<i>OsMPK6</i>plays a critical role in cell differentiation during early embryogenesis in<i>Oryza sativa</i>

Yi, Jakyung; Lee, Yang-Seok; Lee, Dong-Yeon; Cho, Man-Ho; Jeon, Jong‐Seong; An, Gynheung

doi:10.1093/jxb/erw052

Cited by 36 publications

(45 citation statements)

References 78 publications

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“…We therefore performed genome editing using CRISPR/Cas9 in the wild-type FAZ1 background. Unfortunately, we were not able to isolate the related loss-of-function mutants for OsMPK6 due to embryo lethality, which is consistent with the results of a previous study (Yi et al, 2016); instead, we used gene knockdown mutants produced based on RNA interference (RNAi). We found that the OsMPK6 RNAi single mutant, which showed reduced expression of OsMPK6, had reduced grain size and increased grain number per panicle compared with the wild-type FAZ1, strongly suggesting that OsMPK6 is a positive regulator of grain size and is required for panicle and spikelet development ( Figures 6A, 6B, and 6G).…”

Section: Gsn1 Acts As a Negative Regulator Of The Mapk Cascade In Coosupporting

confidence: 54%

GRAIN SIZE AND NUMBER1 Negatively Regulates the OsMKKK10-OsMKK4-OsMPK6 Cascade to Coordinate the Trade-off between Grain Number per Panicle and Grain Size in Rice

et al. 2018

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Grain number and size are interactive agronomic traits that determine grain yield. However, the molecular mechanisms responsible for coordinating the trade-off between these traits remain elusive. Here, we characterized the rice () () mutant, which has larger grains but sparser panicles than the wild type due to disordered localized cell differentiation and proliferation. encodes the mitogen-activated protein kinase phosphatase OsMKP1, a dual-specificity phosphatase of unknown function. Reduced expression of resulted in larger and fewer grains, whereas increased expression resulted in more grains but reduced grain size. GSN1 directly interacts with and inactivates the mitogen-activated protein kinase OsMPK6 via dephosphorylation. Consistent with this finding, the suppression of mitogen-activated protein kinase genes ,, and separately resulted in denser panicles and smaller grains, which rescued the mutant phenotypes. Therefore, OsMKKK10-OsMKK4-OsMPK6 participates in panicle morphogenesis and acts on a common pathway in rice. We confirmed that GSN1 is a negative regulator of the OsMKKK10-OsMKK4-OsMPK6 cascade that determines panicle architecture. The GSN1-MAPK module coordinates the trade-off between grain number and grain size by integrating localized cell differentiation and proliferation. These findings provide important insights into the developmental plasticity of the panicle and a potential means to improve crop yields.

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Section: Gsn1 Acts As a Negative Regulator Of The Mapk Cascade In Coosupporting

confidence: 54%

GRAIN SIZE AND NUMBER1 Negatively Regulates the OsMKKK10-OsMKK4-OsMPK6 Cascade to Coordinate the Trade-off between Grain Number per Panicle and Grain Size in Rice

et al. 2018

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“…While this manuscript was in preparation, a study by Yi et al . () further confirmed this embryo lethality by analyzing heterozygous T‐DNA mutants (there referred to as OsMPK6 ). The mpk1 mutant exhibited defects in cell differentiation and arrestment of embryonic development at the globular stage (Yi et al ., ).…”

Section: Discussionmentioning

confidence: 71%

“…We demonstrated the embryo lethality of MPK1 loss of function based on abnormally small embryos and reduced germination rate (Figure 2; Tables 2 and S1). While this manuscript was in preparation, a study by Yi et al (2016) further confirmed this embryo lethality by analyzing heterozygous T-DNA mutants (there referred to as OsMPK6). The mpk1 mutant exhibited defects in cell differentiation and arrestment of embryonic development at the globular stage (Yi et al, 2016).…”

Section: Discussionmentioning

confidence: 80%

“…While this manuscript was in preparation, a study by Yi et al (2016) further confirmed this embryo lethality by analyzing heterozygous T-DNA mutants (there referred to as OsMPK6). The mpk1 mutant exhibited defects in cell differentiation and arrestment of embryonic development at the globular stage (Yi et al, 2016). In agreement, previous studies could only analyze cell lines with the mpk1 mutation, and failed to report viable mpk1 mutant plants (Kurusu et al, 2005;Kishi-Kaboshi et al, 2010); however, knock-out mutants of AtMPK6, the Arabidopsis ortholog to MPK1, show no obvious growth or developmental defects (Wang et al, 2007).…”

Section: Discussionmentioning

confidence: 80%

“…During the preparation of this manuscript, a study further confirmed that MPK1 loss of function is embryo‐lethal by analyzing heterozygous T‐DNA insertion mutants (there referred to as OsMPK6 with the use of a different nomenclature system; Yi et al ., ). The authors found that mpk1 embryos were affected in cell differentiation, which arrested the embryonic development and led to unviable and abnormally small embryos (Yi et al ., ). We therefore examined the seeds of our heterozygous mpk1 and mpk6 mutants to compare the embryo phenotypes (Figure b; Table S1).…”

Section: Resultsmentioning

confidence: 97%

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Discovery of rice essential genes by characterizing a CRISPR‐edited mutation of closely related rice MAP kinase genes

2017

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SUMMARYThe clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 nuclease (Cas9) system depends on a guide RNA (gRNA) to specify its target. By efficiently co-expressing multiple gRNAs that target different genomic sites, the polycistronic tRNA-gRNA gene (PTG) strategy enables multiplex gene editing in the family of closely related mitogen-activated protein kinase (MPK) genes in Oryza sativa (rice). In this study, we identified MPK1 and MPK6 (Arabidopsis AtMPK6 and AtMPK4 orthologs, respectively) as essential genes for rice development by finding the preservation of MPK functional alleles and normal phenotypes in CRISPR-edited mutants. The true knock-out mutants of MPK1 were severely dwarfed and sterile, and homozygous mpk1 seeds from heterozygous parents were defective in embryo development. By contrast, heterozygous mpk6 mutant plants completely failed to produce homozygous mpk6 seeds. In addition, the functional importance of specific MPK features could be evaluated by characterizing CRISPR-induced allelic variation in the conserved kinase domain of MPK6. By simultaneously targeting between two and eight genomic sites in the closely related MPK genes, we demonstrated 45-86% frequency of biallelic mutations and the successful creation of single, double and quadruple gene mutants. Indels and fragment deletion were both stably inherited to the next generations, and transgene-free mutants of rice MPK genes were readily obtained via genetic segregation, thereby eliminating any positional effects of transgene insertions. Taken together, our study reveals the essentiality of MPK1 and MPK6 in rice development, and enables the functional discovery of previously inaccessible genes or domains with phenotypes masked by lethality or redundancy.

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Genomic Designing of Abiotic Stress Tolerance in Banana

Ravi

Vaganan²,

Anithasree³

et al. 2022

Genomic Designing for Abiotic Stress Resistant Fruit Crops

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OsMPK6plays a critical role in cell differentiation during early embryogenesis inOryza sativa

Abstract: HighlightThe MAP kinase cascade controls early embryogenesis prior to axis formation.

Cited by 36 publications

References 78 publications

GRAIN SIZE AND NUMBER1 Negatively Regulates the OsMKKK10-OsMKK4-OsMPK6 Cascade to Coordinate the Trade-off between Grain Number per Panicle and Grain Size in Rice

GRAIN SIZE AND NUMBER1 Negatively Regulates the OsMKKK10-OsMKK4-OsMPK6 Cascade to Coordinate the Trade-off between Grain Number per Panicle and Grain Size in Rice

Discovery of rice essential genes by characterizing a CRISPR‐edited mutation of closely related rice MAP kinase genes

Genomic Designing of Abiotic Stress Tolerance in Banana

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