特早熟春性甘蓝型油菜sBnFLD基因的克隆及表达

【目的】克隆特早熟春性甘蓝型油菜A基因组上的sBnFLD基因,并对其进行表达研究,为该基因的功能及其在成花转变中的作用研究奠定基础。【方法】根据GenBank中已报道的拟南芥和白菜型油菜FLD同源基因的保守序列设计引物,采用PCR和RT-PCR扩增特早熟春性甘蓝型油菜86号品系(光周期不敏感)的FLD同源基因,用qRT-PCR技术检测sBnFLD基因在86号品系不同发育时期茎、叶和茎尖中的表达情况。【结果】克隆出了sBnFLD基因,命名为sBnFLD,在GenBank中的登录号为KR003079.1。sBnFLD基因cDNA全长2 376bp,有3个内含子,4个外显子,编码791个氨基酸残基,分子量86.5ku,等电点8.5;sBnFLD为非分泌蛋白和非膜蛋白;sBnFLD蛋白N端有2个保守的结构域α螺旋结构域(SWIRM)和NAD(P)-binding-8结构域,该蛋白有多个α螺旋和β折叠。生物信息学分析显示,sBnFLD蛋白与已报道的甘蓝型油菜未知蛋白(CDX73929.1)和电子克隆的白菜型油菜FLD(XP_009135110.1)氨基酸序列相似性达99%,与拟南芥FLD氨基酸序列相似性达87%。qRT-PCR分析结果显示,sBnFLD基因在油菜苗期和现蕾初期茎、叶、茎尖中均有表达,但在蕾期茎尖中表达量最高。【结论】克隆出的sBnFLD基因为甘蓝型油菜的FLD同源基因,该基因在春性特早熟甘蓝型油菜开花调控中可能起着重要的调节作用。     

甜樱桃PaMADS4基因克隆及其表达分析

为研究甜樱桃在高温下胚囊发育的分子机理,以甜樱桃品种‘拉宾斯’为试材,采用RT-PCR结合RACE方法克隆胚囊发育相关的MADS-box SEP类基因c DNA全长,使用半定量PCR法进行特异性表达分析,通过温室和露地环境PaMADS4实时荧光定量分析不同温度下的表达。克隆得到一个基因全长999 bp,命名为PaMADS4(Gen Bank登录号:JQ686726),包含一个735 bp的开放阅读框,编码244个氨基酸;序列比对和进化分析表明,PaMADS4与拟南芥SEP基因家族亲缘关系最近;组织特异性分析表明PaMADS4在四轮花器官中均表达;开花后PaMADS4在露地中的表达量明显大于温室。     

太行山丘陵区不同龄级栓皮栎人工林养分循环特征

为定量分析植被恢复进程的变化特征,以太行山丘陵区的栓皮栎人工林为研究对象,在样方调查的基础上,对其各器官N、P、Ca 3种营养元素的含量、积累、分配和养分循环变化情况进行研究。结果表明：1）不同植物器官中,营养元素含量差异明显;各龄级中,营养元素含量最多的器官是树叶,树干含量最低;营养元素含量的排序为Ca〉N〉P。2）林地土壤有机质、N和P含量随龄级增加呈明显增长趋势,植被恢复能明显加快土壤肥力的提高。3）20-40 a栓皮栎人工林营养元素积累量为2764.58-3689.38 kg/hm^2,年存留量为89.88-138.17 kg/（hm^2·a）,归还量为229.80-262.06 kg/（hm^2·a）。随龄级增加,营养元素储存总量随之升高,而存留量反而降低;吸收量和归还量呈现先升高后降低的趋势;不同营养元素的储存量、吸收量、归还量和存留量的大小排列次序均为Ca〉N〉P。4）循环系数和周转时间随龄级增加而升高,利用系数则呈现下降趋势;各元素之间Ca的周转时间最长（12.73-16.19a）,利用系数最小。     

Use of quantitative real‐time PCR to assess the in vitro survival of specific DNA gene sequences of rumen microbes under simulated abomasal conditions

The use of specific DNA sequences (DS) as a microbial marker in post‐rumen digesta requires their persistence and integrity throughout gastric digestion. The aim of this study was to evaluate in vitro the survival of microbial DS during gastric digestion and the factors involved. Gastric pH had a highly significant effect on the integrity of DS. pH 4.2 allows for a significant growth of microbes in the medium, but at pH 1.2, almost all of the DS were hydrolysed. In the presence of carboxymethylcellulose, the effect of pH was reduced, pepsin activity was inhibited and gene survival increased considerably. In the simulated abomasal conditions (pH = 2.3, 2 g/l of carboxymethylcellulose, and 40‐min retention time), almost all of the bacterial genes and around 78% of the protozoa gene sequences retained their molecular integrity throughout gastric digestion, although factors such as acidity and viscera retention time might compromise the utilisation of DS as a microbial marker.     

十二烷基苯磺酸钠对五氯酚钠光解体系的影响

研究了不同浓度的阴离子表面活性剂十二烷基苯磺酸钠(SDBS)对五氯酚钠在水溶液中光解的影响。结果表明,表面活性剂不直接参与五氯酚钠的光反应;在(ρSDBS)小于其临界胶束浓度(CMC)时,SDBS对五氯酚钠的光反应有抑制作用,而ρ(SDBS)大于其CMC时,SDBS胶束的形成对五氯酚钠的光反应有加速的趋势。推测是由于ρ(SDBS)在达到CMC之前,对光解的影响以光吸收作用为主,对五氯酚钠光解有一定的抑制;而达到CMC后,胶束内部类似有机相的环境,对五氯酚钠有富集作用,从而加速五氯酚钠的光反应。     

扦插时间对四倍体刺槐嫩枝插穗内源激素和氧化酶的影响

为了解四倍体刺槐嫩枝扦插生根机制和选择适宜的扦插时间,2010年在西北农林科技大学林学院教学实验苗圃,研究了不同扦插时间对嫩枝插穗生根过程中内源激素含量和氧化酶活性的影响。结果表明：8月和7月扦插的插穗IAA含量明显高于5月和6月,较高含量的IAA是抑制生根的主要因子;ABA含量在生根过程中没有明显的抑制作用,而是起协...     

Assessing the within-herd prevalence of Coxiella burnetii milk-shedder cows using a real-time PCR applied to bulk tank milk

Thirty-seven bulk tank milk (BTM) and individual milk samples of all contributing cows were tested for Coxiella burnetii detection by a real-time PCR assay and used to assess the relationship between the BTM PCR-response and (i) the within-herd prevalence of milk-shedder cows and (ii) the proportion of heavy milk-shedder cows. The within-herd prevalence of milk-shedder cows (i) was found to be significantly higher in herds with a positive BTM and (ii) increased significantly with the estimated titre in Coxiella burnetii obtained in positive BTM. The proportion of heavy milk-shedder cows among the milk-shedder cows increased significantly with an increased estimated titre in Coxiella burnetii in positive BTM. Therefore, a real-time PCR assay applied to BTM samples collected repeatedly over time appears to be a valuable tool to assess on a larger scale the status of herds towards Coxiella shedding, and to evaluate the efficiency of control actions aimed at controlling and/or preventing Coxiella shedding in dairy herds.     

长壮蝎蝽假死行为的初步研究*

 对长壮蝎蝽(Laccotrephes robustusStl)成虫在水体内外的假死行为以及控制假死行为的关键部位进行了研究。结果表明：(1)水体外假死时长明显高于水体中;(2) 对不同捕食压力的适应导致蝎蝽在水体内外假死行为的差别,并能够提高自身的适合度;(3) 蝎蝽的假死行为并不完全受脑或某一神经节控制,胸部各体节均有独立完成假死动作的可能。     

实时荧光定量RT-PCR检测禽白血病病毒J亚群

将禽白血病病毒J亚群(ALV-J)NX0101毒株接种1日龄和7日龄SPF雏鸡并设阴性对照组,采用实时荧光定量RT-PCR方法,定期检测病毒在体内的复制情况。根据GenBank发表的ALV-Jenv基因保守序列(AY897227)设计1对特异性引物扩增目的基因;根据鸡的3-磷酸甘油醛脱氢酶(GAPDH)基因序列(K01458),在保守区内设计1对引物扩增内参照基因,分别克隆入质粒作为标准品制作标准曲线,采用SYBR GreenⅠ染料建立荧光定量PCR法,并对方法的特异性、敏感性和重复性进行评价。结果显示,标准曲线的Ct值与标准品浓度的对数值之间存在线性关系;最低每个反应可检测到60个拷贝的病毒数,比常规PCR灵敏度高1 000倍。检测结果分别采用绝对定量法和相对定量法进行分析,都达到了良好的效果。通过对病毒含量变化的检测发现,在雏鸡4周龄时,2个接毒组ALV-J病毒突然呈对数式增长。据此分析ALV-J病毒在体内经过3~4周潜伏后,突然呈暴发式增长,这种情况可能和临床表现的免疫抑制直接相关。结果表明,本试验建立了一种特异性强、敏感性高、可定量分析ALV-J病毒增殖的方法,为进一步相关研究奠定了基础。     

Inorganic nitrogen movement in tropical soils

A field experiment was conducted to follow the transformation of ammonium fertilizer applied in rainfed-upland fields. The study was done during the rainy season on a heavy-textured, poorly rained soil. Ammonium fertilizer was applied in rows, and soil samples from different depths were analyzed to check the vertical distribution of inorganic nitrogen. In order to learn the relation between nitrogen loss and nitrification, the effect of nitrification inhibitor was also checked. Ice variety IR5 was used to check nitrogen uptake.

The ammonium content decreased by about 40 per cent up to I month after fertilizing and about 90% up to 2 months after fertilizing in the unplanted plots. Up to 5 months it decreased to a level equal to or lower than that before fertilizer application. The accumulation of nitrate was observed in the lower soil layers-after 3 months, but the accumulated amount was too low to account for the loss of ammonia from the surface. The overall loss of mineral nitrogen within 60 cm of the surface was observed after 5 months in unplanted plots. The addition of nitrification inhibitor retarded the decrease of ammonium nitrogen up to 2 months. After 3 months, the nitrifying activity began to recover.

Only 23 to 27 per cent of the fertilizer nitrogen was taken up by the rice plants, indicating low efficiency.