不同生育期转基因抗虫棉根际土壤细菌群落特征

为深入理解转Bt基因抗虫棉对根际土壤微生物群落的影响,基于大田试验,采用高通量测序技术,研究了不同生育期(播种前期、苗期和吐絮期)转基因抗虫棉和亲本根际土壤微生物群落组成和多样性.结果表明:①转基因抗虫棉及亲本根际土壤优势菌门均为变形菌门(Proteobacteria,相对丰度为0.433 8~0.499 3)、拟杆菌门(Bacteroidetes,相对丰度为0.106 0~0.136 6)、放线菌门(Actinobacteria,相对丰度为0.074 9~0.146 4)、芽单胞菌门(Gemmatimonadetes,相对丰度为0.054 2~0.072 3)、酸杆菌门(Acidobacteria,相对丰度为0.053 8~0.089 9).不同生育期转基因抗虫棉对根际土壤细菌优势菌门相对丰度产生显著影响(P < 0.05).②不同生育期转基因抗虫棉与亲本根际土壤细菌群落的Shannon-Wiener指数、Simpson指数、Sobs指数、PD指数均无显著差异(P>0.05),但PCoA分析发现,苗期和吐絮期转基因抗虫棉和亲本的土壤微生物群落出现空间分异(P < 0.05).③转基因抗虫棉根际土壤Bt蛋白含量在不同生育期具有显著差异(P < 0.05),且苗期和吐絮期转基因抗虫棉根际土壤Bt蛋白含量显著高于亲本(P < 0.05).④根际土壤Bt蛋白含量与细菌优势菌门相对丰度呈显著相关(P < 0.05).研究显示,不同生育期转基因抗虫棉通过根际Bt蛋白对土壤细菌相对丰度及群落结构产生影响,但不影响土壤细菌的alpha多样性.      

转Cry1Ac基因抗虫棉与其亲本棉花根际真菌多样性的比较

在温室条件下,利用PCR-DGGE技术和Q-PCR技术分析了两种转Cry1Ac基因抗虫棉及其亲本棉花在不同取样时期的根际真菌多样性和生物量的动态变化情况,以评价其生态安全.结果表明,转Bt基因棉与对照棉根际真菌优势条带组成在不同的取样时期极为相似,三者间差异条带多为不清晰的条带.转Bt棉SGK321根际真菌的生物量在苗期明显低于亲本对照,转Bt棉XP188根际真菌的生物量在花铃期较对照有轻微降低,其它时期两种转Bt棉根际真菌生物量没有显著降低,甚至还有增加的趋势.不过根际真菌群落结构的改变和生物量的差异不仅存在于转基因棉和其亲本对照棉之间,它同样存在于对照棉SHIYUAN321和JM20之间以及转基因棉SGK321和XP188之间.可见Bt并不是造成转基因棉和非转基因棉根际真菌群落结构差异、多样性改变和生物量减少的唯一诱因,品种的不同也对它们也有影响.     

澎溪河消落带典型植物群落根际土壤无机氮形态及氮转化酶活性

三峡库区消落带是典型的生态脆弱带,其土壤N循环因受到植物根际效应和季节性淹水的影响而具有特殊性.本研究以三峡库区一级支流澎溪河消落带为例,选择4种植被(狗牙根、香附子、苍耳以及玉米)覆盖区,采集植物根际、非根际土壤,分析根际土壤与非根际土壤理化性质、无机氮形态以及7种N素转化相关酶,并比较了4种植物根际效应强度,以反映不同植物覆盖对消落带土壤N循环过程的影响.研究表明:供试植物根际土壤pH值均低于非根际,有机质、全氮、全磷含量均高于非根际,表明植物根际对消落带土壤养分有富集作用;4种植物根际土壤硝态氮、铵态氮、亚硝态氮及14d可矿化氮含量均高于非根际,且土壤硝态氮、亚硝态氮以及14d可矿化氮含量呈现香附子＞狗牙根＞苍耳/玉米;总体上根际土壤N转化酶活性高于非根际,且狗牙根和香附子覆盖区脲酶、亚硝酸还原酶、谷氨酰胺酶、脱氢酶显著高于玉米和苍耳覆盖区;蛋白酶、脲酶、谷氨酰胺酶活性与4种N形态均呈显著相关性,是消落带土壤N转化的主要参与酶类;根际效应分析结果香附子和狗牙根对消落带土壤N转化的根际效应强度大于苍耳和玉米,有利于土壤N素的固定和保持.植物根际效应对消落带土壤N素循环的影响可为消落带植被恢复工程中植被选择提供参考,也为改善消落带土壤退化相关研究提供科学支撑.     

胶质芽胞杆菌对印度芥菜根际土壤镉含量及土壤酶活性影响

采用根袋法盆栽试验,研究分别接种活菌量浓度为1×1010CFU.kg-1(处理A)和2×1010CFU.kg-1(处理C)的胶质芽胞杆菌,对根际土壤镉含量和土壤酶活性的影响.结果表明,处理A、C对根际土壤镉的去除率分别达到37.62%、38.27%,是对照(24.47%)的1.54、1.56倍.胶质芽胞杆菌处理后土壤酶活性均有所增强,土壤脲酶、磷酸酶、过氧化氢酶活性在根际土壤大于非根际,随着时间延长土壤磷酸酶活性逐渐升高,脲酶和过氧化氢酶呈现先增高后降低的变化趋势,而脱氢酶活性在非根际土壤大于根际土壤,也呈现先增高后降低的变化趋势.对照组及处理A、C组中根际Cd含量与土壤磷酸酶、脲酶呈现显著或极显著负相关.研究表明,施入胶质芽胞杆菌不仅可以提高超富集植物对土壤Cd的净化效果而且对土壤脲酶、磷酸酶、过氧化氢酶及脱氢酶均有一定的积极作用,其结果可为该菌辅助植物修复镉污染土壤的机制性研究提供理论指导.     

丛枝菌根真菌在矿区生态环境修复中应用及其作用效果

针对煤矿区生态环境修复中存在的主要问题,研究丛枝菌根真菌与紫穗槐共生状况,丛枝菌根真菌对紫穗槐根系发育的影响及其对煤炭开采塌陷区退化土壤的改良效应.结果表明,接种菌根5个月后,接种丛枝菌根促进紫穗槐地上部分和根系生长,提高了紫穗槐根系侵染率;接种区紫穗槐根际土壤中球囊霉素、易提取球囊霉素含量显著增加;接种菌根提高了紫穗槐根际土壤有效磷和有机质含量,微生物数量明显提高,取得较好的菌根生态效应.接种菌根有利于对矿区根际土壤的改良,促进了矿区生态系统稳定,对维持矿区生态系统的持续性具有重要意义.     

玉米和籽粒苋镉累积特征与根际微生物的关系

该研究针对北方典型污灌区农田开展大田作物玉米与镉富集植物籽粒苋的田间种植试验。通过分析土壤及植株的镉含量特征,并利用高通量测序技术,对比2种植物的根际土壤及非根际土壤的细菌多样性与群落结构,探究土壤不同镉浓度对细菌群落的影响。结果表明,研究区根际土壤总镉及有效态镉含量高于非根际土壤,玉米作物的根、茎、叶部位保留了从土壤中吸收的大部分镉,籽粒部位镉含量最低;土壤镉浓度对土壤细菌多样性与群落结构没有产生显著性影响;植物的根际微生物和土壤镉之间存在一定的交互作用;不同土壤生境细菌的物种组成存在一定差异,变形菌门(19.33%～25.73%)、奇古菌门(16.09%～24.67%)和放线菌门(15.82%～20.86%)为研究区土壤中的优势菌群。研究结果可对中国大面积污灌区镉污染农田修复效果评估及土壤健康评价提供科学依据。     

野鸭湖湿地芦苇根际微生物多样性与磷素形态关系

以野鸭湖湿地芦苇(Phragmites communis)根际微生物为研究对象,采用化学连续提取法分析了芦苇根际/非根际土壤中各形态磷随植物生长(4、7、10月)的变化规律,同时基于细菌的16S rRNA高通量测序技术,分析了芦苇根际/非根际土壤微生物多样性,进一步采用CCA法分析了土壤样品中特定微生物与磷素形态转化关系.结果表明,无机磷含量的总体顺序大小为:钙磷(Ca-P)闭蓄态磷(Oc-P)铁磷(Fe-P)交换态磷(Ex-P)铝磷(Al-P),无机磷含量变化主要受芦苇生长状况影响,在芦苇旺盛期达到最低,且根际土壤中的总无机磷含量普遍低于非根际.有机磷含量分布为高稳定性有机磷(HR-OP)中稳定性有机磷(MR-OP)中活性有机磷(ML-OP)活性有机磷(L-OP),各组分含量均随芦苇生长先降低后升高.芦苇根际/非根际土壤微生物中的优势菌门均为变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、绿弯菌门(Chloroflexi)、放线菌门(Actinobacteria),但在芦苇生长过程中,其根际和非根际微生物群落结构受季节影响变化显著,且根际和非根际的变化有所差异.同时发现了芽孢杆菌属(Bacillus)、肠杆菌属(Enterobacter)、假单胞菌属(Pseudomonas)、伯克霍尔德氏菌属(Burkholderia)、不动杆菌属(Acinetobacter)等可能是与磷形态转化有关的主要功能菌属,这些菌属能利用大部分的有机磷和无机磷,在湿地土壤磷素转化中起着关键作用.     

外源稀土在根际与非根际土壤中的存在形态

采用根际袋法研究了稀土在春小麦根际土壤与非根际土壤中存在形态的差别.结果表明,外源稀土进入土壤后,在根际土壤与非根际土壤中主要以残渣态形式存在,各形态含量的大小顺序为:残渣态>碳酸盐结合态>可交换态>铁锰氧化物结合态>有机质结合态.小麦根系活动明显影响根际土壤中的稀土存在形态,根际土壤中的可交换态和铁锰氧化物结合态含量明显大于非根际土壤中的含量,而碳酸盐结合态有所下降,有机质结合态无明显变化.     

水稻不同生育期根际及非根际土壤砷形态迁移转化规律

选取湖南某矿区污染稻田土,以盆栽试验的方式,采用美国TCLP(toxicity characteristic leaching procedure)和砷形态分析分级测定法,研究了水稻(Oryza sativa L.)各生育期(分蘖期、拔节期、孕穗期、灌浆期、成熟期)根际和非根际土壤中砷的赋存形态及转化规律.结果表明:1随着水稻生育期的延长,根际及非根际土壤p H值和砷TCLP提取态含量均逐渐上升,且非根际土壤p H值和砷TCLP提取态含量均高于同时期根际土壤.2水稻各生育期根际和非根际土壤中交换态砷(AE-As)含量均低于水稻种植前,并随着水稻生育期的延长逐渐上升.与水稻种植前相比,铝型砷(Al-As)、铁型砷(Fe-As)和钙型砷(Ca-As)含量在水稻种植后逐渐上升,但并不显著.残渣态砷(O-As)和总砷(T-As)含量在水稻种植后逐渐下降,在根际土壤中下降了37.30%和14.69%,在非根际土壤中分别下降了31.38%、8.67%.3在水稻不同生育期,土壤中各形态砷含量均表现为:残渣态铁型砷铝型砷钙型砷交换态砷.在p H值为5.0~5.8的范围内,各砷形态和砷TCLP提取态含量与p H值之间大都呈极显著或显著正相关,但根际土壤中钙型砷与p H值的相关关系较差.     

5种矿区土著植物对铅污染土壤的修复潜力研究

为评估我国西北某矿区茵陈蒿、蒲公英、苜蓿、大叶苦菜和车前草5种土著作物对含铅（Pb）土壤的修复潜力,采用盆栽实验,设置4种不同Pb含量水平（0、2‰、3‰和5‰,质量分数）,测定作物不同组织器官中和种植前后土壤中Pb含量,以及根际与非根际土壤微生物生物量碳含量、过氧化氢酶活性等指标。结果表明:茵陈蒿和车前草适合种植于铅含量为2‰、3‰、5‰的土壤中,可去除12%~32%的土壤Pb,车前草根系和茵陈蒿茎叶对Pb的累积量最高分别达到3617,720 mg/kg,显著高于其他植物,作为土壤铅污染修复植物的潜力较大。根际土壤微生物生物量碳含量比非根际土壤微生物生物量碳含量高2.37%~13.89%。土壤Pb抑制了根际与非根际土壤过氧化氢酶活性,使其活性低于对照组0.44%~22.3%,根际土壤过氧化氢酶活性比非根际过氧化氢酶活性高0.89%~8.09%。研究结果可为Pb污染矿区废弃地植物修复和土壤环境质量评价提供理论依据。     

菲、芘污染土壤中丛枝菌根真菌对土壤酶活性的影响

采用温室盆栽试验方法,研究了丛枝菌根真菌(AMF)对菲、芘复合污染土壤中微生物数量和酶活性的影响.结果表明,接种AMF显著增加了三叶草根际和菌丝际土壤中细菌、真菌和放线菌的数量,并对微生物区系有选择性.在供试菲、芘污染浓度范围内,低浓度菲、芘对土壤多酚氧化酶、酸性磷酸酶和过氧化氢酶活性有激活作用;但当菲、芘浓度升高时,3种酶活性受到抑制.与无植物对照土壤相比,接种AMF后三叶草菌根际酸性磷酸酶活性降低了2.4%~23.1%,过氧化氢酶活性增加了12.6%~20.3%,除高浓度处理外多酚氧化酶活性均增加;菌丝际多酚氧化酶活性比菌根际低12.9%~62.9%,酸性磷酸酶活性比菌根际高3.3%~24.0%,过氧化氢酶活性则高于菌根际.     

The role of arbuscular mycorrhiza on change of heavy metal speciation in rhizosphere of maize in wastewater irrigated agriculture soil

To understand the roles of mycorrhiza in metal speciation in the rhizosphere and the impact on increasing host plant tolerance against excessive heavy metals in soil, maize ( Zea mays L. ) inoculated with arbuscular mycorrhizal fungus ( Glomus mosseae) was cultivated in heavy metal contaminated soil. Speciations of copper, zinc and lead in the soil were analyzed with the technique of sequential extraction. The results showed that, in comparison to the bolked soil, the exchangeable copper increased from 26% to 43% in non-infected and AM-infected rhizoshpere respectively; while other speciation (organic, carbonate and Fe-Mn oxide copper) remained constant and the organic bound zinc and lead also increased but the exchangeable zinc and lead were undetectable. The organic bound copper, zinc end lead were higher by 15%, 40% and 20%, respectively, in the rhizosphere of arbuscular mycorrhiza infected maize in comparison to the non-infected maize. The results might indicate that mycorrhiza could protect its host plants from the phytotoxicity of excessive copper, zinc and lead by changing the speciation from bio-available to the non-bio-available form. The fact that copper and zinc accumulation in the roots and shoots of mycorrhia infected plants were significantly lower than those in the non-infected plants might also suggest that mycorrhiza efficiently restricted excessive copper and zinc absorptions into the host plants. Compared to the non-infected seedlings, the lead content of infected seedlings was 60% higher in shoots. This might illustrate that mycorrhiza have a different mechanism for protecting its host from excessive lead phytotoxicity by chelating lead in the shoots.     

丛枝菌根真菌-植物联合修复石油污染土壤研究进展

丛枝菌根(Arbuscular mycorrhiza,AM)是由丛枝菌根真菌与植物根系所建立的一种互惠共生体,在自然界分布广泛,具有降解复杂有机污染物的能力.石油污染土壤中,利用土著的或外接的丛枝菌根真菌可以明显改善植物根际环境,提高土壤生物活性,从而加速土壤中石油污染物的降解.现有研究已证明丛枝菌根真菌修复石油污染土...     

蚯蚓-菌根相互作用对土壤-植物系统中Cd迁移转化的影响

以灰化土(Aquods soil)为供试土壤,分别加入4种含量的Cd2 (0、5、10、20 mg·kg-1)模拟土壤污染,设置单独加8条蚯蚓(Pheretima sp.)、单独接种菌根(Inoculum Endorize-Mix2)、同时接种蚯蚓和菌根的3种处理,以不加蚯蚓和菌根为对照,各处理均种植黑麦草(Lolium multiflorum),研究蚯蚓、菌根相互作用对土壤-植物系统中Cd迁移转化的影响.结果表明:(1)菌根对土壤pH无明显影响,加蚯蚓可使土壤pH比对照约降低0.2,蚯蚓和菌根同时作用对土壤pH降低没有协同作用.(2)蚯蚓或菌根的加入均能显著增加土壤中可溶性有机碳(DOC)含量,蚯蚓的影响大于菌根,同时加入蚯蚓和接种菌根对土壤中DOC的增加有一定的拮抗作用.(3)蚯蚓活动增加了黑麦草根部Cd的积累,菌根则能促进Cd从黑麦草根部向地上部转移,二者具有促进Cd向地上部分转移的协同作用.(4)蚓粪和土壤中DTPA提取态Cd含量与黑麦草吸收Cd量呈显著相关(p＜0.01),而蚓粪中DTPA提取态Cd含量均显著高于土壤中的含量(p＜0.05).因此,蚓粪中有效态Cd是植物吸收Cd的重要供源.     

根际土壤中汞甲基化与去甲基化作用双同位素示踪研究

根际环境特殊的理化性质可能显著促进汞(Hg)的甲基化.为证实上述假设,本研究采集了三峡库区消落带狗牙根根际土和非根际土,分别测定了根际土与非根际土中总汞(THg)和甲基汞(MeHg)含量及主要理化性质,并采用双同位素示踪法(~(199)Hg~(2+)和Me~(201)Hg)进行室内模拟培养实验,探究其Hg的甲基化与去甲基化速率.结果表明,根际土壤中的Fe~(2+)、Mn~(2+)、有机质含量、过氧化氢酶活性显著高于非根际土(p0.05),MeHg含量、细菌、真菌数极显著大于非根际土(p0.01);根际土壤中MeHg的含量与Mn~(2+)、SO_4~(2-)、有机质含量存在显著正相关性(p0.05),与过氧化氢酶活性、细菌、真菌数存在显著正相关性(p0.01);根际土壤中Hg甲基化、去甲基化速率、净甲基化潜力均显著大于非根际土(p0.05),有菌根际土的甲基化与去甲基化速率显著大于无菌根际土(p0.05).本研究证实了根际是Hg发生甲基化的一个活跃区域,其中,微生物在根际土壤中Hg的甲基化与去甲基化过程中均起着主要作用.     

Effects of arbuscular mycorrhizal fungi inoculation on arsenic accumulation by tobacco (Nicotiana tabacum L.)

A pot experiment was conducted to study the e ects of arbuscular mycorrhizal (AM) fungi (from contaminated or uncontaminated soils) on arsenic (As) uptake of tobacco (Nicotiana tabacum L.) in As-contaminated soil. Mycorrhizal colonization rate, dry weight, As and P uptake by plants, concentrations of water-extractable As and As fractions were determined. A low mycorrhizal colonization rate (< 25%) was detected. Our research indicated that AM fungi isolated from polluted soils were no more e ective than those from unpolluted soils when grown in symbiosis with tobacco. No significant di erences were observed in roots and stalks dry weights among all treatments. Leaves and total plant dry weights were much higher in Glomus versiforme treatment than that in control treatment. As contents in roots and stalks from mycorrhizal treatments were much lower than that from control treatment. Total plant As content exhibited the same trend. P concentrations in tobacco were not a ected by colonization, nor were stalks, leaves and total plant P contents. Roots P contents were remarkably lower in HN treatments than in other treatments. Meanwhile, decreased soil pH and lower water-extractable As concentrations and higher levels of As fraction bound to well-crystallized hydrous oxides of Fe and Al were found in mycorrhizal treatments than in controls. The protective e ect of mycorrhiza against plant As uptake may be associated with changes in As solubility mediated by changing soil pH. These results indicated that under As stress, proper mechanisms employed by AM fungi can protect tobacco against As uptake. Results confirmed that AM fungi can play an important role in food quality and safety.     

Rhizophagus irregularis influences As and P uptake by alfafa and the neighboring non-host pepperweed growing in an As-contaminated soil

It was documented that arbuscular mycorrhiza fungi(AMF) play an important role in protecting host plants against arsenic(As) contamination. However, most terrestrial ecosystems contain a considerable number of nonmycorrhizal plants. So far little information is available for the interaction of such non-host plants with AMF under As contaminations. By using a dual compartment cultivation system with a plastic board or a nylon mesh separating roots of non-host pepperweed from roots of the AM-host alfafa plants, avoiding direct root competition, the two plant species were grown separately or partially separated(with rhizosphere effects) in the presence or absence of the AMF Rhizophagus irregularis in As-contaminated soil. The results indicated that mycorrhiza caused phosphorus(P) concentration decrease in the non-host pepperweed, but promoted the P concentration of the AM host alfafa. Mycorrhiza is potentially helpful for non-host pepperweed to adapt to As contamination by decreasing root As concentration and showing no suppressing effect on biomass production. The study provides further evidence for the protective effects of AMF on non-host plants against As contamination, and improved our understanding of the potential role of AMF for non-host plant adaptation to As contaminated soils.