药用真菌桑黄的系统发育分析

采用PCR直接测序法,对供试菌株朝鲜桑黄(Phellinus linteus)、韩国桑黄(Phellinus linteus)、中国桑黄(Phellinus baumii)、中国桑黄火木针层孔菌(Phellinus igniarius)和斜生褐孔菌(Phaeoponus obliquus)的rDNA ITS区域进行了PCR扩增、测序及BLAST分析比对后,认为中国的药用真菌鲍氏针层孔菌(桑黄)与朝鲜桑黄、韩国桑黄是同一物种,即鲍氏针层孔菌(桑黄)(Phellinus baumii),而火木针层孔菌(Phellinus igniarius)是另一种桑黄。     

兰州大尾羊心脏型脂肪酸结合蛋白（H-FABP） 基因克隆及其同源性比较

【目的】克隆兰州大尾羊心脏型肪酸结合蛋白（H-FABP）基因全长cDNA序列,为研究绵羊H-FABP生物学作用和生产应用提供理论依据。【方法】根据已知哺乳动物H-FABP基因 cDNA 序列,设计5''和3''特异引物,运用cDNA 末端快速扩增（RACE）技术获得兰州大尾羊H-FABP基因全长 cDNA 序列。【结果】 扩增获得兰州大尾羊5''端425 bp、3''端231 bp片段和 177 bp中间片段,拼接获得748 bp兰州大尾羊H-FABP基因全长cDNA 序列（GenBank登录号：JQ780322）。 兰州大尾羊H-FABP基因ORF长 402 bp,编码 133 个氨基酸。核苷酸序列分析显示兰州大尾羊H-FABP基因序列与大多数哺乳动物相似,但其第66位发生的碱基转换（T←→G）引起所编码的第22位天门冬氨酸（N）不同于其它所有物种的赖氨酸（K）。构建的基因进化树分析结果显示兰州大尾羊与山羊亲缘关系最近。预测兰州大尾羊H-FABP蛋白质的空间结构与山羊和牛H-FABP类似,由2个α螺旋和10个反向平行的β折叠组成,10 个折叠片围成一个桶状结构,疏水性残基位于桶内,用于结合脂肪酸。【结论】克隆了兰州大尾羊H-FABP基因,为进一步研究该基因的功能奠定了基础。     

利用基因芯片技术筛选棉纤维伸长相关基因

从回交近交系(backcross inbred lines, BIL)群体中选取纤维长度差异较大的两个系NMGA-062 (33.03 mm)和 NMGA-140 (25.87 mm),利用Affymetrix棉花基因芯片,分析其开花后10 d (DPA, days post anthesis)棉纤维伸长相关基因表达谱。在24 029条转录本中,两材料间差异表达的转录本有7 282条,占总数的30.31%;其中差异表达倍数在2倍或2倍以上的转录本有3 993条,占筛选转录本总数的16.62%,功能分类表明这些转录本主要包括功能预测基因(15.57%)、翻译、核糖体结构相关基因(13.54%)和翻译后修饰、蛋白质转换相关基因(9.29%)3大类。为了验证芯片数据的可信性,8个差异表达显著的基因(Ghi.10655.1.S1_s_at, ACO1, ARF1, SAHH, TUA6, TUA7, β-tub1, β-tub10)被用于实时荧光定量PCR。两种检测手段表现出一致性。随后,利用实时荧光定量PCR对3个与棉纤维相关基因(ARF1, β-tub1, β-tub10)在纤维发育不同时期(5、10、15、20和25 DPA)的表达模式进行了研究,结果表明,3个基因在纤维伸长发育时期(10和15 DPA)大量表达,推测这3个基因可能与棉纤维伸长有重要关系。     

Production and characterization of single chain Fv directed against beta2-agonist clenbuterol

The single chain Fv (scFv) directed against beta2-agonist clenbuterol (CBL) was produced by using phage display technology. The heavy chain and light chain variable region genes (VH) VL) were amplified by the polymerase chain reaction (PCR) from CBL specific hybridoma cell lines 5D1 and assembled as a single chain Fv (scFv) fragment with linker peptide (Gly4Ser)3. Then the scFv DNA fragment was cloned into M13 phagemid vector pCANTAB5E and the anti-CBL antibody libraries were constructed. Phages displaying scFv were enriched by panning with CBL-ovalbumin (CBL-OVA) conjugate. After only one round of panning, antigen-positive recombinant phage clones were successfully selected by ELISA. The positive phage was used to infect Escherichia coli HB2151, and the expression of soluble scFv was then induced by IPTG. The scFv showed an improved sensitivity (with IC50 of 0.78 +/- 0.005 ng/mL (n = 4)) when compared with the parent monoclonal antibody (MAb) (with IC50 of 1.34 +/- 0.006 ng/mL (n = 4)) in competitive indirect ELISA (CI-ELISA). Cross-reactivity studies showed that the specificity of scFv was similar to that of MAb. The recombinant scFv prepared in this study could be potentially used instead of conventional antisera or MAb for development of a rapid and affordable immunoassay for the detection of residual CBL in biological matrices.     

Evaluation of the combination of 1,3-dichloropropene and dazomet as an efficient alternative to methyl bromide for cucumber production in China

BACKGROUND: The combination of 1,3‐dichloropropene (1,3‐D) and dazomet (DZ) offers a potential alternative to methyl bromide (MB) for soil disinfection. MB is scheduled to be withdrawn from routine use by 2015 in developing countries. Combination treatments of 1,3‐D + DZ were evaluated in a laboratory study and in two commercial cucumber fields. RESULTS: Laboratory studies found that nearly all of the tested combinations of 1,3‐D and DZ displayed positive synergistic activity on root‐knot nematodes (Meloidogyne spp.), two major soilborne fungi (Fusarium spp. and Phytophthora spp.) and the seeds of two major weed species (Digitaria sanguinalis and Abutilon theophrasti). Field trials revealed that the combination of 1,3‐D and DZ (at 10 + 25 g m^?2) successfully suppressed Meloidogyne spp. root galling, sharply reduced Fusarium spp. and Phytophthora spp. and maintained high cucumber yields. The combination treatment of 1,3‐D + DZ was more effective than 1,3‐D or DZ alone and provided results similar to methyl bromide with respect to pest control, plant mortality, plant height, yield and income. All of the treatments were significantly better than the non‐treated control. CONCLUSION: The results indicate that the tested combination of 1,3‐D and DZ offers an efficient alternative to methyl bromide for cucumber production. Copyright © 2012 Society of Chemical Industry     

Cloning and characterization of two glutathione S-transferases from pyrethroid-resistant Culex pipiens

BACKGROUND: The Marin strain of Culex pipiens Say is a pyrethroid‐resistant population that was collected in Marin County, California, in 2001 and subsequently maintained in the laboratory under regular permethrin exposure. RESULTS: In this study, two cDNAs, CpGSTd1 and CpGSTd2, encoding glutathione S‐transferase (GST) were cloned from Cx. pipiens Marin. Phylogenetic analysis of the deduced amino acid sequences, CpGSTD1 and CpGSTD2, of these genes indicated that they belong to the Delta class of insect GSTs. The nucleotide and deduced amino acid sequences of CpGSTd1 and CpGSTd2 were 59 and 48% identical respectively. CpGSTD1 and CpGSTD2 were expressed in Escherichia coli and purified by affinity chromatography. The recombinant GSTs exhibited unique selectivity towards the general GST substrates 1‐chloro‐2,4‐dinitrobenzene (CDNB) and 1,2‐dichloro‐4‐nitrobenzene (DCNB), and also differed in their sensitivity to known inhibitors of GSTs. CpGSTD1 exhibited peroxidase activity with cumene hydroperoxide, while CpGSTD2 appeared to lack this activity. CpGSTD1 was able to metabolize 1,1,1‐trichloro‐2,2‐bis(4‐chlorophenyl)ethane (DDT), while DDT metabolism by CpGSTD2 was not detectable. CpGSTD1 and CpGSTD2 showed no detectable metabolism of permethrin. Gene expression of CpGSTd1 and CpGSTd2 in Marin mosquitoes was elevated about twofold in comparison with that found in a pyrethroid‐sensitive mosquito strain. CONCLUSION: The results indicate that CpGSTD1 and CpGSTD2 have unique biochemical characteristics, but they do not appear to play major roles in permethrin resistance in Marin mosquitoes. Copyright © 2012 Society of Chemical Industry     

紫色土坡地不同种植模式下水土和养分流失动态特征

以径流小区定位监测为基础,研究了紫色土坡地不同种植模式对水、土及养分流失特征的影响。种植试验结果表明：紫花苜蓿＋青蒿模式和连翘＋紫苏种植模式能较好地保持水土。各小区养分流失表现为：玉米／红苕模式小区地表径流全氮和有效氮含量高于其他小区,玉米／红苕模式和连翘＋紫苏模式的小区流失泥沙全氮养分富集度高于其他小区,传统农作模式玉米／红苕通过地表径流和泥沙迁移的全氮流失总量最大,可能造成土壤耕作层肥力下降。观测期间,除由于较强的生物固氮作用,紫花苜蓿＋青蒿模式增加了土壤全氮含量外,各种植模式下土壤养分均呈下降趋势,表明各小区均出现较明显的养分流失。试验证明：由于传统农作方式水土流失量最大,导致养分流失量大,对环境有较大影响。因此应注意减少传统农作方式中的地面扰动,并提高雨季地表覆盖率,从而保证农业生产的可持续发展。     

玉米叶片叶绿素含量的发育遗传动态及环境互作效应分析

采用包括基因型×环境互作效应的加性-显性遗传模型及非条件和条件的分析方法,研究了不同环境条件下玉米吐丝后不同发育时期叶片叶绿素含量性状的发育动态及其遗传发育规律.结果表明:玉米叶片叶绿素含量在不同发育时期均受到环境变化的影响.控制玉米叶片叶绿素含量的各遗传效应在不同环境条件下存在表达水平上的差异.吐丝至成熟前4个时期性状的表现是以遗传主效应(加性效应VA和显性效应VD)为主,遗传效应表达在不同环境中较为稳定,玉米籽粒成熟期该性状的表现是以环境互作效应为主,此时的遗传效应表达在不同环境中变化较大.玉米腊熟期至成熟期即玉米吐丝后40～50d是控制玉米吐丝后叶片叶绿素含量表现的微效多基因被激活表达最为活跃的时期.一些遗传效应存在发育时期间断表达的现象.