藻类学 植物生物化学 植物育种 植物细胞生物学 植物发育生物学 植物免疫 植物新陈代谢
植物分子生物学 DNA RNA 植物生理学 植物转化Optimized Protocol for DNA Extraction in Three Theobroma Species
三种可可属植物的DNA提取优化方法
DNA extraction is a crucial step in molecular biology research, particularly for genetic and genomic analyses. These studies require a high concentration of high-quality DNA, which is often a challenge for underexplored species or when the available plant material consists of aged tissue. To address these challenges, the cetyltrimethylammonium bromide (CTAB)-based DNA extraction method has been optimized to improve efficiency and yield. The process begins with an overnight incubation of plant tissue macerated with liquid nitrogen in a solution containing a high concentration of CTAB (4%). Subsequently, the mixture undergoes two washes with chloroform: isoamyl alcohol. The nucleic acids are then precipitated using isopropanol, followed by a wash with 70% ethanol to ensure purity. Finally, the purified DNA is resuspended in ultrapure water. This optimized procedure produces high-quality DNA suitable for various downstream applications, including PCR and sequencing, even from older leaves of the three Theobroma species: T. cacao, T. bicolor, and T. grandiflorum. Additionally, this protocol significantly enhances throughput and allows for the parallel processing of a substantially larger number of samples compared to conventional techniques.
Quantification of Botrytis cinerea Growth in Arabidopsis thaliana
拟南芥灰葡萄孢生长的定量研究
Yield losses attributed to plant pathogens pose a serious threat to plant productivity and food security. Botrytis cinerea is one of the most devastating plant pathogens, infecting a wide array of plant species; it has also been established as a model organism to study plant–pathogen interactions. In this context, development of different assays to follow the relative success of B. cinerea infections is required. Here, we describe two methods to quantify B. cinerea development in Arabidopsis thaliana genotypes through measurements of lesion development and quantification of fungal genomic DNA in infected tissues. This provides two independent techniques that are useful in assessing the susceptibility or tolerance of different Arabidopsis genotypes to B. cinerea.
Key features
• Protocol for the propagation of the necrotrophic plant pathogen fungus Botrytis cinerea and spore production.
• Two methods of Arabidopsis thaliana infection with the pathogen using droplet and spray inoculation.
• Two readouts, either by measuring lesion size or by the quantification of fungal DNA using quantitative PCR.
• The two methods are applicable across plant species susceptible the B. cinerea.
Graphical overview
A simplified overview of the droplet and spray infection methods used for the determination of B. cinerea growth in different Arabidopsis genotypes
Safe DNA-extraction Protocol Suitable for Studying Tree-fungus Interactions
适于研究树-真菌互作的安全DNA提取方法
An Optimized CTAB Method for Genomic DNA Extraction from Freshly-picked Pinnae of Fern, Adiantum capillus-veneris L.
一种从新鲜收取的铁线蕨样品中提取基因组DNA 的优化CTAB法
Rolling Circle Amplification to Screen Yam Germplasm for Badnavirus Infections and to Amplify and Characterise Novel Badnavirus Genomes
滚换扩增筛选山药Badnavirus感染种质并扩增鉴定新型Badnavirus基因组
Determining Genome Size from Spores of Seedless Vascular Plants
无种子维管束植物孢子基因组大小的测定
Experimental Pipeline for SNP and SSR Discovery and Genotyping Analysis of Mango (Mangifera indica L.)
芒果SNP和SSR的发现和基因分型分析的实验方法
Genomic DNA Extraction and Genotyping of Dictyochloropsis Green Algae Strains
Dictyochloropsis绿藻菌株的基因组DNA提取和基因分型
Figure 1. Schematic representation of the analysis pipeline