Two forward genetic screens for vein density mutants in sorghum converge on a cytochrome P450 gene in the brassinosteroid pathway

Rizal, Govinda; Thakur, Vivek; Dionora, Jacqueline; Karki, Shanta; Wanchana, Samart; Acebron, Kelvin; Larazo, Nikki; Garcia, Richard; Mabilangan, Abigail E.; Montecillo, Florencia; Danila, Florence R.; Mogul, Reychelle; Pablico, P. P.; Leung, Hei; Langdale, Jane A.; Sheehy, J. E.; Kelly, Steven; Quick, W. Paul

doi:10.1111/tpj.13007

Cited by 38 publications

(37 citation statements)

References 47 publications

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“…Mutants are also potential materials for breeding new cultivar (Takagi et al, 2015). Moreover, mutants are also powerful tools for gene clone and function analysis by reverse or forward approach (Emmanuel and Levy, 2002; Takagi et al, 2013, 2015; Rizal et al, 2015). …”

Section: Introductionmentioning

confidence: 99%

The Analysis of Physiological Variations in M2 Generation of Solanum melongena L. Mutagenized by Ethyl Methane Sulfonate

Xiou

Lin

et al. 2017

Front. Plant Sci.

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The eggplant was mutagenized with ethyl methane sulfonate (EMS) to enhance its genetic variability in our previous paper. In this article, we further analyzed the phenotype of M2 generation of mutant eggplants. A total of 325 independent M2 families were investigated for phenotypic variation. In addition to the visible phenotypic variation, chlorogenic acid (CGA) concentrations were analyzed in 26 fruits of mutants with High Performance Liquid Chromatography assay. Seventeen fruits exhibited significantly higher concentrations of CGAs than those in wild-type. The anthocyanin concentration of S9-1, the purple black mutant, was higher than WT, meanwhile, the anthocyanin concentration of L6-4 and U36-1 was lower than WT. Furthermore, our RT-PCR result demonstrated that the expression levels of anthocyanin biosynthetic genes, except for SmPAL, were increased in S9-1, and the regulator SmMYB1 was decreased in L6-4 and U36-1 mutants. Together, our data indicated that, M2 generation showed abundant phenotypic variations and the strong potential usage for next step of breeding and molecular genetic mechanisms in eggplant.

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Section: Introductionmentioning

confidence: 99%

The Analysis of Physiological Variations in M2 Generation of Solanum melongena L. Mutagenized by Ethyl Methane Sulfonate

Xiou

Lin

et al. 2017

Front. Plant Sci.

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“…This alignment revealed a few unexpected results. First, the 5′ portion of the ALS gene from the S. bicolor 'BTx623' reference genome, which was used for primer design in PCR-based surveys of ALS codons, was found to be nonhomologous to the 5′ ALS sequences from other panicoid grasses or to the ALS gene that was de novo assembled in this study using the 'BTx623-Rooney' resequencing data generated by Rizal et al (2015). In contrast, the 5′ portion of the ALS gene from the de novo assembled 'BTx623-Rooney' sequence matches well with other panicoid grasses.…”

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confidence: 99%

Independent Evolution of Acetolactate Synthase–inhibiting Herbicide Resistance in WeedySorghumPopulations across Common Geographic Regions

et al. 2017

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Traditional breeding has been used to develop grain sorghum germplasm that is tolerant to acetolactate synthase (ALS)-inhibiting herbicides (Inzen Technology, DuPont). Inzen sorghum carries a double mutation in the ALS gene (Val 560 Ile and Trp 574 Leu), which confers high level of tolerance to ALS-inhibiting herbicides. Overreliance on ALS-inhibiting herbicides for weed control during the 1990s resulted in the evolution of ALS inhibitor-resistant shattercane populations in Nebraska. According to a survey conducted in 2013, ALS inhibitor-resistant weedy Sorghum populations persist in Nebraska. The objectives of this research were to determine whether the ALS mutations present in Inzen sorghum were present in the ALS inhibitor-resistant shattercane and johnsongrass populations detected in Nebraska and northern Kansas, and whether these populations evolved ALS resistance independently. Primers specific to the Val 560 and Trp 574 region of the ALS gene were used to screen the populations with PCR. The Trp 574 Leu mutation was present in one ALS inhibitor-resistant johnsongrass population. The Val 560 Ile was detected in three ALS inhibitor-resistant shattercane, one susceptible shattercane, one ALS inhibitor-resistant johnsongrass, and one susceptible johnsongrass population. Moreover, Val 560 Ile was present in resistant and/or susceptible individuals within johnsongrass and shattercane populations that were segregating for ALS resistance, indicating that by itself the Val 560 Ile mutation does not confer resistance to ALS-inhibiting herbicides. None of the populations presented both mutations simultaneously, as does Inzen sorghum. A shattercane population containing the Ser 653 Thr mutation was also detected. This research indicates that the ALS mutations present in Inzen sorghum already exist individually in weedy sorghum populations. Moreover, our results present strong evidence that ALS resistance in these populations evolved independently. Thus, widespread overreliance on ALS-inhibiting herbicides prior to adoption of glyphosate-tolerant crops in the 1990s exerted sufficient selective pressure on shattercane and johnsongrass populations for resistance to evolve multiple times in the Midwest. Finally, a survey of the 5′ portion of the ALS gene in more diverse wild and weedy Sorghum species was hampered by limited coverage in genomic resequencing surveys, suggesting that refined PCR-based methods will be needed to assess SNP variation in this gene region, which includes the Ala 122 , Pro 197 , and Ala 205 codons known to confer ALS resistance in other species. Nomenclature: Acetolactate synthase (ALS)-inhibiting herbicides, johnsongrass, Sorghum halepense (L.) Pers. SORHA, shattercane, Sorghum bicolor (L.) Moench ssp. drummondii (Nees ex Steud.) de Wet ex Davidse SORVU, sorghum, Sorghum bicolor (L.) Moench ssp. Bicolor SORVU

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“…Hence, the accession was named BTx623-Rooney or WT-R [12]. Cultivation for seed multiplication was done in the fields in IRRI, Los Baños (14° 11” N, 121° 15” E) in the Philippines.…”

Section: Methodsmentioning

confidence: 99%

“…We hypothesized that induced mutations can revert a C 4 plant to a C 3 or an intermediate type and this reversion can be detected from its δ 13 C signatures. The seeds of one of the accessions of the first sequenced sorghum (BTx623-Rooney) [12] were mutagenized using EMS and mutant seedlings screened to find those with altered δ 13 C values. Several mutants, showing reduced (more C 3 -like) δ 13 C were isolated for whole genome sequencing (WGS) [13] to map underlying mutations.…”

Section: Introductionmentioning

confidence: 99%

A sorghum (Sorghum bicolor) mutant with altered carbon isotope ratio

et al. 2017

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Recent efforts to engineer C4 photosynthetic traits into C3 plants such as rice demand an understanding of the genetic elements that enable C4 plants to outperform C3 plants. As a part of the C4 Rice Consortium’s efforts to identify genes needed to support C4 photosynthesis, EMS mutagenized sorghum populations were generated and screened to identify genes that cause a loss of C4 function. Stable carbon isotope ratio (δ13C) of leaf dry matter has been used to distinguishspecies with C3 and C4 photosynthetic pathways. Here, we report the identification of a sorghum (Sorghum bicolor) mutant with a low δ13C characteristic. A mutant (named Mut33) with a pale phenotype and stunted growth was identified from an EMS treated sorghum M2 population. The stable carbon isotope analysis of the mutants showed a decrease of 13C uptake capacity. The noise of random mutation was reduced by crossing the mutant and its wildtype (WT). The back-cross (BC1F1) progenies were like the WT parent in terms of 13C values and plant phenotypes. All the BC1F2 plants with low δ13C died before they produced their 6th leaf. Gas exchange measurements of the low δ13C sorghum mutants showed a higher CO2 compensation point (25.24 μmol CO2.mol-1air) and the maximum rate of photosynthesis was less than 5μmol.m-2.s-1. To identify the genetic determinant of this trait, four DNA pools were isolated; two each from normal and low δ13C BC1F2 mutant plants. These were sequenced using an Illumina platform. Comparison of allele frequency of the single nucleotide polymorphisms (SNPs) between the pools with contrasting phenotype showed that a locus in Chromosome 10 between 57,941,104 and 59,985,708 bps had an allele frequency of 1. There were 211 mutations and 37 genes in the locus, out of which mutations in 9 genes showed non-synonymous changes. This finding is expected to contribute to future research on the identification of the causal factor differentiating C4 from C3 species that can be used in the transformation of C3 to C4 plants.

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Two forward genetic screens for vein density mutants in sorghum converge on a cytochrome P450 gene in the brassinosteroid pathway

Cited by 38 publications

References 47 publications

The Analysis of Physiological Variations in M2 Generation of Solanum melongena L. Mutagenized by Ethyl Methane Sulfonate

The Analysis of Physiological Variations in M2 Generation of Solanum melongena L. Mutagenized by Ethyl Methane Sulfonate

Independent Evolution of Acetolactate Synthase–inhibiting Herbicide Resistance in WeedySorghumPopulations across Common Geographic Regions

A sorghum (Sorghum bicolor) mutant with altered carbon isotope ratio

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