Cold Spring Harbor protocols最新文献

Inoculation of Maize Roots with Globisporangium ultimum var. ultimum to Study Pythium Root Rot. 用最后一种球孢根接种玉米根腐病研究。

Cold Spring Harbor protocols Pub Date : 2025-07-16 DOI: 10.1101/pdb.prot108640

Harrison Hall, Peyton Sorensen, Tiffany Jamann, Santiago Mideros

{"title":"Inoculation of Maize Roots with Globisporangium ultimum var. ultimum to Study Pythium Root Rot.","authors":"Harrison Hall, Peyton Sorensen, Tiffany Jamann, Santiago Mideros","doi":"10.1101/pdb.prot108640","DOIUrl":"10.1101/pdb.prot108640","url":null,"abstract":"Maize is a globally important field crop for food and fuel production. Yield can be affected early in the growing season by oomycete and fungal pathogens that cause root rot or prevent seed germination. The diseases caused by these pathogens are referred to as seedling blights, root rots, or damping off. Pythium root rot is one of the most significant of these diseases. The disease is caused by multiple species of the oomycete genera Globisporangium and Pythium and results in significant yield losses due to reduced seed germination and reduced vigor of surviving seedlings. In this protocol, we mimic the natural infection process by mixing the inoculum into the potting media in which seeds are planted. Then, we flood the seeds daily for several days in large plastic totes to induce flooding conditions. Disease severity is assessed using stand counts and measuring root mass and length. This protocol allows researchers to investigate quantitative differences in disease symptoms, isolate aggressiveness, as well as levels of host resistance. This protocol was developed for the pathogen Globisporangium ultimum var. ultimum, but it can be adapted for other species.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144215157","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

Corrigendum: Inoculation of Maize Roots with Globisporangium ultimum var. ultimum to Study Pythium Root Rot. 勘误：用最后一种球孢菊接种玉米根系研究牙周病。

Cold Spring Harbor protocols Pub Date : 2025-07-16 DOI: 10.1101/pdb.Corr108654

Harrison Hall, Peyton Sorensen, Tiffany Jamann, Santiago Mideros

引用次数: 0

The Drosophila Larval Neuromuscular Junction: Developmental Overview. 果蝇幼虫神经肌肉接头：发育概述

Cold Spring Harbor protocols Pub Date : 2025-07-01 DOI: 10.1101/pdb.top108449

James Ashley, Robert A Carrillo

引用次数: 0

Immunohistochemistry and Morphometric Analysis of Drosophila Larval Body Wall Neuromuscular Junction Preparations. 果蝇幼虫体壁神经肌肉接头制剂的免疫组织化学和形态计量分析

Cold Spring Harbor protocols Pub Date : 2025-07-01 DOI: 10.1101/pdb.prot108500

James Ashley, Robert A Carrillo

引用次数: 0

Using the Proximity Ligation Assay to Visualize Colocalization of Proteins at the Drosophila Larval Neuromuscular Junction. 利用近接测定法观察果蝇幼虫神经肌肉接头处蛋白质的共定位现象

Cold Spring Harbor protocols Pub Date : 2025-07-01 DOI: 10.1101/pdb.prot108502

James Ashley, Robert A Carrillo

{"title":"Using the Proximity Ligation Assay to Visualize Colocalization of Proteins at the Drosophila Larval Neuromuscular Junction.","authors":"James Ashley, Robert A Carrillo","doi":"10.1101/pdb.prot108502","DOIUrl":"10.1101/pdb.prot108502","url":null,"abstract":"In the nearly 50 years since the neuromuscular junction (NMJ) was first established as a model synapse, its molecular composition has been extensively characterized. Early work relied on fluorescent signals to determine whether proteins localized to the pre- and postsynaptic regions. As more synaptic molecules were identified, determining the localization of these proteins relative to each other became important. Conventional microscopy lacks the resolving power to assess whether two proteins are within an appropriate distance to bind directly or be part of a larger complex. Super-resolution and immunoelectron microscopies can improve spatial resolution, but these techniques can be difficult to execute and troubleshoot, and access to these instruments is limiting. However, another approach, proximity labeling, overcomes many of these limitations by using a DNA secondary label that can only be amplified if the two proteins of interest are within 40 nm of each other, which is ∼5× greater than the resolving power of conventional microscopy. In this protocol, we describe the use of the proximity ligation assay, which combines immunohistochemistry with DNA amplification, to reveal protein colocalization in the Drosophila NMJ.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":"pdb.prot108502"},"PeriodicalIF":0.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141310238","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

Drosophila Late Embryonic through Late Larval Stage Body Wall Dissection: Dissection Tools and Techniques. 果蝇胚后期至幼虫后期体壁解剖：解剖工具和技术。

Cold Spring Harbor protocols Pub Date : 2025-07-01 DOI: 10.1101/pdb.prot108499

James Ashley, Robert A Carrillo

{"title":"Drosophila Late Embryonic through Late Larval Stage Body Wall Dissection: Dissection Tools and Techniques.","authors":"James Ashley, Robert A Carrillo","doi":"10.1101/pdb.prot108499","DOIUrl":"10.1101/pdb.prot108499","url":null,"abstract":"One of the challenges of studying synaptic structure and function is accessibility. Some of the earliest readily identifiable and accessible synapses were from the frog and various arthropods. To address questions regarding mechanisms that underlie synaptic development and function, genetically tractable systems were required, and researchers turned to the Drosophila melanogaster embryonic/larval neuromuscular preparation. Drosophila embryos are transparent and can be labeled with antibodies or probes and imaged in whole-mount preparation for structural analysis. Embryos can also be dissected to visualize the entire body wall musculature as well as finer details including live protein trafficking and protein-protein interactions. Whereas younger dissected embryos can be mounted directly onto charged slides, more mature embryos and larvae develop a cuticle that impedes this adherence, so different techniques must be applied. In this protocol, we detail how to manufacture dissection tools and collect embryos, and discuss the individual steps of dissecting late-stage embryos, early first-instar larvae, and late-stage third-instar larvae.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":"pdb.prot108499"},"PeriodicalIF":0.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141310233","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

Cell Ablation Techniques for the Larval Drosophila Neuromuscular System. 果蝇幼体神经肌肉系统的细胞消融技术

Cold Spring Harbor protocols Pub Date : 2025-07-01 DOI: 10.1101/pdb.prot108503

James Ashley, Robert A Carrillo

引用次数: 0

Erratum: Two-Step Bacterial Artificial Chromosome (BAC) Engineering: Preparation and Verification of the Recombinant Shuttle Vector. 两步细菌人工染色体（BAC）工程：重组穿梭载体的制备和验证。

Cold Spring Harbor protocols Pub Date : 2025-07-01 DOI: 10.1101/pdb.err108649

Nathaniel Heintz, Shiaoching Gong

引用次数: 0

Labeling of Cell Surface Proteins at the Drosophila Larval Neuromuscular Junction Using Binding Partner Peptides. 利用结合伴侣肽标记果蝇幼虫神经肌肉接头处的细胞表面蛋白

Cold Spring Harbor protocols Pub Date : 2025-07-01 DOI: 10.1101/pdb.prot108501

James Ashley, Robert A Carrillo

引用次数: 0

How to Harvest and Store Corn (Zea mays). 如何收获和储存玉米（玉米）。

Cold Spring Harbor protocols Pub Date : 2025-06-03 DOI: 10.1101/pdb.prot108639

Miriam Nancy Salazar-Vidal, Melissa A Draves, Sarah L Fitzsimmons, Zachary B Traylor, Sherry Flint-Garcia

{"title":"How to Harvest and Store Corn (Zea mays).","authors":"Miriam Nancy Salazar-Vidal, Melissa A Draves, Sarah L Fitzsimmons, Zachary B Traylor, Sherry Flint-Garcia","doi":"10.1101/pdb.prot108639","DOIUrl":"https://doi.org/10.1101/pdb.prot108639","url":null,"abstract":"Harvest scheduling, seed drying, and good storage practices are essential for maize research to avoid negative impacts on the quality of the seeds and to maximize seed viability. Embryo growth and accumulation of energy reserves in the endosperm are completed ∼40 days after pollination, of which the last 10-20 days are devoted to maturation and desiccation. Seed maturity is affected by many factors including temperature, day length, humidity, and soil moisture. Once seeds are mature, they must be harvested. Hand harvesting, which allows for greater control and minimizes ear damage, is primarily used in genetic nurseries and general research because each genotype is represented by a small number of plants. Hand harvesting is also used where there is a mix of manually pollinated and undesired open pollinated ears, and can be a selective harvest depending on the research objectives. However, hand harvesting is more labor-intensive and time-consuming compared to combine harvesting. If harvesting a yield trial, where the purpose is to collect yield data and identify promising genotypes, the use of a combine (not described in this protocol) is critical to consistently capture grain weight, moisture, and test weight for each plot. Following harvest, materials must be dried to the appropriate moisture content before storage. Corn is typically stored using the \"active collection\" model, with temperatures set between 5°C and 10°C and low relative humidity. This harvest protocol is intended to assist laboratories that are new to maize research and may be modified based on project goals, genetic material, equipment, or available space.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144215152","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