Sequence and translation of the murine coronavirus 5''-end genomic RNA reveals the N-terminal structure of the putative RNA polymerase.

A 28-kilodalton protein has been suggested to be the amino-terminal protein cleavage product of the putative coronavirus RNA polymerase (gene A) (M.R. Denison and S. Perlman, Virology 157:565-568, 1987). To elucidate the structure and mechanism of synthesis of this protein, the nucleotide sequence of the 5' 2.0 kilobases of the coronavirus mouse hepatitis virus strain JHM genome was determined. This sequence contains a single, long open reading frame and predicts a highly basic amino-terminal region. Cell-free translation of RNAs transcribed in vitro from DNAs containing gene A sequences in pT7 vectors yielded proteins initiated from the 5'-most optimal initiation codon at position 215 from the 5' end of the genome. The sequence preceding this initiation codon predicts the presence of a stable hairpin loop structure. The presence of an RNA secondary structure at the 5' end of the RNA genome is supported by the observation that gene A sequences were more efficiently translated in vitro when upstream noncoding sequences were removed. By comparing the translation products of virion genomic RNA and in vitro transcribed RNAs, we established that our clones encompassing the 5'-end mouse hepatitis virus genomic RNA encode the 28-kilodalton N-terminal cleavage product of the gene A protein. Possible cleavage sites for this protein are proposed.     

Effect of anti-mosquito antibodies on the infectivity of the rodent malaria parasite Plasmodium berghei to Anopheles farauti

The effect of mouse anti-mosquito antibodies, present in the bloodmeal, on the infectivity of Plasmodium berghei Vincke to Anopheles farauti Laveran was investigated. Significantly fewer oocysts developed in mosquitoes feeding on mice immunized with sugar-fed mosquito midgut antigens than in mosquitoes feeding on control mice. Mosquitoes feeding on mice immunized with the midgut antigens derived from sugar-fed mosquitoes also showed reduced mortality and had lower infection rates than those fed on unimmunized mice. Blood-fed midgut antigen was less effective in producing these effects than sugar-fed midgut antigen.     

RNA recombination of murine coronaviruses: recombination between fusion-positive mouse hepatitis virus A59 and fusion-negative mouse hepatitis virus 2.

It has previously been shown that the murine coronavirus mouse hepatitis virus (MHV) undergoes RNA recombination at a relatively high frequency in both tissue culture and infected animals. Thus far, all of the recombination sites had been localized at the 5' half of the RNA genome. We have now performed a cross between MHV-2, a fusion-negative murine coronavirus, and a temperature-sensitive mutant of the A59 strain of MHV, which is fusion positive at the permissive temperature. By selecting fusion-positive viruses at the nonpermissive temperature, we isolated several recombinants containing multiple crossovers in a single genome. Some of the recombinants became fusion negative during the plaque purification. The fusion ability of the recombinants parallels the presence or absence of the A59 genomic sequences encoding peplomers. Several of the recombinants have crossovers within 3' end genes which encode viral structural proteins, N and E1. These recombination sites were not specifically selected with the selection markers used. This finding, together with results of previous recombination studies, indicates that RNA recombination can occur almost anywhere from the 5' end to the 3' end along the entire genome. The data also show that the replacement of A59 genetic sequences at the 5' end of gene C, which encodes the peplomer protein, with the fusion-negative MHV-2 sequences do not affect the fusion ability of the recombinant viruses. Thus, the crucial determinant for the fusion-inducing capability appears to reside in the more carboxyl portion of the peplomer protein.     

Disintegration of Rhodospirillum rubrum chromatophore membrane into photoreaction units, reaction centers, and ubiquinone-10 protein with mixture of cholate and deoxycholate.

1. The membrane of Rhodospirillum rubrum chromatophores was disintegrated with mild detergents (cholate and deoxycholate) in order to study the spatial arrangement of the functional proteins in the photochemical apparatus and the electron transport system in the membrane. 2. The components solubilized from the membrane by a mixture of cholate and deoxycholate (C-DOC) were separated into four fractions by molecular-sieve chromatography in the presence of C-DOC; they were designated as F1, F2, F3, and F4 in the order of elution. The fractions were further purified by repeated molecular-sieve chromatography in the presence of C-DOC until each fraction was chromatographically homogeneous. 3. F1 appeared to be conjugated forms of F2. 4. The purified F2 was composed of a rigid complex having a weight of 7 X 10(5) daltons, containing approximately 10 different kinds of protein species with molecular weights of 3.8 X 10(4), 3.6 X 10(4), 3.5 X 10(4), 2.8 X 10(4), 2.7 X 10(4), 2.6 X 10(4), 1.3 X 10(4), 1.2 X 10(4), 1.1 X 10(4), and 1.0 X 10(4). The complex contained 33 bacteriochlorophylls, 4 iron atoms, and 90 phosphates, but no cytochrome, ubiquinone, or phospholipid. It showed the same reaction center activity as chromatophores, indicating that the complex was a unit of the photochemical apparatus (photoreaction unit). Each chromatophore of average size was estimated to possess about 24 photoreaction units. 5. The purified F3 showed an absorbance spectrum characteristic of reaction centers, and contained 3.4 bacteriochlorophylls, 2.0 bacteriopheophytins, and 1.9 acid-labile iron atoms, but no cytochrome or ubiquinone (C-DOC reaction center). It had a weight of 1.2 X 10(5) daltons, and the main components were 4 protein species with molecular weights of 2.8 X 10(4), 2.7 X 10(4), 2.6 X 10(4), and 1.0 X 10(4). 6. The purified F4 showed a molecular weight of about 11,000, and contained one mole of ubiquinone-10 per mole (ubiquinone-10 protein). 7. The reaction center activity of C-DOC reaction centers was stimulated by ubiquinone-10 protein. In addition, the reaction center oxidized reduced cytochrome c2 in the light, provided that ubiquinone-10 protein was present (photo-oxidase activity).     

放牧对祁连山高寒典型草原土壤质量的影响

土壤质量是维持陆地生态系统稳定性与功能多样性的基础。放牧作为草地资源最广泛的利用方式之一,其对草地土壤质量的影响却缺乏量化标准,且两者之间的作用机理尚不明确。以祁连山高寒草原两个季节性牧场为研究对象,结合生态系统耦合与生态系统多功能性,探究了放牧对高寒草原土壤质量的影响与潜在机制。试验结果表明：基于最小数据集,不同放牧率下土壤质量指数差异显著(P<0.05),冬季牧场和春秋季牧场放牧率分别在2.45头月^-1 hm^-2和0.80头月^-1 hm^-2时土壤质量指数最高。土壤速效磷、有机碳、氮磷比和土壤pH是决定冬季牧场土壤质量的关键因子,而春秋季牧场中则是土壤有机碳、碳氮比和土壤pH;两个季节性牧场土壤质量指数与物种丰富度指数(P<0.05)和香浓维纳多样性指数(P<0.0001)呈显著正相关。高寒草原季节性牧场放牧地植物群落物种多样性与土壤因子耦合度在0.67—0.81之间,平均耦合度为0.74,属于中度协调;随着放牧率的增加,生态系统多功能性指数逐渐减低且与土壤质量指数变化趋势相似,...     

黄土高原不同人工林型微生物残体碳对土壤有机碳组分的积累贡献及影响因素

为探究不同人工林型微生物残体碳(Microbial necromass carbon, MNC)对土壤有机碳组分的积累贡献及影响因素,在黄土高原选取刺槐林、山杏林、油松林为研究对象,分析了三种人工林0-60 cm土层真菌残体碳(Fungal necromass carbon, FNC)、细菌残体碳(Bacterial necromass carbon, BNC)、MNC对颗粒态有机碳(Particulate organic carbon, POC)和矿物结合态有机碳(Mineral-associated organic carbon, MAOC)的积累贡献及其影响因素。结果表明:(1)三种人工林POC、MAOC中FNC、BNC、MNC含量均随土层深度的增加而降低;(2)刺槐林和山杏林MNC对MAOC的积累贡献(60.9%,52.0%)高于POC(33.5%,49.5%),其中FNC对MAOC的积累贡献分别是BNC的4.4和2.5倍,油松林在0-10 cm土层MNC对POC的积累贡献(73.8%)高于MAOC(48.2%),其中FNC对POC的积累贡献是BNC的3.5倍,而在10-60 cm土层MNC对MAOC的积累贡献(30.9%)高于POC(24.4%),其中FNC对MAOC的积累贡献是BNC的3.4倍;(3)总有机碳和全氮含量与MNC/POC、MNC/MAOC呈显著正相关关系(P ＜ 0.05),黏粒含量与MNC/MAOC呈显著正相关关系(P ＜ 0.05),pH值、砂粒含量与MNC/MAOC呈显著负相关关系(P ＜ 0.05)。说明黄土高原三种人工林0-60 cm土层MNC主要贡献MAOC的积累,油松林0-10cm土层除外,且与细菌残体碳相比,真菌残体碳在土壤有机碳组分积累中的贡献更大,土壤总有机碳、全氮、黏粒、砂粒含量、pH值是影响该区不同人工林型微生物残体碳贡献土壤有机碳组分积累的主要因素。     

海南五指山热带山地雨林植物物种多样性研究

王指山热带山地雨林的物种种类十分丰富,在１ｈｍ＾２样地中共出现５４个科,１００个属的乔木树种１７７种,１３３７个个体。区系地理分析表明属的分布区类型以热带分布型成分占优,为８９．０％,充分体现了五指山山地雨林的热带性质。多样性指数分析结果为：样地１Ｍａｒｇａｌｅｆ指数１７．８２２,Ｓｈａｎｎｏｎ－Ｗｉｅｎｅｒ指数５．６２１,均匀度０．８２３,Ｓｉｍｐｓｏｎ指数０．０５０,均匀度６．７７５,表明五指     

不同植物群落下酸化尾矿养分状况及土壤酶活性

调查不同植被群落下铜尾矿的养分状况及土壤酶活性的变化动态,通过常规方法测定了尾矿的pH和电导率,以及有机质,总氮,有效磷,速效钾的含量。用底物反应法测定了过氧化氢酶,芳基硫酯酶,脲酶,酸性磷酸化酶和脱氢酶的活性。结果表明,与酸化的裸露尾矿相比,定居其上的芦苇(Phragmites australis)、狗牙根(Cynodon dactylon)和双穗雀稗(Paspalum distichum)均能显著增加尾矿基质pH值(从3.6上升到5.4),降低电导率,减缓尾矿酸化过程。芦苇和狗牙根能够显著提高尾矿基质中N、K和有机质含量(P<0.05),增加尾矿中的养分。3种植物群落下有效磷较裸露尾矿没有显著增加。植物的定居显著提高了尾矿中过氧化氢酶、芳基硫脂酶和脲酶的活性(P<0.05);但酸性磷酸化酶和脱氢酶活性没有显著提高。研究表明,植物对尾矿有明显的改良作用,而且芦苇和狗牙根优于双穗雀稗。在各种植被条件的尾矿中除酸性磷酸化酶以外的其它实验土壤酶活性均与尾矿中有机质和总氮呈极显著正相关(P<0.01)。