甘蔗热带种金属硫蛋白家族基因的克隆及响应重金属胁迫的表达分析

金属硫蛋白是一类富含巯基的低分子量蛋白,在植物的重金属解毒及细胞氧化还原调控等方面起重要的作用。本研究以甘蔗热带种Badila组培苗为材料,分别测定了其在CdCl2、ZnSO4和CuCl2水溶液培养条件下地上部和地下部的重金属含量,结果显示其对上述3种重金属有较强的耐受与富集能力。继而克隆了ScMT1(登录号为KJ504373)、ScMT2-1-5(登录号为MH191346)和ScMT3(登录号为KJ5043704)3个金属硫蛋白家族基因,它们分别属于植物MT亚家族中的MT1、MT2和MT3型基因。ScMT1含有1个内含子和2个外显子,开放阅读框(Open Reading Frame,ORF)长228 bp,编码75个氨基酸;ScMT2-1-5含有2个内含子和3个外显子,ORF长246 bp,编码81个氨基酸;ScMT3含有1个内含子和2个外显子,ORF长198 bp,编码65个氨基酸。RT-qPCR显示,Cd2+胁迫下,在甘蔗地上部和地下部,ScMT2-1-5均连续显著上调表达,而ScMT1的上调应答出现延迟。ScMT3在地上部的上调应答出现延迟,在地下部呈“扬-抑”趋势,提示甘蔗响应Cd^2+胁迫过程中ScMT2-1-5起更积极的作用,ScMT1参与胁迫后期的分子响应,而ScMT3不起主导作用。Cu^2+胁迫下,地上部ScMT1连续显著上调表达,ScMT2-1-5和ScMT3呈总体上调的表达趋势;地下部,ScMT1和ScMT2-1-5的上调表答均出现延迟,仅在胁迫后期显著上调表达,而ScMT3仅在胁迫前期显著上调表达。该结果提示了ScMT1、ScMT2-1-5和ScMT3在Cu2+胁迫响应过程中的协作关系,三者共同参与了地上部的胁迫响应,其中ScMT1起更积极的作用;此外三者还先后参与了地下部对Cu^2+胁迫的分子响应。Zn^2+胁迫下,ScMT1和ScMT3分别仅在地上部和地下部显著上调表达;ScMT2-1-5在地上部和地下部均呈“扬-抑”的应答趋势;提示了在甘蔗响应Cd^2+胁迫应答过程中ScMT1和ScMT3分别在地上部和地下部起主要作用,ScMT2-1-5参与了胁迫前期的分子响应。ScMT1、ScMT2-1-5和ScMT3在甘蔗不同组织中及在重金属(Cd^2+、Zn^2+或Cu^2+)不同累积水平下呈现出相似或互补的应答特性,提示上述甘蔗MT家族不同成员在重金属解毒及细胞氧化还原调控等方面产生了功能分化,且三者在应对过量Cd^2+、Zn^2+或Cu^2+对甘蔗组织造成伤害的过程中存在时空上的协同作用。该研究为深入理解多倍体植物甘蔗中MT家族各成员基因在重金属耐受过程中的协同作用机制奠定了基础。     

Do cover crops change the lability of phosphorus in a clayey subtropical soil under different phosphate fertilizers?

Plants have developed different mechanisms to absorb and solubilize phosphorus (P) in the soil, especially in environments with low P availability. This study evaluated the effects of different winter cover crops on soil P availability in a clayey subtropical (Hapludox) soil receiving soluble P fertilizer and a rock phosphate applied to the summer crop, under no‐tillage. The experiment was carried out over 3 yrs (2009–2011) with five different cover crop species: common vetch, fodder radish, ryegrass, black oat, white clover and fallow as control. The soil was sampled after the third year of cover crop cultivation and analysed for inorganic and organic P forms according to the well‐established Hedley fractionation procedure. Phosphate fertilizers promoted accumulation of both labile and nonlabile P pools in soil in the near surface layer, especially under rock phosphate. Fertilizer applications were not able to change P fractions in deeper layers, emphasizing that the Brazilian clayey soils are a sink of P from fertilizer and its mobility is almost nil. Although the cover crops recycled a great amount of P in tissue, in a short‐term evaluation (3 yrs) they only changed the content of moderately labile P in soil, indicating that long‐term studies are needed for more conclusive results.     

Enhancement of sulfide auto-oxidation by metal ion in artificial seawater and a seawater which is a possible source of aoshio

SUMMARY: Aoshio is hypoxic milky blue-green seawater observed in some eutrophic bays. Previous studies have shown that colloidal sulfur causes the coloration and that the source of aoshio water is attributed to coastal anoxic bottom water. Occurrences of aoshio have been reported in limited areas of coastal seawater, although hypoxic transparent water seems rather universal. Promotion in auto-oxidation of sulfide by metal ions in seawater was investigated to explain the occurrences of aoshio. Artificial seawater containing 10 μM metal ion was assayed for the sulfide oxidation rate. The velocity constant which represents the oxidation rate within the initial 30 min and the amount of reacted sulfide in 2 h were determined by oxygen monitoring and sulfide quantification, respectively. Fe²⁺ and Cu²⁺ enhanced the initial 30 min reaction. Fe²⁺, Fe³⁺ and Ni²⁺ increased the amount of reacted sulfide in 2 h, forming whitish turbid water. Seawater from a suspected source of aoshio water was also assayed for the auto-oxidation rate of sulfide. The oxidation rate in water from 12 m depth was 13–19 times higher than the artificial seawater without an addition of heavy metal ions. More than 15% of the oxidation rate in 12 m deep seawater was explained by dissolved iron in the seawater.     

The Effect of Lead and Zinc on Plant Growth and Chlorophyll Content of Holcus lanatus L.

Root length of Holcus lanatus L. declined rapidly with increasing lead and zinc concentrations in nutrient solution. In all used Pb and Zn concentrations root growth of genotypes coming from a Pb-Zn mine area, was greater than that of the control, suggesting that these genotypes evolved tolerance to Pb and Zn.
Negative correlation was also observed between chlorophyll content and increased heavy metal concentrations. The greater chlorophyll content found in tolerant genotypes, in different Pb and Zn concentrations, in comparison with the control, could be served to distinguish tolerant and non-tolerant genotypes.     

铅、镉和铬在叶类蔬菜中的累积及对其生长的影响

 研究结果表明: 重金属元素铅(Pb) 、镉(Cd) 、铬(Cr) 对几种叶类蔬菜生长和产量都有明显的影响, 且其影响因蔬菜种类、元素种类和浓度的不同而有很大差异, 叶用莴苣> 苋菜> 芥菜> 菜薹>蕹菜, Cr > Cd > Pb ; 不同蔬菜类型可食部位积累重金属元素的顺序为苋菜> 叶用莴苣> 菜薹> 蕹菜> 芥菜;各元素从营养液向蔬菜体内转移的迁移累积率顺序为Cd > Cr > Pb。随着各元素浓度的增大, 对蔬菜生长的影响和其可食部分的富集量随之加大。     

Sewage sludge processing determines its impact on soil microbial community structure and function

Composting and thermal drying are amongst the most commonly used post-digestion processes for allowing sanitation and biological stabilization of sewage sludge from municipal treatment plants, and making it suitable as soil conditioner for use in agriculture. To assess the impact of sludge-derived materials on soil microbial properties, fresh (LAF), composted (LAC) and thermally dried (LAT) sludge fractions, each resulting from a different post-treatment process of a same aerobically digested sewage sludge, were added at 1% (w/w) application rate on two contrasting (a loam and a loamy sand) soils and incubated under laboratory conditions for 28 days. Soil respiration, microbial ATP content, hydrolytic activities and arginine ammonification rate were monitored throughout the incubation period. Results showed that soil biochemical variables, including the metabolic quotient (qCO₂), were markedly stimulated after sludge application, and the magnitude of this stimulatory effect was dependent on sludge type (precisely LAT > LAF > LAC), but not on soil type. This effect was related to the content of stable organic matter, which was lower in LAT. Genetic fingerprinting by PCR–DGGE revealed that compositional shifts of soil bacterial and, at greater extent, actinobacterial communities were responsive to the amendment with a differing sludge fraction. The observed time-dependent changes in the DGGE profiles of amended soils reflected the microbial turnover dependent on the sludge nutrient input, whereas no indications of adverse effects of sludge-borne contaminants were noted. Our findings indicate that composting rather thermal drying can represent a more appropriate post-digestion process to make sewage sludge suitable for use as soil conditioner in agriculture.     

Limited effects of land use on soil dissolved organic matter chemistry as assessed by excitation–emission fluorescence spectroscopy and molecular weight fractionation

Dissolved organic matter (DOM) in soil solution represents a complex mixture of organic molecules and plays a central role in carbon and nitrogen cycling in plant–microbial–soil systems. We tested whether excitation–emission matrix (EEM) fluorescence spectroscopy can be used to characterize DOM and support previous findings that the majority of DOM is of high molecular weight (MW). EEM fluorescence spectroscopy was used in conjunction with MW fractionation to characterize DOM in soil solution from a grassland soil land management gradient in North Wales, UK. Data analysis suggested that three distinct fluorescence components could be separated and identified from the EEM data. These components were identified as being of humic‐like or fulvic‐like origin. Contrary to expectations, the majority of the fluorescence signal occurred in the small MW (<1 kDa) fraction, although differences between soils from the differently managed grasslands were more apparent in larger MW fractions. We conclude that following further characterization of the chemical composition of the fluorophores, EEM has potential as a sensitive technique for characterizing the small MW phenolic fraction of DOM in soils.     

菜用睡莲营养成分及重金属含量测定与评价

对绿梗菜用睡莲(Nymphaea tetragona)及红梗菜用睡莲的15营养成分和4种重金属含量进行了测定与评价。结果表明,1与104~140种(品种)新鲜蔬菜已知数据比较,绿梗和红梗菜用睡莲均有较高含量的维生素C、钠及较低含量的干物质、蛋白质、钾、钙、镁、铜、锌、磷及硒;绿梗菜用睡莲有较高含量的粗纤维、锰及较低含量的铁,红梗菜用睡莲有较高含量的铁及较低含量的粗纤维。2菜用睡莲是一种含钠量较高的蔬菜。3绿梗菜用睡莲营养成分含量总体高于红梗菜用睡莲。4湖北产区菜用睡莲的铅、砷、镉及汞等重金属含量符合食品安全国家标准的规定。     

The combined effects of cover crops and symbiotic microbes on phosphatase gene and organic phosphorus hydrolysis in subtropical orchard soils

P deficiency is a major obstacle for crop production in subtropical red soils in South China, and the hydrolysis of organic P (Po) is of great significance in these soils due to the immobilization of P by Fe and Al. Cover cropping in orchards and symbiotic microbial inoculation are considered to improve soil quality, including P status, however, their effects on the hydrolysis of Po is little known. In this study, five soil managements were established in a guava orchard in South China for two and a half years, including clean culture (CC), cover cropping with Paspalum natatu (PN), PN with arbuscular mycorrhizal fungal inoculation (PNA), cover cropping with Stylosanthes guianensis (SG), SG with rhizobial inoculation (SGR). Soil chemical, biochemical and microbial properties were analyzed. Results indicate that soil pH and SOM content tended to increase following cover cropping alone or with microbial inoculation. Po content was significantly elevated in PNA. Po fractionation revealed that cover cropping alone or with microbial inoculation significantly affected the contents of moderately labile Po (MLPo) and moderately resistant Po (FAPo). Enzyme assay indicated that cover cropping with microbial inoculation increased the activities of acidic phosphomonoesterase (ACP), neutral phosphomonoesterase (NP) and alkaline phosphomonoesterase (ALP), with ALP the most sensitive, although ACP activity dominated in red soils. Correlation analysis suggested a significantly positive relationship between ALP activity and MLPo or FAPo. PCR-DGGE profile of the alp-harboring bacterial community showed that cover cropping with S. guianensis and mycorrhizal inoculation to P. natatu promoted the bacterial diversity and/or species richness. For almost all the measured parameters, PN and SG were comparable, however, PNA was superior to SGR, indicating the stronger additive effect of arbuscular mycorrhizal fungus than that of rhizobia. Cat-PCA indicated that MLPo was the most influential factor on phosphomonoesterase. In general, this study suggests that, in subtropical orchards with red soil, cover cropping with microbial inoculation can improve the Po hydrolysis via the promoted alp-harboring bacterial community and then ALP activity. Our results also suggest that the combination of P. natatu and arbuscular mycorrhizal fungus is better than S. guianensis and rhizobia, which possesses practical significance for sustainable production in these orchards.