Reclamation and revegetation of fly ash disposal sites - Challenges and research needs

Coal-fired power generation is a principal energy source throughout the world. Approximately, 70-75% of coal combustion residues are fly ash and its utilization worldwide is only slightly above 30%. The remainder is disposed of in landfills and fly ash basins. It is desirable to revegetate these sites for aesthetic purposes, to stabilize the surface ash against wind and water erosion and to reduce the quantity of water leaching through the deposit. Limitations to plant establishment and growth in fly ash can include a high pH (and consequent deficiencies of Fe, Mn, Cu, Zn and P), high soluble salts, toxic levels of elements such as B, pozzalanic properties of ash resulting in cemented/compacted layers and lack of microbial activity. An integrated organic/biotechnological approach to revegetation seems appropriate and should be investigated further. This would include incorporation of organic matter into the surface layer of ash, mycorrhizal inoculation of establishing vegetation and use of inoculated legumes to add N. Leaching losses from ash disposal sites are likely to be site-specific but a sparse number of studies have revealed enriched concentrations of elements such as Ca, Fe, Cd, Pb, and Sb in surrounding groundwater. This aspect deserves further study particularly in the longer-term. In addition, during weathering of the ash and deposition of organic matter during plant growth, a soil will form with properties vastly different to that of the parent ash. In turn, this will influence the effect that the disposal site has on the surrounding environment. Nevertheless, the effects of ash weathering and organic matter accumulation over time on the chemical, physical and biological properties of the developing ash-derived soil are not well understood and require further study.     

快生型大豆根瘤菌(Rhizobium fredii)内源质粒在培养及共生过程中的稳定性

以湖北洪湖公离的快生型大豆根瘤菌为材料，考察了它们的内源质粒在培养过程及共生过程中的稳定性．结果表明，所有供试菌株的质粒在培养条件下比较稳定，传代50次后没有明显变异．对其中2个菌株与4种不同的大豆宿主共生过程中质粒的稳定性研究，发现HA12－1－12的共生质粒和隐性质粒高度构定，HA7－2－2的质粒却发生了不同程度的变异．质粒检测的结果及RFLP分析表明，HA7－2－2共生质粒的稳定性高于隐性质粒，而且共生质粒上携带的nodDABC和nifHDK基因相当保守，不易受宿主的影响而产生变异．盆栽试验的结果也说明，虽然HA7－2－2的内源质粒在共生过程中发生了遗传重排，但它的共生性状没有明显的变异．     

从水稻种子表面分离的紫云英根瘤菌与不同水稻品种亲合性的研究

采用紫云英琼脂管结瘤试验，在10个供试的不同水稻品种中，仅从“汕优63”和“珍A一代”2个品种的种子表面各分离筛选到在紫云英上结瘤较早且较多的1个菌株，分别命名为SY63－4和SA1D－3。采用无菌盆栽试验分别将这2个菌株接种于4个不同水稻品种“汕优63”、“珍A一代”、“马协63”及“博湛优19”，发现菌株ZA1D－3对4种水稻均表现出显著的促生作用，SY63－4仅对“汕优63”和“珍A一代”表现出明显的促生作用。利用MPN法检测不同水稻品种根内外根瘤菌ZA1D－3和SY63－4的数量，发现ZA1D－3和SY63－4在4种水稻根部的定殖数量也不相同。SY63－4在“汕优63”根部定殖的数量log(ncfumFW^-1/g^-1)为5．20，比在其余3种水稻根部高出2个数量级；ZA1D－3在“珍A一代”根部定殖的最大数量log(ncfumFW^-1/g^-1)为4．54，比在其余3种水稻根部高出1个数量级。这说明SY63－4与“汕优63”，ZA1D－3与“珍A一代”间存在着一定的亲和性。图1表2参12     

多氯联苯污染土壤菌根真菌-紫花苜蓿-根瘤菌联合修复效应

选用紫花苜蓿（Medicago sativa L.）作为宿主植物,盆栽试验研究了丛枝菌根真菌（Glomus caledonium）和苜蓿根瘤菌(Rhizobium meliloti)单接种及双接种对PCBs复合污染土壤的联合修复效应.结果表明,在紫花苜蓿-菌根真菌-根瘤菌共生体系中,紫花苜蓿对土壤中PCBs的降低起到明显作用,使轻度污染和重度污染土壤中PCBs浓度分别下降了15.8%、 23.5%,紫花苜蓿单接种菌根真菌和苜蓿根瘤菌后轻度污染和重度污染土壤中PCBs浓度分别下降了14.8％、 24.1%和20.6%、 25.5%,双接种后土壤PCBs分别降低了23.2％、26.9%,而且也改变了紫花苜蓿根际土壤微生物群落的碳源利用程度,改善了微生物群落功能多样性.可见,紫花苜蓿豆科植物-菌根真菌-根瘤菌特殊共生体对PCBs污染土壤显示了较好的修复潜力.     

Biodegradation of N,N-bis(carboxymethyl)-L-glutamate and its utilization as sole source of carbon,nitrogen, and energy by a Rhizobium radiobacter strain in seawater

Biodegradation of N,N-bis(carboxymethyl)-L-glutamate and its utilization by micro-organisms in marine environments were studied in Organization of Economic Development and Cooperation (OECD) screening tests and with pure bacterial cultures. Biodegradation in excess of 60% was achieved in closed bottle tests in less than 60 days, suggesting complete degradation. The bacterial strain isolated from marine sandy sediment and utilizing N,N-bis(carboxymethyl)-L-glutamate as sole source of carbon, nitrogen, and energy was identified as Rhizobium radiobacter. The strain was capable to mineralize N,N-bis(carboxymethyl)-L-glutamate in seawater and in synthetic media containing 0–60 g L^?1 sea salts. Growth and adaptation studies indicated that N-carboxymethyl-L-glutamate and L-glutamate are main metabolites.     

紫云英根瘤菌共生基因分布及其共生效应的多样性

从同一植株不同根瘤分离40株紫云英根瘤菌，所有菌株对10种抗生素的抗药性测定表明，该群体分为22个抗药类群，质粒检测显示所有公离株都含有质粒，质粒数1～4条，用快生型大豆根瘤菌USDA205质粒作参考，估测质粒Mr分布范围为83～226MU．根据图谱分析表明，该菌群可分为6个不同质粒型．各类型质粒通过与Dig-nodABC和Dig-nifHDK杂交，结果显示带有1条质粒的菌株其共生基因定位在染色体上．带有2条或2条以上质粒的菌株各拥有1条共生质粒，共生质粒Mr范围有差异，大约为117～220MU．研究结果也显示，不同质粒型的菌株其共生效应存在明显差异，其中第6质粒型的菌株共生固氮率最强，第1质粒型菌株共生固氮率较低．共生固氮能力最强的第6质粒型菌株，只占总菌数7．5％．     

Effects of pyrethroid insecticide seed treatments on Rhizobium japonicum and its symbiotic relationship with,and growth of soybean

Abstract

Two pyrethroid insecticides were compared with two fungicides for their effects on the rhizobial and seedling components of the N₂‐fixing symbiosis in soybeans (Glycine max Merr. variety Olinda). In vitro growth of Rhizobium japonicum RI16, was inhibited in the order of HgCl₂ > thiram > cypermethrin > permethrin. The emergence of the seedlings was stimulated by all chemicals tested. Also, none of the pesticides studied had any significant effect on N₂ fixation (acetylene reduction), at the dosages used. Consequently, yields of the various organs were also not significantly influenced by the chemicals.     

苜蓿根瘤菌对多氯联苯降解转化特性研究

采用溶液摇瓶实验研究了苜蓿根瘤菌(Rhizobium meliloti)对三氯代联苯(2,4,4′-trichlorobiphenyl)单体以及18种多氯联苯(polychlorinated biphenyls,PCBs)混合物的降解转化能力.结果表明,接入苜蓿根瘤菌转化7d后,随着溶液中底物2,4,4′-TCB浓度的增加,该菌株对其的降解能力也逐渐提高,在1.0、5.0、10.0、25.0、50.0 mg.L-1浓度条件下该菌对2,4,4′-TCB的降解率分别为34.0%、48.3%、69.7%、96.0%、98.5%,并且通过气相质谱分析,发现溶液中出现了一些新的代谢产物.同时,苜蓿根瘤菌对不同浓度下18种PCBs同系物混合物的降解能力,随着底物浓度的升高呈现出从低到高,然后降低,并趋于平衡的趋势,其最高降解率可达54.7%,并存在高氯代PCBs向低氯代PCBs的转化过程.     

农药降解质粒向根瘤菌的转移

研究了细菌农药降解质粒P~(JP4)向根瘤菌的转移以及农药降解质粒引入根瘤菌后,对其共生性、固氮能力和根瘤结构的影响。结果表明,三种根瘤菌受体均接受了农药2,4-D降解质粒P~(JP4)。转移频率在4.0×10~(-6)—6.4×10~(-6)之间。其中一个受体菌株在灭菌土壤中接受该质粒,其转移频率为5.1×10~(-7)。接受了农药降解质粒的接合子保持了根瘤菌的共生性状、乙炔还原活性,所形成的根瘤结构与亲本株没有明显差别。从根瘤中分离的接合子仍保持了质粒P~(JP4)。     

含发光酶基因的转座子质粒载体的改造及向根瘤菌HN01的转座

将从自杀性重组质粒pDB30上分离的3．6kb含卡那霉素抗性基因和发光酶基因（luxAB）的BglⅡ酶切片段，克隆到小Mr的高拷贝质粒pIJ2925上，构成新的质粒，pHNLX1011；然后将pHNLX1011经BglⅡ部分酶切，酶连在含蔗糖酶基因（sacB）的自杀性重组质粒载体pMH1701上，载体分别采用BglⅡ酶切和BamHⅠ酶切．连接转化后共得到4种重组质粒类型的菌株，它们具有不同发光活性；将它们分别与快生型大豆根瘤菌HN01进行三亲本杂交，发现它们的转座频率均较pDB30高．对经转座标记后的根瘤菌进行发光活性检测，只有pHNC3质粒所转座的根瘤菌浇具有较强发光活性．所得发光根瘤菌衍生菌株能在野大豆和栽培大豆黑农39根际形成正常根瘤，能用X光片记录下根瘤的发光活性．