{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:36:41Z","timestamp":1760060201811,"version":"build-2065373602"},"reference-count":53,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2025,8,14]],"date-time":"2025-08-14T00:00:00Z","timestamp":1755129600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Natural Sciences Foundation of China","award":["32301828","2025M773994","2023ZD04068","BE2022335","JBGS[2021]001","BE2021334-1"],"award-info":[{"award-number":["32301828","2025M773994","2023ZD04068","BE2022335","JBGS[2021]001","BE2021334-1"]}]},{"name":"China Postdoctoral Science Foundation","award":["32301828","2025M773994","2023ZD04068","BE2022335","JBGS[2021]001","BE2021334-1"],"award-info":[{"award-number":["32301828","2025M773994","2023ZD04068","BE2022335","JBGS[2021]001","BE2021334-1"]}]},{"name":"Biological Breeding-National Science and Technology Major Project","award":["32301828","2025M773994","2023ZD04068","BE2022335","JBGS[2021]001","BE2021334-1"],"award-info":[{"award-number":["32301828","2025M773994","2023ZD04068","BE2022335","JBGS[2021]001","BE2021334-1"]}]},{"name":"Jiangsu Province Government","award":["32301828","2025M773994","2023ZD04068","BE2022335","JBGS[2021]001","BE2021334-1"],"award-info":[{"award-number":["32301828","2025M773994","2023ZD04068","BE2022335","JBGS[2021]001","BE2021334-1"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Grain chalkiness is an undesirable trait that significantly compromises rice quality, attracting considerable attention from both consumers and breeders. In this study, we characterized the role of the autophagy-related gene OsATG4b in rice grain development. OsATG4b was predominantly expressed in the endosperm. Compared with wild-type plants, OsATG4b-overexpressing lines exhibited significantly reduced grain chalkiness, whereas OsATG4b knockout mutants displayed a marked increase in chalkiness. Importantly, OsATG4b had no significant effect on other major agronomic traits. Ultrastructure analysis of the endosperm and evaluation of seed storage components revealed that the chalky endosperm in OsATG4b Knockout mutants contained loosely packed starch granules, aberrant protein bodies, and reduced levels of seed storage proteins. Furthermore, gene expression analysis indicated that OsATG4b regulates the expression of genes involved in storage protein biosynthesis. Together, these findings demonstrate that OsATG4b plays a critical regulatory role in determining grain chalkiness in rice.<\/jats:p>","DOI":"10.3390\/plants14162530","type":"journal-article","created":{"date-parts":[[2025,8,14]],"date-time":"2025-08-14T14:51:46Z","timestamp":1755183106000},"page":"2530","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Functional Analysis of the Autophagy-Related Gene OsATG4b in Rice Grain Chalkiness Regulation"],"prefix":"10.3390","volume":"14","author":[{"given":"Zhi","family":"Hu","sequence":"first","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"},{"name":"Co-Innovation Center for Modern Production Technology of Grain Crops of Jiangsu Province, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-4559-4978","authenticated-orcid":false,"given":"Xiang","family":"Han","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yumeng","family":"Yuan","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruishan","family":"Xing","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongchun","family":"Liu","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chenming","family":"Li","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongli","family":"Shen","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yifan","family":"Guo","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7397-8793","authenticated-orcid":false,"given":"Shengyuan","family":"Sun","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yihao","family":"Yang","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Min","family":"Guo","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Changjie","family":"Yan","sequence":"additional","affiliation":[{"name":"Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding\/Zhongshan Biological Breeding Laboratory\/Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yangzhou 225009, China"},{"name":"Co-Innovation Center for Modern Production Technology of Grain Crops of Jiangsu Province, Yangzhou University, Yangzhou 225009, China"},{"name":"Guangling College, Yangzhou University, Yangzhou 225009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1093\/jxb\/erz364","article-title":"Integrating the dynamics of yield traits in rice in response to environmental changes","volume":"71","author":"Nutan","year":"2020","journal-title":"J. Exp. Bot."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"17031","DOI":"10.1038\/nplants.2017.31","article-title":"Rational design of high-yield and superior-quality rice","volume":"3","author":"Zeng","year":"2007","journal-title":"Nat. Plants"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1016\/j.tifs.2019.07.039","article-title":"Rice quality: How is it defined by consumers, industry, food scientists, and geneticists?","volume":"92","author":"Custodio","year":"2019","journal-title":"Trends Food Sci. Tech."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1860","DOI":"10.1111\/jipb.13334","article-title":"Rapid improvement of rice eating and cooking quality through gene editing toward glutelin as target","volume":"64","author":"Yang","year":"2022","journal-title":"J. Integr. Plant Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"979","DOI":"10.1016\/j.cj.2024.06.005","article-title":"Genes controlling grain chalkiness in rice","volume":"12","author":"Chen","year":"2024","journal-title":"Crop J."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Zhao, D., Zhang, C., Li, Q., and Liu, Q. (2022). Genetic control of grain appearance quality in rice. Biotechnol. Adv., 60.","DOI":"10.1016\/j.biotechadv.2022.108014"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1146\/annurev-food-030212-182644","article-title":"Impacts of preharvest factors during kernel development on rice quality and functionality","volume":"4","author":"Siebenmorgen","year":"2013","journal-title":"Annu. Rev. Food Sci. Technol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3012","DOI":"10.1111\/pbi.14422","article-title":"A CRISPR\/Cas9-mediated mutant library of seed-preferred genes in rice","volume":"11","author":"Zhao","year":"2024","journal-title":"Plant Biotechnol. J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1111\/jipb.13633","article-title":"Orchestrating seed storage protein and starch accumulation toward overcoming yield\u2013quality trade-off in cereal crops","volume":"66","author":"Cao","year":"2024","journal-title":"J. Integr. Plant Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1186\/s12284-021-00476-x","article-title":"Fine mapping of two grain chalkiness QTLs sensitive to high temperature in rice","volume":"14","author":"Yang","year":"2021","journal-title":"Rice"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1186\/s12284-016-0100-y","article-title":"Proteomic and Glycomic Characterization of Rice Chalky Grains Produced Under Moderate and High-temperature Conditions in Field System","volume":"9","author":"Kaneko","year":"2016","journal-title":"Rice"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"100463","DOI":"10.1016\/j.xplc.2022.100463","article-title":"OPAQUE3, encoding a transmembrane bZIP transcription factor, regulates endosperm storage protein and starch biosynthesis in rice","volume":"3","author":"Cao","year":"2022","journal-title":"Plant Commun."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3447","DOI":"10.1105\/tpc.112.101360","article-title":"Opaque1 encodes a myosin XI motor protein that is required for endoplasmic reticulum motility and protein body formation in maize endosperm","volume":"24","author":"Wang","year":"2012","journal-title":"Plant Cell"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"410","DOI":"10.1105\/tpc.113.121376","article-title":"GLUTELIN PRECURSOR ACCUMULATION3 encodes a regulator of post-Golgi vesicular traffic essential for vacuolar protein sorting in rice endosperm","volume":"26","author":"Ren","year":"2014","journal-title":"Plant Cell"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"10842","DOI":"10.1073\/pnas.1613721113","article-title":"Maize endosperm-specific transcription factors O2 and PBF network the regulation of protein and starch synthesis","volume":"113","author":"Zhang","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1146\/annurev-arplant-042817-040606","article-title":"Autophagy: The Master of Bulk and Selective Recycling","volume":"69","author":"Marshall","year":"2018","journal-title":"Annu. Rev. Plant Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1146\/annurev-arplant-042811-105441","article-title":"Autophagy: Pathways for self-eating in plant cells","volume":"63","author":"Liu","year":"2012","journal-title":"Annu. Rev. Plant Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1016\/j.pbi.2017.08.011","article-title":"Autophagy as a mediator of life and death in plants","volume":"40","author":"Ustun","year":"2017","journal-title":"Curr. Opin. Plant Biol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Su, W., Bao, Y., Yu, X., Xia, X., Liu, C., and Yin, W. (2020). Autophagy and Its Regulators in Response to Stress in Plants. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21238889"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1146\/annurev-virology-010220-054709","article-title":"Autophagy in Plant-Virus Interactions","volume":"7","author":"Yang","year":"2020","journal-title":"Annu. Rev. Virol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1355","DOI":"10.1093\/pcp\/pcs099","article-title":"Beginning to Understand Autophagy, an Intracellular Self-Degradation System in Plants","volume":"53","author":"Yoshimoto","year":"2012","journal-title":"Plant Cell Physiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"33105","DOI":"10.1074\/jbc.M204630200","article-title":"The APG8\/12-activating enzyme APG7 is required for proper nutrient recycling and senescence in Arabidopsis thaliana","volume":"277","author":"Doelling","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1111\/j.1365-313X.2010.04166.x","article-title":"ATG8 lipidation and ATG8-mediated autophagy in Arabidopsis require ATG12 expressed from the differentially controlled ATG12A and ATG12B loci","volume":"62","author":"Chung","year":"2010","journal-title":"Plant J."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1379","DOI":"10.1016\/j.febslet.2010.01.018","article-title":"Atg8-family interacting motif crucial for selective autophagy","volume":"584","author":"Noda","year":"2010","journal-title":"FEBS Lett."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Shpilka, T., Weidberg, H., Pietrokovski, S., and Elazar, Z. (2011). Atg8: An autophagy-related ubiquitin-like protein family. Genome Biol., 12.","DOI":"10.1186\/gb-2011-12-7-226"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"926","DOI":"10.4161\/auto.28280","article-title":"Arabidopsis ATG4 cysteine proteases specificity toward ATG8 substrates","volume":"10","author":"Park","year":"2014","journal-title":"Autophagy"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Zhen, X., Li, X., Yu, J., and Xu, F. (2019). OsATG8c-Mediated Increased Autophagy Regulates the Yield and Nitrogen Use Efficiency in Rice. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20194956"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2967","DOI":"10.1105\/tpc.104.025395","article-title":"Processing of ATG8s, ubiquitin-like proteins, and their deconjugation by ATG4s are essential for plant autophagy","volume":"16","author":"Yoshimoto","year":"2004","journal-title":"Plant Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1073\/pnas.1318207111","article-title":"Differential processing of Arabidopsis ubiquitin-like Atg8 autophagy proteins by Atg4 cysteine proteases","volume":"14","author":"Woo","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3902","DOI":"10.1105\/tpc.20.00766","article-title":"Abscisic Acid-Triggered Persulfidation of the Cys Protease ATG4 Mediates Regulation of Autophagy by Sulfide","volume":"32","author":"Aroca","year":"2020","journal-title":"Plant Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.cj.2023.12.006","article-title":"Natural variation of an autophagy-family gene among rice subspecies affects grain size and weight","volume":"12","author":"Huang","year":"2024","journal-title":"Crop J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"D1206","DOI":"10.1093\/nar\/gks1125","article-title":"RiceXPro Version 3.0: Expanding the informatics resource for rice transcriptome","volume":"41","author":"Sato","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1111\/jipb.12941","article-title":"Autophagy in plants: Physiological roles and post-translational regulation","volume":"63","author":"Qi","year":"2021","journal-title":"J. Integr. Plant Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2419","DOI":"10.1111\/nph.19271","article-title":"Dynamic monitoring of TGW6 by selective autophagy during grain development in rice","volume":"240","author":"Liu","year":"2023","journal-title":"New Phytol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Zhen, X., Zheng, N., Yu, J., Bi, C., and Xu, F. (2021). Autophagy mediates grain yield and nitrogen stress resistance by modulating nitrogen remobilization in rice. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0244996"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"878","DOI":"10.4161\/auto.28279","article-title":"OsATG7 is required for autophagy-dependent lipid metabolism in rice postmeiotic anther development","volume":"10","author":"Kurusu","year":"2014","journal-title":"Autophagy"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1307","DOI":"10.1104\/pp.114.254078","article-title":"Establishment of monitoring methods for autophagy in rice reveals autophagic recycling of chloroplasts and root plastids during energy limitation","volume":"167","author":"Izumi","year":"2015","journal-title":"Plant Physiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1137","DOI":"10.1016\/j.molp.2022.05.006","article-title":"Autophagy targets Hd1 for vacuolar degradation to regulate rice flowering","volume":"15","author":"Hu","year":"2022","journal-title":"Mol. Plant"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3249","DOI":"10.1093\/jxb\/eru175","article-title":"Fusion, rupture, and degeneration: The fate of in vivo-labelled PSVs in developing barley endosperm","volume":"65","author":"Ibl","year":"2014","journal-title":"J. Exp. Bot."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1403","DOI":"10.1093\/jxb\/ery012","article-title":"Autophagy controls resource allocation and protein storage accumulation in Arabidopsis seeds","volume":"69","author":"Marmagne","year":"2018","journal-title":"J. Exp. Bot."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e51918","DOI":"10.7554\/eLife.51918","article-title":"Reticulon proteins modulate autophagy of the endoplasmic reticulum in maize endosperm","volume":"9","author":"Zhang","year":"2020","journal-title":"eLife"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1038\/ng.2923","article-title":"Chalk5 encodes a vacuolar H+-translocating pyrophosphatase influencing grain chalkiness in rice","volume":"46","author":"Li","year":"2014","journal-title":"Nat. Genet."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1912","DOI":"10.1093\/plcell\/koac057","article-title":"Natural variation in WHITE-CORE RATE 1 regulates redox homeostasis in rice endosperm to affect grain quality","volume":"34","author":"Wu","year":"2022","journal-title":"Plant Cell"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"6653","DOI":"10.1038\/s41467-025-61683-4","article-title":"Natural variation of an E3 ubiquitin ligase encoding gene Chalk9 regulates grain chalkiness in rice","volume":"16","author":"Hu","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2985","DOI":"10.1111\/pbi.14420","article-title":"Natural variation of WBR7 confers rice high yield and quality by modulating sucrose supply in sink organs","volume":"22","author":"Shi","year":"2024","journal-title":"Plant Biotechnol. J."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1694","DOI":"10.1104\/pp.106.082826","article-title":"Synergism between RPBF Dof and RISBZ1 bZIP activators in the regulation of rice seed expression genes","volume":"141","author":"Yamamoto","year":"2006","journal-title":"Plant Physiol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"908","DOI":"10.1111\/j.1365-313X.2009.03925.x","article-title":"Compensation and interaction between RISBZ1 and RPBF during grain filling in rice","volume":"59","author":"Kawakatsu","year":"2009","journal-title":"Plant J."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"890","DOI":"10.1111\/pbi.13994","article-title":"Transcription factors NAC20 and NAC26 interact with RPBF to activate albumin accumulations in rice endosperm","volume":"21","author":"Wu","year":"2023","journal-title":"Plant Biotechnol. J."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2947","DOI":"10.1093\/bioinformatics\/btm404","article-title":"Clustal W and Clustal X version 2.0","volume":"23","author":"Larkin","year":"2007","journal-title":"Bioinformatics"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"3022","DOI":"10.1093\/molbev\/msab120","article-title":"MEGA11: Molecular Evolutionary Genetics Analysis Version 11","volume":"38","author":"Tamura","year":"2021","journal-title":"Mol. Biol. Evol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1120","DOI":"10.1093\/bioinformatics\/btu743","article-title":"CRISPRdirect: Software for designing CRISPR\/Cas guide RNA with reduced off-target sites","volume":"31","author":"Naito","year":"2015","journal-title":"Bioinformatics"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1431","DOI":"10.1016\/j.molp.2015.05.009","article-title":"DSDecode: A Web-Based Tool for Decoding of Sequencing Chromatograms for Genotyping of Targeted Mutations","volume":"8","author":"Liu","year":"2015","journal-title":"Mol. Plant"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1949","DOI":"10.1038\/s41467-019-09919-y","article-title":"Natural variation of OsGluA2 is involved in grain protein content regulation in rice","volume":"10","author":"Yang","year":"2019","journal-title":"Nat. Commun."}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/14\/16\/2530\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:27:26Z","timestamp":1760034446000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/14\/16\/2530"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,14]]},"references-count":53,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2025,8]]}},"alternative-id":["plants14162530"],"URL":"https:\/\/doi.org\/10.3390\/plants14162530","relation":{},"ISSN":["2223-7747"],"issn-type":[{"type":"electronic","value":"2223-7747"}],"subject":[],"published":{"date-parts":[[2025,8,14]]}}}