增氧方式对水芹菜-微生物联合作用处理冬季养殖废水的影响

为探究不同增氧方式对水芹菜-微生物联合作用处理养殖废水消化液及微生物群落结构变化的影响,设置循环和曝气两组试验,以静置处理作为对照,共运行35 d,间隔7 d取样,测定常规水质指标,在试验开始和结束时分别测定水芹菜生长情况,并采用高通量测序技术对微生物群落结构进行分析。结果显示,经过35 d的处理,静置组、曝气组和循环组的DO（溶解氧）分别达到0.60、10.38 mg·L^-1和10.85 mg·L^-1;静置培养的水芹菜长势最好,相对生长速率为0.03 g·g^-1·d^-1;三种处理方式对水质的净化效果差异显著,循环组的水质净化效果最好,COD（化学需氧量）、NH₃-N（氨氮）和TN（总氮）的去除率分别达89.89%、69.40%和77.65%;黄杆菌属（Flavobacterium）的相对丰度从43.21%下降到0.16%,克里斯滕森菌属（Christensenellaceae）和瘤胃球菌属（Ruminococcaceae）的相对丰度则大幅增加。水芹菜在冬季于养殖废水消化液中可以正常生长,曝气和循环处理可以明显提高净化效率;黄杆菌属和假单胞菌属（Pseudononas）是脱氮的功能菌属,梭菌纲（Clostridia）的大量繁殖有利于COD的去除。     

不同磷肥用量对水稻土有机碳矿化和细菌群落多样性的影响

以水稻田田间定位试验为研究对象,利用三维荧光光谱技术(3D EEMs)和454测序技术,采集4个施磷水平(0、30、60、90 kg hm-2a-1)土壤,测定其有机碳矿化、溶解有机碳(DOC)组成和结构特征及细菌群落结构和丰度的变化。结果显示:水田施磷增加了土壤速效磷(Olsen-P),从而提高了土壤DOC含量,加速了有机碳的矿化速率和累积矿化量。3D EEMs结果表明,施磷分别显著增加了荧光指数和鲜度指数值1%~10%和3%~21%,而降低了腐殖化指数,且与土壤生化性质(Olsen-P、DOC和β-葡萄糖苷酶)具有显著相关性。说明施磷通过提高Olsen-P,促进了微生物源DOC的生成,同时降低了DOC的芳香化程度、分子量及腐殖化程度,从而提高了DOC生物可降解性。同时,施磷提高了细菌群落的丰度和多样性,特别是磷诱导了多种具有碳降解功能细菌的增加,从而加速了复杂有机碳的降解和甲烷氧化。此外,主成分分析表明稻田磷素施用量在30~60 kg hm-2a-1时对土壤有机碳矿化及细菌群落多样性的提高作用最为明显。因此,适度施磷能显著提高涉碳降解微生物的活性,从而提高DOC的生物可降解性,加速有机碳的矿化速率,促进稻田土壤有机碳循环。     

不同程度重金属污染对稻田土壤真菌群落结构的影响

为了研究土壤真菌群落结构在不同程度重金属污染中的变化,本文用Illumina Hi Seq高通量测序技术分析了苏南地区某金属冶炼厂和加工产业区的重金属污染水稻土的真菌群落结构,发现不同程度重金属污染对水稻季土壤真菌丰度和群落结构均有显著影响。经过真菌主成分分析发现,PC1影响因素对样品处理差异的贡献率是35.96%,PC2影响因素对样品处理差异的贡献率是21.48%;通过真菌冗余度分析发现,重金属Pb和Cu污染对土壤真菌群落结构的影响显著;通过对真菌属水平的相对丰度分析表明,重金属污染会显著降低敏感真菌的丰度,如被孢霉属相对丰度最高降低了87.50%、木霉属最高降低了99.46%、离壳菌属和菇属最高降低了100.00%,同时耐性真菌的相对丰度会提高,如类球囊霉属的相对丰度最高增加了98倍、四枝孢霉属最高增加了56倍、根囊壶菌属最高增加了2.62倍。综上所述,不同程度重金属污染对稻田土壤真菌群落结构有显著影响,且随着污染程度的增加,抗逆真菌相对数量和种类显著增加,敏感真菌的相对数量急剧减少,真菌群落结构随着重金属污染程度增加进一步分化。     

辐照对瓜实蝇遗传区性品系成虫肠道微生物的影响

为研究辐照对瓜实蝇肠道微生物群落多样性和结构组成的影响,以100 Gy137Cs辐照来自瓜实蝇遗传区性品系(GSS)的5、8、11和13日龄的雌、雄成虫,并采用Illumina MiSeq测序技术分别对其肠道菌群进行测序分析。结果表明,瓜实蝇雌雄成虫的肠道中存在19个主要的菌属,包括17个属名已知,1个属名未知和1个属不可培养。其中,17个已知属分别属于4个细菌门,即变形菌门(Proteobacteria)(占总量的50%~98%)、厚壁菌门(Firmicutes)(占总量的2%~47%)、拟杆菌门(Bacteroidetes)(占总量的1%~16%)和1个至今微生物界还未确定的稀少菌门(unclassified rare phyla)(总量1%)。此外,辐照降低了瓜实蝇肠道中的微生物多样性以及优势菌的丰富度,且这种影响与瓜实蝇的性别和成虫日龄无关。本研究结果为评估和修复辐照对瓜实蝇的损伤提供了参考。     

不同时期的柑橘果实中真菌种类分析

柑橘采后真菌病害发生普遍,对柑橘产业造成了极大的威胁。柑橘体内存在丰富的真菌类群,为了了解柑橘成熟和衰老过程中果实内生真菌的多样性和变化动态,本论文通过MiSeq技术解读真菌ITS区域序列,分别对生长期、成熟期和贮藏期的晚蜜一号果实的果皮和果肉的内生真菌种类进行了分析。发现相比于果肉,果皮中真菌在目和属水平的多样性更为丰富,而成熟期果肉中真菌种类多于果皮。不同生长阶段的两种组织中优势的真菌属也不同,果皮在生长期以Medicopsis和刺盘孢属(Colletotrichum)占绝对优势,成熟期为刺盘孢属,贮藏期以葡萄孢属(Botrytis)、Erythrobasidium和Strelitziana较为常见;而果肉中生长期主要为青霉属(Penicillium)和枝孢菌属(Cladosporium),约占1/4,成熟期半数以上为葡萄孢属,及至贮藏期约90%为链格孢属(Alternaria)和青霉属,其中链格孢属约占2/3。常见植物病原真菌如枝孢菌属(Cladosporium)、巨座壳属(Magnaporthe)、核盘菌属(Sclerotinia)、葡萄孢属(Botrytis)、白粉菌属(Erysiphe)、青霉属(Penicillium)、链格孢属(Alternaria)和镰孢霉属(Fusarium)等在果肉中的数量高于果皮。果肉中存在较大数量的常见植物病原真菌,不同属真菌在不同样品中呈不同的变化动态,暗示采后病变是多种真菌交互作用的结果,采后病害的防治可以提前至开花期。     

利用DNA池技术提高基于InDel标记的种子纯度鉴定效率

为提高基于InDel-Pyrosequencing的黄瓜杂交种纯度检测通量,降低检测成本,本研究模拟10种DNA Pooling进行PCR、Pyrosequencing及等位基因频率分析。通过TTest分析不同Pooling间等位基因频率的差异性,确定3 Pooling为种子纯度检测最适Pooling数;建立3 Pooling-InDel-Pyrosequencing标准曲线,其R2达0.999 1;根据该标准曲线,检测黄瓜杂交品种"园中王"30粒杂交种种子纯度,结果为96.67%。本研究丰富了基于InDel-Pyrosequencing的黄瓜杂交种纯度检测技术体系。     

4个不同苯甲醇含量茶树品种的转录组分析

香气是茶叶品质决定因子之一,苯甲醇作为一种芳香族醇,参与茶叶香气物质形成。本研究选用4个苯甲醇含量不同的茶树品种,分别是‘薮北’(苯甲醇含量低)、‘湘妃翠’、‘牛皮茶’(苯甲醇含量中等)和‘铁香茗’(苯甲醇含量高),通过Illumina Hiseq^TM2000高通量测序技术对上述4种茶树一芽二叶的转录组进行测序,通过Blast搜索比对,共有151 198条Unigene获得了基因注释。4种茶树在新陈代谢路径中注释的基因数最多,其中次生代谢物生物合成和萜类和聚酮化合物代谢途径可能与茶树苯甲醇合成有关,通过对4种茶树样品的Unigene与NR数据库比对,发现5条可能编码茶树苯甲醇合成途径的关键酶β-葡萄糖苷酶基因,这些相关研究为分析茶树香气功能基因提供理论指导,为香气相关候选基因的发掘提供重要依据。     

A comprehensive analysis of the response of the fungal community structure to long-term continuous cropping in three typical upland crops

Certain agricultural management practices are known to affect the soil microbial community structure; however, knowledge of the response of the fungal community structure to the long-term continuous cropping and rotation of soybean, maize and wheat in the same agroecosystem is limited. We assessed the fungal abundance, composition and diversity among soybean rotation, maize rotation and wheat rotation systems and among long-term continuous cropping systems of soybean, maize and wheat as the effect of crop types on fungal community structure. We compared these fungal parameters of same crop between long-term crop rotation and continuous cropping systems as the effect of cropping systems on fungal community structure. The fungal abundance and composition were measured by quantitative real-time PCR and Illumina MiSeq sequencing. The results revealed that long-term continuous soybean cropping increased the soil fungal abundance compared with soybean rotation, and the fungal abundance was decreased in long-term continuous maize cropping compared with maize rotation. The long-term continuous soybean cropping also exhibited increased soil fungal diversity. The variation in the fungal community structure among the three crops was greater than that between long-term continuous cropping and rotation cropping. Mortierella, Guehomyces and Alternaria were the most important contributors to the dissimilarity of the fungal communities between the continuous cropping and rotation cropping of soybean, maize and wheat. There were 11 potential pathogen and 11 potential biocontrol fungi identified, and the relative abundance of most of the potential pathogenic fungi increased during the long-term continuous cropping of all three crops. The relative abundance of most biocontrol fungi increased in long-term continuous soybean cropping but decreased in long-term continuous maize and wheat cropping. Our results indicate that the response of the soil fungal community structure to long-term continuous cropping varies based upon crop types.     

Microbial Community Changes Along a Land-Use Gradient of Desert Soil Origin

Soil harbors remarkably stabilize bacterial communities at the phylum level.However,no two soils have exactly the same structure of bacterial phyla.The structure of microbial community is strongly influenced by the type of land-use through changes in soil attributes.Using high-throughput pyrosequencing and quantitative polymerase chain reaction techniques,soil microbial community structures were investigated along a land-use gradient of 100- and 27-year farmlands,a 33-year Pinus forest,a 28-year poplar forest,and a 21-year shrubland,as well as a native desert from which all cultivated systems were converted.The results revealed that the dominant phylotypes in the native soil comprised primarily of Alphaproteobacteria,Actinobacteria,Bacteroidetes,and Firmicutes,accounting for >71.4%of the total bacterial 16S rRNA sequence reads.Changes in land-use led to a significant decrease in these dominant phylotypes down to 33.4%.In contrast,the phylotypes with low abundance,such as Acidobacteria,Chloroflexi,Nitrospira,and Gammaproteobacteria,increased sharply from 4.5%-5.9%in the native soil to 20.9%-30.2%of the total 16S rRNA gene sequences in the cultivated soils except for the soil from the shrubland.These contrasting changes in the major taxa appear to be correlated with the changes in soil attributes.For instance,bacterial and archaeal amoA genes were found to be 960-and 3 800-fold more abundant in the soil from the 100-year farmland than the native soil.The changes in numerically less dominant nitrifying phylotypes are consistent with soil inorganic nitrogen dynamics.Quantification of the 16S rRNA genes demonstrated that bacteria and archaea were about two to three orders of magnitude more abundant in the cultivated soil than in the native soil.Hence,land-use type affects the soil bacterial community structure,which has profound consequences on ecosystem function.