High Enrichment [13C]-Labeling of Plants Grown Hydroponically from Seed to Seed in a Controlled 13C-Carbon Dioxide Atmosphere Enclosure

Q1 Agricultural and Biological Sciences 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
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引用次数: 2

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.

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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
体内同位素标记使蛋白质组学和代谢组学分析能够解决生物分子组成、环境和表型之间的关系。碳-13对植物标记特别有用,因为它可以通过13CO2气体引入,并通过自然固碳很容易被植物代谢系统吸收。虽然短期标记实验可以在简单的密封环境中进行,但在孤立的环境中长期生长会带来许多挑战,而不仅仅是营养物质的可用性和代谢废物的积累。必须通过不损害碳-13富集完整性的手段来维持可行的生长条件。为了解决这些问题,一种配备了对策的自动化生长室已经开发出来,以中和压力并确保在植物的整个生命周期中都有高同位素富集。下面介绍了这个生长室及其在一个例子中的应用,在130天的生长中,10株大豆植株完全成熟,实现了100%的新种子组织碳-13富集。©2018 by John Wiley &儿子,Inc。
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Current protocols in plant biology
Current protocols in plant biology Agricultural and Biological Sciences-Plant Science
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期刊介绍: Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.
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Issue Information Isolation, Library Preparation, and Bioinformatic Analysis of Historical and Ancient Plant DNA Isolation of Plant Root Nuclei for Single Cell RNA Sequencing Selective Enrichment Coupled with Proteomics to Identify S-Acylated Plasma Membrane Proteins in Arabidopsis In-Plate Quantitative Characterization of Arabidopsis thaliana Susceptibility to the Fungal Vascular Pathogen Fusarium oxysporum.
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