Chemical Mutagenesis of Seed and Vegetatively Propagated Plants Using EMS

Jankowicz-Cieślak, Joanna; Till, Bradley J.

doi:10.1002/cppb.20040

Cited by 48 publications

(23 citation statements)

References 40 publications

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“…Two gamma dosages were applied towards the banana populations, 20 Gy and 40 Gy. Genetic mosaicism resulting from mutagenizing multicellular tissues was removed through repeated rounds of longitudinal bisection of the shoot apical meristem as previously described (Jankowicz‐Cieslak and Till, , ).…”

Section: Resultsmentioning

confidence: 99%

Induction and recovery of copy number variation in banana through gamma irradiation and low‐coverage whole‐genome sequencing

Datta

Jankowicz-Cieślak

Nielen

et al. 2018

Plant Biotechnology Journal

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SummaryTraditional breeding methods are hindered in bananas due to the fact that major cultivars are sterile, parthenocarpic, triploid and thus clonally propagated. This has resulted in a narrow genetic base and limited resilience to biotic and abiotic stresses. Mutagenesis of in vitro propagated bananas is one method to introduce novel alleles and broaden genetic diversity. We previously established a method for the induction and recovery of single nucleotide mutations generated with the chemical mutagen EMS. However, officially released mutant banana varieties have been created using gamma rays, a mutagen that can produce large genomic insertions and deletions (indels). Such dosage mutations may be important for generating observable phenotypes in polyploids. In this study, we establish a low‐coverage whole‐genome sequencing approach in triploid bananas to recover large genomic indels caused by treatment with gamma irradiation. We first evaluated the commercially released mutant cultivar ‘Novaria’ and found that it harbours multiple predicted deletions, ranging from 0.3 to 3.8 million base pairs (Mbp). In total, predicted deletions span 189 coding regions. To evaluate the feasibility of generating and maintaining new mutations, we developed a pipeline for mutagenesis and screening for copy number variation in Cavendish bananas using the cultivar ‘Williams’. Putative mutations were recovered in 70% of lines treated with 20 Gy and 60% of the lines treated with 40 Gy. While deletion events predominate, insertions were identified in 20 Gy‐treated material. Based on these results, we believe this approach can be scaled up to support large breeding projects.

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Section: Resultsmentioning

confidence: 99%

Induction and recovery of copy number variation in banana through gamma irradiation and low‐coverage whole‐genome sequencing

Datta

Jankowicz-Cieślak

Nielen

et al. 2018

Plant Biotechnology Journal

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“…EMS solution was prepared as 0.1 M in phosphate buffer at pH 7.0. One hundred embryogenic calli of FN39 and ROC22 were harvested and treated with different concentrations of EMS (0%, 0.05%, 0.1%, 0.2%, and 0.5%) at different time intervals (5,10,15,20, and 25 h) [20]. Then, callus tissues were cut at a uniform diameter (3 mm), immersed into EMS phosphate buffer, and further placed on a shaker at 20 • C, 130 rpm/min.…”

Section: Measurement Of Optimal Ems Concentration and Mutagenic Treatmentioning

confidence: 99%

“…In this regard, chemical mutagenesis provides an easy and cost-effective way to produce a high density of novel nucleotide diversity within plants having a complex genome. Ethyl methanesulfonate (EMS) is widely used as a chemical mutagen, in order to induce hundreds to thousands of heritable single base mutations in a single plant line [15]. After mutagenesis, EMS-mutated plant populations can be screened against environmental stresses on a phenotypic and molecular basis [16].…”

Section: Introductionmentioning

confidence: 99%

Screening of EMS-Induced Drought-Tolerant Sugarcane Mutants Employing Physiological, Molecular and Enzymatic Approaches

et al. 2018

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Drought stress is one of the major agronomic concerns that lead towards a sharp decline in sugarcane yield. An urgent demand to overcome drought is critical to ensure sugarcane production. Mutation breeding is one of the promising tools available to produce stress-resistant plants, with the induction of new alleles due to point mutation within existing sugarcane germplasm. The current study was directed to chemically mutagenize the calli of two sugarcane cultivars (ROC22 and FN39) via 0.1% EMS, with focus on inducing mutations in their genome. The 1644 regenerated plants of ROC22 and 1398 of FN39 were exposed to 28% PEG-6000 stimulated osmotic stress. Eighteen plants of ROC22 and 2 plants of FN39, that survived after in vitro osmotic stress treatment, were then subjected to preliminary greenhouse pot trials to confirm drought tolerance by analyzing them using various physiological parameters, including photosystem II (PSII) photochemical efficiency (Fv/Fm), leaf chlorophyll content, and photosynthetic rate. The genetic diversity among drought-resistant mutant lines was further assessed by 15 pairs of simple sequence repeat (SSR) markers amplification and CEL (Celery) I endonuclease digestion, to investigate the mutated sites. Mutant lines of ROC22 (i.e., MR22-15 and MR22-20) were found to be promising for future drought resistance breeding, due to better physiological adaptation under drought stress.

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“…Mutant isolation by TILLING using an EMS population (Jankowicz-Cieslak & Till, 2016) is also available for Medicago. Because mutated genes are not tagged in these populations, they are less popular than the Tnt1 insertion mutants.…”

Section: Tilling In Medicagomentioning

confidence: 99%

Medicago truncatula: Genetic and Genomic Resources

et al. 2017

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Medicago truncatula was chosen by the legume community, along with Lotus japonicus, as a model plant to study legume biology. Since then, numerous resources and tools have been developed for M. truncatula. These include, for example, its genome sequence, core ecotype collections, transformation/regeneration methods, extensive mutant collections, and a gene expression atlas. This review aims to describe the different genetic and genomic tools and resources currently available for M. truncatula. We also describe how these resources were generated and provide all the information necessary to access these resources and use them from a practical point of view. © 2017 by John Wiley & Sons, Inc.

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Chemical Mutagenesis of Seed and Vegetatively Propagated Plants Using EMS

Cited by 48 publications

References 40 publications

Induction and recovery of copy number variation in banana through gamma irradiation and low‐coverage whole‐genome sequencing

Induction and recovery of copy number variation in banana through gamma irradiation and low‐coverage whole‐genome sequencing

Screening of EMS-Induced Drought-Tolerant Sugarcane Mutants Employing Physiological, Molecular and Enzymatic Approaches

Medicago truncatula: Genetic and Genomic Resources

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