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

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

Physical health care monitoring for people with serious mental illness

Background Current guidance suggests that we should monitor the physical health of people with serious mental illness and there has been a significant financial investment over recent years to provide this. Objectives To assess the effectiveness of physical health monitoring as a means of reducing morbidity, mortality and reduction in quality of life in people with serious mental illness. Search methods We searched the Cochrane Schizophrenia Group Trials Register (October 2009) which is based on regular searches of CINAHL, EMBASE, MEDLINE and PsycINFO. Selection criteria All randomised or quasi-randomised clinical trials focusing on physical health monitoring versus standard care or comparing i) self monitoring vs monitoring by health care professional; ii) simple vs complex monitoring; iii) specific vs non-specific checks iv) once only vs regular checks or v) comparison of different guidance. Data collection and analysis The authors (GT, AC, SM) independently screened search results and identified three studies as possibly fulfilling the review's criteria. On examination, however, all three were subsequently excluded. Main results We did not identify any randomised trials which assessed the effectiveness of physical health monitoring in people with serious mental illness. Authors' conclusions There is no evidence from randomised trials to support current guidance and practice. Guidance and practice are based on expert consensus, clinical experience and good intentions rather than high quality evidenc

General physical health advice for people with serious mental illness

There is currently much focus on provision of general physical health advice to people with serious mental illness and there has been increasing pressure for services to take responsibility for providing this. For the comparison of physical healthcare advice versus standard care we identified five studies (total n = 884) of limited quality. For measures of quality of life one trial found no difference (n = 54, 1 RCT, MD Lehman scale 0.00 CI -0.67 to 0.67) but another did (n = 407, 1 RCT, MD Quality of Life Medical Outcomes Scale - mental component 3.7 CI 1.7 to 5.6). There was no difference between groups for the outcome of death (n = 407, 1 RCT, RR 1.3 CI 0.3 to 6.0), for the outcome of uptake of ill-health prevention services, one study found percentages significantly greater in the advice group (n = 363, 1 RCT, MD 36.9 CI 33.1 to 40.7). Economic data were equivocal. Attrition was large (> 30%) but similar for both groups (n = 884, 5 RCTs, RR 1.18 CI 0.97 to 1.43). Comparisons of one type of physical healthcare advice with another were grossly underpowered and equivocal