青翠可人的青苹果竹芋

<正>翠叶青枝银饰链,和露带雨惹人怜。不慕颜色不争香,只留清气在人间。——咏青苹果竹芋种了这么多年的花,青苹果竹芋是我最偏爱的盆栽观叶植物。这种模样可爱的观叶植物叶片浑圆硕大,四季常绿,时尚自然,颇有特色:夏日炎炎,它青翠欲滴,给家里添了许多凉意;数九寒冬,它仍绿意盎然,让房间里充满生机。因此,青苹果竹芋成了我家客厅里的常客。     

盆栽竹芋的养护要点及老苗快速复壮技术

针对竹芋对环境因子敏感、栽培技术要求高,易出现叶片生理病害及叶斑病、根茎腐病等病害的情况,介绍了竹芋小苗、大苗期盆栽不同生长时期栽培管理的要点.针对盆花养护与栽培中出现的秋季、冬春季普遍存在大量失去美感的盆栽竹芋老苗,通过大量养护复壮工作,总结了秋季盆栽竹芋适当重剪、夏季高温后盆栽加强遮荫与叶面喷水、长期室内摆放后的老苗适当增光控水迅速复壮技术;春季分盆换土、保湿、保温促新芽萌发的低温冬季后的竹芋盆花彻底复壮技术.     

5个观赏竹芋品种在昆明的引种初报

对5个观赏竹芋品种猫眼竹芋（Calathea_Veitchiana）、青苹果（Calat hea rotundifolia cv.Fasciata）、紫背竹芋（Stromanthe sanguinea）、双线竹芋（Calathea ornate cv.Roses-lineata）和孔雀竹芋（Calathea makoyana）进行了为期二年的引种观察试验。结果表明：5个品种在昆明学院温室年生长过程中,猫眼竹芋、紫背竹芋综合性状最好,冬季表现出较强的抗冻性,双线竹芋和青苹果抗冻性最差,当室内温度低于10℃时,叶片逐渐萎蔫枯萎,有效观赏期较短。     

一月花卉主要病虫害案例诊断与处方

一、一月主要病害案例的诊断与处方 案例一：青苹果竹芋叶斑病 诊断：为害多种竹芋类观叶植物,造成盆栽竹芋观赏价值降低。该病初呈直径不到1mm水渍状的小斑点,然后转变为直径约2mm红褐色斑点,病斑周围无任何晕圈。后期病斑扩大、合并,周围有较宽且明显的黄色晕圈。病菌以菌丝体或分生孢子在土壤中、病残体上越冬．成为翌年初侵染源。     

怎样养好盆栽含笑

含笑原产福建和广东等地的常绿灌木,为木兰科含笑属.花朵新颖别致,盛开时花开不满,具有"未尝逢露齿,只恐欲倾城",每逢花开时,芳香四溢,令人陶醉.听说飘散空气中的花香,具有杀菌作用,特别对结核菌,肺炎球菌杀灭具有明显的效果.所以北方室内盆栽含笑,正值春季从春节前后开始至4、5月份开花,此时室内香气宜人养肺,是值得一养的盆栽室内花卉.     

红豆杉的养护

自从红豆杉在2010年上海"世博会"亮相以来,立即掀起栽植"红豆杉热",在室内盆栽养护需做好如下五方面.     

植物激素对青苹果竹芋继代培养的影响

以青苹果竹芋带芽的茎段为材料进行继代增殖培养,以MS培养基为基本培养基,添加不同浓度的6-BA、NAA或不同浓度的6-BA、IBA进行培养。结果表明:青苹果竹芋茎段的继代培养基以MS+6-BA 2mg/L+NAA 0.2mg/L和以MS+6-BA4mg/L+IBA 0.5mg/L为最适培养基。     

固体酸调配营养液对竹芋生长和地上部对养分吸收的影响

以"青苹果"竹芋为试材,采用盆栽种植与室内分析相结合的方法,分析了在固体酸调配营养液不同pH条件下对竹芋生长及地上部对养分吸收的影响。结果表明:在自来水配制和固体酸调配营养液pH的植株可以正常生长。与CK相比,各固体酸处理下叶面积不存在显著性差异;第106天时,除处理N1+SPA1和N2+SPA2外,CK与其它处理株高差异不显著,固体磷酸处理下植株叶绿素含量均呈增长趋势;Mo、Zn含量最高的N3+ASA3和N3+SPA3处理植株的2种元素含量相对处理CK显著降低了46.50%、23.13%,N3+SPA3处理的植株Mo含量在所有固体酸处理中最高,但积累量显著降低了45.09%。综上,该试验选取N3+SPA3为自来水配制配方肥料营养液的最佳养分浓度和固体磷酸调酸剂。     

南方夏季设施大棚圆叶竹芋盆花高效栽培技术

通过多批次对圆叶竹芋规模化栽培,对其南方夏季设施大棚的适应性、生长因子进行研究,总结出南方圆叶竹芋的配套栽培措施及技术要点,以期为圆叶竹芋的夏季南方栽培提供技术依据。     

盆栽观赏竹芋配套栽培技术

通过连续多年对竹芋引种及规模化栽培,对其适应性、生长因子以及市场接受程度进行研究,总结出竹芋的配套栽培措施及技术要点,以期为观赏竹芋的推广栽培提供技术依托。     

Ligation of the HCV 5’ NCR,C, E1, E2/NS1 sequences andits cloning into pLNSX

AIM: To construct retrovirus recombinant HCV/pLNSX (pLHC), which is used to investigate new ways to control HCV infection and gene therapy. METHODS and RESULTS: Primers with their sequences homologous to multiple strains of HCV had been designed, synthesized and used in RT-PCR to amplify 5 fragments from 4 regions, 5’ NCR, C, E1 and E2/NS1. The products had been cloned, restrictively cut aside, and overlappingly amplified to ligate a consecutive sequence of 2547 bp in length that contains the whole HCV 5’ NCR and all the structural protein coding regions. The sequence was then inserted into the vector pGEM-3Zf (+) yielding a recombinant pHC2547, and into the retrovirus pLNSX yielding a pLHC. Both plasmid DNA were analyzed by PCR, Southern blot, and enzymatic cut. CONCLUSION: Retrovirus recombinant HCV/pLNSX (pLHC) constructed successfully is useful to study both the regulation of intracellular HCV gene expression and the pathogenesis of HCV infection, as well as the molecular foundation of related transgene animals and gene therapy.     

Effects of hepatitis C virus core protein on cell cycle of HepG2 cells

AIM: To investigate the molecular mechanism of hepatitis C virus (HCV) chronic infection by studying the effect of its core protein on cell growth and the expression of cell cycle regulators such as cyclin D1 and pRb/p130 in HepG2 cells. METHODS: A eukaryotic expression vector that carried a gene encoding HCV-core-1b was constructed. The cDNA of HCV core protein was subcloned into pBabe-Flag-puro vector to generate pBabe-Flag-HCV-core-1b. The plasmid was transfected into Pheonix 293T packaging cells to produce retroviruses. The virus-containing supernatant collected from the cell culture was used to infect HepG2 cells and subsequently the cell line that stably expressed the core protein of HCV was obtained.The cell cycle was analyzed by flow cytometry. The expression of cyclin D1 and pRb/p130 was examined by Western blotting. RESULTS: The pBabe-Flag-HCV-core-1b vector was confirmed by DNA sequencing. The expression of HCV gene type 1b core protein was verified by Western blotting. The overexpression of HCV gene type 1b core protein impaired the cell cycle progression in G₀/G₁ phase and significantly reduced the levels of cyclin D1 and pRb/p130 in the cells. CONCLUSION: A eukaryotic expression plasmid that contains the cDNA of HCV core protein is successfully constructed, and a HepG2 cell line which stably expressed the core protein of HCV is also established. HCV gene type 1b core protein inhibits the cell cycle possibly through down-regulation of cyclin D1 and pRb/p130 proteins in the cells.     

HBsAg/截短型HCV核心蛋白融合基因植物表达载体的构建及对番茄的遗传转化

探索乙肝病毒表面抗原/截短型HCV核心蛋白融合基因在植物中表达乙肝丙肝双价抗原的可能性.以期为进一步研制植物乙肝丙肝双价口服疫苗打下基础。利用重组PCR技术将乙肝病毒(HBV)S基因连接在丙肝病毒(HCV)截短型核心蛋白序列的5′端,二者通过柔性肽(Gly4Ser)2序列相连,构建成-融合基因BC,测序结果正确。将融合基因BC克降到植物表达载体pBin438上,获得pBin438BC,然后用冻融法将其转入根癌农杆菌EHA105中,采用叶盘法转化番茄。获得了15株抗卡那霉素的番茄植株。对抗性植株进行PCR及Southern检测,8株获得阳性信号,表明目的基因已整合到了番茄基因组中。提取阳性植株的总蛋白.经透析后.将适当浓度蛋白质点在PVDF膜上,用Dot-ELISA方法验证番茄中表达蛋白的活性。结果表明,转基因番茄中有重组蛋白的表达。     

矮牵牛花发育相关基因PhTs2的克隆及功能分析

从矮牵牛‘幻想’中克隆了花发育相关基因PhTs2，该基因CDS长855 bp，编码284个氨基酸，具有保守的adh-short结构域。进化分析表明，PhTs2蛋白与同科植物烟草和番茄的同源蛋白亲缘关系较近。实时荧光定量PCR分析发现该基因在矮牵牛花药发育的早期特异性表达。为分析其功能，构建PhTs2超量表达载体，转化了烟草，并对转基因烟草进行了鉴定：表型观测发现其花药形态异常，花粉数量减少；花粉活力和萌发率极显著降低；扫描电镜观测发现其花粉粒畸形，外壁纹饰不规则；半薄切片显示其绒毡层发育过程紊乱。该研究为培育雄性不育系提供了潜在的基因资源。     

葡萄病毒B外壳蛋白原核表达及抗血清制备

根据已报道的葡萄病毒B(Grapevine virus B,GVB)核苷酸序列设计引物,采用RT-PCR方法从葡萄中扩增获得GVB外壳蛋白基因(cp),大小为594 bp。通过序列测定和分析,选取优势分离物GVB-JFL cp基因克隆到原核表达载体pET-28a,转化大肠杆菌BL21(DE3),IPTG诱导表达并纯化融合蛋白pET-GVB CP。SDS-PAGE电泳结果显示,融合蛋白获得过量表达,分子量约26 kD。以纯化蛋白为抗原免疫新西兰大耳白兔制备抗血清。采用间接ELISA和dot-ELISA对抗血清特异性进行检测,结果表明,抗血清可与感染GVB的西方烟和葡萄样品发生阳性反应,与健康植株及感染同属另一种病毒葡萄病毒A(Grapevine virus A,GVA)的样品不反应,阳性样品检测效价达1︰4 000,16个田间样品摩擦接种至烟草后有9个样品ELISA检测为阳性,表明制备的抗血清可用于GVB的特异性检测。     

筇竹CBF1基因的原核表达和多克隆抗体的制备

以筇竹（Qiongzhuea tumidinoda Hsueh et Yi）叶片为试材,采用RT-PCR方法克隆其CBF1基因,并将CBF1连接到原核表达载体pET32a（+）上,经克隆测序确定所构建的重组载体pET32-QZ开放阅读框正确。将重组载体pET32-QZ转化大肠杆菌Rosetta2（DE3）菌株,经IPTG诱导表达,SDS-PAGE凝胶电泳,考马斯亮蓝染色,证明CBF1蛋白得到了高效表达,所表达蛋白是大小约为45 kD的融合蛋白。经镍柱纯化后作为抗原免疫家兔,制备CBF1蛋白特异性抗血清。所制备的多克隆抗体能够与融合蛋白和经冷诱导的筇竹叶片总蛋白在25 kD处出现杂交条带。上述结果表明,表达的目的蛋白可用于免疫组织化学、蛋白质印迹检测。     

Construction and expression analysis of idiotype TCR Vβ2 DNA vaccine in vitro

AIM:To develop an anti-lymphoblastic leukemia TCR idiotypic DNA vaccine, analyze its transfer activity into K562 cells and to detect its expression in vitro. METHODS:The TCR Vβ2 gene segment, which was identified from an idiotypic TCR Vβ2 clone-Molt4 cell line, was amplified using RT-PCR, and the PCR products were then cloned into pIRES vector. The recombinant plasmids were transferred into K562 cells. The condition of idiotypic protein expression was tested by indirect immunophenotyping fluorescein dyeing, SDS-PAGE and Western blotting. RESULTS:The recombinant DNA plasmid, pIRES-TCR Vβ2, was developed successfully. The expression of TCR Vβ2 was identified on the surface of K562 cells. A 15 kD protein, which bound to TCR Vβ2 antibody specifically, were identified from pIRES-TCR Vβ2 transfected K562 cells by Western blotting, indicating that TCR Vβ2 protein was expressed in vitro. CONCLUSION:The recombinant plasmid pIRES-TCR Vβ2 DNA vaccine was developed successfully, which was expressed TCR Vβ2 protein specifically in transfected K562 cells.     

Cloning expression and toxic effect on the host bacterial viability of an outer envelope protein from the strong virulent leptospira lai in China

AIM: Construction of an eukaryote-E. coli shuttle expressing recombinant plasmid which expresses OmpL1 envelope protein of pathogenic Leptospira, serovar Lai strain 017. METHODS: The OmpL 1 gene was amplified by PCR from the leptospiral genome. Then it was cut with restriction enzymes and ligated to the plasmid pBK-CMV. The correct recombinant plasmid was screened out with analysis of restriction enzymes and PCR. After inducing the E. coli baring recombinant plasmid with IPTG,the complete protein of the bacteria was extracted for SDS-PAGE. At the same time, OD600 of the host bacteria was examined at different time after inducing or uninducing with IPTG. RESULTS: Five strains E. coli containing proper recombinant plasmids were screened out. Four strains E. coli expressed a new protein with a weight of 37 kD among them. With the expression of the heterogenous protein,the OD600 of the host bacteria decreased. CONCLUSION: The shuttle expressing plasmid of the OmpL 1 gene of strong virulent Leptospira strain 017 was successfully constructed. Furthermore,the recombinant plasmid expressed the expected OmpL 1 fusion protein in E. coli and the expression of the heterogenous protein had toxic effect on the host bacteria. This work was important for the future research of OmpL1 protein which relates to the diagnosis,new vaccine preparing and the pathogenic mechanism of leptospirosis.     

百合无症病毒16kD基因的克隆、原核表达及抗血清制备

以感染百合无症病毒(LSV)的百合叶片为试材,克隆LSV16 k D基因,连接到原核表达载体p ET-28a(+)上。将获得的重组质粒p ET-28a(+)+16 k D转化大肠杆菌BL21(DE3),经IPTG诱导得到了高效表达的16 k D蛋白,融合蛋白分子量约为20 k D。融合蛋白经过镍柱纯化后作为抗原免疫注射小鼠,制备得到16 k D蛋白抗血清。Western blot分析显示所制备的抗血清与诱导表达的融合蛋白发生特异性反应;通过ELISA检测和RT-PCR检测百合样品,证实制备的抗血清与LSV侵染的百合叶片发生了相同的特异性反应。结果表明,目的蛋白表达成功,所制备的抗血清具有特异性,可用于LSV的快速检测、免疫组织化学以及16 k D蛋白功能研究。     

Construction and expression of recombinant adenovirus vector encoding human somatostatin receptor type 2

AIM: To construct recombinant adenovirus vector carrying the gene of human somatostatin receptor type 2 （SSTR2） for gene therapy of pancreatic carcinoma.METHODS: SSTR2 cDNA was inserted into adenovirus shuttle plasmid pDC316, named pDC316-SSTR2.pDC316-SSTR2 was cotransfected with rescue plasmid pBHGlox (delta) E1, 3Cre into 293 cells by liposome reagent.Ad-SSTR2 was generated by site-specific recombination and confirmed by PCR.Ad-SSTR2 was propagated in 293 cells and purified.The titer of viral stock was determined by end-point dilution assay.Western blotting was used to determine the expression of SSTR2 protein after human pancreatic carcinoma cell capan-2 was infected with recombinant adenovirus.RESULTS: pDC316-SSTR2 was successfully constructed.Recombinant adenovirus Ad-SSTR2 was acquired by pDC316-SSTR2 and pBHGlox (delta) E1, 3Cre cotransfected into 293 cells.Ad-SSTR2 was characterized by PCR.The virus titer was 6.0×1012 pfu/L.SSTR2 protein was detected after adenovirus infected capan-2 48 h with Western blotting.CONCLUSION: The recombinant adenovirus vector encoding human SSTR2 is successfully constructed and correctly expressed in pancreatic carcinoma cells.This investigation provides the basis for study of gene therapy of pancreatic carcinoma.