FZP determines grain size and sterile lemma fate in rice

Ren, Deyong; Hu, Jiang; Xu, Qiankun; Cui, Yuanjiang; Zhang, Yu; Zhou, Tingting; Rao, Yuchun; Xue, Dawei; Zeng, Dali; Zhang, Guangheng; Gao, Zhenyu; Zhu, Li; Shen, Lan; Chen, Guang; Guo, Longbiao

doi:10.1093/jxb/ery264

Cited by 54 publications

(37 citation statements)

References 44 publications

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“…The recombinant vector was then introduced into calli of Nipponbare to generate transgenic plants. Different tissues of transgenic plants, involving roots, leaves, leaf sheaths, culms and panicles were used for GUS assay performed based on method of Jefferson et al (1987) and Ren et al (2018).…”

Section: Histochemical Gus Assaymentioning

confidence: 99%

OsACL‐A2 negatively regulates cell death and disease resistance in rice

Ruan

Hua

Zhao

et al. 2019

Plant Biotechnology Journal

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Summary ATP ‐citrate lyases ( ACL ) play critical roles in tumour cell propagation, foetal development and growth, and histone acetylation in human and animals. Here, we report a novel function of ACL in cell death‐mediated pathogen defence responses in rice. Using ethyl methanesulphonate ( EMS ) mutagenesis and map‐based cloning, we identified an Oryza sativa ACL ‐A2 mutant allele, termed spotted leaf 30‐1 ( spl30‐1 ), in which an A‐to‐T transversion converts an Asn at position 343 to a Tyr (N343Y), causing a recessive mutation that led to a lesion mimic phenotype. Compared to wild‐type plants, spl30‐1 significantly reduces ACL enzymatic activity, accumulates high reactive oxygen species and increases degradation rate of nuclear deoxyribonucleic acids. CRISPR /Cas9‐mediated insertion/deletion mutation analysis and complementation assay confirmed that the phenotype of spl30‐1 resulted from the defective function of Os ACL ‐A2 protein. We further biochemically identified that the N343Y mutation caused a significant degradation of SPL 30 N343Y in a ubiquitin‐26S proteasome system ( UPS )‐dependent manner without alteration in transcripts of Os ACL ‐A2 in spl30‐1 . Transcriptome analysis identified a number of up‐regulated genes associated with pathogen defence responses in recessive mutants of Os ACL ‐A2, implying its role in innate immunity. Suppressor mutant screen suggested that Os SL , which encodes a P450 monooxygenase protein, acted as a downstream key regulator in spl30‐1 ‐mediated pathogen defence responses. Taken together, our study discovered a novel role of Os ACL ‐A2 in negatively regulating innate immune responses in rice.

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Section: Histochemical Gus Assaymentioning

confidence: 99%

OsACL‐A2 negatively regulates cell death and disease resistance in rice

Ruan

Hua

Zhao

et al. 2019

Plant Biotechnology Journal

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“…First, the coding regions of the AH2 cDNA and the transcriptional activator FZP cDNA were fused to the GAL4 DNA‐binding domain (BD). The empty pGBKT7 and FZP‐BD vectors were regarded as negative and positive controls, respectively (Ren et al ., 2018b). Constructs producing AH2‐BD, FZP‐BD and pGBKT7 were expressed in yeast cells, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…Paraffin section and SEM analyses were performed according to Ren et al . (,b). In brief, the samples were embedded in paraffin and cut in 8‐μm thick sections, and stained, then observed with an ENIKON ECLIPSE 90i microscope.…”

Section: Methodsmentioning

confidence: 99%

“…The coding frames of AH2 and FZP were cloned into the pGBKT7 vector. All resulting constructs were transformed into yeast AH109 and the clones were grown on SD/ÀTrp plates and selective medium lacking adenine, histidine, and tryptophan (Ren et al, 2018b). A fragment amplified from the AH2 cDNA was cloned into a GAL4DB vector with the Cauliflower mosaic virus 35S promoter and a GAL4 DNA-BD to generate a construct expressing AH2-GAL.…”

Section: Transcriptional Activity Analysismentioning

confidence: 99%

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AH2 encodes a MYB domain protein that determines hull fate and affects grain yield and quality in rice

Ren

Cui

et al. 2019

The Plant Journal

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Summary The palea and lemma (hull) are grass‐specific organs, and determine grain size and quality. In the study, AH2 encodes a MYB domain protein, and functions in the development of hull and grain. Mutation of AH2 produces smaller grains and alters grain quality including decreased amylose content and gel consistency, and increased protein content. Meantime, part of the hull lost the outer silicified cells, and induces a transformation of the outer rough epidermis to inner smooth epidermis cells, and the body of the palea was reduced in the ah2 mutant. We confirmed the function of AH2 by complementation, CRISPR‐Cas9, and cytological and molecular tests. Additionally, AH2, as a repressor, repress transcription of the downstream genes. Our results revealed that AH2 plays an important role in the determination of hull epidermis development, palea identity, and grain size.

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“…The average grain length of the wild type was 7.5 mm, while it was 8.6 mm of gs3-9 and 8.2 mm of gs3-21 ( Figure 3B). In recent years, increasing numbers of genes have been cloned and studied, most of which regulate the grain size by controlling cell proliferation and expansion in spikelet hulls, such as FZP, GS5, and GS2 (Li et al, 2011;Hu et al, 2015;Ren et al, 2018). Two genes, CYCT1 and H1, involved in the cell cycle, were selected to measure the effects of the CRISPR/Cas9 system on the GS3 target sites according to previous studies.…”

Section: The Grain Size Was Strikingly Enlarged In the Gs3 Edited Linesmentioning

confidence: 99%

Rational Improvement of Rice Yield and Cold Tolerance by Editing the Three Genes OsPIN5b, GS3, and OsMYB30 With the CRISPR–Cas9 System

et al. 2020

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Significant increases in rice yield and stress resistance are constant demands for breeders. However, high yield and high stress resistance are often antagonistic to each other. Here, we report several new rice mutants with high yield and excellent cold tolerance that were generated by simultaneously editing three genes, OsPIN5b (a panicle length gene), GS3 (a grain size gene) and OsMYB30 (a cold tolerance gene) with the CRISPR-Cas9 (clustered regularly interspaced short palindromic repeatsassociated protein 9) system. We edited two target sites of each gene with high efficiency: 53% for OsPIN5b-site1, 42% for OsPIN5b-site2, 66% for GS3-site1, 63% for GS3-site2, 63% for OsMYB30-site1, and 58% for OsMYB30-site2. Consequently, the ospin5b mutants, the gs3 mutants, and the osmyb30 mutants exhibited increased panicle length, enlarged grain size and increased cold tolerance, respectively. Then nine transgenic lines of the ospin5b/gs3, six lines of ospin5b/osmyb30 and six lines of gs3/ osmyb30 were also acquired, and their yield related traits and cold tolerance corresponded to the genes being edited. Additionally, we obtained eight ospin5b/gs3/ osmyb30 triple mutants by editing all three genes simultaneously. Aside from the ospin5b/ gs3/osmyb30-4 and ospin5b/gs3/osmyb30-25 mutants, the remaining six mutants had off-target events at the putative off-target site of OsMYB30-site1. The results also showed that the T 2 generations of these two mutants exhibited higher yield and better cold tolerance compared with the wild type. Together, these results demonstrated that new and excellent rice varieties with improved yield and abiotic stress resistance can be generated through gene editing techniques and may be applied to rice breeding. Furthermore, our study proved that the comprehensive agronomic traits of rice can be improved with the CRISPR-Cas9 system.

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FZP determines grain size and sterile lemma fate in rice

Abstract: This study revealed that FZP functions in grain size and sterile lemma identity, and supported the hypothesis that the lemma, rudimentary glume, and sterile lemma are homologous organs.

Cited by 54 publications

References 44 publications

OsACL‐A2 negatively regulates cell death and disease resistance in rice

OsACL‐A2 negatively regulates cell death and disease resistance in rice

AH2 encodes a MYB domain protein that determines hull fate and affects grain yield and quality in rice

Rational Improvement of Rice Yield and Cold Tolerance by Editing the Three Genes OsPIN5b, GS3, and OsMYB30 With the CRISPR–Cas9 System

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