核黄素诱导番茄抗病性与Pti蛋白激酶基因表达

研究了核黄素诱导番茄对丁香假单胞菌番茄致病变种（Pseudom onas syringaepv.tomato）的抗性及相关防卫反应.核黄素诱导处理后,番茄对P.syringaepv.tomato的抗性增强,蛋白激酶基因Pti4,Pti5,Pti6和抗病防卫基因PR-1a,GluA,GluB,ChtA,ChtB表达量提高,水杨酸含量明显升高.蛋白激酶抑制剂K252 a可以部分抑制这些基因的表达,取消核黄素对水杨酸的诱导作用,可减弱诱导抗病性的程度.这些结果表明,核黄素诱导的抗病性受植物细胞内多种信号传导因子控制.     

PuFAD8基因过表达载体的构建及其在番茄中的遗传转化

脂肪酸去饱和酶8(fatty acid desaturase 8,FAD8)是FAD基因家族成员,可参与催化不饱和脂肪酸的合成进而影响果实香气。为进一步研究PuFAD8基因的功能及其与番茄果实挥发性物质的关系,文章构建了模式植物番茄PuFAD8基因的过表达载体,通过农杆菌介导遗传转化、植物组织培养技术获得番茄PuFAD8基因过表达植株。结果表明,番茄PuFAD8基因过表达载体构建成功,并获得番茄过表达植株,以便进一步研究该基因的分子功能,为明确其在番茄果实成熟过程中挥发性物质的调控机制奠定了基础。     

TeLCYB基因的番茄遗传转化分析

万寿菊(Tagetes erecta L.)中含有类胡萝卜素,不仅可用于天然色素提取,还是研究类胡萝卜素合成代谢的植物材料.番茄红素β-环化酶(lycopene beta cyclase,LCYB)是类胡萝卜素合成代谢途径中的一个关键酶,文章通过对万寿菊中TeLCYB基因克隆、植物表达载体构建、并利用农杆菌介导法进行番...     

农杆菌介导的抗病基因对番茄的遗传转化

通过农杆菌介导法将含有几丁质酶基因和β-1,3-葡聚糖酶基因的双价植物表达载体pBLGC转入番茄子叶外植体。经过共培养、卡那霉素筛选和分化再生,获得了23株卡那霉素抗性植株。PCR检测和Southern Blot检测结果表明,有7株呈阳性检测反应。通过病原真菌接种试验证明,有3株转基因植株表现出较强的抗病性反应。实验结果为进一步研究番茄抗病性和培育抗病番茄新品种奠定了重要基础。     

乙肝表面抗原小蛋白S基因导入番茄的遗传转化体系的建立

以丽春番茄无菌苗的子叶为外植体，以农杆菌菌株EHA105(包含质粒pBin438S)为载体，将乙肝表面抗原小蛋白S基因导入番茄组织中，基本培养基采用MS 6—BA1．0mg／L IAA0．2mg／L，在共培养时加入乙酰香酮100μmol／L作为诱导剂，经过组织培养得到转基因植株，对转化植株基因组进行的分子学PCR检测表明，外源基因有可能已整合到番茄基因组中，该转基因遗传体系的建立为利用转基因植物生产乙肝口服疫苗奠定了基础。     

番茄Pti基因瞬时表达与互作检测

丁香假单胞菌所导致的细菌性斑点病是影响番茄产量和品质的重要病害,而番茄色氨酸苏氨酸激酶(Pto)是植物识别、防御这一病原菌的重要抗性蛋白。文章通过克隆番茄Pti(Pto interaction protein)基因Pti4、Pti5和Pti6,分别构建植物表达载体,利用农杆菌介导方法在烟草叶片中进行瞬时表达和western blotting检测,并进一步利用免疫共沉淀技术,在植物体系中检测了它们与番茄Pto蛋白的相互作用。实验结果表明,利用所构建Pti4、Pti5和Pti6基因植物表达载体,在烟草中获得了预期大小Pti4、Pti5和Pti6蛋白,并在植物体系中验证了它们与Pto蛋白的相互作用。该工作为深入研究Pti基因在番茄分子免疫途径的功能奠定了基础。     

农杆菌介导的双价抗虫基因对番茄遗传转化的研究

采用农杆菌介导法将含有苏云金芽孢杆菌毒蛋白基因(CryIAc)与半夏凝集素抗虫基因(Pta)的高效植物表达载体pCAMBIA3300转入番茄品系Micro Tom的子叶外植体中。经过共培养、除草剂筛选和分化再生,获得了24个具有除草剂抗性的株系。再将转化后的番茄植株经过PCR检测和Southern Blot检测,确定检测后呈阳性反应的株系为8个。通过小菜蛾幼虫初步抗性试验证明,转基因株系表现出较强的抗虫性。实验结果为进一步研究番茄抗虫性和培育抗虫番茄新品种奠定了重要基础。     

6-苄基腺嘌呤对长根菇酶活性的影响

6—苄基腺嘌呤(6-BA)在适当浓度下，可促进长根菇的生长，通过测定长根菇相关酶类的活性变化，了解6-BA促进长根菇生长的一些机理．     

番茄CDT2同源基因RNA干涉载体的构建及遗传转化研究

真核生物CDT2是CUL4-DDB1 E3泛素复合体的组成部分,在细胞周期调控、DNA复制与损伤修复中起到重要作用.本研究克隆了番茄CDT2同源基因片段并构建了CDT2基因RNA干涉植物表达载体pBI121-CDT2-RNAi.通过根癌农杆菌介导转入番茄子叶,经组织培养成功获得转基因植株.半定量RT-PCR分析显示,转基因植株叶片内CDT2的表达量明显低于野生型植株.转基因植株叶片叶绿素含量比野生型明显升高.该研究结果揭示番茄CDT2基因的功能做出了新的尝试.     

Structural analysis of the promoter of tomato 1-aminocyclopropane-1-carboxylate synthase 6 gene(Le-ACS6)

Ethylene plays an important role in the regulation of many growth and developmental processes of higher plants. In tomato,Le-ACS6,a member of the ACC synthase multigene family involved in system 1 ethylene biosynthesis during fruit ripening,is subject to negative feedback regulation by ethylene. To identify the cis-elements that are responsible for the negative feedback control,we established an in vitro transient assay system employing particle bombardment on mature-green tomato fruit pericarp to examine the expression of a luciferase(LUC) reporter gene driven by a 5′-serially deleted Le-ACS6 promoter. The results localized putative cis-elements required for negative ethylene-response between -347 and -266 upstream from the translational start site ATG. Several lines of stable transformation of the Le-ACS6 promoter and GUS reporter fusion gene containing internal deletion from -347 to -266 were generated. The expression pattern of the GUS reporter showed that removal of the nucleotides from -347 to -266 completely eliminated the response of the Le-ACS6 promoter to exogenous ethylene.     

Genetic background of phenotypic variation

A noteworthy feature of the living world is its bewildering variability. A key issue in several biological disciplines is the achievement of an understanding of the hereditary basis of this variability. Two opposing, but not necessarily irreconcilable conceptions attempt to explain the underlying mechanism. The gene function paradigm postulates that phenotypic variance is generated by the polymorphism in the coding sequences of genes. However, comparisons of a great number of homologous gene and protein sequences have revealed that they predominantly remained functionally conserved even across distantly related phylogenic taxa. Alternatively, the gene regulation paradigm assumes that differences in the cis-regulatory region of genes do account for phenotype variation within species. An extension of this latter concept is that phenotypic variability is generated by the polymorphism in the overall gene expression profiles of gene networks. In other words, the activity of a particular gene is a system property determined both by the cis-regulatory sequences of the given genes and by the other genes of a gene network, whose expressions vary among individuals, too. Novel proponents of gene function paradigm claim that functional genetic variance within the coding sequences of regulatory genes is critical for the generation of morphological polymorphism. Note, however, that these developmental genes play direct regulatory roles in the control of gene expression.     

Genetic background of phenotypic variation

A noteworthy feature of the living world is its bewildering variability. A key issue in several biological disciplines is the achievement of an understanding of the hereditary basis of this variability. Two opposing, but not necessarily irreconcilable conceptions attempt to explain the underlying mechanism. The gene function paradigm postulates that phenotypic variance is generated by the polymorphism in the coding sequences of genes. However, comparisons of a great number of homologous gene and protein sequences have revealed that they predominantly remained functionally conserved even across distantly related phylogenic taxa. Alternatively, the gene regulation paradigm assumes that differences in the cis-regulatory region of genes do account for phenotype variation within species. An extension of this latter concept is that phenotypic variability is generated by the polymorphism in the overall gene expression profiles of gene networks. In other words, the activity of a particular gene is a system property determined both by the cis-regulatory sequences of the given genes and by the other genes of a gene network, whose expressions vary among individuals, too. Novel proponents of gene function paradigm claim that functional genetic variance within the coding sequences of regulatory genes is critical for the generation of morphological polymorphism. Note, however, that these developmental genes play direct regulatory roles in the control of gene expression.     

Structural analysis of the promoter of tomato 1-aminocyclopropane-l-carboxylate synthase 6 gene （Le-ACS6）

Ethylene plays an important role in the regulation of many growth and developmental processes of higher plants. In tomato, Le-ACS6, a member of the ACC synthase multigene family involved in system 1 ethylene biosynthesis during fruit ripening, is subject to negative feedback regulation by ethylene. To identify the cis-elements that are responsible for the negative feedback control, we established an in vitro transient assay system employing particle bombardment on mature-green tomato fruit pericarp to examine the expression of a luciferase （LUC） reporter gene driven by a 5＇-serially deleted Le-ACS6 promoter. The results localized putative cis-elements required for negative ethylene-response between -347 and -266 upstream from the translational start site ATG Several lines of stable transformation of the Le-ACS6 promoter and GUS reporter fusion gene containing internal deletion from -347 to -266 were generated. The expression pattern of the GUS reporter showed that removal of the nucleotides from -347 to -266 completely eliminated the response of the Le-ACS6 promoter to exogenous ethylene.     

番茄DNA甲基转移基因SlDRM2L的克隆及表达分析

DNA甲基化主要是通过在甲基转移酶的催化下修饰基因组DNA来响应外界环境的胁迫,是表观遗传学的重要手段之一,参与植物中DNA甲基化起始的主要DNA甲基转移酶是DRM2.文章通过美国国家生物技术信息中心(National Center for Biotechnology Information,NCBI)数据库,在番茄中...     

Genetic analysis and fine mapping of the pubescence gene GL6 in rice (Oryza sativa L.)

The pubescence of the leaf blade surface is an important agronomic characteristic for rice morphology and significantly influences rice growth as well as physiological characteristics. This characteristic was analyzed in F1 and F2 plants derived by crossing cultivar 75-1-127 with the indica cultivar Minghui 63, as well as the glabrous cultivar Lemont and indica cultivar 9311. Results indicated that the pubescence of the leaf blade surface was a dominant trait and controlled by a single gene. The GL6 gene was primarily mapped on rice chromosome 6 with recessive F2 population derived from 75-1-127/Minghui 63 by combining bulked segregation analysis and recessive class analysis using the Mapmaker3.0/MapDraw software. The genetic distances between the simple sequence repeat markers RM20491 and RM20547 were 7.2 and 2.2 cM, respectively. The GL6 gene was fine mapped in the interval between InDel-106 and InDel-115 at genetic distances of 0.3 and 0.1 cM, respectively. The large, recessive F2 population was derived from 75-1-127/Minghui 63. A high-resolution genetic and physical map of GL6 was constructed. Derived from the map-based sequences published by the International Rice Genome Sequencing Project, the GL6 gene was localized at an interval of 79 (japonica) and 116.82 kb (9311) bracketed by InDel-106 and InDel-115 within the BAC accession numbers AP008403 and AP005760. Seven annotated genes (japonica) and eight annotated genes (9311) were present. The basis was further set for GL6 cloning and function analysis.     

衣藻的核基因组转化及外源基因转录分析

通过珠磨法将衣藻表达载体pSP108转入到莱茵衣藻细胞壁缺陷型CC-400藻株中,经抗性筛选及PCR鉴定,获得了24个转化藻株.在抗性基因ble编码序列的中间部位和末端部位分别设计两对不同的探针引物,运用实时荧光定量PCR对转基因藻株进行外源基因转录水平的分析.结果发现:不同转化藻株外源基因的转录水平有明显差异;同一转化藻株两对探针引物,外源基因的转录水平也存在差异.说明莱茵衣藻在转化过程中,外源基因整合到基因组上的片段数量及片段长度和完整性具有不确定性.