Direct amplification of nodD from community DNA reveals the genetic diversity of Rhizobium leguminosarum in soil

Sequences of nodD , a gene found only in rhizobia, were amplified from total community DNA isolated from a pasture soil. The polymerase chain reaction (PCR) primers used, Y5 and Y6, match nodD from Rhizobium leguminosarum biovar trifolii , R. leguminosarum biovar viciae and Sinorhizobium meliloti . The PCR product was cloned and yielded 68 clones that were identified by restriction pattern as derived from biovar trifolii [11 restriction fragment length polymorphism (RFLP) types] and 15 clones identified as viciae (seven RFLP types). These identifications were confirmed by sequencing. There were no clones related to S. meliloti nodD . For comparison, 122 strains were isolated from nodules of white clover ( Trifolium repens ) growing at the field site, and 134 from nodules on trap plants of T. repens inoculated with the soil. The nodule isolates were of four nodD RFLP types, with 77% being of a single type. All four of these patterns were also found among the clones from soil DNA, and the same type was the most abundant, although it made up only 34% of the trifolii -like clones. We conclude that clover selects specific genotypes from the available soil population, and that R. leguminosarum biovar trifolii was approximately five times more abundant than biovar viciae in this pasture soil, whereas S. meliloti was rare.     

Bacteria from wheat and cucurbit plant roots metabolize PAHs and aromatic root exudates: Implications for rhizodegradation

The chemical interaction between plants and bacteria in the root zone can lead to soil decontamination. Bacteria that degrade polycyclic aromatic hydrocarbons (PAHs) have been isolated from the rhizospheres of plant species with varied biological traits; however, it is not known what phytochemicals promote contaminant degradation. One monocot and two dicotyledon plants were grown in PAH-contaminated soil from a manufactured gas plant (MGP) site. A phytotoxicity assay confirmed greater soil decontamination in rhizospheres when compared to bulk soil controls. Bacteria were isolated from plant roots (rhizobacteria) and selected for growth on anthracene and chrysene on PAH-amended plates. Rhizosphere isolates metabolized 3- and 4-ring PAHs and PAH catabolic intermediates in liquid incubations. Aromatic root exudate compounds, namely flavonoids and simple phenols, were also substrates for isolated rhizobacteria. In particular, the phenolic compounds—morin, caffeic acid, and protocatechuic acid—appear to be linked to bacterial degradation of 3- and 4-ring PAHs in the rhizosphere.     

The isolation of Yersinia intermedia strains with a plasmid of 82-megadalton molecular weight in natural populations of Yersinia

For the first time Y. intermedia strains containing plasmids with a molecular weight of 82 MD have been detected in natural populations of urease-positive Yersinia. Such populations have been isolated from two species of birds and from the soil in the area where they have been killed (the Maritime Territory), as well as from washings from the surface of onions in a vegetable store (Chita Province). The strains, administered orally to white mice, proved to be nonpathogenic. Plasmids with a molecular weight of 82 MD are supposed to occur in natural populations of other Yersinia species.     

The occurrence of Achlya recurva (Saprolegniales, Oomycetes) in hydrocarbon-polluted soil from Argentina

Saprolegniaceae are ubiquitous filamentous water molds. They occur as saprotrophs or parasitic on aquatic and terrestial plants and on aquatic animals, and a number of interesting fungi belonging to the Saprolegniaceae have been isolated from the soil. In the course of an investigation of the water molds in Argentina, an unusual species of Achlya was isolated once from soil collections. Brassica napus seeds were used as bait as well as for the maintenance of this fungus in gross water cultures. Achlya recurva Cornu (Saprolegniales, Oomycetes), was isolated from a hydrocarbon-polluted soil (crude oil) in the neighborhood of an oil tank from a distillery (Ensenada, Buenos Aires Province). This species is reported, described and illustrated for the first time from Argentina. Remarks on the habitat are also made because it was never found in polluted areas.     

Engineering plants for the phytoremediation of RDX in the presence of the co-contaminating explosive TNT

The explosive compounds hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and 2,4,6-trinitrotoluene (TNT) are widespread environmental contaminants commonly found as co-pollutants on military training ranges. TNT is a toxic carcinogen which remains tightly bound to the soil, whereas RDX is highly mobile leaching into groundwater and threatening drinking water supplies. We have engineered Arabidopsis plants that are able to degrade RDX, whilst withstanding the phytotoxicity of TNT. Arabidopsis thaliana (Arabidopsis) was transformed with the bacterial RDX-degrading xplA, and associated reductase xplB, from Rhodococcus rhodochrous strain 11Y, in combination with the TNT-detoxifying nitroreductase (NR), nfsI, from Enterobacter cloacae. Plants expressing XplA, XplB and NR remove RDX from soil leachate and grow on soil contaminated with RDX and TNT at concentrations inhibitory to XplA-only expressing plants. This is the first study to demonstrate the use of transgenic plants to tackle two chemically diverse organic compounds at levels comparable with those found on contaminated training ranges, indicating that this technology is capable of remediating concentrations of RDX found in?situ. In addition, plants expressing XplA and XplB have substantially less RDX available in aerial tissues for herbivory and potential bioaccumulation.     

Molecular and evolutionary analysis of a plant Y chromosome.

Plants have evolved a great diversity of sex determination systems. Among these, the XY system, also found in mammals, is one of the most exciting since it gives the opportunity to compare the evolution of sex chromosomes in two different kingdoms. Whereas genetic and molecular mechanisms controlling sex determination in drosophila and mammals, have been well studied, very little is known about such processes in plants. White campion (Silene latifolia) is an example of plant with X and Y chromosomes. What is the origin of the X and Y chromosomes? How did they evolve from a pair of autosomes? In our laboratory, we have isolated the first active genes located on a plant Y chromosome. We are using them as markers to trace the origin and evolution of sex chromosomes in the Silene genus.     

Dynamic gene order on the <Emphasis Type="Italic">Silene latifolia</Emphasis> Y chromosome

Dioecious Silene latifolia evolved heteromorphic sex chromosomes within the last ten million years, making it a species of choice for studies of the early stages of sex chromosome evolution in plants. About a dozen genes have been isolated from its sex chromosomes and basic genetic and deletion maps exist for the X and Y chromosomes. However, discrepancies between Y chromosome maps led to the proposal that individual Y chromosomes may differ in gene order. Here, we use an alternative approach, with fluorescence in situ hybridization (FISH), to locate individual genes on S. latifolia sex chromosomes. We demonstrate that gene order on the Y chromosome differs between plants from two populations. We suggest that dynamic gene order may be a general property of Y chromosomes in species with XY systems, in view of recent work demonstrating that the gene order on the Y chromosomes of humans and chimpanzees are dramatically different.     

A TaqMan polymerase chain reaction method for monitoring RDX-degrading bacteria based on the xplA functional gene

Hexahydro-1,3,5-trinitro-1,3,5,-triazine (RDX) is a cyclic nitramine explosive that is a major component in many military high-explosive formulations. In this study, we developed a real-time TaqMan polymerase chain reaction (PCR) that targets the xplA functional gene involved in the breakdown/transformation of RDX. The xplA gene, described previously [Seth-Smith, H.M., Rosser, S.J., Basran, A., Travis, E.R., Dabbs, E.R., Nicklin S., Bruce, N.C., 2002. Cloning, sequencing, and characterization of the hexahydro-1,3,5-trinitro-1,3,5-triazine degradation gene cluster from Rhodococcus rhodochrous. Appl. Environ. Microbiol. 68, 4764-4771.], was isolated from Rhodococcus rhodochrous 11Y and codes for a fused flavodoxin-cytochrome P450 protein. We applied the xplA TaqMan PCR assay to detect and monitor strain 11Y in soil microcosms that had been amended with strain 11Y and RDX as well as soil microcosms in which soils had been subjected to heat-sterilization prior to the addition of strain 11Y and RDX. The specificity of the assay was tested against a number of genomic bacterial templates and surprisingly found to cross react with other RDX degrading bacteria. Two of these strains, Gordonia sp. KTR9 and Williamsia sp. KTR4, were previously isolated in our laboratory and were not known to possess xplA homologs. Southern blot analysis confirmed the presence of xplA gene homologs in both of these strains. The sensitivity of the xplA TaqMan PCR primer/probes set was evaluated using 11Y cell standards as well as 11Y cell standards spiked in soils that mimicked conditions found in the experimental soil microcosms. While the assay was found to be linear over a range of 6 orders of magnitude for both sets of standards, sensitivity of the assay was reduced between one and two logs for cells spiked in soil. The capacity to monitor the presence of specific microorganisms and/or genes coding enzymes involved in RDX transformation/breakdown in complex environmental samples will be critical for bioremediation strategies targeting explosives that rely on in situ bioaugmentation and monitored natural attenuation.     

华北棉区第三代棉铃虫的经济阈值

1980—1982、1984—1985年在河北省饶阳县的不同土壤肥力水平、不同棉花品种上进行了模拟为害和自然为害试验,旨在确定第三代棉铃虫 Heliothis armigera(Hubner)虫口密度与棉花产量之间的关系.结果表明,棉花对于第三代棉铃虫为害的补偿能力较弱.较好的土壤肥力亦不能促使棉株完全补偿三代期间的受害损失.每亩皮棉减产斤数Y与第三代百株累计卵量E₃的关系为:Y=-0.2801+0.0643E₃.根据防治的直接收益与直接代价,算出了第三代棉铃虫的经济阈值为百株累计卵量35粒,这一阈值可适用于不同肥力水平的地块.     

Somatic Crossing over in GLYCINE MAX (L.) Merrill: Mutagenicity of Sodium Azide and Lack of Synergistic Effect with Caffeine and Mitomycin C

Glycine max (soybean) is one angiosperm which lends itself to the study of somatic crossing over. This is made possible because some varieties have gene combinations Y(11)Y(11), Y(11)y(11) and y(11)y(11) in the segregating populations from Y(11)y(11) plants. The gene in question is responsible for chlorophyll synthesis. The Y(11)Y(11) plants have dark green leaves, Y(11)y(11) are light green and y(11)y(11) plants are golden yellow. The heterozygous plants have dark green, yellow and dark green-yellow (double) spots on the leaves of the untreated control material, whereas the two homozygotes are almost always devoid of somatic sectoring. Application of caffeine, or mitomycin C, to the seeds increased the frequency of double, dark green and yellow spots on the Y(11)y(11) background. Possibly, some dark green or yellow spots originate by failure of one of the two components of what might start as a double spot due to somatic crossing over. The application of NaN(3) increases the frequency of dark green or yellow spots, almost exclusively. The two spots increase in equal frequency. The y(11)y(11) plants so treated do not have any light green sectors, but dark green, Y(11)Y(11), plants do develop a few light green or very dark green spots. The data indicate that NaN(3) is capable of inducing nondisjunction, but does not cause mutations (at this locus), chromosome fragmentations (segmental losses) or somatic crossing over to an appreciable degree. It has previously been shown that caffeine-induced chromosome rejoining in Vicia faba can be inhibited by treating the roots with NaN(3). In the present experiments NaN(3) did not affect the processes of somatic crossing over as induced by caffeine or mitomycin C. The effect was additive. This system offers advantages for studying chemical mutagens in that somatic crossing over, point mutations, segmental losses through chromosome breakage and nondisjunction can all be studied in a single treatment to the seeds.     

广西蜘蛛抱蛋中一种甾体皂甙定性定量研究

为了进一步开发和利用广西甾体药源植物奠定基础 ,对广西蜘蛛抱蛋 (Aspidistra retusa K.Y.Langet.S.Z.Huang)块茎甾体皂甙化合物进行了提取分离。在分离得化合物中含量最高的甾体皂甙化合物 4经鉴定为 :偏诺甾体四糖皂甙 ,即 pennogenin-3-β-O-β-D-glucopyranosyl[-α-L-rhamno-pyranosyl(1 -2 ) ]-α-L-rham-no-pyanosyl(1 -4 ) -α-L-rhamnopyranosyl(1 -4 )。该化合物为首次从蜘蛛抱蛋属植物中分离得到。经 HPLC测定其在广西蜘蛛抱蛋中的含量为 1 .5 95 %。     

Isolation of Yersinia group in human infections, animals and environment factors

The presence of Y. enterocolitica and Y. pseudotuberculosis was studied in 4479 enteritis cases, 430 children, presenting appendicular syndrome, and 60 hospitalized patients with arthritis and Reiter syndrome. Y. enteritis was detected in 41 (0.9%) enteritis cases, 15 (3.4%) appendectomized children and 5 (8.3%) arthritis cases. Antibodies to Y. pseudotuberculosis were detected in 2 (3.3%) arthritis patients. Y. enterocolitica was isolated in swine, fish and environment factors (water, soil, food). Y. pseudotuberculosis was isolated in soil. The isolated strains belonged to biotypes 1, 2, 4 and serotypes 0:3; 0:5; 0:5.27; 0:5, 6, 7, 8; 0:6; 0:9; some were non-typable and polyagglutinable. The strains were sensitive to bacteriophages for Yersinia, obtained in our laboratory.     

Sex Determination by Sex Chromosomes in Dioecious Plants

Abstract: Sex chromosomes have been reported in several dioecious plants. The most general system of sex determination with sex chromosomes is the XY system, in which males are the heterogametic sex and females are homogametic. Genetic systems in sex determination are divided into two classes including an X chromosome counting system and an active Y chromosome system. Dioecious plants have unisexual flowers, which have stamens or pistils. The development of unisexual flowers is caused by the suppression of opposite sex primordia. The expression of floral organ identity genes is different between male and female flower primordia. However, these floral organ identity genes show no evidence of sex chromosome linkage. The Y chromosome of Rumex acetosa contains Y chromosome-specific repetitive sequences, whereas the Y chromosome of Silene latifolia has not accumulated chromosome-specific repetitive sequences. The different degree of Y chromosome degeneration may reflect on evolutionary time since the origination of dioecy. The Y chromosome of S. latifolia functions in suppression of female development and initiation and completion of anther development. Analyses of mutants suggested that female suppressor and stamen promoter genes are localized on the Y chromosome. Recently, some sex chromosome-linked genes were isolated from flower buds of S. latifolia.     

紫花苜蓿根际氢氧化细菌的分离及其对小麦种子促生作用的研究

从大豆植物根际分离的氢氧化细菌对植物的生长有促进作用,但是关于其他的豆科植物根际分离的氢氧化细菌是否也有促生作用的研究甚少。从紫花苜蓿根际土壤分离氢氧化细菌,并进行其对小麦种子促生实验的研究,判断氢氧化细菌是否有促生作用,从而丰富具有促生作用的根际微生物资源。采用MSA培养基,从铜川新区紫花苜蓿根际土壤中分离得到氢氧化细菌疑似菌株,对其进行TTC法检测菌株氢化酶活性和自养能力的特性,以获得氢氧化细菌;通过小麦种子的萌发进行促生实验验证。结果表明,16株菌株处理过的小麦根长分别增加25%～128%,芽长增长27%～73%,鲜重增加48%～103%。从苜蓿根际土壤分离出的氢氧化细菌均具有较明显的促生作用。     

具细菌群体感应抑制活性海洋细菌的筛选鉴定

目的:从海洋环境中筛选对细菌群体感应有抑制作用的活性菌株,为以致病菌群体感应系统为靶点的新型疗法提供新的药用资源。方法:从海水中分离纯化细菌菌株,采用根癌农杆菌平板筛选模型筛选细菌群体感应抑制活性细菌,对筛选出的海洋细菌进行生理生化和16S rDNA序列测定,根据《伯杰氏手册》进行菌种分类鉴定。结果:从217株海洋细菌中筛选出1株能显著抑制细菌群体感应效应的海洋细菌Y2,该海洋细菌具有蜡样芽孢杆菌(Bacillus cereus)的典型特征,其16S rDNA序列与GenBank中蜡样芽孢杆菌16S rDNA的部分序列有100%的同源性。结论:海洋环境中也存在具有抑制细菌群体感应活性的微生物。     

Degradation of estrogens by Rhodococcus zopfii and Rhodococcus equi isolates from activated sludge in wastewater treatment plants

We have isolated four strains of Rhodococcus which specifically degrade estrogens by using enrichment culture of activated sludge from wastewater treatment plants. Strain Y 50158, identified as Rhodococcus zopfii, completely and rapidly degraded 100 mg of 17beta-estradiol, estrone, estriol, and ethinyl estradiol/liter, as demonstrated by thin-layer chromatography and gas chromatography-mass spectrometry analyses. Strains Y 50155, Y 50156, and Y 50157, identified as Rhodococcus equi, showed degradation activities comparable with that of Y 50158. Using the random amplified polymorphism DNA fingerprinting test, these three strains were confirmed to have been derived from different sources. R. zopfii Y 50158, which showed the highest activity among these four strains, revealed that the strain selectively degraded 17beta-estradiol during jar fermentation, even when glucose was used as a readily utilizable carbon source in the culture medium. Measurement of estrogenic activities with human breast cancer-derived MVLN cells showed that these four strains each degraded 100 mg of 17beta-estradiol/liter to 1/100 of the specific activity level after 24 h. It is thus suggested that these strains degrade 17beta-estradiol into substances without estrogenic activity.     

Mesophyll protoplasts from Solanum melongena var depressum bailey regenerate to fertile plants

Mesophyll protoplasts have been isolated from soil grown plants of Solanum melongena var depressum Bailey and protoplast-derived colonies regenerated in liquid DPD culture medium. Protoplast-derived colonies proliferated on agar-solidified MS culture medium and regenerated shoots which grew to fertile plants in potting compost.on leave from Landzhou University, Peoples Republic of China     

Genome sequence of Micromonospora lupini Lupac 08, isolated from root nodules of Lupinus angustifolius

Micromonospora strains have been isolated from diverse niches, including soil, water, and marine sediments and root nodules of diverse symbiotic plants. In this work, we report the genome sequence of Micromonospora lupini Lupac 08 isolated from root nodules of the wild legume Lupinus angustifolious.     

Role of dissimilatory fermentative iron-reducing bacteria in Fe uptake by common bean (Phaseolus vulgaris L.) plants grown in alkaline soil

Iron (Fe) is an essential element for plant growth and development. Some plant growth-promoting rhizobacteria can increase Fe uptake by plants through reduction of Fe(III) to Fe(II) at the root surface. The aim of this work was to identify novel bacterial strains with high Fe(III) reduction ability and to evaluate their role in plant Fe uptake. Four bacterial strains (UMCV1 to UMCV4) showing dissimilatory Fe-reducing activity were isolated from the rhizosphere of bean and maize plants and further identified by 16S rDNA amplification and sequence analysis. From these analyses, UMCV1 and UMCV2 isolates were identified as Bacillus megaterium and Arthrobacter spp., respectively, whereas UMCV3 and UMCV4 were identified as Stenotrophomonas maltophilia. All four isolates showed Fe reduction in a nonflooded soil and when associated with roots of bean plants grown in alkaline soil or in mineral medium. In addition, the bacterial isolates were able to stimulate plant growth in vitro and on a broad level, plants grown in inoculated soil were generally bigger and with higher Fe content than those grown in sterilized soil. These results indicate that bacterial species isolated from the rhizosphere of bean and maize plants contribute significantly to Fe uptake by plants likely through increased Fe(III) reduction in the rhizosphere.