The C-terminal motif of SiAGO1b is required for the regulation of growth, development and stress responses in foxtail millet (<i>Setaria italica</i>(L.) P. Beauv)

Liu, Xiaotong; Tang, Sha; Jia, Guanqing; Schnable, James C.; Su, Hongyan; Tang, Chanjuan; Zhi, Hui; Diao, Xianmin

doi:10.1093/jxb/erw135

Cited by 40 publications

(27 citation statements)

References 66 publications

(70 reference statements)

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“…() developed a millet protoplast isolation protocol. Millet protoplasts have been used for protoplast transfection and for BiFC investigation (Liu et al ., ). Using protoplast transfection, we demonstrate that the millet genome can be mutated using the CRISPR/Cas9 system.…”

Section: Discussionmentioning

confidence: 97%

Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single‐cell mutation detection to mutant plant regeneration

Lin

Hsu

Yang

et al. 2018

Plant Biotechnology Journal

253

176

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SummaryPlant protoplasts are useful for assessing the efficiency of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein 9 (Cas9) mutagenesis. We improved the process of protoplast isolation and transfection of several plant species. We also developed a method to isolate and regenerate single mutagenized Nicotianna tabacum protoplasts into mature plants. Following transfection of protoplasts with constructs encoding Cas9 and sgRNAs, target gene DNA could be amplified for further analysis to determine mutagenesis efficiency. We investigated N. tabacum protoplasts and derived regenerated plants for targeted mutagenesis of the phytoene desaturase (NtPDS) gene. Genotyping of albino regenerants indicated that all four NtPDS alleles were mutated in amphidiploid tobacco, and no Cas9 DNA could be detected in most regenerated plants.

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Section: Discussionmentioning

confidence: 97%

Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single‐cell mutation detection to mutant plant regeneration

Lin

Hsu

Yang

et al. 2018

Plant Biotechnology Journal

253

176

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“…A bulked segregant analysis, followed by map‐based cloning, is a powerful tool for gene isolation in model plants, such as Arabidopsis and rice. In Setaria, this approach was found to be efficient for rapid mapping of favorable loci (Li et al, 2016; Liu et al, 2016; Xue et al, 2016). A MutMap method was developed on the basis of high‐throughput genome resequencing techniques (Abe et al, 2012; Fekih et al, 2013; Takagi et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

“…The ethyl methanesulfonate (EMS) treatment is an effective approach to create genetic variation in crop species (Mizuno et al, 2014; Patel et al, 2014; Tagu et al, 2014), which has been applied in the Setaria genus (Li et al, 2016; Liu et al, 2016). Here, an EMS‐induced Setaria dwarfing mutant was characterized, and the dwarfing gene was mapped using forward‐genetic approach.…”

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confidence: 99%

Identification and Fine Mapping of SiDWARF3 (D3), a Pleiotropic Locus Controlling Environment‐Independent Dwarfism in Foxtail Millet

et al. 2017

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The dwarfing of crop species could enhance lodging resistance in elite cultivars and thus increase cereal grain yields. Foxtail millet [Setaria italica (L.) P. Beauv.] is an ancient grain crop originating from China, and it served as staple food during the development of agricultural‐based civilization in South and East Asia, where it is still grown today. Breeding for dwarfism of foxtail millet has great potential in fulfilling the food needs of an increasing world population. This study was undertaken to characterize an ethyl methanesulfonate‐induced dwarf mutant of foxtail millet (Sidwarf3) derived from ‘Yugu1’. The dwarf character of the mutant was latitude independent. The mutant had fewer and shorter internodes because of smaller cells compared with the wild‐type. Furthermore, the mutant showed increased seed length and panicle tightness; however, it exhibited decreased drought tolerance. Endogenous accumulations of gibberellin, auxin, brassinosteroid, abscisic acid, and zeatin‐riboside in the mutant were changed at the jointing and heading stages of the growth period, and the plant height of Sidwarf3 could not be fully restored by applying gradient concentrations (0–5 × 10−4 M) of gibberellic acid solution. Using combined map‐based approaches and a MutMap single‐nucleotide polymorphism index analysis, the dwarfing gene D3 was mapped to chromosome 8 and located in a genomic interval spanning 296.7 kb, which annotated 20 candidate genes for further investigation. This study provides essential information for the molecular breeding of lodging‐resistant foxtail millet cultivars through an ethyl methanesulfonate‐induced mutation approach.

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“…RNA extractions and qRT–PCR assays were carried out as described previously [7], with three independent biological replications. The primers used in qRT-PCR are listed in S1 Table.…”

Section: Methodsmentioning

confidence: 99%

Loose Panicle1 encoding a novel WRKY transcription factor, regulates panicle development, stem elongation, and seed size in foxtail millet [Setaria italica (L.) P. Beauv.]

et al. 2017

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Panicle development is an important agronomic trait that aids in determining crop productivity. Foxtail millet and its wild ancestor green foxtail have recently been used as model systems to dissect gene functions. Here, we characterized a recessive mutant of foxtail millet, loose-panicle 1 (lp1), which showed pleiotropic phenotypes, such as a lax primary branching pattern, aberrant branch morphology, semi-dwarfism, and enlarged seed size. The loose panicle phenotype was attributed to increased panicle lengths and decreased primary branch numbers. Map-based cloning, combined with high-throughput sequencing, revealed that LP1, which encodes a novel WRKY transcription factor, is responsible for the mutant phenotype. A phylogenetic analysis revealed that LP1 belongs to the Group I WRKY subfamily, which possesses two WRKY domains (WRKY I and II). A single G-to-A transition in the fifth intron of LP1 resulted in three disorganized splicing events in mutant plants. For each of these aberrant splice variants, the normal C2H2 motif in the WRKY II domain was completely disrupted, resulting in a loss-of-function mutation. LP1 mRNA was expressed in all of the tissues examined, with higher expression levels observed in inflorescences, roots, and seeds at the grain-filling stage. A subcellular localization analysis showed that LP1 predominantly accumulated in the nucleus, which confirmed its role as a transcriptional regulator. This study provides novel insights into the roles of WRKY proteins in regulating reproductive organ development in plants and may help to develop molecular markers associated with crop yields.

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The C-terminal motif of SiAGO1b is required for the regulation of growth, development and stress responses in foxtail millet (Setaria italica(L.) P. Beauv)

Abstract: HighlightThe C-terminus of SiAGO1b is an essential motif for the interaction between SiAGO1b and SiHYL1, and plays a key role in regulating growth, development and stress responses in Setaria italic.

Cited by 40 publications

References 66 publications

Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single‐cell mutation detection to mutant plant regeneration

Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single‐cell mutation detection to mutant plant regeneration

Identification and Fine Mapping of SiDWARF3 (D3), a Pleiotropic Locus Controlling Environment‐Independent Dwarfism in Foxtail Millet

Loose Panicle1 encoding a novel WRKY transcription factor, regulates panicle development, stem elongation, and seed size in foxtail millet [Setaria italica (L.) P. Beauv.]

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