The B3 transcription factors(TFs)in plants play vital roles in numerous biological processes.Although B3 genes have been broadly identified in many plants,little is known about their potential functions in mediating s...The B3 transcription factors(TFs)in plants play vital roles in numerous biological processes.Although B3 genes have been broadly identified in many plants,little is known about their potential functions in mediating seed development and material accumulation.Castor bean(Ricinus communis)is a non-edible oilseed crop considered an ideal model system for seed biology research.Here,we identified a total of 61 B3 genes in the castor bean genome,which can be classified into five subfamilies,including ABI3/VP1,HSI,ARF,RAV and REM.The expression profiles revealed that RcABI3/VP1 subfamily genes are significantly up-regulated in the middle and later stages of seed development,indicating that these genes may be associated with the accumulation of storage oils.Furthermore,through yeast one-hybrid and tobacco transient expression assays,we detected that ABI3/VP1 subfamily member RcLEC2 directly regulates the transcription of RcOleosin2,which encodes an oil-body structural protein.This finding suggests that RcLEC2,as a seed-specific TF,may be involved in the regulation of storage materials accumulation.This study provides novel insights into the potential roles and molecular basis of B3 family proteins in seed development and material accumulation.展开更多
目的探讨ABI家族成员3结合蛋白(ABI family member 3-binding protein,ABI3BP)在血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导内皮祖细胞功能障碍中的作用及机制。方法为探讨ABI3BP在AngⅡ诱导内皮祖细胞功能障碍中的作用,将细胞分为4组,sh-N...目的探讨ABI家族成员3结合蛋白(ABI family member 3-binding protein,ABI3BP)在血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导内皮祖细胞功能障碍中的作用及机制。方法为探讨ABI3BP在AngⅡ诱导内皮祖细胞功能障碍中的作用,将细胞分为4组,sh-NC组[转染阴性对照短发夹RNA(LV-scramble-shRNA)+磷酸盐缓冲液(phosphate buffered saline,PBS)]、sh-ABI3BP组[转染ABI3BP shRNA(LV-ABI3BP-shRNA)+PBS]、sh-NC+AngⅡ组(LV-scramble-shRNA+AngⅡ)和sh-ABI3BP+AngⅡ组(LV-ABI3BP-shRNA+AngⅡ)。采用Transwell实验检测细胞迁移能力,黏附实验检测细胞黏附能力,Matrigel检测细胞成管能力,原位末端标记法检测细胞凋亡。Western blot检测整合素β1-黏着斑激酶(focal adhesion kinase,FAK)-P53信号通路变化情况。结果与sh-NC组比较,sh-NC+AngⅡ组迁移细胞数量、黏附细胞数量、小管形成数量显著降低,细胞凋亡率、整合素β1、磷酸化FAK(p-FAK)/FAK及P53蛋白表达显著增高,差异有统计学意义(P<0.05)。与sh-NC+AngⅡ组比较,sh-ABI3BP+AngⅡ组迁移细胞数量[(88.67±8.33)个vs(62.33±7.37)个]、黏附细胞数量[(104.33±6.03)个vs(68.33±10.05)个]、小管形成数量[(36.33±3.21)个vs(19.33±3.06)个]显著增高,细胞凋亡率、整合素β1、p-FAK/FAK及P53蛋白表达水平显著降低,差异有统计学意义(P<0.05)。结论AngⅡ可上调ABI3BP表达,敲低ABI3BP基因表达可改善AngⅡ诱导的内皮祖细胞功能障碍,其机制可能与抑制整合素β1-FAK-P53信号通路有关。展开更多
FUSCA3(FUS3)is a member of B3-domain transcription factor family and master regulator of seed development.It has potential roles in hormone biosynthesis and signaling pathways and therefore plays diverse roles in plan...FUSCA3(FUS3)is a member of B3-domain transcription factor family and master regulator of seed development.It has potential roles in hormone biosynthesis and signaling pathways and therefore plays diverse roles in plant life cycle,especially in seed germination,dormancy,embryo formation,seed and fruit development,and maturation.However,there is limited information about its functions in seed and fruit development of grapevine.In this study,we expressed VvFUS3 in tomato for its functional characterization.Overexpression of VvFUS3 in tomato led to a reduction in seed number and seed weight without affecting the fruit size.Histological analysis found that both cell expansion and cell division in transgenic seed and fruit pericarp have been affected.However,there were no obvious differences in pollen size,shape,and viability,suggesting that VvFUS3 affects seed development but not the pollen grains.Moreover,the expression of several genes with presumed roles in seed development and hormone signaling pathways was also influenced by VvFUS3.These results suggest that VvFUS3 is involved in hormonal signaling pathways that regulate seed number and size.In conclusion,our study provides novel preliminary information about the pivotal roles of VvFUS3 in seed and fruit development and these findings can potentially serve as a reference for molecular breeding of seedless grapes.展开更多
Repression of embryonic traits during the seedto-seedling phase transition requires the inactivation of master transcription factors associated with embryogenesis.How the timing of such inactivation is controlled is u...Repression of embryonic traits during the seedto-seedling phase transition requires the inactivation of master transcription factors associated with embryogenesis.How the timing of such inactivation is controlled is unclear.Here,we report on a novel transcriptional corepressor,Arabidopsis thaliana SDR4L,that forms a feedback inhibition loop with the master transcription factors LEC1 and ABI3 to repress embryonic traits post-imbibition.LEC1 and ABI3 regulate their own expression by inducing AtSDR4L during mid to late embryogenesis.AtSDR4L binds to sites upstream of LEC1 and ABI4,and these transcripts are upregulated in Atsdr4l seedlings.Atsdr4 l seedlings phenocopy a LEC1 overexpressor.The embryonic traits of Atsdr4l can be partially rescued by impairing LEC1 or ABI3.The penetrance and expressivity of the Atsdr4l phenotypes depend on both developmental and external cues,demonstrating the importance of AtSDR4L in seedling establishment under suboptimal conditions.展开更多
文摘密码子偏性是影响外源基因表达的重要因素,也是反映基因家族进化的关键线索之一。兰科植物是植物保护中的“旗舰”类群,且多数物种具有药用和观赏价值,但种子不易萌发,而ABI3(abscisic acid insensitive 3)基因是ABA信号转导的中心调控因子,是调控植物种子萌发的关键因子。因此,深入研究兰科ABI3基因密码子偏性,可为解析该基因功能、优化密码子组成奠定基础。本研究以拟南芥ABI3基因为探针,从公共数据库(NCBI)筛选出45条兰科植物ABI3基因,通过CodonW、SPSS、MEGA等生物信息学程序对其进行密码子使用模式和偏好性分析。相关参数分析结果表明,该基因的ENC(effective number of codon)值范围分布为40.84~58.46,其中大于52的占82.2%,CAI(codon adaptation index)平均值为0.203,远小于1,表明ABI3基因的密码子偏性相对较弱;中性绘图分析显示,GC12与GC3的回归曲线的斜率为0.6103,R2=0.7928,P<0.05,有极高的相关性,这暗示着碱基组成对密码子偏性具有重要的影响;ENC-plot分析中,所有基因ENC值都低于期望值,但相差不大,说明兰科ABI3基因密码子主要受碱基突变的影响,同时也受选择压力等其他因素影响。奇偶偏好性分析显示,大多数基因分布在平面图的右侧区域,密码子第3位A/T(U)的使用频率高于G/C。通过RSCU和ΔRSCU分析确定了26个最优密码子,其中以A/T(U)结尾的密码子有14个,以T(U)结尾的密码子有10个,说明兰科植物ABI3基因相对偏好使用A/U结尾的密码子,尤其偏好使用T(U)结尾的密码子,这与大多数单子叶植物密码子使用偏好不同。本研究可为兰科植物ABI3基因的系统发育、功能解析、提高ABI3基因表达效率提供理论基础,同时可为促进植物密码子生物学研究及外源基因改良提供参考。
基金National Natural Science Foundation of China(31661143002,81760507,31571709,31771839,31701123 and 31501034)Yunnan Applied Basic Research Projects(2016FA011,2016FB060 and 2016FB040)+1 种基金the National R&D Infrastructure and Facility development Program of China"Fundamental Science Data Sharing Platform(DKA 201712-02-16)the 13th Five-year informatization Plan of Chinese Academy of Sciences(No.XXH13506)。
文摘The B3 transcription factors(TFs)in plants play vital roles in numerous biological processes.Although B3 genes have been broadly identified in many plants,little is known about their potential functions in mediating seed development and material accumulation.Castor bean(Ricinus communis)is a non-edible oilseed crop considered an ideal model system for seed biology research.Here,we identified a total of 61 B3 genes in the castor bean genome,which can be classified into five subfamilies,including ABI3/VP1,HSI,ARF,RAV and REM.The expression profiles revealed that RcABI3/VP1 subfamily genes are significantly up-regulated in the middle and later stages of seed development,indicating that these genes may be associated with the accumulation of storage oils.Furthermore,through yeast one-hybrid and tobacco transient expression assays,we detected that ABI3/VP1 subfamily member RcLEC2 directly regulates the transcription of RcOleosin2,which encodes an oil-body structural protein.This finding suggests that RcLEC2,as a seed-specific TF,may be involved in the regulation of storage materials accumulation.This study provides novel insights into the potential roles and molecular basis of B3 family proteins in seed development and material accumulation.
基金This work was supported by the Joint Funds of the National Natural Science Foundation of China(Grant No.U1603234)the Program for Innovative Research Team of Grape Germplasm Resources and Breeding(Grant No.2013KCT-25).
文摘FUSCA3(FUS3)is a member of B3-domain transcription factor family and master regulator of seed development.It has potential roles in hormone biosynthesis and signaling pathways and therefore plays diverse roles in plant life cycle,especially in seed germination,dormancy,embryo formation,seed and fruit development,and maturation.However,there is limited information about its functions in seed and fruit development of grapevine.In this study,we expressed VvFUS3 in tomato for its functional characterization.Overexpression of VvFUS3 in tomato led to a reduction in seed number and seed weight without affecting the fruit size.Histological analysis found that both cell expansion and cell division in transgenic seed and fruit pericarp have been affected.However,there were no obvious differences in pollen size,shape,and viability,suggesting that VvFUS3 affects seed development but not the pollen grains.Moreover,the expression of several genes with presumed roles in seed development and hormone signaling pathways was also influenced by VvFUS3.These results suggest that VvFUS3 is involved in hormonal signaling pathways that regulate seed number and size.In conclusion,our study provides novel preliminary information about the pivotal roles of VvFUS3 in seed and fruit development and these findings can potentially serve as a reference for molecular breeding of seedless grapes.
基金funded by the National Science Foundation of China(31921001)the Beijing Outstanding University Discipline Program to Z.G.+4 种基金UBC Four Year Doctoral Fellowship to M.A.BC Graduate Scholarship to R.H.Bio Talent/SWPP funding to R.H.and E.L.UBC Kay Earle Memorial Botany Fund Fellowship to B.L.NSERC RGPIN-2019-05039 and CFI JELF/BCKDF 38187 to L.S.
文摘Repression of embryonic traits during the seedto-seedling phase transition requires the inactivation of master transcription factors associated with embryogenesis.How the timing of such inactivation is controlled is unclear.Here,we report on a novel transcriptional corepressor,Arabidopsis thaliana SDR4L,that forms a feedback inhibition loop with the master transcription factors LEC1 and ABI3 to repress embryonic traits post-imbibition.LEC1 and ABI3 regulate their own expression by inducing AtSDR4L during mid to late embryogenesis.AtSDR4L binds to sites upstream of LEC1 and ABI4,and these transcripts are upregulated in Atsdr4l seedlings.Atsdr4 l seedlings phenocopy a LEC1 overexpressor.The embryonic traits of Atsdr4l can be partially rescued by impairing LEC1 or ABI3.The penetrance and expressivity of the Atsdr4l phenotypes depend on both developmental and external cues,demonstrating the importance of AtSDR4L in seedling establishment under suboptimal conditions.
