Evaluation of fluorochromes for imaging bacteria in soil

The distribution of inoculated Escherichia coli cells and indigenous bacteria in a silt loam soil and a sandy soil was studied using fluorescence microscopy techniques. Ethidium bromide stained inoculated cells against the soil and resin background. Satisfactory results were also obtained with epifluorescence microscopy of samples stained with calcofluor white M2R (CFW) and 4′,6-diamidino-2-phenylindole (DAPI). Indigenous soil bacteria were visualized in soil thin sections, after staining with ethidium bromide, using fluorescence microscopy. The density of these bacteria was estimated to be 10⁷-10⁸ cells/cm³. Inoculated E. coli cells, stained with one of the green fluorochromes (fluorescein isothiocyanate (FITC), 5-(4,6-dichlorotriazinyl)aminofluorescein, or eosin Y), could be clearly distinguished in sandy soil thin sections. Confocal laser scanning microscopy with 3D image reconstruction was also successfully applied to characterize distribution of E. coli introduced to soil. The fate of introduced bacteria and the location of indigenous bacteria in soil can be confirmed using the microscopic techniques described in this paper.     

基因芯片技术检测5种马病毒

通过分子克隆技术获得马疱疹病毒1型(Equineherpesvirus1,EHV1)、马动脉炎病毒(Equinearteritisvirus,EAV)、马流感病毒(Equineinfluenzavirus,EIV)、马传染性贫血病毒(Equineinfectiousanaemiavirus,EIAV)和东部马脑脊髓炎病毒(East-ernequineencephalomyelitisvirus,EEEV)等5种病毒各一段高度保守的特异性基因片段,用芯片点样仪逐点分配到处理过的玻片上,制备成检测芯片。提取样品中的RNA,进行反转录和荧光标记后滴加到芯片上进行特异性杂交,对杂交结果进行扫描检测和计算机软件分析。结果显示,制备的基因芯片可同时检测和鉴别上述5种病毒,可检测到阳性杂交信号的最高稀释度为10-6的病毒液,约25个病毒DNA拷贝,但其它病毒材料未见红色荧光信号,证明了本方法的特异性。在进口马的隔离检疫期间,采集马鼻肺炎、马动脉炎中和抗体阳性但病毒分离阴性马匹的白细胞悬液,分别在EHV1和EAV位点处可检测到阳性杂交信号。证明基因芯片技术不但快速、准确和敏感,而且可同时进行多种病毒的检测。     

~(60)Coγ射线诱变与杂交相结合选育花生新品种——花育22号

花育22号是山东省花生研究所采用60Coγ射线诱变处理与杂交相结合的方法育成的高产、优质、多抗及适应性广的传统出口型大花生新品种。该品种内在品质与鲁花10号、花17、8130相当,外观品质特别是籽仁色泽优于同类品种,产量水平高于大花生主栽高产品种鲁花11号、海花1号、鲁花9号等品种。该品种的生育期比同类品种短10~15d,且抗旱耐涝性及抗病性等综合抗逆性状较为平衡,是传统出口型大花生新品种选育的一个突破,可在我国北方大花生产区广泛推广利用。     

Grazing of a common species of soil protozoa (Acanthamoeba castellanii) affects rhizosphere bacterial community composition and root architecture of rice (Oryza sativa L.)

We performed a controlled experiment with rice seedlings (Oryza sativa L.) growing in Petri dishes on homogeneous nutrient agar containing a simple rhizosphere food web consisting of a diverse bacterial community and a common soil protozoa, Acanthamoeba castellanii, as bacterial grazer. Presence of amoebae increased bacterial activity and significantly changed the community composition and spatial distribution of bacteria in the rhizosphere. In particular, Betaproteobacteria did benefit from protozoan grazing. We hypothesize that the changes in bacterial community composition affected the root architecture of rice plants. These effects on root architecture affect a fundamental aspect of plant productivity. Root systems in presence of protozoa were characterized by high numbers of elongated (L-type) laterals, those laterals that are a prerequisite for the construction of branched root systems. This was in sharp contrast to root system development in absence of protozoa, where high numbers of lateral root primordia and short (S-type) laterals occurred which did not grow out of the rhizosphere region of the axile root. As a consequence of nutrient release from grazed bacteria and changes in root architecture, the nitrogen content of rice shoots increased by 45% in presence of protozoa. Our study illustrates that interactions over three trophic levels, i.e. between plants, bacteria and protozoa significantly modify root architecture and nutrient uptake by plants.     

microRNA-let-7家族在约克夏猪甲状腺生长发育过程中的表达分析

miR-let-7广泛参与动物组织器官发育调控,是研究猪甲状腺发育的重要候选miRNAs。为了研究miR-let-7家族在约克夏猪(Sus scrofa)甲状腺生长发育过程中的作用,本实验一方面通过对不同发育阶段(胚龄105,出生后60、90、150日龄)的猪甲状腺组织切片进行苏木精-伊红染色(hematoxylin-eosin staining,HE),研究甲状腺发育形态学的变化;另一方面,采用原位杂交(in situ hybridization,ISH)和荧光定量PCR技术鉴定miR-let-7家族在不同生长阶段(胚龄105,出生后60、90、150日龄)约克夏猪甲状腺中的表达。HE染色结果发现,甲状腺滤泡随着日龄的增加逐渐增大。原位杂交结果显示,miR-let-7主要位于甲状腺的滤泡上皮细胞中,并且let-7的表达在出生后有先减少后增加的态势。通过荧光定量PCR结果发现,在生长期随着日龄的增加,let-7d-5p、let-7f和let-7g的相对表达量逐渐上升,let-7c、let-7d-3p、let-7e和let-7i的表达先升后降。研究结果提示,约克夏猪甲状腺miR-let-7的表达直接或间接影响甲状腺激素水平的变化,在甲状腺生长发育中发挥了重要的作用,为深入研究猪的生长发育提供了一个新的思路。     

转基因大麦的gfp基因表达及染色体定位

为了深入探讨转基因的整合位置对基因表达的影响，对转绿色荧光蛋白基因（gfp）大麦的荧光表达和gfp基因的染色体位置进行了研究。结果表明，绿色荧光蛋白（gfp）在大麦的根尖和花粉中均有表达，四倍体的荧光表达强度大于二倍体，表现出基因的剂量效应。gfp基因插入到第6染色体（6H）短臂和另一染色体的短臂近末端。     

辣椒种间杂交花粉母细胞减数分裂行为研究

研究辣椒种间杂交花粉母细胞减数分裂行为,有助于育种工作者了解辣椒的微观发育行为并掌握辣椒种间杂交不育的具体原因。通过改良的染色体制片方法共观察了从前期I至四分孢子时期的486个花粉母细胞,发现辣椒花粉母细胞减数分裂行为中细胞质分裂类型为同时型,且染色体有多种异常行为。中期Ⅱ的细胞中有23.8%出现染色体滞后现象,另外,在前期Ⅰ、中期Ⅰ、后期Ⅰ中也存在少量的染色体异常行为。末期Ⅱ中产生微核的概率为7.0%。这一结果可以较好地解释辣椒种间杂交花粉育性和结实率低的现象,为进一步创制新的种质提供一定的理论依据。     

试论宁夏肉羊产业的发展方向

针对宁夏养羊业存在的主要问题，通过分析入世后宁夏养羊业面临的机遇与挑战，认为宁夏发展肉羊业必须实现四个转变，同时，提出了宁夏肉羊业的发展思路和对策。     

Functional analysis of cold-inducible cDNA clones in the legume Ammopiptanthus mongolicus

Functional analysis of cold-inducible cDNA clones in the legume Ammopiptanthus mongolicus