近年来全球极端低温天气频发,严重影响了茶树的产量和品质。ICE(Inducer of CBF expression)基因家族主要参与植物的低温胁迫响应,但在茶树领域中的相关研究还不够全面。本研究从茶树基因组中鉴定出51个茶树CsICEs基因,对其理化性质、...近年来全球极端低温天气频发,严重影响了茶树的产量和品质。ICE(Inducer of CBF expression)基因家族主要参与植物的低温胁迫响应,但在茶树领域中的相关研究还不够全面。本研究从茶树基因组中鉴定出51个茶树CsICEs基因,对其理化性质、基因结构和启动子顺式作用元件展开生物信息学分析。茶树CsICEs基因的启动子区域富含光响应、植物激素、生长发育及非生物胁迫相关顺式作用元件,其可能参与多种逆境胁迫响应。转录组分析和RT-qPCR验证结果发现,低温下CsICE43基因的表达量上升了4.24倍,其可能与茶树低温响应相关。以茶树品种‘保靖黄金茶1号’的cDNA为模板,克隆获得了CsICE43基因,其在不同组织中的表达模式存在差异,在顶芽和嫩叶中特异性高表达。蛋白氨基酸序列和系统进化树分析表明,CsICE43基因包含与ICE家族其他成员一致的S-rich、bHLH、ACT等保守结构域,且与毛花猕猴桃(Actinidiaeriantha)的亲缘关系较近。在STRING在线网站中以拟南芥AtICEs为模型,推测茶树CsICE43蛋白与HOS1、MYB15、DREB1/2存在潜在的互作关系。亚细胞定位试验表明CsICE43定位于细胞核,与跨膜结构分析结果一致。综上所述,本研究发现CsICE43基因可能与茶树低温响应关联,为深入挖掘其基因功能与抗寒分子机理提供了一定的理论基础。展开更多
Objective:To anatomically and phenotypically characterize the insular cortex(IC)-nucleus tractus soli-tari(NTS)neural pathway.Methods:Adult male Sprague-Dawley rats were divided into three experimental cohorts for neu...Objective:To anatomically and phenotypically characterize the insular cortex(IC)-nucleus tractus soli-tari(NTS)neural pathway.Methods:Adult male Sprague-Dawley rats were divided into three experimental cohorts for neural circuit tracing.Anterograde labeling was achieved by injecting anterograde self-complementary adeno-associated viruses(scAAVs)into the IC.Retrograde tracing involved NTS injections of either retrograde scAAVs or FluoroGold(FG),combined with immunofluorescence histochemical staining to identify IC-originating projection neurons.For postsynaptic neurochemical phenotype characterization,IC was injected with AAV2/1-CaMKII-Cre,while a mixture of AAV2/9-Syn-DIO-mCherry and AAV2/9-VGAT1-EGFP was injected into the NTS.The rats were allowed to survive for one week following scAAVs or FG injection or four weeks after recombinase-dependent systems injection.Then the rats were sacrificed,and serial brain sections were prepared for immunofluorescence histochemical staining(brain section containing FG)and subsequent fluorescence/confocal microscopic analysis.Results:(1)Anterograde viral tracing re-vealed dense axonal terminals from the IC projecting to the medial subnucleus of the NTS,while retrograde tracing re-vealed that IC neurons projecting to the NTS were predominantly localized within the dysgranular layer;(2)IC-NTS projection neurons were exclusive glutamatergic(100%,n=3);(3)NTS neurons receiving IC inputs were mainly lo-calized in the medial subnucleus,and were predominantly GABAergic(79.8±3.2%,n=3).Conclusion:The pres-ent results indicate that a descending pathway from excitatory neurons of the IC terminates onto inhibitory neurons of the NTS,which might represent a potential neuromodulatory target for visceral pain disorders.展开更多
文摘近年来全球极端低温天气频发,严重影响了茶树的产量和品质。ICE(Inducer of CBF expression)基因家族主要参与植物的低温胁迫响应,但在茶树领域中的相关研究还不够全面。本研究从茶树基因组中鉴定出51个茶树CsICEs基因,对其理化性质、基因结构和启动子顺式作用元件展开生物信息学分析。茶树CsICEs基因的启动子区域富含光响应、植物激素、生长发育及非生物胁迫相关顺式作用元件,其可能参与多种逆境胁迫响应。转录组分析和RT-qPCR验证结果发现,低温下CsICE43基因的表达量上升了4.24倍,其可能与茶树低温响应相关。以茶树品种‘保靖黄金茶1号’的cDNA为模板,克隆获得了CsICE43基因,其在不同组织中的表达模式存在差异,在顶芽和嫩叶中特异性高表达。蛋白氨基酸序列和系统进化树分析表明,CsICE43基因包含与ICE家族其他成员一致的S-rich、bHLH、ACT等保守结构域,且与毛花猕猴桃(Actinidiaeriantha)的亲缘关系较近。在STRING在线网站中以拟南芥AtICEs为模型,推测茶树CsICE43蛋白与HOS1、MYB15、DREB1/2存在潜在的互作关系。亚细胞定位试验表明CsICE43定位于细胞核,与跨膜结构分析结果一致。综上所述,本研究发现CsICE43基因可能与茶树低温响应关联,为深入挖掘其基因功能与抗寒分子机理提供了一定的理论基础。
文摘Objective:To anatomically and phenotypically characterize the insular cortex(IC)-nucleus tractus soli-tari(NTS)neural pathway.Methods:Adult male Sprague-Dawley rats were divided into three experimental cohorts for neural circuit tracing.Anterograde labeling was achieved by injecting anterograde self-complementary adeno-associated viruses(scAAVs)into the IC.Retrograde tracing involved NTS injections of either retrograde scAAVs or FluoroGold(FG),combined with immunofluorescence histochemical staining to identify IC-originating projection neurons.For postsynaptic neurochemical phenotype characterization,IC was injected with AAV2/1-CaMKII-Cre,while a mixture of AAV2/9-Syn-DIO-mCherry and AAV2/9-VGAT1-EGFP was injected into the NTS.The rats were allowed to survive for one week following scAAVs or FG injection or four weeks after recombinase-dependent systems injection.Then the rats were sacrificed,and serial brain sections were prepared for immunofluorescence histochemical staining(brain section containing FG)and subsequent fluorescence/confocal microscopic analysis.Results:(1)Anterograde viral tracing re-vealed dense axonal terminals from the IC projecting to the medial subnucleus of the NTS,while retrograde tracing re-vealed that IC neurons projecting to the NTS were predominantly localized within the dysgranular layer;(2)IC-NTS projection neurons were exclusive glutamatergic(100%,n=3);(3)NTS neurons receiving IC inputs were mainly lo-calized in the medial subnucleus,and were predominantly GABAergic(79.8±3.2%,n=3).Conclusion:The pres-ent results indicate that a descending pathway from excitatory neurons of the IC terminates onto inhibitory neurons of the NTS,which might represent a potential neuromodulatory target for visceral pain disorders.
