TILLING to detect induced mutations in soybean

Cooper, Jennifer L.; Till, Bradley J.; Laport, Robert G.; Darlow, Margaret C; Kleffner, Justin M; Jamai, Aziz; El-Mellouki, Tarik; Liu, Shiming; Ritchie, Rae; Nielsen, Niels Chr.; Bilyeu, Kristin; Meksem, Khalid; Comai, Luca; Henikoff, Steven

doi:10.1186/1471-2229-8-9

Cited by 261 publications

(212 citation statements)

References 35 publications

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“…This raises the possibility that gene functional redundancy due to the presence of homeologous gene copies could limit the ability to obtain informative phenotypes for single transposon insertions. However, clearly, phenotypes can be obtained by chemical or radiation mutagenesis (Cooper et al, 2008;Bolon et al, 2011). Moreover, we previously identified a male-sterile mutant of soybean using Ac/Ds mutagenesis (Mathieu et al, 2009).…”

Section: Tnt1: An Insertion Mutagen In Soybean?mentioning

confidence: 99%

Tnt1 Retrotransposon Mutagenesis: A Tool for Soybean Functional Genomics

et al. 2012

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Insertional mutagenesis is a powerful tool for determining gene function in both model and crop plant species. Tnt1, the transposable element of tobacco (Nicotiana tabacum) cell type 1, is a retrotransposon that replicates via an RNA copy that is reverse transcribed and integrated elsewhere in the plant genome. Based on studies in a variety of plants, Tnt1 appears to be inactive in normal plant tissue but can be reactivated by tissue culture. Our goal was to evaluate the utility of the Tnt1 retrotransposon as a mutagenesis strategy in soybean (Glycine max). Experiments showed that the Tnt1 element was stably transformed into soybean plants by Agrobacterium tumefaciens-mediated transformation. Twenty-seven independent transgenic lines carrying Tnt1 insertions were generated. Southernblot analysis revealed that the copy number of transposed Tnt1 elements ranged from four to 19 insertions, with an average of approximately eight copies per line. These insertions showed Mendelian segregation and did not transpose under normal growth conditions. Analysis of 99 Tnt1 flanking sequences revealed insertions into 62 (62%) annotated genes, indicating that the element preferentially inserts into protein-coding regions. Tnt1 insertions were found in all 20 soybean chromosomes, indicating that Tnt1 transposed throughout the soybean genome. Furthermore, fluorescence in situ hybridization experiments validated that Tnt1 inserted into multiple chromosomes. Passage of transgenic lines through two different tissue culture treatments resulted in Tnt1 transposition, significantly increasing the number of insertions per line. Thus, our data demonstrate the Tnt1 retrotransposon to be a powerful system that can be used for effective large-scale insertional mutagenesis in soybean.

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Section: Tnt1: An Insertion Mutagen In Soybean?mentioning

confidence: 99%

Tnt1 Retrotransposon Mutagenesis: A Tool for Soybean Functional Genomics

et al. 2012

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“…Two EMS mutant M2 populations (FM2011 and FM2013) were developed from the SCN-resistant soybean cv Forrest at Southern Illinois University in Carbondale according to the methods described by Cooper et al (2008) and Meksem et al (2008). The FM2011 and FM2013 populations contained 1,588 and 2,827 lines, respectively.…”

Section: Mutant Screening and Phenotypingmentioning

confidence: 99%

Systematic Mutagenesis of Serine Hydroxymethyltransferase Reveals an Essential Role in Nematode Resistance

et al. 2017

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Rhg4 is a major genetic locus that contributes to soybean cyst nematode (SCN) resistance in the Peking-type resistance of soybean (Glycine max), which also requires the rhg1 gene. By map-based cloning and functional genomic approaches, we previously showed that the Rhg4 gene encodes a predicted cytosolic serine hydroxymethyltransferase (GmSHMT08); however, the novel gain of function of GmSHMT08 in SCN resistance remains to be characterized. Using a forward genetic screen, we identified an allelic series of GmSHMT08 mutants that shed new light on the mechanistic aspects of GmSHMT08-mediated resistance. The new mutants provide compelling genetic evidence that Peking-type rhg1 resistance in cv Forrest is fully dependent on the GmSHMT08 gene and demonstrates that this resistance is mechanistically different from the PI 88788-type of resistance that only requires rhg1. We also demonstrated that rhg1-a from cv Forrest, although required, does not exert selection pressure on the nematode to shift from HG type 7, which further validates the bigenic nature of this resistance. Mapping of the identified mutations onto the SHMT structural model uncovered key residues for structural stability, ligand binding, enzyme activity, and protein interactions, suggesting that GmSHMT08 has additional functions aside from its main enzymatic role in SCN resistance. Lastly, we demonstrate the functionality of the GmSHMT08 SCN resistance gene in a transgenic soybean plant.

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“…These mutations have been isolated from a heavily mutagenized population created for the reverse genetics TILLING approach and carry a number of base changes at additional sites [13]. As with all variants created using a mutation breeding strategy, it will therefore be necessary to introgress these loci into elite cultivars for several generations to eliminate potentially deleterious mutations at other loci.…”

Section: Discussionmentioning

confidence: 99%

“…The mutant population consists of N-nitroso-N-methylurea (NMU) mutagenized Glycine max cv. Williams-82 (W82) and was described previously [13,14]. To screen for new, and as-yet unidentified, mutations that contribute to the fatty acid composition of mature soybean seeds, a forward genetic screen was conducted for seed oil composition.…”

Section: Plant Materials and Growthmentioning

confidence: 99%

Soybean Oil-Quality Variants Identified by Large-Scale Mutagenesis

Hudson

2012

International Journal of Agronomy

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To identify genetic variation for fatty acid composition in mature soybean seeds, 4566 M 3 generation seed samples from a chemically mutagenized population were subjected to fatty acid profiling by gas chromatography. In the population, a wide range of variation in the content for each of the five major fatty acids was observed. Seventy-nine lines were identified which contained significantly high or low levels of one of the five major soybean fatty acids. These lines were advanced to the subsequent generation. Of the 79 lines showing a variant fatty acid profile in the M 3 , 52 showed clear heritability for the oil composition in the seeds of the subsequent generation. These lines are likely to represent 52 distinct genetic mutations. These mutants may represent new loci involved in the determination of soybean seed oil content or could be new isolates or alleles of previously identified genetic variants for soybean oil composition.

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TILLING to detect induced mutations in soybean

Cited by 261 publications

References 35 publications

Tnt1 Retrotransposon Mutagenesis: A Tool for Soybean Functional Genomics

Tnt1 Retrotransposon Mutagenesis: A Tool for Soybean Functional Genomics

Systematic Mutagenesis of Serine Hydroxymethyltransferase Reveals an Essential Role in Nematode Resistance

Soybean Oil-Quality Variants Identified by Large-Scale Mutagenesis

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