Autophagy is a highly conserved cellular program in eukaryotic cells which mediates the degradation of cytoplasmic components through the lysosome,also named the vacuole in plants.However,the molecular mechanisms underlying the fusion of autophagosomes with the vacuole remain unclear.Here,we report the functional characterization of a rice(Oryza sativa)mutant with defects in storage protein transport in endosperm cells and accumulation of numerous autophagosomes in root cells.Cytological and immunocytochemical experiments showed that this mutant exhibits a defect in the fusion between autophagosomes and vacuoles.The mutant harbors a loss-of-function mutation in the rice homolog of Arabidopsis thaliana MONENSIN SENSITIVITY1(MON1).Biochemical and genetic evidence revealed a synergistic interaction between rice MON1 and AUTOPHAGY-RELATED 8a in maintaining normal growth and development.In addition,the rice mon1 mutant disrupted storage protein sorting to protein storage vacuoles.Furthermore,quantitative proteomics verified that the loss of MON1 function influenced diverse biological pathways including autophagy and vacuolar transport,thus decreasing the transport of autophagic and vacuolar cargoes to vacuoles.Together,our findings establish a molecular link between autophagy and vacuolar protein transport,and offer insights into the dual functions of the MON1–CCZ1(CAFFEINE ZINC SENSITIVITY1)complex in plants.
转基因玉米MON87411是孟山都远东有限公司研发的抗虫耐除草剂玉米转化体,该转化体已获得进口加工原料的农业转基因生物安全证书。以转基因玉米MON87411品系特异性序列为靶标设计引物和探针,经特异性测试、体系优化、灵敏度测试及检出限测试,建立了转基因玉米MON87411的实时荧光PCR定性检测方法。结果表明,该方法能检测出转基因玉米MON87411的转化体成分,检出限(limit of detection,LOD)可达0.05%,具有稳定性好、特异性强、灵敏度高的特点。国内8家转基因生物安全检测机构对本方法进行了特异性测试、检出限测试和再现性测试,循环验证报告显示该方法符合国家标准方法的各项要求,可在检测行业推广应用。研究建立的方法可为我国对转基因玉米MON87411品系的安全监管提供有效的技术支撑。
