江淮地区不同轮茬作物对苜蓿产量及根际土壤质量的影响

为阐明江淮地区不同轮茬作物对苜蓿产量及根际土壤质量的影响,以苜蓿生长4年翻耕灭茬重新种植1年为对照(CK),研究了不同轮茬处理(3年苜蓿-玉米-苜蓿记作T 1,3年苜蓿-高粱-苜蓿记作T 2),利用传统法测定苜蓿产量及不同土层土壤理化性质,使用试剂盒测定根际土壤酶活性,通过高通量测序法对根际土壤细菌群落多样性进行了分析。结果表明:轮茬玉米(T 1)及轮茬高粱(T 2)后苜蓿产量分别是苜蓿连种(CK)的1.27和1.13倍;轮茬高粱使土壤pH、有机质含量显著增加(P<0.05),轮茬玉米土壤总氮含量显著增加(P<0.05),轮茬玉米及轮茬高粱显著提高了土壤速效钾含量(P<0.05);不同轮茬均能减轻土壤容重,且轮茬高粱更为显著;与苜蓿连种相比,轮茬玉米与轮茬高粱土壤过氧化氢酶及土壤脲酶活性均显著高于苜蓿连种土壤(P<0.05);借助高通量测序法对根际土壤细菌群落结构多样性分析表明,各处理土壤前3位高丰度表达优势细菌门均为变形菌、拟杆菌和厚壁菌;轮茬玉米与轮茬高粱处理根际土壤细菌种群数量显著高于CK,OTU数量分别是苜蓿连种土壤的1.25和1.39倍,且轮茬高粱处理的土壤细菌Shannon、Chao、Sobs多样性指数显著高于苜蓿连作土壤(P<0.05);而在细菌相对丰度上,苜蓿连作土壤中变形菌及厚壁菌相对丰度显著高于轮茬处理(P<0.05),而轮茬土壤中绿湾菌及浮霉菌要显著高于连作土壤(P<0.05)。综上表明,江淮地区轮茬较苜蓿连作更有助于提高土壤肥力及土壤酶活性,稳定细菌群落结构,改良土壤,提高土地生产力进而提高产量。     

健康和患病凡纳滨对虾幼虾消化道菌群结构的比较

凡纳滨对虾养殖过程中,早期阶段是病害易感阶段,而消化道菌群结构与对虾健康关系密切。因此,探讨幼虾的消化道菌群尤其是弧菌类细菌与对虾发病的关系对病害有效防治具有重要意义。本实验采用Illumina测序研究了凡纳滨对虾健康和患病幼虾的消化道细菌群落结构,并基于纯培养和16S r DNA,rec A和pyr H基因序列比对,分析了幼虾消化道中弧菌的主要种类组成。结果发现,健康幼虾消化道中α-变形菌纲和厚壁菌门丰度较高,而患病幼虾中γ-变形菌纲、拟杆菌门、放线菌门和β-变形菌纲较高,其中放线菌门丰度差异显著。基于科水平的响应比分析,发现患病幼虾消化道中动性球菌科和噬菌弧菌科的丰度显著降低,而弧菌科的丰度显著升高。相似度分析发现,驱动群落变异的OTU主要来源于弧菌属、海洋杆状菌属、冷杆菌属、假交替单胞菌属以及未分类至属的红杆菌科和微杆菌科。健康幼虾消化道弧菌组成以锡那罗亚州弧菌为主,而患病幼虾消化道弧菌组成以坎氏弧菌为主。尽管健康和患病幼虾消化道内细菌群落结构整体差异不显著,但一些重要类群丰度变化显著,其特征与已知的生态功能一致。     

长期施用尿素对东北黑土中氨氧化古菌群落的影响

【目的】表征34年施肥条件下东北黑土中氨氧化古菌（AOA）群落特征,明确不同尿素水平对其丰度和群落结构的影响,并与土壤化学性质进行关联分析,探讨引起影响东北黑土中AOA群落变化的主效环境因子,为进一步揭示黑土土壤硝化作用过程和机制,以及改良施肥方式提供依据。【方法】依托黑龙江省农业科学院34年长期定位试验,选取3组不同施肥处理：无尿素组（对照CK和磷钾处理PK）、一倍尿素组（单施一倍氮肥处理N₁,氮磷处理NP,氮磷钾处理NPK和氮钾处理NK）和二倍尿素组（单施二倍氮肥处理N₂）共7个处理的耕层土壤为研究对象,借助454高通量测序和Real-time PCR技术,以AOA Arch-amoA基因为分子标靶,解析不同尿素水平对黑土中AOA群落组成和丰度的影响,并对AOA群落结构与环境因子进行相关性分析。【结果】随着尿素施用量的增加,黑土中AOA 的Arch-amoA拷贝数由2.64×10⁷/g土壤显著降低至8.34×10⁵/g土壤,且黑土pH是引起其数量降低的直接原因;OTU水平上的聚类和非度量多维度分析（NMDS）结果均表明,相同尿素水平的各处理AOA群落结构相似,不同尿素水平处理间的AOA群落差异显著;冗余分析表明,土壤pH、水溶性有机碳和硝态氮浓度是影响土壤AOA群落变化的主效环境因子（P＜0.05）;系统发育分析表明,东北黑土中AOA主要有Nitrososphaera和Nitrosotalea两个属：无尿素组中99.3%和一倍尿素组中90.1%的Arch-amoA序列归类于Nitrososphaera,二倍尿素组中67.9%的Arch-amoA序列归类于Nitrosotalea。【结论】本研究表征了长期不同施肥下黑土中的AOA群落特征,通过与土壤化学性质耦合分析,确定了引起黑土中AOA群落变化的主要因素。长期施用氮肥的黑土中,AOA Arch-amoA拷贝数和群落结构受尿素施用量的影响显著,低倍尿素提高其多样性,而高倍尿素则使之降低,土壤pH、有机碳和硝态氮浓度是导致AOA群落差异主要因素。     

Influence of different dietary carbohydrate sources on the growth and intestinal microbiota of Litopenaeus vannamei at low salinity

Because of the increasing market potential of Pacific white shrimp (Litopenaeus vannamei) and limited availability of coastal areas for production, culture of L. vannamei at low salinity is a growing trend throughout the world. Dietary manipulation could improve the growth performance of L. vannamei at low salinity. This study reared L. vannamei with glucose, sucrose and corn starch as dietary carbohydrate sources, respectively, at a low salinity. The results indicated that the sucrose and glucose feeding groups showed better growth performance. An Illumina‐based sequencing method was used to examine the intestinal bacterial composition and the results indicated that Proteobacteria were the most prevalent members, but abundance of Actinobacteria decreased while Firmicutes increased in the corn starch‐fed group. Furthermore, bacteria related to complex carbohydrate degradation were in lower abundance, whereas the abundance of opportunistic pathogenic bacteria increased in corn starch‐fed group than the other two groups, suggesting that the diet imposes selective pressure on the intestinal microbiota. Complex carbohydrates were not the ideal energy sources for L. vannamei at low salinity because the host has higher energy demand in the stressful conditions while the complex carbohydrate degradation efficiency of the gut microbiota in L. vannamei is limited.     

基于Illumina MiSeq技术比较二种多脂鱼在腌干过程中的菌相变化

通过比较研究多脂红肉鱼(蓝圆鲹)和白肉鱼(带鱼)腌干加工中菌相的变化规律,以探讨加工过程对菌相的影响并寻找具有抗氧化作用的优势菌。在腌干加工过程中采用Illumina平台的MiSeq技术比较分析了两种多脂鱼的菌相变化情况。结果显示,两种鱼的菌相主要分布在拟杆菌门、变形菌门;在科水平上,初始原料的蓝圆鲹和带鱼分别含7个和15个科的细菌,带鱼包括了蓝圆鲹的所有菌群,肠杆菌科作为共同的优势菌,在蓝圆鲹和带鱼中分别占47%和26%。从腌制开始,两种鱼的菌群数都大量减少,弧菌和芽孢杆菌科作为共同优势菌,前者平均占蓝圆鲹和带鱼的40.3%和42.2%,后者则平均占16.7%和13.3%。原料中,蓝圆鲹和带鱼都包含了肠杆菌科、假单胞菌、弧菌科和希瓦氏菌科这4种腐败菌,加工阶段,两种鱼的优势腐败菌都为弧菌科。乳酸菌包括链球菌科和乳杆菌科,仅出现在带鱼中。研究表明,在腌干加工中,带鱼的细菌减少程度大于蓝圆鲹,总体上均呈现下降趋势,两种鱼含共同的菌群和优势菌,却表现出明显的差异。腌干后两种鱼的腐败菌大大减少,说明腌干加工有利于降低鱼类腐败的可能性。可选择带鱼作乳酸菌的分离以进行后续的抗氧化研究。     

利用Illumina高通量测序对复合重金属污染下根际和非根际稻田土壤细菌的研究

There is an increasing concern about rice (Oryza sativa L.) soil microbiomes under the influence of mixed heavy metal contamination.We used the high-throughput Illumina MiSeq sequencing approach to explore the bacterial diversity and community composition of soils in four paddy fields,exhibiting four degrees of mixed heavy metal (Cd,Pb and Zn) pollution,and examined the effects of these metals on the bacterial communities.Our results showed that up to 2 104 to 4 359 bacterial operational taxonomic units (OTUs) were found in the bulk and rhizosphere soils of the paddy fields,with the dominant bacterial phyla (greater than 1％ of the overall community) including Proteobacteria,Actinobacteria,Firmicutes,Acidobacteria,Gemmatimonadetes,Chloroflexi,Bacteroidetes and Nitrospirae.A number of rare and candidate bacterial groups were also detected,and Saprospirales,HOC36,SC-I-84 and Anaerospora were rarely detected in rice paddy soils.Venn diagram analysis showed that 174 bacterial OTUs were shared among the bulk soils with four pollution degrees.Rice rhizosphere soils displayed higher bacterial diversity indices (ACE and Chao 1) and more unique OTUs than bulk soils.Total Cd and Zn in the soils were significantly negatively correlated with ACE and Chao 1,respectively,and the Mantel test suggested that total Pb,total Zn,pH,total nitrogen and total phosphorus significantly affected the community structure.Overall,these results provided baseline data for the bacterial communities in bulk and rhizosphere soils of paddy fields contaminated with mixed heavy metals.     

水稻叶际细菌群对二氧化碳升高的不同响应特征

A vast number of microorganisms colonize the leaf surface of terrestrial plants, known as the phyllosphere, and these microorganisms are thought to be of critical importance in plant growth and health. However, the taxonomic identities and ecological functions of the microorganisms inhabiting the rice phyllosphere remain poorly understood. Using a massive, parallel pyrosequencing technique, we identified the phyllosphere bacterial taxa of four different rice varieties and investigated the microbial response to elevated CO2 （eCO2） in a rice field of a free-air CO2 enrichment （FACE） facility located in Jiangsu Province, China. The results showed that the dominant phylotype, the Enterobacteriaceae family of Gammaproteobacteria~ accounted for 70.6%-93.8% of the total bacterial communities in the rice phyllosphere. The dominant phylotype was stimulated by eCO2, with its relative abundance increasing from 70.6%-75.2% at ambient CO2 （aCO2） to 86.5%-93.8% at eCO2 in the phyllosphere of rice varieties IIYou084 （TY-084）, YangLiangYou6 （YLY-6）, and ZhenXian96 （ZX-96）. The rare phylotypes, including the bacterial taxa of Sphingobacteriaceae, Xanthomonadaceae, Oxalobacteraceae, Clostridiaceae, and Pseudomonadaceae, were suppressed and their relative abundance decreased from 13.4%-23.0% at aCO2 to 1.47% 6.11% at eGO2. Furthermore, the bacterial diversity indices decreased at eCO2 in the phyllosphere of the rice varieties TY-084, YLY-6, and ZX-96. In contrast, an opposite response pattern was observed for the rice variety of YangDao8 （YD-8）. In the phyllosphere of this variety, the relative abundance of the dominant phylotype, Enterobacteriaceae, decreased from 94.1% at aCO2 to 81.4% at eCO2, while that of the rare phylotypes increased from 3.37% to 6.59%. In addition, eCO2 appeared to stimulate bacterial diversity in the rice variety YD-8. Our results suggest that the phyllosphere microbial response to eCO2 might be relative abundance-dependent in paddy fields.     

泰山白首乌根际土壤真菌多样性分析

为了解泰山白首乌根际土壤微生物群落结构和组成,充分利用根际微生物资源,采用Illumina MiSeq高通量测序技术,分析测定来自山东省莱芜市莲花山（LW）和济南市历下区（LX）、长清区（CQ）3个产地的泰山白首乌根茎10和20 cm深度根际土壤真菌群落的丰富度和多样性。结果表明,6个样品共得到真菌1 589个OTUs,分属于11门35纲84目142科237属。子囊菌门（Ascomycota）和被孢霉门（Mortierellomycota）是6个样品共同的优势菌门。样品CQ20的群落丰度和多样性最高,样品LX20的真菌群落分布最均匀;随着地下根茎深度加深,真菌微生物群落的丰度和多样性增大。不同产地和根茎深度样品间土壤真菌群落存在显著差异,上述结果为优选根际促生菌、提高仿野生种植技术提供了理论依据。     

甘蔗根际土壤 AM 真菌群落季节性变化研究

为了探明甘蔗根际土壤 AM 真菌群落季节性变化规律及其影响因素,2016 年 4 月、8 月、10 月和 12 月在里建甘蔗试验基地,采集 4 个施肥水平（0%、25%、50%、100%）下的甘蔗根际土壤,采用高通量测序技术分析不同施肥水平下 AM 真菌群落季节变化规律。结果表明：4 个季节共检测到 10 属 AM 真菌,AM 真菌 Chaol、Shannon-Wiener 指数 Shannon-Wiener 和 Invsimpson index 指数从春季到冬季呈先降后升的趋势,夏季最低;4 个季节共检测出 281 个 AM 真菌 OTUs（operational taxonomic units）,春季、夏季、秋季、冬季的 OTUs 数目分别为160、68、103、198 个。4 个季节甘蔗根际土壤中球囊霉属（Glomus）占有绝对的优势,之后是类球囊霉属（Paraglomus）和多样孢囊霉属（Diversispora）。此外,也有少量的原囊霉属（Archaeospora）、巨孢囊霉属（Gigaspora）、无梗囊霉属（Acaulospora）、盾巨孢囊霉属（Scutellospora）和两性球囊霉属（Ambispora）。不同季节 AM 真菌群落组成差异显著,但总体来说冬季和春季种群结构相似,夏季和秋季相似,而夏季和秋季的 AM 真菌种类比春季和冬季的 AM 真菌种类多。