生物有机肥对板栗土壤微生物群落代谢活性的影响

2011年在陕西省柞水县下梁镇沙坪村板栗园,设置对照、施用复合肥和施用生物有机肥3个处理,于板栗收获期采集土壤样品,并运用Biolog方法研究了生物有机肥对土壤微生物群落代谢能力的影响.结果表明:生物有机肥处理的微生物群落碳源利用率(AWCD)、Shannon均匀度、丰富度指数和McIntosh指数均显著高于对照和复合肥处理;与对照相比,施用生物有机肥处理的土壤微生物对糖类和多聚物类碳源的利用能力增强,而施用复合肥处理对各类碳源的利用能力均较弱.主成分分析表明,不同施肥处理的土壤微生物群落明显不同,起分异作用的碳源主要为糖类和氨基酸类.     

三种不同土壤改良剂对黄瓜根际微生物数量和群落代谢功能多样性的影响

【背景】土壤微生物在调节植物生长、提高土壤养分利用率及维持生态系统稳定性等方面发挥着重要作用。【目的】探讨蚯蚓原位处理废弃物及脱硫石膏改良土壤后的微生物群落结构多样性特征,深入研究黄瓜根际微生物群落数量与微生物指数之间的相关性。【方法】通过平板培养法测定可培养真菌、细菌、放线菌群落数量与Biolog-ECO微平板法分析OD590值微生物碳源利用率相结合,研究蚯蚓处理废弃物原位生成蚯蚓粪替代土壤(T1)、脱硫石膏(3 t/hm2)(T2)、蚯蚓处理废弃物原位生成蚯蚓粪与脱硫石膏混施(T3)这3种土壤处理下栽培黄瓜,其根际微生物群落多样性特征。【结果】T1、T3处理的可培养放线菌数量显著高于CK、T2;细菌与真菌菌落数量无显著性差异。各处理的平均每孔颜色变化率(average well color development,AWCD)呈“厂”型趋势变化,在0-96h上升迅速,96-144h缓慢上升至最高值,T1、T3处理的AWCD值高于CK、T2。与对照相比,各处理的AWCD值、Shannon指数、Simpson指数、Pielou指数和McIntosh指数存在显著性差异。与CK相比,T1与T...     

不同地下滴灌制度下黄瓜根际微生物活性及功能多样性

采用微生物培养、BIOLOG碳素利用法和土壤酶活性测定等方法,分析了日光温室不同地下灌溉制度下黄瓜根际土壤中微生物活性及功能多样性.结果表明: 根际土壤微生物生物量C、N含量、基础呼吸、代谢熵、AWCD值、Shannon指数和McIntosh指数随灌水量的增加呈先升高后下降的趋势;在0.8E_p(E_p为20 cm标准蒸发皿蒸发量)灌溉水平下,I2处理(灌水周期8 d)根际土壤微生物生物量C、N含量、基础呼吸、代谢熵、AWCD值、Shannon指数和McIntosh指数显著高于I₁处理(灌水周期4 d).0.8E_p处理下,细菌、放线菌、自生固氮菌数量及脲酶、磷酸酶、蔗糖酶、过氧化氢酶和多酚氧化酶活性显著高于其他2个灌水量处理（0.6E_p和1.0E_p）;I₂处理的细菌和自生固氮菌数量、脲酶、磷酸酶和蔗糖酶活性显著高于I₁处理,放线菌数量、过氧化氢酶和多酚氧化酶活性与I₁处理差异不显著,而真菌数量显著低于I₁处理.I 20.8E_p处理使黄瓜根际土壤中微生物代谢活性和微生物群落功能多样性升高,微生物区系得以改善,土壤酶活性提高,促进黄瓜生长.     

不同施肥处理与地膜覆盖对土壤微生物群落功能多样性的影响

微生物多样性是表征土壤质量变化的敏感指标。应用Biolog技术探讨了不同施肥处理与地膜覆盖对土壤微生物功能多样性的影响,从微生物功能多样性的角度评价施肥与地膜覆盖对土壤质量的影响。试验结果表明：裸地条件下,肥料合理配施可以增强微生物对碳源的利用程度（AWCD）,显著增加微生物功能多样性（Shannon指数）。地膜覆盖和施肥的交互作用降低了微生物对碳源的利用率,降低微生物的丰富度,改变其均匀度。土壤微生物碳源利用的聚类和主成分分析表明,各施肥处理在碳源的利用上存在较大差异,覆膜加剧了各处理之间的分异程度。糖类和氨基酸类碳源是微生物利用的主要碳源。土壤微生物对碳源利用受到土壤pH、速效钾的显著影响。此外,有机碳、速效氮含量和土壤碳氮比与土壤微生物群落功能多样性密切相关。     

长期施肥对茶园土壤微生物群落功能多样性的影响

利用BIOLOG研究长期施肥后茶园土壤微生物群落功能多样性变化,结果表明:与清耕(CK)相比,各长期施肥处理AWCD值的变化速度(斜率)和最终能达到的AWCD值均有不同程度提高,其大小顺序为:DWY>DY>W>CK,可见施肥能不同程度提高微生物整体活性和丰富度,以有机无机肥配施效果最好。分析多样性指数表明:各处理对土壤常见微生物种类影响并不大,施有机肥使微生物群落均匀度有所降低。对碳源利用主成分起分异作用的主要是糖类和羧酸类物质。对AWCD值、多样性指数与土壤养分因子的相关性分析表明,在红壤茶园中土壤肥力提高有利于土壤微生物活性的提高。     

玉米连作及其施肥对土壤微生物群落功能多样性的影响

采用Biolog技术,借助吉林农业大学1984年建立的长期定位试验,以撂荒和非玉米连作(当季作物为芸豆)为对照,研究玉米连作及其不同施肥措施对土壤微生物功能多样性的影响。结果表明,反映土壤微生物活性的平均颜色变化率(AWCD)呈现出以下变化规律:撂荒(UC)非玉米连作(NCC)玉米连作不施肥(CK);玉米连作配施秸秆(S、SN、SNPK)玉米连作配施NPK(NPK)玉米连作不施肥(CK)玉米连作单施N(N)。微生物培养72h活性旺盛,各处理AWCD在0.395—0.732之间,其中撂荒AWCD显著高于非玉米连作和玉米连作不施肥(P0.05);玉米连作配施秸秆明显提高微生物活性,玉米连作配施秸秆(S、SN、SNPK)AWCD是玉米连作不施肥和单施化肥(N、NPK)的1.26—1.62倍。玉米连作不施肥土壤微生物群落多样性指数(H、E、S)低于撂荒处理,但高于非玉米连作处理,非玉米连作提高了土壤微生物优势度指数。玉米连作配施秸秆土壤微生物物种丰富度指数和均匀度指数高于玉米连作不施肥和玉米连作施化肥,玉米连作单施氮肥优势度指数较高,其它多样性指数降低。主成分分析结果表明,不同处理土壤微生物碳源利用特征出现分异:撂荒和玉米连作配施秸秆处理集中在第1主成分正方向,得分系数在2.39—4.17之间,土壤微生物碳源利用特征相似;玉米连作不施肥、玉米连作单施化肥和非玉米连作处理分布在第1主成分负方向,得分系数在-5.43—-1.59之间。土壤微生物利用的碳源主要是糖类、羧酸类、氨基酸和聚合物。玉米连作配施秸秆和撂荒有利于提高土壤微生物代谢活性和土壤微生物群落功能多样性,玉米连作单施化肥尤其单施氮肥土壤微生物活性和功能多样性下降。     

EG03菌剂对辣椒青枯病的防治效果及对根围土壤微生物群落的影响

EG03菌剂由多种抗青枯劳尔氏菌(Ralstonia solanacearum)的拮抗细菌复配而成,田间施用能有效防治辣椒青枯病,平均防效达85.8%.采用稀释平板法、最大或然数(MPN)法以及Biolog微孔板培养系统研究了EG03菌剂对田间辣椒根围土壤微生物群落特征的影响.结果表明:EG03菌剂对根围土壤微生物群落的影响随时间变化而不同,施用后不同程度提高了真菌和芽孢杆菌的数量,并显著增加了自生固氮细菌的数量.根围土壤微生物群落平板每孔颜色平均变化率(AWCD)的变化随培养时间呈现典型的"S"型曲线,辣椒生长后期根围土壤AWCD值高于前期.6类碳源利用分析表明,EG03菌剂的施用在短期内会降低根围土壤微生物的碳源利用率,辣椒生长后期根围土壤中以糖类物质作为碳源的微生物占主导地位.微生物多样性指数分析发现,EG03菌剂施用前期会不同程度降低根围土壤微生物各项多样性指数,但施用后期会提高各项多样性指数,尤其在Simpson指数和McIntosh均匀度两项指标上差异显著.     

长期不同施肥对稻田土壤微生物群落功能多样性的影响

微生物群落功能多样性对土壤管理具有重要指示作用,施肥措施对土壤微生物群落功能多样性产生重要的影响,从而影响土壤有机质的含量,引起土地碳贮量变化,进而影响到陆地生态系统的碳汇.以位于湖南省桃江县国家级稻田肥力长期定位试验点的土样为研究对象,采用BIOLOG测试板对不同施肥处理下土壤微生物功能多样性进行了研究.研究结果显示,土壤微生物的碳源利用率因长期不同的施肥处理而发生分异.Shannon和Simpson指数的结果显示所有施肥处理均有利于维持微生物群落多样性,但秸秆还田和习惯施肥使群落均匀度(McIntosh指数)降低.主成分分析表明,试验点土壤微生物群落利用的主要碳源为氨基酸类和糖类,但不同施肥处理碳源利用类型有差异.结果可以得出,不同的施肥对土壤微生物功能多样性产生了不同影响,从而影响土壤有机质中碳、氮含量.这些信息可以为深入研究施肥对全球碳氮循环以及全球气候变迁提供依据.     

不同施肥处理对玉米-小麦轮作土壤微生物群落功能多样性的影响

土壤微生物多样性能反应土壤的肥力,不同的施肥措施对土壤微生物的种群和功能多样性也会产生重要的影响。以山东德州连续两年小麦季和玉米季收获后土壤为研究对象,利用Biolog技术研究了6种不同施肥处理对土壤微生物群落功能多样性的影响。结果表明:其中各个施肥处理的平均颜色变化率(average well color development,AWCD)差异显著,常规氮磷钾肥+全量秸秆还田+秸秆腐熟剂(FS)处理代谢活性最高;物种丰富度指数(H)和均匀度指数(E)也表明各施肥方式均能够维持微生物种群的多样性,其中FS和30%猪粪+70%常规氮磷钾肥(OF)处理物种丰富度指数(H)和均匀度指数(E)最高;PCA及RDA分析显示,OF和FS处理微生物功能多样性相似,且其微生物功能多样性与有机质(Soil organic matter,SOM)、全氮(Total N,TN)、速效磷(Available P,AP)和速效钾(Available K,AK)密切相关。猪粪堆肥有机无机复合肥3 600 kg/hm2(OI2)处理与猪粪堆肥有机无机复合肥1 800 kg/hm2(OI1)处理相似,其功能多样性比常规施肥(CF)处理稍高。综上所述,OF处理和FS处理的土壤微生物群落功能多样性程度高于其他处理,说明秸秆还田+秸秆腐熟剂和有机肥部分替代氮磷钾肥能够显著提高土壤微生物功能多样性,有利于保护土壤微生态。     

有机物料种类及腐熟水平对土壤微生物群落的影响

应用Biolog方法研究了温室盆栽番茄条件下,施用不同种类及不同腐熟水平的有机物料对土壤微生物群落的影响,施用有机物料60d后取土分析土壤微生物群落多样性及土壤微生物对Biolog微平板中胺、氨基酸、糖、羧酸、聚合物和其它类碳源的利用情况,结果表明,施用有机物料可提高土壤微生物群落多样性,施用新鲜酒糟的多样性指数略高于施用腐熟10d酒糟,牛粪不同腐熟水平对多样性影响显著,且对多样性具有正向或负向的影响;对照和施用酒糟的土壤微生物对聚合物的利用率高于施用牛粪处理,施用新鲜物料处理的土壤微生物对聚合物的利用率高于施用腐熟物料处理。     

Ultrastructure of the Mature Egg and Fertilization in the Fern Ceratopteris thalictroides

The ultrastructure of the mature egg and fertilization in the fern Ceratopteris thalictroides (L.) Brongn. were observed by transmission electron microscopy. The results revealed that the mature egg possesses an obvious egg membrane at the periphery of the egg. Furthermore, a fertilization pore was identified in the upper egg membrane of the mature egg. The structure of the pore is described for the first time. The fertilization experiment indicated that spermatozoids crowd into the cavity above the egg through the neck canal of the archegonium; however, only one of these can penetrate into the egg through the fertilization pore. Immediately on penetration of the spermatozoid, the egg begins to shrink. The volume of the fertilized egg decreases to almost one-half that of the unfertilized egg. As a result, the protoplasm of the fertilized egg becomes dense and opaque, which may lead to a situation where the organelles of both the egg and the fertilizing spermatozoid become indistinguishable. Simultaneously, abundant vesicles containing concentric membranes or opaque materials appear near the fertilization pore in the cytoplasm of the fertilized egg. These vesicles are considered to act as a barrier that prevents polyspermy. The present study provides a new insight into the ultrastructure of the mature egg and the cytological mechanism of fertilization in ferns.     

Mode of pollen tube growth in pistils of Ticodendron incognitum (Ticodendraceae,Fagales) and the evolution of chalazogamy

Ticodendron incognitum is the sole species of the Ticodendraceae, which was established as a new family in the Fagales less than 20 years ago. Considering the diverse modes of pollen tube growth observed in other Fagales, we investigated the growth of pollen tubes in the pistil of Ticodendron. At the time of pollination, T. incognitum had four immature ovules in a bilocular ovary, thus exhibiting delayed fertilization, as in other Fagales. During the period when fertilization was delayed, pollen tube growth in the pistil was intermittent, consisting of five steps associated with development of the ovules and embryo sacs. Four cessation sites occurred: in the style, in the tissue of the upper part of the ovary, inside and outside of the funicle and at the chalaza. A single pollen tube eventually reaches a mature embryo sac through the chalaza in one of the four ovules. While both delayed fertilization and intermittent pollen tube growth play a role in male and female gametophyte selection, as in other Fagales, the five‐step process of pollen tube growth through the chalaza (i.e. chalazogamy) is characteristic of lineages of the Casuarinaceae, Ticodendraceae and Betulaceae (the latter with the loss of one step). © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 157 , 621–631.     

Ultrastructure of the Mature Egg and Fertilization in the Fern Ceratopteris thalictroides

The ultrastructure of the mature egg and fertilization in the fern Ceratopteris thafictroides （L.） Brongn. were observed by transmission electron microscopy. The results revealed that the mature egg possesses an obvious egg membrane at the periphery of the egg. Furthermore, a fertilization pore was identified in the upper egg membrane of the mature egg. The structure of the pore is described for the first time. The fertilization experiment indicated that spermatozoids crowd into the cavity above the egg through the neck canal of the archegonium; however, only one of these can penetrate into the egg through the fertilization pore. Immediately on penetration of the spermatozoid, the egg begins to shrink. The volume of the fertilized egg decreases to almost one-half that of the unfertilized egg. As a result, the protoplasm of the fertilized egg becomes dense and opaque, which may lead to a situation where the organelles of both the egg and the fertilizing spermatozoid become indistinguishable. Simultaneously, abundant vesicles containing concentric membranes or opaque materials appear near the fertilization pore in the cytoplasm of the fertilized egg. These vesicles are considered to act as a barrier that prevents polyspermy. The present study provides a new insight into the ultrastructure of the mature egg and the cytological mechanism of fertilization in ferns.     

Ultrastructure of the Mature Egg and Fertilization in the Fern Ceratopteris thafictroides

The ultrastructure of the mature egg and fertilization in the fern Ceratopteris thalictroides (L.) Brongn. were observed by transmission electron microscopy. The results revealed that the mature egg possesses an obvious egg membrane at the periphery of the egg. Furthermore, a fertilization pore was identified in the upper egg membrane of the mature egg. The structure of the pore is described for the first time. The fertilization experiment indicated that spermatozoids crowd into the cavity above the egg through the neck canal of the archegonium; however, only one of these can penetrate into the egg through the fertilization pore. Immediately on penetration of the spermatozoid, the egg begins to shrink. The volume of the fertilized egg decreases to almost one-half that of the unfertilized egg. As a result, the protoplasm of the fertilized egg becomes dense and opaque, which may lead to a situation where the organelles of both the egg and the fertilizing spermatozoid become indistinguishable. Simultaneously, abundant vesicles containing concentric membranes or opaque materials appear near the fertilization pore in the cytoplasm of the fertilized egg. These vesicles are considered to act as a barrier that prevents polyspermy. The present study provides a new insight into the ultrastructure of the mature egg and the cytological mechanism of fertilization in ferns.     

不测土条件下半定量施肥原理和模型评述

1　不测土条件下半定量推荐施肥的需求矿质营养学说是施肥最基本原理 ,它的诞生标志着人类主动施肥时期的开始。从早期的定性施肥阶段 ,发展到今天的定量施肥阶段 ,如何确定施肥量一直是施肥技术的核心和难点。测土是定量施肥或精确施肥的前提。然而 ,测土本身存在各种偏差或精度、测土需要费用 ,本文仅对代表性的不测土施肥模型及其原理进行评述。即使有了土壤有效养分的测定结果 ,如何将其转化为一季内土壤提供的养分量一直是个理论难题 ,特别是Ｎ素土壤供应量更为难测或难以估算。因此 ,如何在使用定量施肥模型的同时 ,而又将土壤养分因…     

水稻双受精过程的细胞形态学及时间进程的观察

应用常规石蜡切片和荧光显微镜观察水稻（Oryza sativa）受精过程中雌雄性细胞融合时的形态特征及时间进程,确定合子期,为花粉管通道转基因技术的实施提供理论依据。结果表明：授粉后,花粉随即萌发,花粉管进入羽毛状柱头分支结构的细胞间隙,继续生长于花柱至子房顶部的引导组织的细胞间隙中,而后进入子房,在子房壁与外珠被之间的缝隙中向珠孔方向生长,花粉与花粉管均具有明显的绿色荧光。花粉管经珠孔及珠心表皮细胞间隙进入一个助细胞,释放精子。精子释放前,两极核移向卵细胞的合点端：两精子释放于卵细胞与中央细胞的间隙后,先后脱去细胞质,然后分别移向卵核和极核,移向卵核的精核快于移向极核的精核：精核与两极核在向反足细胞团方向移动的过程中完成雌雄核融合。大量图片显示了雌雄性核融合的详细过程以及多精受精现象。水稻受精过程经历的时间表如下：授粉后,花粉在柱头萌发：花粉萌发至花粉管进入珠孔大约需要0．5小时：授粉后0．54,时左右,花粉管进入一个助细胞,释放精子：授粉后0．5—2．5小时,精卵融合形成合子：授粉后约10．0小时,合子第1次分裂,合子期为授粉后2．5-10．04,时：授粉后1．0-3．04,时,精核与两极核融合：授粉后约5．0小时,初生胚乳核分裂。’     

Expression in Escherichia coli and immunological characterization of three zona pellucida proteins (ZP1, ZP2, and ZP3) from a marsupial,the brushtail possum (Trichosurus vulpecula)

The brushtail possum (Trichosurus vulpecula) zona pellucida (ZP) is composed of three major glycoproteins, designated ZP1, ZP2, and ZP3 based on their size and homology with eutherian ZP proteins. These proteins are candidate antigens for the development of an immunocontraceptive vaccine to control the fertility of the brushtail possum in New Zealand, where it is an introduced pest. In order to further their immunological and functional characterization, recombinant possum ZP proteins were produced in Escherichia coli (E. coli) strain JM109, M15, SG13009, or BL21 codon plus. Each of the proteins produced possessed a N-terminal six histidine tag (His)(6) to facilitate purification and consisted of amino acid (aa) residues 18-471 of possum ZP1, aa residues 40-311 of ZP2 (ZP2-N), aa residues 305-634 of ZP2 (ZP2-C), and aa residues 23-342 of ZP3. Immunoblot using anti-RGS(His)(4) antibodies and polyclonal rabbit anti-porcine ZP antibodies detected major bands at 54 kDa for ZP1, 32 kDa for ZP2-N, 39 kDa for ZP2-C, and 40 kDa for ZP3. Immunization of male and female rabbits with ZP2-N, ZP2-C, and ZP3 purified on Ni-NTA resin under denaturing conditions generated antibodies reactive with recombinant ZP proteins on Western blot and with native ZP proteins in possum ovarian sections using immunofluorescence. Antibodies generated against ZP1 in the same way were reactive with recombinant ZP proteins on Western blot only. The recombinant possum ZP proteins and specific antibodies produced in this study give an indication of the antigenic relationship of the possum ZP proteins and are vital tools for future studies of sperm-ZP binding in marsupials and for the evaluation of ZP-based contraceptive vaccines in possums and other marsupials.     

Sperm-egg interaction in the mouse using live and glutaraldehyde-fixed eggs

A study of interaction between gametes in the mouse, using live and glutaraldehyde-fixed eggs, showed that, while in live eggs the binding is a two-step process (“early” and “late” binding), the process is one step when the spermatozoa interact with fixed eggs. The second point that emerged from this study is that uncapacitated and capacitated spermatozoa bind the zonae pellucidae of live and fixed eggs in the same way, but only the capacitated spermatozoa bound to live eggs undergo a complete acrosome reaction and penetrate the zona pellucida. Moreover, it has been shown that binding to fixed eggs, just as to live eggs, is Ca²⁺-dependent, and it can be reversed by EGTA. Fixed eggs thus are a good model to study only one step of the sperm-egg interaction removed from all the other events of the fertilization process.     

Incidence of chromosome anomalies in first-cleavage mouse embryos obtained from frozen-thawed oocytes fertilized in vitro

To assess the effect of low temperature storage on mouse oocytes we (1) examined the capacity for normal development of embryos derived from frozen oocytes fertilized in vitro after transfer to pseudopregnant foster mothers and (2) analyzed the chromosome complement at the first cleavage division. Fewer frozen than control oocytes were fertilized (36% vs 66%), but after embryo transfer the proportion of fertilized eggs that implanted (67–68%) and formed normal foetuses (50–53%) was similar in the two groups. Freezing did not affect the observed incidence of aneuploidy (1.5–3.3%). The frequency of polyploid embryos derived from frozen oocytes was almost double that of controls (15.8% vs 8.5%), but it is unclear whether this is a real effect of freezing or is an artifact produced by the chromosome preparation technique.