Green Fluorescent Protein- and Discosoma sp. Red Fluorescent Protein-Tagged Organelle Marker Lines for Protein Subcellular Localization in Rice

Chen, Ziqiang; Zheng, Wenhui; Chen, Longhai; Li, Chenlu; Liang, Tingmin; Chen, Zaijie; Xu, Huibing; Han, Yijuan; Kong, Lan; Zhao, Xu; Wang, Feng; Wang, Zonghua; Chen, Songbiao

doi:10.3389/fpls.2019.01421

Cited by 19 publications

(12 citation statements)

References 35 publications

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“…To analyze the subcellular localization of CiMADS43 , the 35S::CiMADS43-GFP (Green fluorescent protein) fusion vector and 35S::GFP empty vector (control) were transiently expressed in epidermal cells of tobacco leaves. Red fluorescent protein (RFP) labeled was used as a nuclear marker to indicate the location of the nucleus [ 55 ]. The green fluorescence of the CiMADS43-GFP fusion proteins was observed in the nucleus, whereas the signals of the control were distributed on the whole cell ( Figure 1 c).…”

Section: Resultsmentioning

confidence: 99%

A MADS-Box Gene CiMADS43 Is Involved in Citrus Flowering and Leaf Development through Interaction with CiAGL9

Zhang

Hou

et al. 2021

IJMS

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MADS-box genes are involved in various developmental processes including vegetative development, flower architecture, flowering, pollen formation, seed and fruit development. However, the function of most MADS-box genes and their regulation mechanism are still unclear in woody plants compared with model plants. In this study, a MADS-box gene (CiMADS43) was identified in citrus. Phylogenetic and sequence analysis showed that CiMADS43 is a GOA-like Bsister MADS-box gene. It was localized in the nucleus and as a transcriptional activator. Overexpression of CiMADS43 promoted early flowering and leaves curling in transgenic Arabidopsis. Besides, overexpression or knockout of CiMADS43 also showed leaf curl phenotype in citrus similar to that of CiMADS43 overexpressed in Arabidopsis. Protein–protein interaction found that a SEPALLATA (SEP)-like protein (CiAGL9) interacted with CiMADS43 protein. Interestingly, CiAGL9 also can bind to the CiMADS43 promoter and promote its transcription. Expression analysis also showed that these two genes were closely related to seasonal flowering and the development of the leaf in citrus. Our findings revealed the multifunctional roles of CiMADS43 in the vegetative and reproductive development of citrus. These results will facilitate our understanding of the evolution and molecular mechanisms of MADS-box genes in citrus.

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Section: Resultsmentioning

confidence: 99%

A MADS-Box Gene CiMADS43 Is Involved in Citrus Flowering and Leaf Development through Interaction with CiAGL9

Zhang

Hou

et al. 2021

IJMS

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“…To explore the subcellular localization of OsMBD707, we performed transient expression of a GFP-OsMBD707 fusion construct in rice protoplasts. Microscopy revealed that the fluorescence signal of GFP-OsMBD707 was inside the nucleus region, which was merged with the signal of the nucleus DsRed-OsH2B marker [ 29 ] (Fig. 3 b).…”

Section: Resultsmentioning

confidence: 99%

“…Rice protoplasts were prepared from the sheath and stem tissues of 2-week-old etiolated seedlings. The GFP alone and GFP-OsMBD707 fusion constructs were, respectively, co-transfected with a nucleus marker construct DsRed-OsH2B [ 29 ] into rice protoplasts via a PEG-mediated method as previously described [ 55 ]. Rice stable transformation was conducted as previously described [ 56 ].…”

Section: Methodsmentioning

confidence: 99%

Overexpression of a methyl-CpG-binding protein gene OsMBD707 leads to larger tiller angles and reduced photoperiod sensitivity in rice

Zhang

Liang

et al. 2021

BMC Plant Biol

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Background Methyl-CpG-binding domain (MBD) proteins play important roles in epigenetic gene regulation, and have diverse molecular, cellular, and biological functions in plants. MBD proteins have been functionally characterized in various plant species, including Arabidopsis, wheat, maize, and tomato. In rice, 17 sequences were bioinformatically predicted as putative MBD proteins. However, very little is known regarding the function of MBD proteins in rice. Results We explored the expression patterns of the rice OsMBD family genes and identified 13 OsMBDs with active expression in various rice tissues. We further characterized the function of a rice class I MBD protein OsMBD707, and demonstrated that OsMBD707 is constitutively expressed and localized in the nucleus. Transgenic rice overexpressing OsMBD707 displayed larger tiller angles and reduced photoperiod sensitivity—delayed flowering under short day (SD) and early flowering under long day (LD). RNA-seq analysis revealed that overexpression of OsMBD707 led to reduced photoperiod sensitivity in rice and to expression changes in flowering regulator genes in the Ehd1-Hd3a/RFT1 pathway. Conclusion The results of this study suggested that OsMBD707 plays important roles in rice growth and development, and should lead to further studies on the functions of OsMBD proteins in growth, development, or other molecular, cellular, and biological processes in rice.

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“…The full-length ZmSRL5 coding sequence without the termination codon was obtained by PCR using the PEGAD-SRL5-F/R primers (Table S1), which was inserted into the PEGAD-GFP vector to generate a ZmSRL5 -GFP fusion protein. RFP-OsPIP2 was used as plasma membrane marker (Chen et al 2019). The fusion construct was introduced into the tobacco leaf epidermis, and an Olympus FluoView™ FV1200 confocal microscope (Olympus, Tokyo, Japan) was used to detect the GFP and RFP (red plasma membrane marker) signals.…”

Section: Subcellular Localization Of Zmsrl5mentioning

confidence: 99%

ZmSRL5is involved in drought tolerance by maintaining cuticular wax structure in maize

Pan

Liu

Zhao

et al. 2020

JIPB

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Cuticular wax is a natural barrier on terrestrial plant organs, which protects plants from damages caused by a variety of stresses. Here, we report the identification and functional characterization of a cuticular‐wax‐related gene, Zea mays L. SEMI‐ROLLED LEAF 5 (ZmSRL5). The loss‐of‐function mutant srl5, which was created by a 3,745 bp insertion in the first intron that led to the premature transcript, exhibited abnormal wax crystal morphology and distribution, which, in turn, caused the pleiotropic phenotypes including increased chlorophyll leaching and water loss rate, decreased leaf temperature, sensitivity to drought, as well as semi‐rolled mature leaves. However, total wax amounts showed no significant difference between wild type and semi‐rolled leaf5 (srl5) mutant. The phenotype of srl5 was confirmed through the generation of two allelic mutants using CRISPR/Cas9. ZmSRL5 encodes a CASPARIAN‐STRIP‐MEMBRANE‐DOMAIN‐LIKE (CASPL) protein located in plasma membrane, and highly expressed in developing leaves. Further analysis showed that the expressions of the most wax related genes were not affected or slightly altered in srl5. This study, thus, primarily uncovers that ZmSRL5 is required for the structure formation of the cuticular wax and could increase the drought tolerance by maintaining the proper cuticular wax structure in maize.

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Green Fluorescent Protein- and Discosoma sp. Red Fluorescent Protein-Tagged Organelle Marker Lines for Protein Subcellular Localization in Rice

Cited by 19 publications

References 35 publications

A MADS-Box Gene CiMADS43 Is Involved in Citrus Flowering and Leaf Development through Interaction with CiAGL9

A MADS-Box Gene CiMADS43 Is Involved in Citrus Flowering and Leaf Development through Interaction with CiAGL9

Overexpression of a methyl-CpG-binding protein gene OsMBD707 leads to larger tiller angles and reduced photoperiod sensitivity in rice

ZmSRL5is involved in drought tolerance by maintaining cuticular wax structure in maize

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