Soybean (<i>Glycine max</i>) Mutant and Germplasm Resources: Current Status and Future Prospects

Campbell, Ben; Stupar, Robert M.

doi:10.1002/cppb.20015

Cited by 11 publications

(5 citation statements)

References 153 publications

(170 reference statements)

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“…Since the announcement of the soya bean draft genome, functional characterization of all 46–56 thousand annotated genes has been the primary goal in soya bean genetic studies (O'Rourke et al ., ; Schmutz et al ., ). Genetic mutagenesis, including chemical, radiation (X‐rays, gamma rays and fast neutrons) and transformation‐induced (T‐DNA and transposon insertion) methods (Bolon et al ., ; Campbell and Stupar, ; Cui et al ., ; Espina et al ., ; Li et al ., ; Mathieu et al ., ; Tsuda et al ., ), is routinely used as forward genetic strategies. However, the complexity of the soya bean genome limits the success of these approaches.…”

Section: Introductionmentioning

confidence: 99%

Generation of a multiplex mutagenesis population via pooled CRISPR‐Cas9 in soya bean

Bai

Yuan

Kuang

et al. 2019

Plant Biotechnology Journal

149

112

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Summary The output of genetic mutant screenings in soya bean [Glycine max (L.) Merr.] has been limited by its paleopolypoid genome. CRISPR‐Cas9 can generate multiplex mutants in crops with complex genomes. Nevertheless, the transformation efficiency of soya bean remains low and, hence, remains the major obstacle in the application of CRISPR‐Cas9 as a mutant screening tool. Here, we report a pooled CRISPR‐Cas9 platform to generate soya bean multiplex mutagenesis populations. We optimized the key steps in the screening protocol, including vector construction, sgRNA assessment, pooled transformation, sgRNA identification and gene editing verification. We constructed 70 CRISPR‐Cas9 vectors to target 102 candidate genes and their paralogs which were subjected to pooled transformation in 16 batches. A population consisting of 407 T0 lines was obtained containing all sgRNAs at an average mutagenesis frequency of 59.2%, including 35.6% lines carrying multiplex mutations. The mutation frequency in the T1 progeny could be increased further despite obtaining a transgenic chimera. In this population, we characterized gmric1/gmric2 double mutants with increased nodule numbers and gmrdn1‐1/1‐2/1‐3 triple mutant lines with decreased nodulation. Our study provides an advanced strategy for the generation of a targeted multiplex mutant population to overcome the gene redundancy problem in soya bean as well as in other major crops.

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Section: Introductionmentioning

confidence: 99%

Generation of a multiplex mutagenesis population via pooled CRISPR‐Cas9 in soya bean

Bai

Yuan

Kuang

et al. 2019

Plant Biotechnology Journal

149

112

View full text Add to dashboard Cite

show abstract

“…Mutant populations are valuable genetic resources for identifying genetic variations and studying gene function in plants. In soybean, several mutant populations have been created using various mutagenic agents such as chemical, irradiation, or transposon (Campbell & Stupar, 2016). More recently, fast neutron (FN) mutant populations were utilized to identify and characterize several causative genes or genetic loci for important seed composition phenotypes in soybean such as increased production of vitamin E (Stacey et al ., 2016), sucrose (Dobbels et al ., 2017), stearic acid (Gillman et al ., 2014), reduced seed phytic acid (Vincent et al ., 2015) or altered plant morphology (Bolon et al ., 2011; Hwang et al ., 2015).…”

Section: Introductionmentioning

confidence: 99%

GmKIX8‐1 regulates organ size in soybean and is the causative gene for the major seed weight QTL qSw17‐1

et al. 2020

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Seed weight is one of the most important agronomic traits in soybean for yield improvement and food production. Several quantitative trait loci (QTLs) associated with the trait have been identified in soybean. However, the genes underlying the QTLs and their functions remain largely unknown. Using forward genetic methods and CRISPR/Cas9 gene editing, we identified and characterized the role of GmKIX8-1 in the control of organ size in soybean. GmKIX8-1 belongs to a family of KIX domain-containing proteins that negatively regulate cell proliferation in plants. Consistent with this predicted function, we found that loss-of-function GmKIX8-1 mutants showed a significant increase in the size of aerial plant organs, such as seeds and leaves. Likewise, the increase in organ size is due to increased cell proliferation, rather than cell expansion, and increased expression of CYCLIN D3;1-10. Lastly, molecular analysis of soybean germplasms harboring the qSw17-1 QTL for the bigseeded phenotype indicated that reduced expression of GmKIX8-1 is the genetic basis of the qSw17-1 phenotype.

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“…Several soybean mutagenesis platforms have been developed for functional characterization of soybean genes (reviewed by Campbell and Stupar 2016 ). These platforms include chemical mutagenesis (Cooper et al 2008 ; Gillman et al 2014 ), transposon tagging (Palmer et al 2008a , b ; Mathieu et al 2009 ; Hancock et al 2011 ; Cui et al 2013 ; Raval et al 2013 ), and irradiation mutagenesis (Men et al 2002 ; Bolon et al 2011 ; Gillman et al 2014 ).…”

Section: Introductionmentioning

confidence: 99%

Fast neutron-induced structural rearrangements at a soybean NAP1 locus result in gnarled trichomes

et al. 2016

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Key messageThree adjacent and distinct sequence rearrangements were identified at a NAP1 locus in a soybean mutant. Genetic dissection and validation revealed the function of this gene in soybean trichome development.AbstractA soybean (Glycine max (L.) Merr.) gnarled trichome mutant, exhibiting stunted trichomes compared to wild-type, was identified in a fast neutron mutant population. Genetic mapping using whole genome sequencing-based bulked segregant analysis identified a 26.6 megabase interval on chromosome 20 that co-segregated with the phenotype. Comparative genomic hybridization analysis of the mutant indicated that the chromosome 20 interval included a small structural variant within the coding region of a soybean ortholog (Glyma.20G019300) of Arabidopsis Nck-Associated Protein 1 (NAP1), a regulator of actin nucleation during trichome morphogenesis. Sequence analysis of the candidate allele revealed multiple rearrangements within the coding region, including two deletions (approximately 1–2 kb each), a translocation, and an inversion. Further analyses revealed that the mutant allele perfectly co-segregated with the phenotype, and a wild-type soybean NAP1 transgene functionally complemented an Arabidopsis nap1 mutant. In addition, mapping and exon sequencing of NAP1 in a spontaneous soybean gnarled trichome mutant (T31) identified a frame shift mutation resulting in a truncation of the coding region. These data indicate that the soybean NAP1 gene is essential for proper trichome development and show the utility of the soybean fast neutron population for forward genetic approaches for identifying genes.Electronic supplementary materialThe online version of this article (doi:10.1007/s00122-016-2735-x) contains supplementary material, which is available to authorized users.

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Soybean (Glycine max) Mutant and Germplasm Resources: Current Status and Future Prospects

Cited by 11 publications

References 153 publications

Generation of a multiplex mutagenesis population via pooled CRISPR‐Cas9 in soya bean

Generation of a multiplex mutagenesis population via pooled CRISPR‐Cas9 in soya bean

GmKIX8‐1 regulates organ size in soybean and is the causative gene for the major seed weight QTL qSw17‐1

Fast neutron-induced structural rearrangements at a soybean NAP1 locus result in gnarled trichomes

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