Cold Spring Harbor protocols最新文献_第2页

The Rolled Towel Method for Hormone Response Assays in Maize. 玉米激素反应测定的卷纸法

Cold Spring Harbor protocols Pub Date : 2024-10-30 DOI: 10.1101/pdb.prot108623

Clarice F Gonzales, Craig L Cowling, Dior R Kelley

引用次数: 0

Exogenous Hormone Treatments in Maize. 玉米的外源激素处理。

Cold Spring Harbor protocols Pub Date : 2024-10-30 DOI: 10.1101/pdb.top108526

José Alfredo Guzmán-López, Rodrigo Muñoz-Javier, Dior R Kelley, María Jazmín Abraham-Juárez

{"title":"Exogenous Hormone Treatments in Maize.","authors":"José Alfredo Guzmán-López, Rodrigo Muñoz-Javier, Dior R Kelley, María Jazmín Abraham-Juárez","doi":"10.1101/pdb.top108526","DOIUrl":"https://doi.org/10.1101/pdb.top108526","url":null,"abstract":"Plant hormones have key functions in plant morphology, physiology, and stress responses. Studies on the biology of hormones and their effect on plant physiology and metabolism are greatly facilitated by the exogenous application of these compounds. In general, methods for exogenous hormone application are easy and fast, and provide useful information about their effects in planta. Although hormone effects have been studied in several plant species, the used methods need to be tailored specifically to each species to get robust data. Maize is an established model for basic and applied research, and an excellent system for studying the effects of hormones on developmental and stress responses in a cereal crop. Different methods have been reported for the exogenous application of plant growth regulators in maize, including watering, spraying, immersion, and application to the apical whorl. These various methods are useful to analyze hormone responses at different developmental stages, in specific organs, and within tissues. As with all exogenous application assays, suitable experimental design and the inclusion of proper controls are critical factors in these methods, to obtain reliable and reproducible results. Here, we provide an overview of various methods for hormone exogenous application in maize, and technical considerations to get successful results.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142544159","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

Foliar Spray Treatment for Exogenous Application of Hormones in Maize. 玉米外源应用激素的叶面喷洒处理。

Cold Spring Harbor protocols Pub Date : 2024-10-30 DOI: 10.1101/pdb.prot108621

Enrique Pola-Sánchez, Rodrigo Muñoz-Javier, José Alfredo Guzmán-López, María Jazmín Abraham-Juárez

引用次数: 0

Apical Whorl Treatment for Exogenous Application of Hormones in Maize. 玉米外源应用激素的顶轮处理

Cold Spring Harbor protocols Pub Date : 2024-10-30 DOI: 10.1101/pdb.prot108622

Rodrigo Muñoz-Javier, Enrique Pola-Sánchez, José Alfredo Guzmán-López, María Jazmín Abraham-Juárez

引用次数: 0

Semisynthetic Phage Display Library Construction: Generation of Single-Chain Variable Fragment Secondary Libraries. 半合成噬菌体展示文库构建：单链可变片段二级文库的生成。

Cold Spring Harbor protocols Pub Date : 2024-10-16 DOI: 10.1101/pdb.prot108616

Juan C Almagro, Mary Ann Pohl

{"title":"Semisynthetic Phage Display Library Construction: Generation of Single-Chain Variable Fragment Secondary Libraries.","authors":"Juan C Almagro, Mary Ann Pohl","doi":"10.1101/pdb.prot108616","DOIUrl":"https://doi.org/10.1101/pdb.prot108616","url":null,"abstract":"Display of antibody fragments on the surface of M13 filamentous bacteriophages is a well-established approach for the identification of antibodies binding to a target of interest. Here, we describe the third and final step of a three-step method to construct Antibody Libraries for Therapeutic Antibody Discovery (ALTHEA) Libraries. The three-step method involves (1) primary library construction, (2) filtered library (FL) construction, and (3) secondary library (SL) construction. In the third step, described here, the nucleotide sequences encoding the single-chain variable fragments (scFvs) of FLs are amplified by PCR and combined with the heavy- chain CDR3 region (HCDR3) and joining fragments (H3J) obtained from a pool of donors to maximize diversity (\"natural H3J fragments\"). These natural H3J fragments are amplified with a set of primers designed to capture >95% of the natural H3J repertoire. The resultant fragments replace the neutral H3J fragments of the FLs, resulting in the final semisynthetic secondary libraries. The quality of these libraries is assessed by sequencing clones chosen at random from the libraries, typically 96 clones. These libraries are then ready to be used for phage selections on targets of interest, providing a robust antibody discovery platform.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142459809","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

Doubled Haploid Technology: Generation of Doubled Haploid Maize Lines Using Haploid Inducers. 双单倍体技术：使用单倍体诱导剂生成双倍单倍体玉米品系。

Cold Spring Harbor protocols Pub Date : 2024-10-16 DOI: 10.1101/pdb.prot108624

Vencke K Grüning, Thomas Lübberstedt, Ursula K Frei

{"title":"Doubled Haploid Technology: Generation of Doubled Haploid Maize Lines Using Haploid Inducers.","authors":"Vencke K Grüning, Thomas Lübberstedt, Ursula K Frei","doi":"10.1101/pdb.prot108624","DOIUrl":"https://doi.org/10.1101/pdb.prot108624","url":null,"abstract":"Doubled haploid (DH) technology allows for the development of completely homozygous lines from heterozygous plants in only two generations. This approach has been widely adopted in maize breeding programs, as it expedites the generation of inbred lines compared to traditional methods. The DH approach is based on the use of maize genotypes that have the ability to induce haploid seeds when used as the pollen parent. The most common method for producing maize haploid plants for the generation of DH lines is in vivo maternal haploid induction. The process involves pollination with a haploid inducer maize line to generate haploid seeds. Then, haploids are screened for and identified (typically via the expression of a particular marker gene), germinated, treated with an exogenous doubling agent to induce genome duplication, and transplanted to the field. Following successful self-pollination, seeds harvested from the ear represent fully homozygous lines. The seed set at this stage, however, is often low, necessitating one or two additional rounds of self-pollination to increase the number of fully homozygous inbred lines. Here, we describe a protocol for the generation of maize DH lines using maternal haploid-inducing maize lines. We outline the steps for setting up the donor material, performing induction crosses, selecting haploids based on two different marker alleles, treating seedlings with colchicine to double the genome, transplanting the treated seedlings to the field, and self-pollinating the treated plants.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142459795","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

Doubled Haploid Technology: Opportunities and Challenges for the Rapid Generation of Maize Homozygous Lines. 双单倍体技术：快速生成玉米同源系的机遇与挑战。

Cold Spring Harbor protocols Pub Date : 2024-10-16 DOI: 10.1101/pdb.top108437

Vencke K Grüning, Thomas Lübberstedt, Ursula K Frei

引用次数: 0

Semisynthetic Phage Display Library Construction: Generation of Filtered Libraries. 半合成噬菌体展示文库构建：过滤文库的生成。

Cold Spring Harbor protocols Pub Date : 2024-10-16 DOI: 10.1101/pdb.prot108615

Juan C Almagro, Mary Ann Pohl

引用次数: 0

Considerations for Using Phage Display Technology in Therapeutic Antibody Drug Discovery. 在治疗性抗体药物研发中使用噬菌体展示技术的注意事项。

Cold Spring Harbor protocols Pub Date : 2024-10-16 DOI: 10.1101/pdb.top107757

Mary Ann Pohl, Juan C Almagro

引用次数: 0

Maize Abiotic Stress Treatments in Controlled Environments. 受控环境中的玉米非生物胁迫处理。

Cold Spring Harbor protocols Pub Date : 2024-10-16 DOI: 10.1101/pdb.prot108620

Katherine M Murphy, Anna L Casto, Leonardo Chavez, Leonardo W Lima, Alejandra Quiñones, Malia A Gehan, Cory D Hirsch

{"title":"Maize Abiotic Stress Treatments in Controlled Environments.","authors":"Katherine M Murphy, Anna L Casto, Leonardo Chavez, Leonardo W Lima, Alejandra Quiñones, Malia A Gehan, Cory D Hirsch","doi":"10.1101/pdb.prot108620","DOIUrl":"https://doi.org/10.1101/pdb.prot108620","url":null,"abstract":"Maize (Zea mays) is one of the world's most important crops, providing food for humans and livestock and serving as a bioenergy source. Climate change and the resulting abiotic stressors in the field reduce crop yields, threatening food security and the global economy. Water deficit (i.e., drought), heat, and insufficient nutrients (e.g., nitrogen and phosphorus) are major environmental stressors that affect maize yields, and impact growth and development at all stages of the plant life cycle. Understanding the biological processes underlying these responses in maize has the potential to increase yields in the face of abiotic stress. Optimizing individual or combined abiotic stress treatments in controlled environments reduces potential noise in data collection that can be present under less controlled growth conditions. Here, we describe methods and conditions for controlled abiotic stress treatments and associated controls during early vegetative growth of maize, conducted in greenhouses or growth chambers. This includes the environmental conditions, equipment, soil preparation, and intensity and duration of heat, drought, nitrogen deficiency, and phosphorous deficiency. Controlled experiments at early growth stages are informative for future in-field studies that require greater labor and inputs, saving researchers time and growing space, and thus research funds, before testing plants across later stages of development. We suggest that stress treatments be severe enough to result in a measurable phenotype, but not so severe that all plants die prior to sample collection. This protocol is designed to set important standards for replicable research in maize.","PeriodicalId":10496,"journal":{"name":"Cold Spring Harbor protocols","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142459803","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