S to produce OsFWL7-YN and OsFWL7-YC constructs, respectively. The coding sequences in the two prohibitin genes have been cloned into the p2YC vector. The constructs were transformed into Agrobacterium tumefaciens strain EHA105 and transfected Nicotiana benthamiana leaves. The fluorescence was monitored using the LSM 700 confocal laser scanning microscope (Carl Zeiss). The PCR primers utilized for the building of BiFC vectors are listed in Table S1. four.six. Measurement of Heavy Metal Levels Heavy metal levels in Butenafine Purity diverse tissues have been determined in line with the methods described by Zhang et al. [56]. In brief, the roots of hydroponically cultured seedlings were soaked in 20 mM EDTA for 15 min and washed with deionized water. The root and shoot samples were dried at 80 C for three days and ground to a fine powder working with an analytical mill (Cole-Parmer, Vernon Hills, IL, USA). Then, the samples (0.five g) had been digested with HNO3 and H2 O2 in a microwave digestion instrument (MARS 5; CEM, Matthews, NC, USA). Cd and micronutrient metal levels had been measured using inductively coupled plasma optical emission spectrometry (ICP-OES, Thermo Scientific iCAP 6300; Thermo Fisher Scientific, Grand Island, NY, USA).Int. J. Mol. Sci. 2021, 22,13 of4.7. Gossypin Purity Statistical Analysis All data are presented as imply and typical deviation of replicates. The statistical significance of differences in suggests was evaluated working with Student’s t-test. Column charts were made employing SigmaPlot 10.0. five. Conclusions In summary, OsFWL7 encodes a plasma membrane protein that regulates Cd and micronutrient metal accumulation in rice. The rice FWL proteins interact with themselves and one a further and may perhaps be situated in membrane microdomains.Supplementary Components: The following are accessible online at mdpi/article/10 .3390/ijms222212583/s1. Author Contributions: H.Z. (Hao Zhang) and X.Z. developed the study; Q.G., L.L., H.Z. (Haiying Zhou), X.L., W.L., Y.M., Y.Y. and J.J. performed the experiments; Q.G. and L.L. analyzed the data; and Q.G., H.Z. (Hao Zhang) and X.Z. wrote the manuscript. All authors have study and agreed to the published version with the manuscript. Funding: This analysis was funded by the Six Talent Peaks Project in Jiangsu Province (SWYY151), the Priority Academic Plan Improvement of Jiangsu Higher Education Institutions, the Natural Science Foundation of Jiangsu Provincial Division of Education (19KJB180011), the All-natural Science Analysis Program of Huai’an Municipality (HAB202155), and the Innovation and Entrepreneurship Instruction Plan of College Students in Jiangsu Province (202110323012Z). Data Availability Statement: All information are presented as figures and Supplementary Components, that are incorporated in this paper. Acknowledgments: and ideas. We thank the anonymous reviewers for their constructive commentsConflicts of Interest: The authors declare no conflict of interest. The funders had no function within the style of the study; in the collection, analyses, or interpretation of data; inside the writing of the manuscript, or in the choice to publish the results.Academic Editor: Julia Costa Received: 19 October 2021 Accepted: 20 November 2021 Published: 22 NovemberPublisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.Copyright: 2021 by the authors. Licensee MDPI, Basel, Switzerland. This short article is an open access short article distributed under the terms and conditions with the Creative Commons Attribution (CC BY).
