Molecular Cloning and Its Transformation of ACC Synthase NtACS1Gene from Narcissus tazetta var.Yunxiang'

为探究多花水仙ACS基因的序列特征及功能,以云香'水仙盛花期花瓣为试验材料,根据云香'水仙花朵转录组数据信息,通过RT-PCR方法克隆出1个AC; S基因,命名为NtACS1(GenBank; KX082936);NtACS1开放阅读框(ORF)长度为552bp,编码183个氨基酸。编码蛋白质分子量约为20.6KDa,理论等电点为6.3; 0,不稳定系数为65.49,属于不稳定的疏水性蛋白。通过qRT-PCR对云香'水仙不同时期花瓣和副冠中的NtACS1基因进行了表达分析,得到与云; 香'水仙花朵转录组数据中相同的结果:NtACS1基因在云香'水仙花瓣和副冠中的表达都是随着花衰老过程呈现逐渐下降的趋势,且NtACS1基因在花瓣; 和副冠中的表达峰值都在花苞期,表明; NtACS1基因编码的蛋白是在乙烯生物合成途径的系统1发挥催化作用的ACC合成酶。成功构建了NtACS1基因的正义植物表达载体,并通过农杆菌介导; 法获得8株转基因烟草,PCR和RT-PCR检测显示其中有6株为阳性植株,初步证实NtACS1基因已导入烟草基因组中且在烟草中已表达。该研究结果为; 进一步分析NtACS1基因的功能和后续转化水仙延长其花期研究奠定了基础。Aimed to study the characteristics and functions of ACSgene,in the; present study,we cloned a 1-aminocyclopropane-1-carboxylate synthetase; gene named NtACS1(GeneBank KX082936)based on the RNA-Seq database from; the flower of Narcissus tazetta var.Yunxiang'using RT-PCR method.The; length of the open reading frame(ORF)of ACSis 552bp,encoding 183amino; acids coupled with a molecular weight of 20.6kDa and theoretical; isoelectric point of 6.30.qRT-PCR analysis showed that the relative; expression level of NtACS1both in petals and coronas are decreased; gradually along with the aging of flower.Moreover,the expression data of; NtACS1gene were consistent with those obtained by RNA-Seq, implied that; the NtACS1protein as an ACC synthetase might play a role in the; catalytic system 1 of ethylene biosynthesis.Furthermore,sense plant; expression vectors of NtACS1were successfully constructed with; agrobacterium mediated transformation,and 6positive transgenic tobacco; plants were ultimately obtained. Our current study will lay an; experimental foundation for the future application of the genetic; transformation to prolong florescence ofYunxiang'.福建省种业创新与产业化工程项

Isolation and Function Analysis of NtWRKYY1Transcription Factor Gene in Narcissus tazetea var.Yunxiang

为明确WRKY转录因子在云香水仙中的功能,该研究以云香水仙为材料,克隆了WRKY基因,命名为NtWRKYY1(GenBank登录号为KX0564; 95)。序列分析显示,NtWRKYY1基因开放阅读框(ORF)长度为510; bp,编码169个氨基酸。多序列比对和系统进化树分析显示,NtWRKYY1编码蛋白含1个WRKY结构域和C_2H_2锌指结构(Cx_4Cx_(2; 3)HxH),与AtWRKY57聚在一起,属于第Ⅱc类WRKY转录因子。组织表达和时空表达分析显示,NtWRKYY1基因在花中表达量高于根和叶,; 且在花瓣及副冠中的表达量随开花过程(花蕾期、始花期、盛花期、衰败期)呈上升趋势。植物激素和非生物胁迫分析显示,NtWRKYY1基因受脱落酸(AB; A)、高温、干旱和盐诱导,受茉莉酸甲酯(JA)抑制,表明NtWRKYY1基因可能在云香水仙花朵的衰老过程中起正调节作用,同时参与云香水仙ABA、; JA等激素信号转导及高温、干旱、盐碱等非生物胁迫过程的调控。利用In-Fusion克隆技术成功构建过表达载体pMDC140-NtWRKYY1,并; 采用农杆菌介导叶盘法转化烟草。RT-PCR和GUS染色结果显示,目的基因已成功导入烟草基因组中。WRKY transcription factors play an important adjusting role in the; process of plant growth and development,hormone signal transduction and; abiotic stress response.To clarify the gene Narcissus tazetea; var.Yunxiang,we cloned the NtWRKYY1gene(GenBank accession; No.KX056495)based on Yunxiang,analyzed the gene sequence; features,evolutionary relationship and expression characteristics,; constructed the expression vector before transformed into; tobacco.Sequence analysis revealed that the length of NtWRKYY1gene open; reading frame(ORF)is 510bp,encoding apolypeptide of 169amino; acids.Multiple alignments and phylogenetic analysis showed that; NtWRKYY1protein containing one WRKY consecutive structural domain and; C_2H_2type zinc finger(Cx_4Cx_(23)HxH)belong toⅡc sub-group of WRKY; transcription factor together with Arabidopsis AtWRKY57.Tissue-specific; expression and temporal and spatial expression showed that; NtWRKYY1inYunxianghad a much higher expression in flowers than that in; roots and leaves,and a rising express trend in petal and corona of; alabastrum stage,early flowering stage,full bloom stage and faded; stage.NtWRKYY1can be induced by abscisic acid(ABA), high; temperature,drought and saline,and restrained by methyl; jasmonate(JA)through hormones and abiotic stresses analyzing.We can; concluded that NtWRKYY1gene may play a regulating role during the flower; senescence process inYunxiang,involve in the hormone signal transduction; of ABA,JA and the abiotic stress regulation of high temperature,drought; and saline at the same time.In addition,we constructed overexpressing; vector pMDC140-NtWRKYY1using In-Fusion cloning technique,transformed; into tobacco by the method of Agrobacteriumthough leaf disc; transformation.The carrier of PCR and GUS staining results indicated the; resistant plantlets were positive.This study will make a good foundation; for further exploring the function of the WRKY transcription factor; inYunxiang.福建省种业创新与产业化工程项

中国典型地区人文—经济地理研究进展与展望[J]

城市密集区、老工业基地、陆疆、山区是中国主要的地域类型,也是人文—经济地理学研究"人—地关系地域系统"形成规律、驱动力和作用机制、演化过程与空间格局的重要对象。伴随全球化、工业化、城市化、信息化的快速发展,不同地区的社会经济与自然系统之间的匹配和适应关系正发生着深刻而剧烈的变化,加强上述典型地区的研究既是人文—经济地理学科发展的内在要求,也是强化学科在国家和区域发展中的服务职能的必然要求。本文分析了中国城市密集区的空间格局及其演化,城市老工业区改造,边境城镇集聚扩散的动力机制,山区聚落空间、文化景观及其环境与山区资源环境承载力等方面的最新研究进展;总结了在国家重大地域空间规划和战略咨询中发挥的..