Seed oil accumulation requires precise coordination of multiple factors,and transcription factors(TFs)play crucial roles in governing this process.However,details of how TFs and crosstalk between different TF families...Seed oil accumulation requires precise coordination of multiple factors,and transcription factors(TFs)play crucial roles in governing this process.However,details of how TFs and crosstalk between different TF families coordinate the transcriptional network associated with seed oil accumulation remain to be clarified.Here,we report that basic helix-loop-helix7(bHLH7)and PROTODERMAL FACTOR2(PDF2)TFs,both highly expressed in developing embryos,negatively control seed oil accumulation.bHLH7 and PDF2 physically interact and cooperatively inhibit the transcription of MYB DOMAIN PROTEIN 96(MYB96),PYRUVATE DEHYDROGENASE E1 BETA1(PDH-E1β1),PDH-E1β2,and BIOTIN CARBOXYL CARRIER PROTEIN1(BCCP1)by directly binding to their promoters.Additional genetic evidence demonstrated that bHLH7 and PDF2 act in an additive manner to repress seed oil accumulation,and this effect is partially dependent on PDH-E1β1,which significantly promotes seed oil deposition.Collectively,our results provide insights into the mechanisms by which the bHLH7-PDF2 module mediates transcriptional regulation to control oil accumulation in A.thaliana seeds.展开更多
【目的】通过不同生理状态牛卵泡高通量测序筛选与卵泡发育相关的基因。【方法】母牛同期发情后,B超声波连续监测并适时采集第一卵泡波出现偏差前的最大卵泡(the largest follicle at predeviation,PDF1)和第二大卵泡(the second larges...【目的】通过不同生理状态牛卵泡高通量测序筛选与卵泡发育相关的基因。【方法】母牛同期发情后,B超声波连续监测并适时采集第一卵泡波出现偏差前的最大卵泡(the largest follicle at predeviation,PDF1)和第二大卵泡(the second largest follicle at predeviation,PDF2),构建RNA文库后Illumina平台测序,经数据库比对,设定参数筛选高表达基因、差异表达基因并进行GO分析,Genecards基因功能查询进一步筛选与卵泡发育直接相关的调控基因,q RT-PCR对筛选的基因进行表达量验证分析。【结果】两个转录本中共获得263个差异表达基因;GO功能聚类分析共分为三大类90组:其中生物学过程占64.4%,细胞组分占17.8%,分子功能占17.8%;获得一些重要的功能富集通路,如调控信号转导和细胞因子应答的生物途径;基因表达量分析筛选出10个高表达的上调和下调基因,获得参与雌激素合成和胎儿性别发育的CYP17A1、参与类固醇激素合成的LOC785462、细胞发育过程中调节细胞凋亡的DACH1以及调节ERK和MEK1/2信号通路的MAP3K3。Genecards功能查询共获得6个基因与卵泡发育关系较为密切,其中上调基因分别为PRSS35,PTGRF,ARID4B,GPR116;下调基因为APOA1和CPXM1;q RT-PCR结果显示,PRSS35和ARID4B在DF中的表达量显著高于SF(P<0.05),CPXM1在SF中的表达量极显著高于DF(P<0.01),PTGRF,GPR116和APOA1在DF和SF中的表达量不存在显著差异(P>0.05),但其表达变化趋势与高通量测序结果相一致。【结论】转录组测序结果真实可靠,PRSS35和ARID4B在卵泡发育过程中发挥正调控作用,CPXM1在卵泡发育过程中发挥负调控作用,获得的牛卵泡发育相关基因和重要调节途径,对后期深入探讨卵泡发育调控机理的研究具有重要意义。展开更多
基金supported by the National Key Research and Development Program of China(grant no.2022YFD1200400)the Scientific and Technological Innovation Team of Shaanxi Province(grant no.2024RSCXTD-69)a grant from the Yang Ling Seed Industry Innovation Center(grant no.K3031122024).
文摘Seed oil accumulation requires precise coordination of multiple factors,and transcription factors(TFs)play crucial roles in governing this process.However,details of how TFs and crosstalk between different TF families coordinate the transcriptional network associated with seed oil accumulation remain to be clarified.Here,we report that basic helix-loop-helix7(bHLH7)and PROTODERMAL FACTOR2(PDF2)TFs,both highly expressed in developing embryos,negatively control seed oil accumulation.bHLH7 and PDF2 physically interact and cooperatively inhibit the transcription of MYB DOMAIN PROTEIN 96(MYB96),PYRUVATE DEHYDROGENASE E1 BETA1(PDH-E1β1),PDH-E1β2,and BIOTIN CARBOXYL CARRIER PROTEIN1(BCCP1)by directly binding to their promoters.Additional genetic evidence demonstrated that bHLH7 and PDF2 act in an additive manner to repress seed oil accumulation,and this effect is partially dependent on PDH-E1β1,which significantly promotes seed oil deposition.Collectively,our results provide insights into the mechanisms by which the bHLH7-PDF2 module mediates transcriptional regulation to control oil accumulation in A.thaliana seeds.
文摘【目的】通过不同生理状态牛卵泡高通量测序筛选与卵泡发育相关的基因。【方法】母牛同期发情后,B超声波连续监测并适时采集第一卵泡波出现偏差前的最大卵泡(the largest follicle at predeviation,PDF1)和第二大卵泡(the second largest follicle at predeviation,PDF2),构建RNA文库后Illumina平台测序,经数据库比对,设定参数筛选高表达基因、差异表达基因并进行GO分析,Genecards基因功能查询进一步筛选与卵泡发育直接相关的调控基因,q RT-PCR对筛选的基因进行表达量验证分析。【结果】两个转录本中共获得263个差异表达基因;GO功能聚类分析共分为三大类90组:其中生物学过程占64.4%,细胞组分占17.8%,分子功能占17.8%;获得一些重要的功能富集通路,如调控信号转导和细胞因子应答的生物途径;基因表达量分析筛选出10个高表达的上调和下调基因,获得参与雌激素合成和胎儿性别发育的CYP17A1、参与类固醇激素合成的LOC785462、细胞发育过程中调节细胞凋亡的DACH1以及调节ERK和MEK1/2信号通路的MAP3K3。Genecards功能查询共获得6个基因与卵泡发育关系较为密切,其中上调基因分别为PRSS35,PTGRF,ARID4B,GPR116;下调基因为APOA1和CPXM1;q RT-PCR结果显示,PRSS35和ARID4B在DF中的表达量显著高于SF(P<0.05),CPXM1在SF中的表达量极显著高于DF(P<0.01),PTGRF,GPR116和APOA1在DF和SF中的表达量不存在显著差异(P>0.05),但其表达变化趋势与高通量测序结果相一致。【结论】转录组测序结果真实可靠,PRSS35和ARID4B在卵泡发育过程中发挥正调控作用,CPXM1在卵泡发育过程中发挥负调控作用,获得的牛卵泡发育相关基因和重要调节途径,对后期深入探讨卵泡发育调控机理的研究具有重要意义。
