Determination of trace alkaline phosphatase by solid-substrate room-temperature phosphorimetry based on Triticum vulgare lectin labeled with fullerenol

Fullerenol (F) shows a strong and stable room-temperature phosphorescence (RTP) signal on the surface of nitrocellulose membrane (NCM) at lambda ex max/ lambda em max =542.0/709.4 nm. When modified by dodecylbenzenesulfonic acid sodium salt (DBS), fullerenol emits a stronger signal. It was also found that quantitative specific affinity-adsorption reaction can be carried out between Triticum vulgare lectin (WGA) labeled with DBS-F and alkaline phosphatase (ALP) on the surface of NCM, and the product obtained (WGA-ALP-WGA-F-DBS) emits a strong and stable RTP signal. Furthermore, the content of ALP was proportional to the DeltaI(p) value. Based on the facts above, a new method for the determination of trace amounts of ALP by affinity-adsorption solid-substrate room-temperature phosphorimetry (AA-SS-RTP) was established, using fullerenol modified with DBS to label WGA. The detection limit was 0.011 fg spot(-1) (corresponding concentration: 2.8x10(-14) g ml(-1), namely 2.8x10(-16) mol l(-1)). This method with high sensitivity, accuracy, and precision has been successfully applied to the determination of the content of ALP in human serum survey and forecast human disease, and the results are tallied with those using alkaline phosphatase kits. The mechanism for the determination of ALP using AA-SS-RTP was also discussed.     

菠萝果实cDNA文库的构建

以菠萝幼果为研究材料,采用Creator^TMSMART^TM cDNA Library Construction Kit技术构建菠萝幼果cDNA文库,其文库的库容量为1．01×10^6,重组率为92％,PCR检测表明大多数插入片段均大于1000bp。     

西双版纳热带季节雨林优势树种树干呼吸特征

 采用红外气体分析法（IRGA）原位监测了西双版纳热带季节雨林11种优势树种树干呼吸速率、1 cm深树干温度以及林内空气变化情况。研究发 现,11种优势树种的树干呼吸具有相同的季节规律,并且雨季均大于干季时的树干呼吸。树种间树干呼吸速率差异显著,在0. 823～2.727 μmol&;#8226;m^-2&;#8226;s^-1。树干1.3 m处所测南北方向树干呼吸无显著性差异。树干呼吸与树干温度显著相关(0.552<0.92),呈良好的自然指数回归关 系,Q₁₀值为1.90～3.03。20 ℃时各树种的RT（总树干呼吸）速率为0.771～2.570μmol&;#8226;m^-2&;#8226;s^-1。     

通过根癌农杆菌介导的共转化获得具有蜡质基因反义片段的转基因水稻

在构建剔除潮霉素抗性基因的反义蜡质基因重组质粒p13W8的基础上,用分别携带p13W8质粒和有潮霉素抗性基因的pCAMBIA1300质粒的根癌农杆菌,在不同处理条件下对粳稻3个品种进行共转化.试验结果显示,共感染混合液中pCAMBIA1300菌液比例高,抗性愈伤组织的转化频率也较高;抗性愈伤组织的PCR检测结果表明,潮霉素抗性基因的阳性检测率为98%,反义蜡质基因的阳性检测率为68%.在105株转基因植株中,反义蜡质基因阳性株为30株,占转基因植株的28.6%.乙酰丁香酮处理组的平均转化频率略高于未处理组;在乙酰丁香酮的各种处理中,直接浸泡愈伤组织的转化频率稍高于其它处理方式.     

中国戈壁针茅草原的分布、群落特征和分类

戈壁针茅(Stipa tianschanica var. gobica)草原主要分布于荒漠草原区的石质丘陵或石质山坡, 向东可以分布在典型草原区的石质丘陵或山地, 向西也可以分布在荒漠区的石质山坡上。该研究调查了中国戈壁针茅草原的主要植被类型, 通过对115个样地的样方数据分析, 量化描述了这一草原类型的主要植物群落特征。结果表明, 中国戈壁针茅草原共有维管植物272种, 隶属于38科127属; 其中裸子植物3种, 隶属于1科1属; 被子植物269种, 隶属于37科126属, 物种数大于等于10种的科为: 菊科、禾本科、豆科、蔷薇科、百合科、藜科、石竹科; 戈壁针茅草原植物区系的生活型谱以地面芽植物为主, 共178种植物, 占总种数的65.44%; 水分生态类型以旱生植物为主, 共120种, 占总种数的44.12%; 植物区系地理成分上以东古北极分布种和亚洲中部分布种(含其变型)最多, 分别为82种和80种, 占总种数的30.14%和29.41%; 盖度等级划分上以0.1%-1.0%的盖度最多, 共174种, 占总种数的63.97%; 恒有度等级划分上以I级(0-20%)的物种数最多, 共258种植物, 占所有植物种数的94.85%。基于群落调查数据, 按照群落-外貌分类原则, 将戈壁针茅群系划分为8个群丛组106个群丛。     

赣南柑橘木虱体内黄龙病菌psy62株系原噬菌体类型多样性的研究

为探明柑橘木虱体内黄龙病菌psy62株系中原噬菌体遗传多样性及与柑橘株系的遗传差异,本研究将利用3对引物在psy62基因组中2个噬菌体FP1和FP2的同源序列区域内对赣南地区阳性样品进行PCR扩增和测序。结果显示,赣南地区阳性样品在此基因区域内有28头柑橘木虱检测到原噬菌体,共有5种类型,分别为A,A2,B,C,C1。其中A,B的检测率相对较高;C1检测率最低,为7.1%。由此可见,在此基因位点不同地理位置的柑橘木虱原噬菌体种群有较大的遗传差异,且与美国佛罗里达的"Ca.L.asiaticus"株系,中国柑橘叶片"Ca.L.asiaticus"株系均有显著的遗传差异,这有助于进一步研究分析原噬菌体的种类差异对柑橘木虱传毒机制的影响。     

Induction of autoreactive CD8+ cytotoxic T cells during Theiler's murine encephalomyelitis virus infection: implications for autoimmunity

Theiler's murine encephalomyelitis virus (TMEV) belongs to the family Picornaviridae and causes demyelinating disease in the spinal cords of infected mice. Although immune responses have been shown to play an important role in demyelination, the precise effector mechanism(s) is unknown. Potentially autoreactive cytotoxic cells could contribute to the destruction. We tested whether an autoreactive cell induced by TMEV infection mediated cytotoxicity by using a 5-h (51)Cr release assay in SJL/J mice. Spleen cells from TMEV-infected mice were stimulated with irradiated TMEV antigen-presenting cells and used as effector cells. The effector cells differed from conventional cytotoxic T cells since these cells could kill both TMEV-infected and uninfected syngeneic or semisyngenic cell lines (PSJLSV and BxSF11gSV) but could not kill an allogeneic cell line (C57SV). The TMEV-induced autoreactive cells were also different from conventional natural killer (NK) cells or lymphokine-activated killer (LAK) cells, because they could kill neither NK cell-sensitive YAC-1 nor NK cell-resistant P815 and EL4 cells. Induction of autoreactive cells was not detected in vaccinia virus infection. The autoreactive killing required direct cell-to-cell contact and was mediated by a Fas-FasL pathway but not by a perforin pathway. The phenotype of the killer cells was CD3(+) CD4(-) CD8(+). Intracerebral inoculation of the effector cells into naive mice caused meningitis and perivascular cuffing not only in the brain parenchyma but also in the spinal cord, with no evidence of viral antigen-positive cells. This is the first report demonstrating that TMEV can induce autoreactive cytotoxic cells that induce central nervous system pathology.     

内蒙古冰草属（禾本科）一新变种——毛稃沙芦草

报道了内蒙古冰草属一新变种——毛稃沙芦草(Agropyron mongolicum Keng var. helinicum L.Q.Zhao et J. Yang)。该变种外稃密被长柔毛,颖光滑无毛不同于沙芦草（A. mongolicum Keng）和毛沙芦草（A. mongolicum Keng var. villosum H.L.Ying）。     

FISH技术在栉孔扇贝和虾夷扇贝杂交子代染色体识别中的运用初探

核糖体间隔序列(ITS)是扇贝基因组中进化较快的基因片段,研究利用虾夷扇贝(Patinopecten yessoen-sis)与栉孔扇贝(Chlamys farreri)在核糖体间隔区ITS-1基因的序列差异,构建了虾夷扇贝的种特异性荧光原位杂交(Fluorescence In Situ Hybridization,FISH)探针,并成功地将其运用于两种扇贝杂交子代的染色体识别和遗传变异分析中.研究结果表明,在适当的杂交和洗脱条件下,两扇贝ITS-1基因20.3%的差异已足够使虾夷扇贝ITS-1探针产生明显种特异性,探针只在虾夷扇贝分裂相第3对和第5对亚端部染色体或间期细胞上产生稳定的4个杂交信号,但在栉孔扇贝上并不产生明显的杂交信号;采用该探针对两扇贝的正反交子代染色体进行分析,结果表明所有杂交子代染色体分裂相均产生半数的杂交信号,与杂交种含半数的虾夷扇贝亲本染色体相符,杂交信号在杂交种染色体卜的数目和位置保持恒定,并未产生明显的变异;该结果证实了两扇贝的杂交为真正的精卵结合意义的杂交,子代对双亲遗传物质的继承仍以单倍染色体组的叠加为主;同时研究也表明ITS-1探针可有效地运用于杂交子代中特定染色体识别和跟踪过程中,对今后扇贝遗传变异系的识别和鉴定及进一步育种具有重要意义.