[Cited FineTest Antibody] Current Research on Genetics and Plant Biology

FineTest antibody contributes to the research on genetics and plant biology. The immunoblot is designed to measure 6-His and GST tags in endosperm sample.

Publication Details
Article Title: Natural variation in WHITE-CORE RATE 1 regulates redox homeostasis in rice endosperm to affect grain quality
Journal Title: The Plant Cell
DOI: 10.1093/plcell/koac057
IF: 12.796
PMID: 35171272

Abstract:   Grain chalkiness reduces the quality of rice (Oryza sativa) and is a highly undesirable trait for breeding and marketing. However, the underlying molecular cause of chalkiness remains largely unknown. Here, we cloned the F-box gene WHITECORE RATE 1 (WCR1), which negatively regulates grain chalkiness and improves grain quality in rice. A functional A/G variation in the promoter region of WCR1 generates the alleles WCR1A and WCR1G, which originated from tropical japonica and wild rice Oryza rufipogon, respectively. OsDOF17 is a transcriptional activator that binds to the AAAAG cis-element in the WCR1A promoter. WCR1 positively affects the transcription of the metallothionein gene MT2b and interacts with MT2b to inhibit its 26S proteasome-mediated degradation, leading to decreased reactive oxygen species production and delayed programmed cell death in rice endosperm. This, in turn, leads to reduced chalkiness. Our findings uncover a molecular mechanism underlying rice chalkiness and identify the promising natural variant WCR1A, with application potential for rice breeding.

Keywords: Genetics, Gene Expression Regulation, Homeostasis, Metabolism, Oxidation-Reduction, Plant Biology

Immunoblot

Validated Image

Figure Source: Plant Cell. 2022 Apr 26;34(5):1912-1932. doi: 10.1093/plcell/koac057.

Figure 6.   WCR1 interacts with MT2b to scavenge ROS and delay PCD in developing endosperm. A, WCR1 interacts with MT2b in yeast cells. The N terminus of WCR1 including the F-box domain and the C terminus including the MYND domain are shown. SD-2 and SD-4 represent SD/– Trp–Leu and SD/–Trp–Leu–His–Ade þ X-Gal selection medium, respectively. Negative, empty vectors pGBKT7 and pGADT7. Positive, the OsMADS1 gene, which formed a homodimer, was linked to pGBKT7 (BD-OsMADS1) and pGADT7 (AD-OsMADS1), respectively, and yeast harboring these two vectors grew on SD-4 medium. B, Pull-down assay. Proteins were pulled down by GST agarose beads and immunoblotted using His and GST antibodies. C, Split-LUC assay. nLUC-tagged WCR1 and cLUC-tagged MT2b were co-transformed into N. benthamiana leaves. Color bar shows the intensity of LUC signals. D–F, Detection of ROS, H2O2, and O 2 contents in endosperm at 7, 14, and 21 DAF (7E, 14E, and 21E). Each endosperm sample was obtained from 30 grains with three repeats. G, Trypan blue and FDA staining of transverse sections of grains at different developmental stages for NIL-WCR1-BL and NIL-WCR1-J. Blue areas and green fluorescence represent areas of dead and living cells, respectively. Scale bars, 500 lm. H, Relative expression levels of PCD-related OsMC genes in 9 DAF endosperm. Endosperm samples were taken from at least four plants. In (D–F and H), significant differences were determined by two-tailed t tests, *P0.05, **P0.01. Error bars, SEM. I, Dynamic changes in mean dry weights of caryopses. Data show the total weights of 30 dry dehulled caryopses with three biological repeats from the upper parts of panicles from different plants. Error bars, SEM.