Ethyl methanesulfonate mutant library construction in Gossypium hirsutum L. for allotetraploid functional genomics and germplasm innovation

Abstract

SUMMARY As the gene pool is exposed to both strain on land resources and a lack of diversity in elite allotetraploid cotton, the acquisition and identification of novel alleles has taken on epic importance in facilitating cotton genetic improvement and functional genomics research. Ethyl methanesulfonate (EMS) is an excellent mutagen that induces genome‐wide efficient mutations to activate the mutagenic potential of plants with many advantages. The present study established, determined and verified the experimental procedure suitable for EMS‐based mutant library construction as the general reference guide in allotetraploid upland cotton. This optimized method and procedure are efficient, and abundant EMS mutant libraries (approximately 12 000) in allotetraploid cotton were successfully obtained. More than 20 mutant phenotypes were observed and screened, including phenotypes of the leaf, flower, fruit, fiber and plant architecture. Through the plants mutant library, high‐throughput and high‐resolution melting technology‐based variation evaluation detected the EMS‐induced site mutation. Additionally, based on overall genome‐wide mutation analyses by re‐sequencing and mutant library assessment, the examination results demonstrated the ideal quality of the cotton EMS‐treated mutant library constructed in this study with appropriate high mutation density and saturated genome. What is more, the collection is composed of a broad repertoire of mutants, which is the valuable resource for basic genetic research and functional genomics underlying complex allotetraploid traits, as well as cotton breeding.

Highly Efficient and Comprehensive Identification of Ethyl Methanesulfonate-Induced Mutations in Nicotiana tabacum L. by Whole-Genome and Whole-Exome Sequencing

“…The mutation density of the 19 tobacco mutants tested in this study was one mutation per 19.3 kb and 18.0 kb of target regions in the WES and WGS datasets, respectively. These mutation densities are slightly higher than those reported for allotetraploid cotton and hexaploid wheat (one mutation per 26 kb and 24 kb, respectively; Slade et al, 2005 ; Lian et al, 2020 ), and more than 10-fold higher than those reported for diploid rice and tomato (one mutation per 294 kb and 367 kb, respectively; Till et al, 2007 ; Gupta et al, 2017 ). In allotetraploid tobacco, most genes have two functional copies referred to as homeologs.…”

Construction of Gossypium barbadense Mutant Library Provides Genetic Resources for Cotton Germplasm Improvement

“…Genome-wide analyses of EMS-induced SNPs in virescent mutant line YL01 by resequencing revealed the ideal quality of the cotton EMS mutant library with the high and appropriate mutation density ( Table 1 ). Recently published EMS mutant library of Upland cotton [ 13 ] showed 85% of EMS SNPs occurred in intergenic region compared to 81.8% of our Sea island mutant library. In addition, our library quality was further assessed by taking into account 9.16% of the effective functional mutation in genic region compared to 8.3% of Upland cotton mutant library.…”

“…EMS-induced mutations in Upland cotton has been used to generate new valuable alleles and traits, leading to development of commercial cultivated varieties [ 6 , 9 , 10 , 11 , 12 ]. Recently, EMS-induced mutant libraries have been constructed in Upland cotton [ 13 ] representing beneficial genetic variations. Such collection of mutants represented a broad repertoire of mutants, that may serve as a valuable resource for functional genomic research on complex allotetraploid traits.…”

Metabolization and Sequestration of Plant Specialized Metabolites in Insect Herbivores: Current and Emerging Approaches

“…Mutants of Brachypodium, an emerging plant model, are available at the Joint Genome Institute (T-DNA, chemical, and radiation mutants (Fitzgerald et al, 2015;Raissig and Woods, 2022). Additional mutant libraries were further produced for numerous plants, including tobacco (Julio et al, 2008;Lewis et al, 2010;Lewis et al, 2015Takakura et al, 2018Udagawa et al, 2020;Zhao et al, 2020), tomato (Saito et al, 2011), rice (Meng et al, 2017), or cotton (Lian et al, 2020).…”