following describes this growth chamber and its use in an example 130-day growth of ten soybean plants to full maturity, achieving 100% carbon-13 enrichment of new seed tissue. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":10932,"journal":{"name":"Current protocols in plant biology","volume":"3 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cppb.20069","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36243858","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2