一株敌敌畏降解菌DDV-1(Ochrobactrium sp.)的分离鉴定及其降解特性的研究

Soil organic carbon （C） and total nitrogen （N） pools of a Chinese fir （Cunninghamia lanceolata （Lamb.） Hook.） （CF） forest, and an evergreen broadleaf （EB） forest located in mid-subtropical, southeastern China, were compared before clearcutting, with the effect of slash burning on organic C and total N in the top 10 cm of soil before and after burning also being evaluated. Prior to clearcutting CF forest had significantly lower （P 〈0.05） organic C and total N in the soil （0-100 cm） compared to EB forest with approximately 60% of the C and N at the two forest sites stored at the 0 to 40 cm soil. In post-burn samples of the 0-10 cm depth at 5 days, 1 year, and 5 years for CF and EB forests, significantly lower levels （P 〈0.05） of organic C and total N than those in the pre-burn samples were observed. Compared to the pre-burn levels, at post-burn year 5, surface soil organic C storage was only 85% in CF forest and 72% in EB forest, while total N storage was 77% for CF forest and 73% for EB forest. Slash burning caused marked long-term changes in surface soil C and N in the two forest types.     

接种固氮菌Klebsiella sp.120对甘蔗光合特性和主要矿质营养元素含量的影响

【目的】甘蔗是最重要的糖料和能源作物,生物量大,产量高,但大量氮肥的使用不仅增加了甘蔗的种植成本,而且对农田环境造成了污染。发挥与甘蔗联合固氮的生物固氮菌的作用,是降低甘蔗氮肥施用量的方法之一。Klebsiella sp.120是从广西种植的甘蔗体内分离到的具有固氮能力的菌株。为了探讨该菌对甘蔗的促生效应及其在甘蔗上的应用潜能,本文研究了接种该固氮菌对甘蔗光合特性及矿质元素含量的影响。【方法】采用桶栽方法,在甘蔗的分蘖期用接种菌株淋浇4个不同甘蔗品种的根际,在处理后的30 d和60 d取样分析该菌对甘蔗的光合生理和矿质营养元素含量的影响。【结果】接种菌株对4个甘蔗品种的叶绿素含量和净光合速率有一定的促进作用,但不同品种之间存在差异。处理30 d后,淋浇固氮菌处理的甘蔗品种ROC22的叶绿素含量明显提高,比对照增加23%。另外3个甘蔗品种(B8、GT21和GT28)处理组的叶绿素含量均有一定的提高,但与对照相比差异不显著。处理30 d后,只有ROC22和B8品种的净光合速率高于对照,且ROC22品种处理与对照间的差异达到了显著水平。处理60 d后,4个甘蔗品种的净光合速率均高于处理30 d,而且各品种接种处理均高于对照,其中以对ROC22和GT21的促进效果最为明显,达到了显著水平。接种处理对不同甘蔗品种的营养元素含量影响不同,接种处理对4个甘蔗品种叶片中的氮元素含量有促进作用,其中对ROC22的促进效果最为明显,接种处理比对照增加59.8%。接种固氮菌对甘蔗叶片磷和钾元素含量的影响不明显。接种固氮菌对4个甘蔗品种叶片的铁、锰元素含量的影响不明显;ROC22叶片中的锌含量及ROC22和B8叶片中的铜含量接种处理与对照相比显著提高。【结论】接种Klebsiella sp.120固氮菌对甘蔗光合特性及矿质元素含量的影响与甘蔗品种有关,其中对ROC22品种的促生效果最为明显。研究结果可为固氮菌株Klebsiella sp.120在甘蔗上的进一步应用提供参考依据。     

Isolation, identification, and characterization of an aluminum-tolerant bacterium Burkholderia sp. SB1 from an acidic red soil

Aluminum (Al) toxicity usually occurs in acidic soils worldwide, which is detrimental to the growth of organisms. An Al-tolerant bacterium, SB1, was isolated from an acidic red soil of Chingkang Mountain, located in Jiangxi Province of China. Polyphasic analysis, including a 16S rDNA phylogenetic tree as well as morphological and physicochemical properties, revealed that the isolate was a gramnegative, rod-shaped bacterium, which was recognized as Burkholderia sp. SB1 and had extreme acidity tolerance (pH 2.2) and excellent Al resistance (270 mg L^-1 Al³⁺). It could remove Al by up to 97.7% at a concentration of 54 mg L^-1 Al³⁺. SB1 behavior under different temperatures and antibiotics was also examined. SB1 preferred moderate temperature conditions, ranging from 25 to 37℃, and exhibited notable resistance to multiple antibiotics (including ampicillin, streptomycin, and tetracycline), except for being sensitive to chloramphenicol. Therefore, as the first reported bacterium to possess favorable Al resistance and excellent Al removal, Burkholderia sp. SB1 can potentially be used as an agent for bioremediation of Al-contaminated acidic red soils.     

荧光原位杂交技术检测土壤中博德特氏菌探针的设计与应用

根据博德特氏菌(Bordetellasp.)的16S rRNA基因序列,设计荧光原位杂交(FISH)检测博德特氏菌的寡核苷酸探针FW_iso_62和FW_iso_761,在20%～60%甲酰胺均有很强的荧光信号。采用探针FW_iso_62及其竞争探针,结合Nycodenz和DAPI技术,建立定量检测土壤中博德特氏菌的DAPI-FISH方法。该方法可排除土壤颗粒的自动荧光对细菌信号的掩盖,保证图片中有大量微生物供统计分析,还能有效保存微生物的原位信息。应用该方法分析土壤中1,2,4-三氯苯降解菌-博德特氏菌,结果未受氯苯污染的农田土壤中没有检测到博德特氏菌,而氯苯污染土壤中检测到大量的博德特氏菌,每克土壤含3.78×106个。将该污染土壤中分离的博德特氏降解菌及其降解菌群接种至农田土壤中,降解菌的数量均随培养而增加,一个月后分别占DAPI计数的1.7%和3.8%。本研究设计的探针可有效用于复杂环境样品中博德特氏菌的定性与定量检测。     

Vigna radiata var. GM4 Plant Growth Enhancement and Root Colonization by a Multi-Metal-Resistant Plant GrowthPromoting Bacterium Enterobacter sp. C1D in Cr(VI)-Amended Soils

Contamination of agricultural soils by heavy metals has become a major concern due to their toxic effects on plant growth, symbiosis and consequently the yields of crops. In the present study, to enhance plant growth in Cr(VI)-amended soils, novel metalresistant plant growth-promoting bacteria (PGPB) were isolated from a soil contaminated with industrial waste effluent. One of the bacterial isolates, identified as Enterobacter sp. C1D by 16S rRNA gene sequencing, was found to be multi-metal resistant in nature with excellent plant growth-promoting (PGP) traits. Mung bean (Vigna radiata var. GM4) inoculation with Enterobacter sp. C1D significantly (P < 0.01) increased root and shoot length, shoot and root weight, and chlorophyll content in a range of Cr(VI) treatments. Plant tolerance towards Cr(VI) measured as effective concentration showed higher values with Enterobacter sp. C1Dtreated plants compared to un-inoculated plants. Root colonization study was also carried out using green fluorescence protein-labeled Enterobacter sp. C1D under a hydroponic system. Confocal laser scanning microscopy of the plant roots showed heavy bacterial loads on the surface of the plant root specifically at the root tip and the point of root hair/lateral root formation. The results of PGP traits showed that elevated indole acetic acid levels and 1-aminocyclopropane-1-carboxylate deaminase activity enabled Enterobacter sp. C1D to enhance V. radiata growth in Cr(VI)-amended soils, whereby it significantly increased plant tolerance towards elevated Cr(VI) concentrations.     

Methane oxidation activity and bacterial community composition in a simulated landfill cover soil is influenced by the growth of Chenopodium album L.

Oxygen availability in landfill cover soil is a major limitation to the growth and activity of methanotrophs as methane oxidation is an aerobic microbial process. Plants tolerant to high concentrations of landfill gas (LFG) may play an important role in improving methane oxidation within landfill cover soil and reducing emission of methane, a greenhouse gas, from it. In this study, the effect of an LFG tolerant plant Chenopodium album L. on methane oxidation activity (MOA) and bacterial community composition in landfill cover soil was investigated. Soil samples from four simulated lysimeters with and without LFG and plant vegetation were taken at 4 stages during the plant's development cycle. Results showed that the total number of culturable bacteria in soil could be significantly increased (P < 0.05) by the growth of C. album. The total number of methanotrophs and MOA in soils with LFG was significantly higher (P < 0.05) than in soils without LFG on sampling days 90, 150 and 210. The total number of methanotrophs and MOA in lysimeters with LFG added increased in the presence of C. album when the plant entered the seed setting stage. Polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) gel patterns of 16S rDNA gene fragment and band sequencing analyses showed apparent differences in soil bacterial communities in the presence of LFG and plant vegetation. Members of the genus Methylosarcina were found to be the active and dominant methanotrophs in rhizosphere soil of C. album with LFG, while Methylococcus, Methylocystis, and Methylosinus were the primary methanotroph genera in LFG soil without C. album. Thus, C. album appears to select for specific methanotrophic bacteria in the presence of LFG. Soil MOA and microbial diversity can also be significantly affected by the presence of this plant.     

Nitrite production by Nitrosomonas europaea and Nitrosospira sp. AV in soils at different solution concentrations of ammonium

Although it remains unclear why NH₃-oxidizing bacteria (AOB) of the genus Nitrosospira dominate soil environments, and why Nitrosomonas spp. are less common, virtually no studies have compared their behavior in soil. In this study, the NH₃ oxidation rates of Nitrosomonas europaea (ATCC 19718) and Nitrosospira sp. AV were compared in three differently textured soils containing a range of extractable contents (2-11 μg soil). Soils were adjusted to pH 7.0-7.4 with CaCO₃ and sterilized with γ-radiation. Cell suspensions of each bacterium were inoculated into the soils to bring them to two-third of water-holding capacity and cell densities ∼2.5×10⁶ g⁻¹ soil. In virtually all cases, rates of production for both N. europaea and Nitrosospira sp. AV were linear over 48 h, and represented between 13 and 75%, respectively, of the maximum rates achieved in soil-free bacterial suspensions. Soil solution concentrations that supported these rates ranged between 0.2 and 1.5 mM. Addition of 21-36 μg soil raised soil solution levels to 1.8-2.5 mM and stimulated production to a greater extent in N. europaea (3.3-6.6-fold) than in Nitrosospira sp. AV (1-2.1-fold). Maximum rates of production were obtained by raising soil solution levels to 3-4 mM with a supplement of ∼80-90 μg soil. K_s values in soil for Nitrosospira sp. AV and N. europaea were estimated as 0.14 and 1.9 mM , respectively, and estimates of V_max were about 3.5-times higher for N. europaea (0.007 pmol h⁻¹ cell⁻¹) than for Nitrosospira sp. AV (0.002 pmol h⁻¹ cell⁻¹). The cell density of N. europaea increased in sterile Steiwer soil independent of supplemental . In the case of treatments receiving supplemental , growth yields of N. europaea calculated from either produced or consumed were similar to those reported in literature (3.5×10⁶-6×10⁶ cells μmol⁻¹). A higher growth yield was measured in the case of zero added (2.7×10⁷ cells μmol⁻¹), indicating that use of organic carbon compounds might have occurred and resulted in some energy sparing. Our results suggest that Nitrosospira spp. with a K_s similar to Nitrosospira sp. AV may have an advantage for survival in soil environments where soil solution levels are less than 1 mM. However, it is apparent that AOB like N. europaea are poised to take advantages of modest increases in extractable that raise soil solution levels to about 2.0-2.5 mM.