Short-term population dynamics of ammonia oxidizing bacteria in an agricultural soil

Ammonia oxidizing bacteria (AOB) control the rate limiting step of nitrification, the conversion of ammonia (NH₄⁺) to nitrite (NO₂⁻). The AOB therefore have an important role to play in regulating soil nitrogen cycling. Tillage aerates the soil, stimulating rapid changes in soil N cycling and microbial communities. Here we report results of a study of the short term responses of AOB and net nitrification to simulated tillage and NH₄⁺ addition to soil. The intensively farmed vegetable soils of the Salinas Valley, California, provide the context for this study. These soils are cultivated frequently, receive large N fertilizer inputs and there are regional concerns about groundwater N concentrations. An understanding of N dynamics in these systems is therefore important. AOB population sizes were quantified using a real-time PCR approach. In a 15 day experiment AOB populations, increased rapidly following tillage and NH₄⁺ addition and persisted after the depletion of soil NH₄⁺. AOB population sizes increased to a similar degree, over a 1.5-day period, irrespective of the amount of NH₄⁺ supplied. These data suggest selection of an AOB community in this intensively farmed and C-limited soil, that rapidly uses NH₄⁺ that becomes available. These data also suggest that mineralization may play an especially important role in regulating AOB populations where NH₄⁺ pool sizes are very low. Methodological considerations in the study of soil AOB communities are also discussed.     

The influence of soil properties on the structure of bacterial and fungal communities across land-use types

Land-use change can have significant impacts on soil conditions and microbial communities are likely to respond to these changes. However, such responses are poorly characterized as few studies have examined how specific changes in edaphic characteristics do, or do not, influence the composition of soil bacterial and fungal communities across land-use types. Soil samples were collected from four replicated (n = 3) land-use types (hardwood and pine forests, cultivated and livestock pasture lands) in the southeastern US to assess the effects of land-use change on microbial community structure and distribution. We used quantitative PCR to estimate bacterial–fungal ratios and clone libraries targeting small-subunit rRNA genes to independently characterize the bacterial and fungal communities. Although some soil properties (soil texture and nutrient status) did significantly differ across land-use types, other edaphic factors (e.g., pH) did not vary consistently with land-use. Bacterial–fungal ratios were not significantly different across the land-uses and distinct land-use types did not necessarily harbor distinct soil fungal or bacterial communities. Rather, the composition of bacterial and fungal communities was most strongly correlated with specific soil properties. Soil pH was the best predictor of bacterial community composition across this landscape while fungal community composition was most closely associated with changes in soil nutrient status. Together these results suggest that specific changes in edaphic properties, not necessarily land-use type itself, may best predict shifts in microbial community composition across a given landscape. In addition, our results demonstrate the utility of using sequence-based approaches to concurrently analyze bacterial and fungal communities as such analyses provide detailed phylogenetic information on individual communities and permit the robust assessment of the biogeographical patterns exhibited by soil microbial communities.     

仿蛛网农田无线传感器网络抗毁性量化指标体系构建

为解决传统抗毁性量化指标无法准确描述网络组件失效的耦合关系和全局作用,难以有效归纳、继承蛛网抗毁性机制与规律的问题,该文提出了一套基于节点平均路径数和节点、链路平均使用次数的人工蛛网模型抗毁性量化指标体系,评测失效网络组件的全网影响度和权重等指标。仿真试验表明该指标体系可有效量化评价不同规模的人工蛛网模型的抗毁性,测评各网络组件的抗毁性权重占比,其中,节点、弦链、辐链分别占50%、39.44%、10.56%,同时与传统抗毁性量化指标相比,该文提出的指标具有独特的优势。田间试验结果表明节点遭受不同程度损坏时,仿蛛网部署仍可通过备用链路进行数据传输,相较非交叠分簇部署、栅格部署具有更优的抗毁性。人工蛛网模型抗毁性量化分析可为优化农田无线传感器网络部署,实现规模化可靠应用提供参考。     

植酸酶基因定性PCR检测方法及阳性质粒分子的构建

植酸酶基因在农业生产中具有很高的应用价值,正被广泛用于农作物的基因工程研究。为了满足转植酸酶基因作物安全监管的需要,通过优化PCR检测条件,建立了植酸酶基因特异性定性PCR检测方法。该方法具有良好的扩增特异性和检测灵敏度,可以满足我国植酸酶转基因作物监管需要。此外,我们将6大作物(小麦、水稻、棉花、大豆、玉米和油菜)的内标准基因和植酸酶基因克隆到同一载体上,构建成了阳性质粒分子pBS Endogenous-phytase。该质粒分子适用于这6大作物中植酸酶基因的筛查检测。本研究为转植酸酶基因作物的安全监管提供了阳性材料和检测方法。     

牡丹类psDHN-YSK2基因全长cDNA的克隆与表达分析

为了获得牡丹类脱水素基因的全长cDNA序列,推测其在休眠解除进程中的生物学功能,以不同低温处理时间的牡丹花芽为供试材料,末端快速扩增方法克隆全长cDNA序列,实时定量PCR分析其表达模式。结果表明牡丹类脱水素基因全长cDNA序列为1187 bp,包括831 bp的开放阅读框,114 bp的5’非编码区和242 bp的3’非编码区。其编码的蛋白具有两个植物脱水素蛋白特征性K片段,根据Close的分类方法,属于YSK2类脱水素蛋白。系统发生分析表明psDHN-YSK2基因与葡萄亲缘关系最近。在花芽内休眠解除前期随着低温处理时间的延长psDHN-YSK2表达呈上调趋势。本研究克隆了牡丹psDHN-YSK2的全长cDNA序列,分析了其在花芽内休眠解除过程的表达趋势,暗示了其参与了牡丹花芽的内休眠过程。     

基于SNP标记的玉米株高及穗位高QTL定位

为进一步弄清玉米株高和穗位高的遗传机理,为育种生产提供服务,本研究以K22×CI7、K22×Dan3402个F₂群体为作图群体,利用覆盖玉米10条染色体的SNP标记构建了2个连锁图谱。并将这2个F₂群体衍生的分别含237和218个家系的F_2:3群体用于田间性状的鉴定。用复合区间作图模型对2个群体的株高、穗位高表型进行QTL定位分析,结果显示,在武汉和南宁两种环境条件下共定位到21个株高QTL和27个穗位高QTL;单个QTL表型变异贡献率的变幅为4.9%~17.9%;株高和穗位高QTL的作用方式以加性和部分显性为主;第7染色体上可能存在控制株高和穗位高的主效QTL。     

春兰AGL6基因的克隆及实时定量表达分析

本研究采用RT-PCR结合RACE技术从春兰(Cymbidium goeringii)中分离到一个AGL6基因。序列分析表明,该基因含有一个729bp长的开放阅读框(ORF),共编码242个氨基酸。系统进化树分析显示,该基因属于MADS-box基因家族AP1/AGL9组的AGL6同源基因,其编码的蛋白与其它植物AGL6类蛋白具有较高的同源性,命名为CgAGL6(基因登录号为HM208533)。实时荧光定量表达分析表明,CgAGL6基因春兰不同组织中均有表达,其中在唇瓣、花芽和子房中的表达量较高,在花瓣和萼片中的表达量次之,在根、叶和蕊柱中的表达量最低,显示了CgAGL6基因可能在春兰成花转变和花器官的形成过程中起着重要作用。     

芝麻油纯度快速定性、定量方法的研究及应用

通过发现芝麻油通常特质发生的异常变化和市场反常规贸易情况,确立了全新、快速、行之有效的芝麻油纯度定性方法;根据Villavecehia试验原理,研究建立了简便、灵敏、准确、特别快速的芝麻油纯度定量方法.对定性、定量法的原理、特质判定、测定方法和多年实际应用情况进行了详细的阐述.     

猪圆环病毒Ⅱ型PCR检测方法的研究

【目的】建立快速、简便、特异的检测猪圆环病毒II型的PCR方法;【方法】根据已发表的猪圆环病毒II型基因序列设计合成了一对引物,用酚-氯仿法抽提可疑病料中的病毒DNA,应用PCR技术对猪圆环病毒进行基因扩增,PCR产物进行序列测定;以猪瘟病毒、猪伪狂犬病毒、猪细小病毒为对照,进行特异性试验;取部分病料进行重复性试验;【结果】PCR方法扩增出了长度为702bp的片段,扩增产物经测序和序列分析表明扩增的序列为PCV2序列;猪瘟病毒、猪伪狂犬病毒、猪细小病毒的PCR扩增均为阴性,与预期结果一致;部分病料PCR重复检测5次,检测结果完全一致;【结论】PCR检测方法可以对猪圆环病毒病作出敏感、特异、准确地诊断,该方法检测的基因最低模板量为2×10-5ng/ml。     

富含多糖的转基因石斛基因组DNA提取方法(英文)

在石斛兰转基因研究中,需通过PCR和Southern杂交等手段检测外源基因是否转入并整合到转化植株基因组.由于转化石斛植株生长缓慢,且含有多糖,因此从转化植株中提取高质量的基因组DNA以尽快对转基因苗进行分子检测存在较大困难.本研究旨在通过改良现有的DNA提取法(CTAB法或SDS法1,克服转化石斛植株基因组DNA提取中碰到的产量低,纯度不高而导致难以进行PCR或酶切等问题.在本研究所用的3种改良法中,方法Ⅱ能从少量的转化石斛苗中提取出高产量和高纯度的基因组DNA.研究结果表明方法Ⅱ提取的基因组DNA完全适用于转基因石斛的外源基因PCR扩增,限制性酶切和Southern杂交分析.