拟南芥IQM6突变推迟远轴面表皮毛的发生
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摘要
IQM6 (IQ motif containing 6)由At3g58480编码,是拟南芥IQM家族中第6位成员,其基因的功能未见报道。本研究通过筛选与鉴定获得了iqm6-1的纯合体,并利用CRISPR/Cas9基因编辑技术构建iqm6-2;对突变体的表型分析显示, iqm6-1和iqm6-2的远轴面表皮毛的发生晚于野生型Col-0;利用RT-qPCR对表皮毛发生的关键基因进行转录本水平分析发现,与野生型Col-0相比, iqm6-1和iqm6-2延迟miR156的下调和SPL3的上调。这些结果表明,拟南芥IQM6突变可能介导miR156/SPL3组件的表达,推迟远轴面表皮毛的发生。
IQ motif-containing protein 6(IQM6) is coded by At3 g58480, which is the sixth member of the IQM family in Arabidopsis. However, no functional characterizations have been performed for IQM6 in Arabidopsis.In this study, homozygotes of iqm6-1 were identified and iqm6-2 were constructed using CRISPR/Cas9 gene editing technology. The results of phenotypic analysis showed that iqm6-1 and iqm6-2 delayed the initiation of abaxial trichomes. RT-qPCR results showed that iqm6-1 and iqm6-2 delayed the decrease of miRNA156 and the increase of SPL3 transcription level when compared with Col-0. These results suggest that the mutation of IQM6 changes the expression of miRNA156/SPL3, thus delaying the initiation of abaxial trichomes in Arabidopsis.
IQ motif-containing protein 6(IQM6) is coded by At3 g58480, which is the sixth member of the IQM family in Arabidopsis. However, no functional characterizations have been performed for IQM6 in Arabidopsis.In this study, homozygotes of iqm6-1 were identified and iqm6-2 were constructed using CRISPR/Cas9 gene editing technology. The results of phenotypic analysis showed that iqm6-1 and iqm6-2 delayed the initiation of abaxial trichomes. RT-qPCR results showed that iqm6-1 and iqm6-2 delayed the decrease of miRNA156 and the increase of SPL3 transcription level when compared with Col-0. These results suggest that the mutation of IQM6 changes the expression of miRNA156/SPL3, thus delaying the initiation of abaxial trichomes in Arabidopsis.
引文
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Bender KW,Snedden WA(2013).Calmodulin-related proteins step out from the shadow of their namesake.Plant Physiol,163:486-495
Chien JC,Sussex IM(1996).Differential regulation of trichome formation on the adaxial and abaxial leaf surfaces by gibberellins and photoperiod in Arabidopsis thaliana(L.)Heynh.Plant Physiol,111(4):1321-1328
Gao Y,Guo JQ,Zhao JF(2011).Molecular mechanisms of Arabidopsis trichome development.Chin Bull Bot,46(1):119-127(in Chinese with English abstract)[高英,郭建强,赵金凤(2011).拟南芥表皮毛发育的分子机制.植物学报,46(1):119-127]
Gifford J,Walsh M,Vogel H(2007).Structures and metal-ion-binding properties of the Ca2+-binding helix-loophelix EF-hand motifs.Biochem J,405(2):199-221
Guo CK,Xu YM,Shi M,et al(2017).Repression of miR156by miR159 regulates the timing of the juvenile-to-adult transition in Arabidopsis.Plant Cell,29:1293-1304
Li M,Yang S,Ruan YY,et al(2006).Identification of atsuc3with T-DNA insertion by PCR.Plant Physiol Commun,42(1):91-94(in Chinese with English abstract)[李敏,杨双,阮燕晔等(2006).拟南芥T-DNA插入突变体atsuc3的PCR鉴定.植物生理学通讯,42(1):91-94]
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Ma X,Liu YG(2016).CRISPR/Cas9-based multiplex genome editing in monocot and dicot plants.Curr Protoc Mol Biol,115:16
O’Day DH(2003).CaMBOT:profiling and characterizing calmodulin-binding protein.Cell Signal,15:347-354
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Schulz P,Herde M,Romeis T(2013).Calcium-dependent protein kinases:hubs in plant stress signaling and development.Plant Physiol,163:523-530
Schwab R,Palatnik JF,Riester M,et al(2005).Specific effects of microRNAs on the plant transcriptome.Dev Cell,8(4):517-527
Tian CE,Zhou YP(2013).Research progress in plant IQmotif-containing calmodulin-binding proteins.Chin Bull Bot,48(4):447-460(in Chinese with English abstract)[田长恩,周玉萍(2013).植物具IQ基序的钙调素结合蛋白的研究进展.植物学报,48(4):447-460]
Usami T,Horiguchi G,Yano S,et al(2009).The more and smaller cells mutants of Arabidopsis thaliana identify novel roles for SQUAMOSA PROMOTER BINDINGPROTEIN-LIKE genes in the control of heteroblasty.Development,136(6):955-964
Wang JW(2014).Regulation of flowering time by the miR156-mediated age pathway.J Exp Bot,65(17):4723-4730
Xu HM,Zhang LJ,Liu Chun(2010).Study of infection Arabidopsis thaliana by Agrobacterium dipping flower method.Northern Hortic,14:143-146(in Chinese with English abstract)[许红梅,张立军,刘淳(2010).农杆菌蘸花法侵染拟南芥的研究.北方园艺,14:143-146]
Xu Y,Zhang L,Wu G(2018).Epigenetic regulation of juvenile-to-adult transition in plants.Front Plant Sci,9:1048
Yin QQ,Li M,Ding B,et al(2016).Research advances on plant calmodulin binding protein IQD.Mol Plant Breed,14(11):3224-3231(in Chinese with English abstract)[尹倩倩,李明,丁博等(2016).植物钙调素结合蛋白IQD的研究概况.分子植物育种,14(11):3224-3231]
Zeng HQ,Zhang YX,Wang S,et al(2016).Calcium/calmodulin-mediated signal transduction system in plants.Chin Bull Bot,51(5):705-723(in Chinese with English abstract)[曾后清,张亚仙,汪尚等(2016).植物钙/钙调素介导的信号转导系统.植物学报,51(5):705-723]
Zhou YP,Chen YZ,Yamamoto KT,et al(2010).Sequence and expression analysis of the Arabidopsis IQM family.Acta Physiol Plant,32:191-198
Bender KW,Snedden WA(2013).Calmodulin-related proteins step out from the shadow of their namesake.Plant Physiol,163:486-495
Chien JC,Sussex IM(1996).Differential regulation of trichome formation on the adaxial and abaxial leaf surfaces by gibberellins and photoperiod in Arabidopsis thaliana(L.)Heynh.Plant Physiol,111(4):1321-1328
Gao Y,Guo JQ,Zhao JF(2011).Molecular mechanisms of Arabidopsis trichome development.Chin Bull Bot,46(1):119-127(in Chinese with English abstract)[高英,郭建强,赵金凤(2011).拟南芥表皮毛发育的分子机制.植物学报,46(1):119-127]
Gifford J,Walsh M,Vogel H(2007).Structures and metal-ion-binding properties of the Ca2+-binding helix-loophelix EF-hand motifs.Biochem J,405(2):199-221
Guo CK,Xu YM,Shi M,et al(2017).Repression of miR156by miR159 regulates the timing of the juvenile-to-adult transition in Arabidopsis.Plant Cell,29:1293-1304
Li M,Yang S,Ruan YY,et al(2006).Identification of atsuc3with T-DNA insertion by PCR.Plant Physiol Commun,42(1):91-94(in Chinese with English abstract)[李敏,杨双,阮燕晔等(2006).拟南芥T-DNA插入突变体atsuc3的PCR鉴定.植物生理学通讯,42(1):91-94]
Lu Y,Long H(2015).The effect of leaf number-altered mutants in Arabidopsis thaliana.Chin Bull Bot,50(3):331-336(in Chinese with English abstract)[卢阳,龙鸿(2015).拟南芥叶片数目变化突变体对营养生长时相转变的影响.植物学报,50(3):331-336]
Ma X,Liu YG(2016).CRISPR/Cas9-based multiplex genome editing in monocot and dicot plants.Curr Protoc Mol Biol,115:16
O’Day DH(2003).CaMBOT:profiling and characterizing calmodulin-binding protein.Cell Signal,15:347-354
Poethig RS(2013).Vegetative phase change and shoot maturation in plants.Curr Top Dev Biol,105(3):125-152
Reddy AS,Day IS(2001).Analysis of the myosins encoded in the recently completed Arabidopsis thaliana genome sequence.Genome Biol,2:RESEARCH0024
Schulz P,Herde M,Romeis T(2013).Calcium-dependent protein kinases:hubs in plant stress signaling and development.Plant Physiol,163:523-530
Schwab R,Palatnik JF,Riester M,et al(2005).Specific effects of microRNAs on the plant transcriptome.Dev Cell,8(4):517-527
Tian CE,Zhou YP(2013).Research progress in plant IQmotif-containing calmodulin-binding proteins.Chin Bull Bot,48(4):447-460(in Chinese with English abstract)[田长恩,周玉萍(2013).植物具IQ基序的钙调素结合蛋白的研究进展.植物学报,48(4):447-460]
Usami T,Horiguchi G,Yano S,et al(2009).The more and smaller cells mutants of Arabidopsis thaliana identify novel roles for SQUAMOSA PROMOTER BINDINGPROTEIN-LIKE genes in the control of heteroblasty.Development,136(6):955-964
Wang JW(2014).Regulation of flowering time by the miR156-mediated age pathway.J Exp Bot,65(17):4723-4730
Xu HM,Zhang LJ,Liu Chun(2010).Study of infection Arabidopsis thaliana by Agrobacterium dipping flower method.Northern Hortic,14:143-146(in Chinese with English abstract)[许红梅,张立军,刘淳(2010).农杆菌蘸花法侵染拟南芥的研究.北方园艺,14:143-146]
Xu Y,Zhang L,Wu G(2018).Epigenetic regulation of juvenile-to-adult transition in plants.Front Plant Sci,9:1048
Yin QQ,Li M,Ding B,et al(2016).Research advances on plant calmodulin binding protein IQD.Mol Plant Breed,14(11):3224-3231(in Chinese with English abstract)[尹倩倩,李明,丁博等(2016).植物钙调素结合蛋白IQD的研究概况.分子植物育种,14(11):3224-3231]
Zeng HQ,Zhang YX,Wang S,et al(2016).Calcium/calmodulin-mediated signal transduction system in plants.Chin Bull Bot,51(5):705-723(in Chinese with English abstract)[曾后清,张亚仙,汪尚等(2016).植物钙/钙调素介导的信号转导系统.植物学报,51(5):705-723]
Zhou YP,Chen YZ,Yamamoto KT,et al(2010).Sequence and expression analysis of the Arabidopsis IQM family.Acta Physiol Plant,32:191-198