Fusarium wilt, 1st ed. 2022 Methods and Protocols Methods in Molecular Biology Series, Vol. 2391

This volume provides a collection of molecular protocols detailing the most common and modern techniques on fusarium wilt. Chapters guide readers through methods on initial isolation, molecular-based identification, genome characterization, generation of mutants, and characterization of interactions with other organisms including host plants.  Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

Authoritative and cutting-edge, Fusarium wilt: Methods and Protocols aims to be a valuable resource for mycologists, plant pathologists, microbiologists, geneticists, and other scientists that have an interest in members of the Fusarium oxysporum species complex or closely related fungi.   

1.  Pure culture and DNA sequence-based identification of Fusarium from symptomatic plants and diverse substrates

Kerry O’Donnell, Imana Laraba, and David M. Geiser

2.  Isolation of high-molecular weight (HMW) DNA from Fusarium oxysporum for long-read sequencing

Zunyong Liu, Eun-Gyu No, Catherine Danmaigona Clement, Ping He, and Libo Shan

3.  Karyotyping of Fusarium oxysporum by pulsed field gel electrophoresis and the germ tube burst method

Yu Ayukawa and Masatoki Taga

4.  RNA-seq data analysis for differential expression

Navdeep Gill and Braham Dhillon

5.  Quantification and isolation of spontaneous colony growth variants

Cristina López Díaz, Lucía Gómez-Gil, Elena Pérez-Nadales, Gesabel Navarro Velasco, and Antonio Di Pietro

6.  Generation of a T-DNA insertion mutant library of Fusarium oxysporum using Agrobacterium tumefaciens

Yan-Hong Dong and Shu-Tong Wang

7.  Targeted gene disruption via CRISPR/Cas9 ribonucleoprotein complexes in Fusarium oxysporum

Ambika Pokhrel, Seungyeon Seo, Qiang Wang, and Jeffrey J. Coleman

8.  CRISPR/Cas9 RNP-mediated gene fusion to assess protein quantification and subcellular localization in Fusarium oxysporum

9.  In vivo monitoring of cytosolic pH using the ratiometric pH sensor pHluorin

Tânia R. Fernandes, Antonio Serrano, and Antonio Di Pietro

10.  Multi-pronged investigation of volatile compound-mediated interactions of Fusarium oxysporum with plants, fungi and bacteria

Ningxiao Li and Seogchan Kang

11.  Isolation of mitochondria from Fusarium species

Michael MacKillop and John C. Kennell

12.  In planta gene expression analysis and colonization of Fusarium oxysporum

Virginia Casado-del Castillo, Ernesto Pérez Benito, and José María Díaz-Mínguez

Hye-Seon Kim, Sook-Young Park, Seogchan Kang, and Kirk Czymmek

14.  High-throughput screening assays to identify plant natural products with antifungal properties against Fusarium oxysporum

Yong Zhang, Pei-Lun Kao, Akaansha Rampal, Sibongile Mafu, Sergey N. Savinov, and Li-Jun Ma

15.  Screening and assessment of pisatin demethylase activity (PDA)

16.  Detection and genotyping of Fusarium oxysporum f. sp. vasinfectum race 4 (VCG0114), the Fusarium wilt pathogen of cotton

Jinggao Liu and Tanya A. Wagner

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible result

Contains key notes and implementation advice from the experts