开发高质量的核隔离以研究植物根-微生物相互作用,用于大豆单核转录基因测序
Leonidas W D'Agostino1, Lenin Yong-Villalobos1, Luis Herrera-Estrella1,2
1Institute of Genomics for Crop Abiotic Stress Tolerance, Department of Plant and Soil Science, Texas Tech University, Lubbock, TX 79409, USA.
Plants (Basel, Switzerland)
|July 14, 2023
概括
研究人员开发了一种新方法,从植物组织中分离出高质量的,无微生物的细胞核,这对于单核RNA测序 (sNucRNA-seq) 和植物中其他单细胞分析至关重要. 这种技术克服了微生物污染带来的挑战,使先进的基因组研究能够得到更广泛的应用.
科学领域:
- 植物生物学 植物生物学
- 基因组学就是基因组学.
- 分子生物学分子生物学
背景情况:
- 单核RNA测序 (sNucRNA-seq) 对于解剖复杂组织中的细胞异质性至关重要.
- 植物组织中的微生物污染对高质量的细胞核隔离构成重大挑战,阻碍了像sNucRNA-seq.这样的下游应用.
- 现有的方法难以从与微生物相关的植物样本中分离纯核,例如根结节.
研究的目的:
- 开发一种简单,有效和可扩展的方法,从植物组织中分离出高质量,无微生物的核,特别是大豆根和结节.
- 解决从微生物殖民植物样本中获得纯核的技术障碍,用于单细胞基因组学.
- 为了促进植物研究中sNucRNA-seq和单核测试用于转移酶可访问的染色体测序 (sNucATAC-seq) 的应用.
主要方法:
- 开发了一种用于从大豆根和结节中隔离核的新方案.
- 包含一个洗步骤来消除细菌污染.
- 通过使用 10× Genomics 微流体平台生成条形码库来验证协议.
主要成果:
- 成功地从大豆根结核中分离出无细菌污染的高质量核.
- 该协议在标准实验室设置中很容易实现,并且可以扩展到多个样本.
- 使用 10× Genomics 平台生成一个有效的条形码库,证明该协议对下游应用程序的有效性.
结论:
- 开发的协议提供了一种可访问和有效的解决方案,用于从与微生物相关的植物组织中分离纯核.
- 这种方法扩大了使用sNucRNA-seq和sNucATAC-seq用于研究复杂的植物微生物相互作用和其他生物过程的可行性.
- 该协议为基于仪器的分类方法提供了一个有价值的替代方案,在植物科学中民主化了先进的单细胞分析.
相关概念视频
01:16In-vitro Mutagenesis
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
01:16In vitro Mutagenesis
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
02:50Transgenic Plants
Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically modified organisms are called transgenic organisms.
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...