Pre-harvest sprouting(PHS)describes the germination of physiologically mature grains in spikes prior to harvest in cereal crops.PHS could seriously decrease grain yield and quality,which makes it a major constraint to...Pre-harvest sprouting(PHS)describes the germination of physiologically mature grains in spikes prior to harvest in cereal crops.PHS could seriously decrease grain yield and quality,which makes it a major constraint to cereal production worldwide.A number of PHS-associated genes in cereals have been reported;however,the molecular mechanisms underlying PHS remain largely elusive.Here,we report a CRISPRCas9 mutant with severe PHS in a paddy field.The mutated gene OsMFT2 encodes a phosphatidylethanolamine-binding protein(PEBP).Intriguingly,the OsMFT1,in the same PEBP family,had the opposite effect in controlling rice PHS as does OsMFT2.Germination tests of seeds of chimeric protein-expressing plants revealed that the fourth exon conferred the antagonistic activity of OsMFT1 and OsMFT2 in rice PHS.Additionally,two lines of these plants showed elevated grain numbers per panicle,implying that chimeric protein has potential to significantly increase yield.Moreover,transcriptome analysis and genetic studies indicated that OsMFT1 and OsMFT2 performed opposing functions in rice PHS owing to three co-regulated genes that being contrastingly affected by OsMFT1 and OsMFT2.Overall,it seemed that the proper combination of PEBP family members could obtain optimal PHS resistance and high yield.展开更多
MOTHER OF FT AND TFL1(MFT)属于磷脂酰乙醇胺结合蛋白(phosphatidyl ethanolamine-binding protein,PEBP)家族,该家族在被子植物中主要起着促进或抑制开花和控制株型的作用。本课题组前期从海岛棉(Gossypium barbadense)中克隆了一个MF...MOTHER OF FT AND TFL1(MFT)属于磷脂酰乙醇胺结合蛋白(phosphatidyl ethanolamine-binding protein,PEBP)家族,该家族在被子植物中主要起着促进或抑制开花和控制株型的作用。本课题组前期从海岛棉(Gossypium barbadense)中克隆了一个MFT-like同源基因GbMFT2,发现其在种子中表达量较高,并且受ABA诱导。为了进一步探究该基因的功能,本研究首先检测了200 mmol/L Na Cl胁迫海岛棉0、1 h、3 h、6 h、12 h后基因的表达量,表明在盐诱导1 h GbMFT2基因的表达量最高。然后构建了过量表达载体p35S:GbMFT2,并通过农杆菌介导的叶盘法将其转化到普通烟草(Nicotiana tabacum cv.NC89)中,半定量RT-PCR表明GbMFT2基因在烟草中异位表达成功。利用0、100 mmol/L、200 mmol/L、300 mmol/L、400 mmol/L和500 mmol/L Na Cl胁迫不同的转基因株系,表明转基因烟草比野生型烟草更耐盐。再分别测量300 mmol/L Na Cl处理前后的野生型烟草和转基因烟草的脯氨酸含量、POD活性,发现处理后的野生型烟草的脯氨酸含量、POD活性均明显降低,而转基因烟草株系中的脯氨酸含量、POD活性均有明显的提升。研究结果表明,GbMFT2基因能够响应盐胁迫,并增加烟草的耐盐性。本研究可为研究植物MFT-like基因的功能提供了新思路,也为进一步解析棉花发育基因的功能提供了研究基础。展开更多
基金supported by the National Natural Science Foundation of China(32172059)Fundamental Research Funds for the Central Universities(SWUXDJH202315).
文摘Pre-harvest sprouting(PHS)describes the germination of physiologically mature grains in spikes prior to harvest in cereal crops.PHS could seriously decrease grain yield and quality,which makes it a major constraint to cereal production worldwide.A number of PHS-associated genes in cereals have been reported;however,the molecular mechanisms underlying PHS remain largely elusive.Here,we report a CRISPRCas9 mutant with severe PHS in a paddy field.The mutated gene OsMFT2 encodes a phosphatidylethanolamine-binding protein(PEBP).Intriguingly,the OsMFT1,in the same PEBP family,had the opposite effect in controlling rice PHS as does OsMFT2.Germination tests of seeds of chimeric protein-expressing plants revealed that the fourth exon conferred the antagonistic activity of OsMFT1 and OsMFT2 in rice PHS.Additionally,two lines of these plants showed elevated grain numbers per panicle,implying that chimeric protein has potential to significantly increase yield.Moreover,transcriptome analysis and genetic studies indicated that OsMFT1 and OsMFT2 performed opposing functions in rice PHS owing to three co-regulated genes that being contrastingly affected by OsMFT1 and OsMFT2.Overall,it seemed that the proper combination of PEBP family members could obtain optimal PHS resistance and high yield.
文摘MOTHER OF FT AND TFL1(MFT)属于磷脂酰乙醇胺结合蛋白(phosphatidyl ethanolamine-binding protein,PEBP)家族,该家族在被子植物中主要起着促进或抑制开花和控制株型的作用。本课题组前期从海岛棉(Gossypium barbadense)中克隆了一个MFT-like同源基因GbMFT2,发现其在种子中表达量较高,并且受ABA诱导。为了进一步探究该基因的功能,本研究首先检测了200 mmol/L Na Cl胁迫海岛棉0、1 h、3 h、6 h、12 h后基因的表达量,表明在盐诱导1 h GbMFT2基因的表达量最高。然后构建了过量表达载体p35S:GbMFT2,并通过农杆菌介导的叶盘法将其转化到普通烟草(Nicotiana tabacum cv.NC89)中,半定量RT-PCR表明GbMFT2基因在烟草中异位表达成功。利用0、100 mmol/L、200 mmol/L、300 mmol/L、400 mmol/L和500 mmol/L Na Cl胁迫不同的转基因株系,表明转基因烟草比野生型烟草更耐盐。再分别测量300 mmol/L Na Cl处理前后的野生型烟草和转基因烟草的脯氨酸含量、POD活性,发现处理后的野生型烟草的脯氨酸含量、POD活性均明显降低,而转基因烟草株系中的脯氨酸含量、POD活性均有明显的提升。研究结果表明,GbMFT2基因能够响应盐胁迫,并增加烟草的耐盐性。本研究可为研究植物MFT-like基因的功能提供了新思路,也为进一步解析棉花发育基因的功能提供了研究基础。
