Preserving Population Allele Frequencies in Ex Situ Conservation Programs

Abstract: Optimization of contributions of parents to progeny by minimizing the average coancestry of the progeny is an effective strategy for maintaining genetic diversity in ex situ conservation programs, but its application on the basis of molecular markers has the negative collateral effect of homogenizing the allelic frequencies at each locus. Because one of the objectives of a conservation program is to preserve the genetic composition of the original endangered population, we devised a method in which markers are used to maintain the allele frequency distribution at each locus as closely as possible to that of the native population. Contributions of parents were obtained so as to minimize changes in allele frequency for a set of molecular markers in a population of reduced size. We used computer simulations, under a range of scenarios, to assess the effectiveness of the method in comparison with methods in which contributions of minimum coancestry are sought, either making use of molecular markers or genealogical information. Our simulations indicated that the proposed method effectively maintained the original distribution of allele frequencies, particularly under strong linkage, and maintained acceptable levels of genetic diversity in the population. Nevertheless, contributions of minimum coancestry determined from pedigree information but ignoring the genealogy previous to the conservation program, was the most effective method for maintaining allelic frequencies in realistic situations.     

集约化柑橘种植抑制土壤磷循环微生物活性

微生物是调节土壤磷循环的关键驱动力.阐明土壤解磷菌的微生物矿化过程对于提高植物养分吸收率和作物产量具有重要意义.通过测定柑橘园与毗邻的自然林地土壤编码碱性磷酸酶基因（phoD）丰度、解磷细菌群落多样性和土壤无机磷组分,探究柑橘种植对土壤微生物获取磷策略的影响机制.结果表明,柑橘种植导致土壤pH下降,土壤有效磷累积,ω（有效磷）平均值高达112 mg·kg^-1,显著高于毗邻的自然林地（3.7 mg·kg^-1）.柑橘种植也会影响土壤磷素组成,柑橘土壤含有较高的可溶态磷（CaCl₂-P）、柠檬酸提取态磷（Citrate-P）和矿物结合态磷（HCl-P）.自然林地土壤各磷组分均显著低于柑橘土壤,而phoD基因丰度和碱性磷酸酶活性显著高于柑橘土壤.高通量测序结果表明,柑橘土壤解磷细菌Shannon指数（4.61）显著低于自然林地（5.35）,群落结构也有别于自然林地.柑橘种植改变了土壤解磷菌的群落组成,自然林地变形菌门的相对丰度显著低于柑橘土壤.土壤有效磷含量与碱性磷酸酶活性呈显著负相关,表明土壤高磷累积抑制土壤解磷细菌的活性.柑橘种植改变了土壤微生物对磷的获取策略,在柑橘园中,土壤微生物主要依赖外源磷,而自然林地土壤微生物主要以微生物分泌碱性磷酸酶矿化有机磷来获取磷的方式满足其生长需求.     

真鲷去毒相关基因cDNA序列的克隆与肝脏组成型表达分析

赤潮发生时产生的一些海洋生物毒素对人类和海洋动物的安全形成潜在的威胁甚至导致死亡.为从分子水平探讨鱼类中海洋藻毒素的去毒分子机理,采用RT-PCR法克隆了真鲷(Pagrus major)肝脏芳香烃受体核转位蛋白(ARNT)和I时相异生素代谢酶细胞色素P4501A(CYP1A)基因cDNA核心序列,同时,应用半定量RT-PCR方法,以β-肌动蛋白作为外参照,在其指数期增长的范围内研究了芳香烃受体(AHR)、ARNT、CYP1A、II时相异生素代谢酶alpha型谷胱甘肽S-转移酶(GSTA1、GSTA2)、rho型谷胱甘肽S-转移酶(GSTR)、热休克蛋白70(HSP70)基因组成型表达水平.结果发现,真鲷ARNT、CYP1A基因cDNA核心序列片段分别长438bp和908bp,分别编码146和302个氨基酸.序列同源性分析发现,真鲷与门齿鲷(Stenotomus chrysops)、石首鱼(Micropogon undulatus)ARNT基因氨基酸序列同源性高达97.2%、95.2%,与斑马鱼(Brachydanio rerio)、人、大鼠、小鼠ARNT同源性较低(77.2%～79.3%).真鲷与门齿鲷、金头鲷(Sparus auratus)、欧洲川鲽(Rhombus maximus)、欧洲海鲈(Dicentrachus labrax)CYP1A基因氨基酸序列同源性较高,为84.8%～94.0%,与斑马鱼、人、小鼠CYP1A同源性较低,为59.6%～77.8%.真鲷肝脏AHR、ARNT、CYP1A、GSTA1、GSTA2、GSTR和HSP70基因组成型表达水平分别为(25.32±6.56)%、(26.22±4.24)%、(146.5±16.06)%、(55.42±3.75)%、(48.82±10.89)%、(79.47±3.13)%、(107.42±14.34)%.     

五氯酚对土壤跳虫代谢转化酶基因和蜕皮相关基因表达的影响

五氯酚(PCP)是一种广泛存在于环境中的有机污染物。利用土壤跳虫(Folsomia candida)作为受试生物,研究了不同浓度的PCP暴露下细胞色素P450(CYP450)基因、谷胱甘肽转移酶(GST)基因和蜕皮相关基因表达水平的变化。结果显示,PCP暴露下,跳虫CYP450基因Fcc01651、Fcc02155、Fcc03650和GST基因Fcc04073、Fcc05260的表达水平未发生显著变化。在PCP浓度为240 mg·kg-1时,跳虫GST基因Fcc00494的表达显著上调。在PCP浓度为120 mg·kg-1和240 mg·kg-1时,几丁质酶基因Fcc00881和几丁质结合域基因Fcc01306的表达受到显著抑制。研究结果可为评价PCP暴露对跳虫的毒性提供一定依据。     

农杆菌介导的抗除草剂基因转入两系杂交稻恢复系的研究

利用农杆菌水稻高效转化系统,成功地将外源ＢＡＲ基因转入实用性两系要交稻恢复系Ｒ１８７和秀水０４,经过除草剂Ｂａｓｔａ田间涂布试验,其阳性率达到９０％以上。对转基因植株同步进行的ＰＣＲ特异扩增和总ＤＮＡ Ｓｏｕｔｈｅｒｎ检测结果也证实抗Ｂａｓｔａ的植株中含有ＢＡＲ基因。讨论了应用转ＢＡＲ基因的方法提高两系杂交水稻制种纯度的条件。图２表１参３     

中药材哈蟆油PCR鉴定的初步研究

从中国林蛙 (Ranachensinensis)、中华大蟾蜍 (Bufogargarizans)以及鳕鱼 (Gadusmacrocephalus)等组织材料中提取DNA ,扩增出约 340bp的 12SrRNA基因片段 ,测定其序列 ,并参考有关序列 ,设计一对哈蟆油的鉴别引物HsmL1、HsmH1,用该鉴别引物扩增蛙类原动物的模板DNA时 ,只有中国林蛙和黑龙江林蛙的模板能得到阳性扩增 ,其余样品为阴性 .因此该对引物能用于哈蟆油药材来源的鉴别 .对购自 5个不同药材商店的哈蟆油PCR鉴定结果表明 ,现市售商品哈蟆油有不同的材料来源 .图 3表 1参 9     

Control factors of partial nitritation for landfill leachate treatment

Anaerobic ammonium oxidation (ANAMMOX) technology has potential technical superiority and economical efficiency for the nitrogen removal from landfill leachate, which contains high-strength ammonium nitrogen (NH4 -N) and refractory organics. To complete the ANAMMOX process, a preceding partial nitritation step to produce the appropriate ratio of nitrite/ammonium is a key stage. The objective of this study was to determine the optimal conditions to acquire constant partial nitritation for landfill leachate treatment, and a bench scale fixed bed bio-film reactor was used in this study to investigate the effects of the running factors on the partial nitritation. The results showed that both the dissolved oxygen (DO) concentration and the ammonium volumetric loading rate (Nv) had effects on the partial nitritation. In the controlling conditions with a temperature of 30±1℃, Nv of 0.2-1.0 kg NH4 -N/(m3d), and DO concentration of 0.8-2.3 mg/L, the steady partial nitritation was achieved as follows: more than 94% partial nitritation efficiency (nitrite as the main product), 60%-74% NH4 -N removal efficiency, and NO2--N/NH4 -N ratio (concentration ratio) of 1.0-1.4 in the effluent. The impact of temperature was related to iVv at certain DO concentration, and the temperature range of 25-30癈 was suitable for treating high strength ammonium leachate. Ammonium-oxidizing bacteria could be acclimated to higher FA (free ammonium) in the range of 122-224 mg/L. According to the denaturing gradient gel electrophoresis analysis result of the bio-film in the reactor, there were 25 kinds of 16S rRNA gene fragments, which indicated that abundant microbial communities existed in the bio-film, although high concentrations of ammonium and FA may inhibit the growth of the nitrite-oxidizing bacteria and other microorganisms in the reactor.     

The Probability of a Horizontal Gene Transfer from Roundup Ready® Soybean to Root Symbiotic Bacteria: A Risk Assessment Study on the GSF Lysimeter Station

The gene transfer from glyphosate tolerant soybean to Bradyrhizobium japonicum was evaluated in a free-air lysimeter experiment under natural conditions and increasing selection pressure, to monitor for the probability of horizontal gene transfer (HGT). A large volume lysimeter study that offers conditions comparable to normal farming was conducted in 2004 and 2005 with Roundup Ready® (RR) soybean and Roundup® application according to agricultural practice. Analysis of nodules showed, as expected, the presence of the transgenic 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). However, in bacteroids that were isolated from nodules and then cultivated for several rounds in the presence of high levels of glyphosate, the EPSPS gene could no longer be detected. This indicates no stable HGT transfer of the whole EPSPS gene under field conditions.     

短小芽孢杆菌A—30耐碱性木聚糖酶基因的分子生物学研究

采用PCR方法对短小芽孢杆菌A－30菌株的耐碱性木聚糖酶基因进行克隆。在木聚糖选择平板上用刚果红染色法筛选出阳性克隆,提取阳性克隆的重组质粒进行酶切鉴定和测序。该基因在大肠杆菌中表达,过夜培养物胞外、胞内和周质空间的木聚糖酶酶活分别为0．159IUmL^-1、0.322IUmL^-1和0.007IUmL^-1。此木聚糖酶表现出较宽的pH作用范围,最适作用pH7左右,在pH9时仍有60％以上的酶活性。图4参14