Transport of cadmium from soil to grain in cereal crops: A review

Due to rapid urbanization and industrialization, many soils for crop production are contaminated by cadmium(Cd), a heavy metal highly toxic to many organisms. Cereal crops such as rice, wheat, maize, and barley are the primary dietary source of Cd for humans, and reducing Cd transfer from soil to their grains is therefore an important issue for food safety. During the last decade, great progress has been made in elucidating the molecular mechanisms of Cd transport, particularly in rice. Inter-and intraspecific variations in Cd accumulation have been observed in cereal crops. Transporters for Cd have been identified in rice and other cereal crops using genotypic differences in Cd accumulation and mutant approaches. These transporters belong to different transporter families and are involved in the uptake, vacuolar sequestration, root-to-shoot translocation, and distribution of Cd. Attempts have been made to reduce Cd accumulation in grains by manipulating these transporters through overexpression or knockout of the transporter genes, as well as through marker-assisted selection breeding based on genotypic differences in Cd accumulation in the grains. In this review, we describe recent progress on molecular mechanisms of Cd accumulation in cereal crops and compare different molecular strategies for minimizing Cd accumulation in grains.     

Effects of plant growth-promoting rhizobacteria on the molecular responses of maize under drought and heat stresses: A review

Drought and heat are major environmental stresses that continually influence plant growth and development. Under field conditions, these stresses occur more frequently in combination than alone, which magnifies corresponding detrimental effects on the growth and productivity of agriculturally important crops. Plant responses to such abiotic stresses are quite complex and manifested in a range of developmental, molecular, and physiological modifications that lead either to stress sensitivity or tolerance/resistance. Maize (Zea mays L.) is known for its sensitivity to abiotic stresses, which often results in substantial loss in crop productivity. Bioaugmentation with plant growth-promoting rhizobacteria (PGPR) has the potential to mitigate the adverse effects of drought and heat stresses on plants. Hence, this is considered a promising and eco-friendly strategy to ensure sustainable and long-term maize production under adverse climatic conditions. These microorganisms possess various plant growth-promoting (PGP) characteristics that can induce drought and heat tolerance in maize plants by directly or indirectly influencing molecular, metabolic, and physiological stress responses of plants. This review aims to assess the current knowledge regarding the ability of PGPR to induce drought and heat stress tolerance in maize plants. Furthermore, the drought and heat stress-induced expression of drought and heat stress response genes for this crop is discussed with the mechanisms through which PGPR alter maize stress response gene expression.     

密集化学农业下热带变性土中的微生物种群、活性及基因丰度

There are increasing concerns on the environmental impacts of intensive chemical agriculture.The effect of high agrochemical inputs used in intensive chemical farming was assessed on soil microbiological,molecular and biochemical properties in tropical Vertisols in India.Farm field sites under normal cultivation of arable crops using high inputs of fertilizers and pesticides in chili(Capsicum annum L.,5.0× dose for fertilizers and 1.5× dose for pesticides over normal inputs) and black gram(Vigna mungo L.Hepper,2.2x dose for fertilizers and 2.3× dose for pesticides over normal inputs) were compared with adjacent sites using normal recommended doses.Organic carbon and basal respiration showed no response to high inputs of fertilizers and pesticides in soils of both crops.Labile carbon decreased by 10% in chili soils and increased by 24% in black gram soils under high input farming system.The proportion of soil labile carbon as a fraction of soil organic carbon was unaffected by high inputs.The labile carbon mineralization coefficient(qM_(LC)) increased by 50.0% in chili soils,indicating that the soil microorganisms were under stress due to high agochemical inputs,whereas qM_(LC) decreased by 36.4% in black gram soils.Copiotrophs increased due to high inputs in soils of both chili(63.1%) and black gram(47.1%).Oligotrophs increased by 10.8% in black gram soils but not in chili soils.The abundance of amoA gene reduced by 39.3% in chili soils due to high inputs and increased significantly by 110.8% in black gram soils.β-Glucosidase also increased by 27.2% and 325.0%,respectively.Acid phosphatase activity reduced by 29.2% due to high inputs in chili soils and increased by 105.0% in black gram soils.The use of high agrochemical inputs thus had adverse consequences on biological health in chili but not in black gram soils.In soils cultivated with black gram,the moderating effect of cultivating legumes and their beneficial effect on soil health were evident from the increase in soil labile carbon,lower qM_(LC),higher amoA gene and enzyme activities.Overall results showed that cultivation of legumes permits intensive chemical farming without deteriorating soil biological health.     

芦苇定居后铜尾矿细菌群落结构的变化

Soil samples were collected from both bare and vegetated mine tailings to study the changes in bacterial communities and soil chemical properties of copper mine tailings due to reed (Phragmites communis) colonization. The structures of bacterial communities were investigated using culture-independent 16S rRNA gene sequencing method. The bacterial diversity in the bare mine tailing was lower than that of the vegetated mine tailing. The former was dominated by sulfur metabolizing bacteria, whereas the latter was by nitrogen fixing bacteria. The bare mine tailing was acidic (pH = 3.78), whereas the vegetated mine tailing was near neutral (pH = 7.28). The contents of organic matter, total nitrogen, and ammonium acetate-extractable potassium in vegetated mine tailings were significantly higher than those in the bare mine tailings (P < 0.01), whereas available phosphorus and electrical conductivity were significantly lower than those in the bare mine tailings (P < 0.01). The results demonstrated that 16S rRNA gene sequencing could be successfully used to study the bacterial diversity in mine tailings. The colonization of the mine tailings by reed significantly changed the bacterial community and the chemical properties of tailings. The complex interactions between bacteria and plants deserve further investigation.     

亚马逊黑土及其黑碳颗粒中的真菌丰度和多样性互不相同

Amazonian Dark Earth (ADE) is a highly fertile soil of anthropogenic origin characterized by high levels of charred black carbon (BC).It is considered a model of fertility;however,knowledge on the fungal community structure and diversity inhabiting ADE and its BC particles is scarce.Fungal community structure and diversity of ADE and its BC from four sites under different land uses (three agricultural systems and a secondary pristine forest) in the Brazilian Central Amazon were evaluated by 18S rRNA gene pyrosequencing.Fungal communities in ADE and BC were dissimilar and showed differential abundances of fungal operational taxonomic units (OTUs).Estimated fungal species richness (abundance-based coverage estimate and Chao-1 index) and diversity estimators (Shannon and Simpson's reciprocal indices) were higher in ADE than in BC in all agricultural areas.No differences were observed in those parameters in ADE and BC samples from the secondary forest.Pezizomycotina fungi and OTUs assigned to Cordyceps confragosa,Acremonium vitellinum,Camarops microspora,and Hirsutella rhossiliensis were more abundant in BC particles than in ADE.These findings represent a breakthrough in our understanding of fungal communities in BC particles from ADE,and will be valuable in future studies considering biochar application in soil.     

沿海滩涂土壤中反硝化细菌群落随盐分梯度的更迭

To better understand the effect of salinity on denitrification communities,soils along a salinity gradient (ranging from 7.32 to 1.70 mS cm 1) in a wetland along the Yellow Sea coastline in Jiangsu Province,China,were studied using both culture-dependent and-independent methods.Culture efforts yielded 82 isolates in total,81.7% of which were close relatives of Bacillus sp.based on partial sequences of their 16S rRNA genes.Denaturing gradient gel electrophoresis (DGGE) analysis based on 16S rRNA sequences suggested possible existence of bacterial community succession along the salinity gradient.Clone library analysis based on nosZ gene sequences (coding nitrous oxide reductase) showed that operational taxonomic units (OTUs) associated with α-proteobacteria dominated in all three soils,whereas those associated with β-and γ-subdivisions showed a clear succession.In the high salinity soil,only the OTUs associated with α-subdivision were found.In the medium salinity soil,small proportions of β-(6.5%) and γ-associated (19.6%) OTUs were found.In the low salinity soil,the proportions were further increased to 33% and 25% for β-and γ-Proteobacteria,respectively.Statistic analysis using Unifrac P test showed that nosZ-communities in different saline soils were significantly different from each other.It could be concluded that α-subdivision of nosZ-community tended to be sustained in high salinity environments whereas β-and γ-subdivisions,especially the former,tended to be sustained in low salinity environments.Salinity was the key determinant of nosZ-community composition in the environment.     

鸡γ-干扰素基因(ChIFN-γ)的瞬时表达及其抗病毒活性检测

采用PCR技术从质粒pET-ChIFN-γ中扩增得到鸡(Gallus gallus)γ-干扰素(chicken interferon gamma,ChIFN-γ)基因,将其亚克隆到真核表达载体pCAGGS中.将鉴定正确的克隆命名为pCAGGS-ChIFN-γ,体外转染鸡胚成纤维(CEF)细胞,24 h后经间接免疫荧光(IFA)和免疫印迹(Western blot)检测,表达蛋白具有良好的免疫原性.然后利用表达绿色荧光蛋白(GFP)的重组水疱口炎病毒(VSV*GFP)在CEF细胞上检测表达的ChIFN-γ抗病毒活性(AVA),经检测其抗病毒活性为2×10~3AU/mL,并且其活性可被抗鸡IFN-γ的多克隆抗体阻断.     

番茄红素β环化酶基因（Lyc-β）RNAi载体构建及表达鉴定

从番茄(Lycopersicun esculentum Mill)提取总RNA,根据GenBank中番茄红素β环化酶基因(Lye-β)序列(X86452),经mRNA反转录扩增出2段Lyc-β基因高度保守的300 bp DNA 片段,从β-葡萄糖苷酶基因(gusA)中扩增出170 bp的内含子片段.分别将两段不同的Lyc-β片段的正、反向序列与内含子连接,构建出两套干扰Lyc-β基因的植物表达载体.经根癌农杆菌(Agrobacterium tumefaciems)介导转化番茄,22 株转基因植株通过荧光定量PCR分析干扰效果,结果显示,2套干扰载体的干扰效率表现出一定的差异,Lyc-β基因mRNA平均含量分别为对照的11.78%和13.86%,进一步用HPLC分析转化株的番茄红素含量,结果表明,转基因植株中番茄红素的含量最高可达13.84 μg/g FW,是对照的4.26倍.     

Evaluation of pinewood biochar as a carrier of bacterial strain Enterobacter cloacae UW5 for soil inoculation

The large-scale production of biochar for carbon sequestration provides an opportunity for using these materials as inoculum carriers to deliver plant growth-promoting rhizobacteria (PGPR) into agricultural soils. Here, we evaluated the suitability of a biochar produced from pinewood pyrolyzed at 300 °C as a carrier for a well-studied PGPR strain, Enterobacter cloacae UW5. This strain was genetically modified to produce a green fluorescent protein marker that enabled tracking of the inoculum. Results from selective plate count assays and quantitative PCR (qPCR) confirmed that cell survival was slightly improved by addition of bacteria to soil using biochar as a carrier for the inoculant, as compared to soil directly inoculated. Total 16S rRNA genes were quantified using qPCR and DNA templates from the same soil treatments to distinguish the impact of biochar on total bacterial abundance from its influence on inoculum survival. Here total bacterial abundance was not influenced by biochar. All treatments resulted in bacterial colonization of roots at population densities of approximately 10⁵ CFU g⁻¹ root mass. Cucumber plants grown in the biochar amended soils had significantly greater biomass and root development than those planted in un-amended soil, regardless of the presence of inoculum. The ability of bacteria to colonize the plant roots and produce a plant growth hormone was not affected by biochar. However, UW5 inoculum did not promote root development in cucumber in any of the soils tested here. Overall, these experiments suggest that the 300 °C pine biochar is effective for evenly distributing inoculum into soil and promotes cucumber development in sandy loams.     

Hydrocarbon degradation potential and activity of endophytic bacteria associated with prairie plants

Phytoremediation systems for organic compounds such as petroleum hydrocarbons rely on a synergistic relationship between plants and their root-associated microbial communities. To determine the probable role of endophytic bacterial communities in these systems, this study examined both rhizosphere and endophytic communities of five different plant species at a long-term phytoremediation field site. Hydrocarbon degradation potential and activity were assessed using MPN assays, PCR analysis of catabolic genes associated with hydrocarbon degradation, and mineralization assays with C-14 labeled hydrocarbons. Microbial community structure in each niche was assessed by DGGE analysis of 16S rRNA gene fragments and subsequent band sequencing. Both endophytic degrader populations and endophytic degrader activity showed substantial inter-species variation, largely independent of that shown by the respective rhizosphere populations. Endophytic hydrocarbon degradation was linked to dominant bacterial endophytes. Pseudomonas spp. dominated endophytic communities exhibited increased alkane hydrocarbon degradation potential and activity, while Brevundimonas and Pseudomonas rhodesiae dominated endophytic communities were associated with increased PAH degradation potential and activity. In one plant species, Lolium perenne, increased endophytic alkane hydrocarbon degradation was associated with increased rhizosphere alkane degradation and decreased rhizosphere PAH degradation. Our results show that diverse plant species growing in weathered-hydrocarbon contaminated soil maintain distinct, heterogeneously distributed endophytic microbial populations, which may impact upon the ability of plants to promote the degradation of specific types of hydrocarbons.     

Comparisons of vesicular-arbuscular mycorrhizal species and inocula formulations in a commercial nursery and on diverse Florida beaches

For efficient use of mycorrhizal inoculum the effectiveness of the isolate used and the rate of application required for maximum colonization must be known. The objectives of this research were to (1) define the lower limit of inoculum density required for maximum colonization of Uniola paniculata in a commercial nursery and (2) evaluate the performance of a selected native dune vesicular-arbuscular mycorrhizal (VAM) isolate versus a commercially available non-dune VAM (foreign) isolate on three diverse Florida beaches. An inoculum-dilution study was conducted in a commercial nursery with cutroot inoculum of a Glomus sp. that had been isolated from a Florida dune. Maximum colonization was achieved with approximately 1 propagule ml^-1 of growth medium. In a separate nursery study, 10 inoculation treatments (combinations of inoculum source and level) were established in the commercial nursery. Treatments included cut-root and sheared-root inoculum of the native dune isolate, and Nutri-Link, a commercial inoculum of G. intraradices. Colonized plants from selected treatments were transplanted to beach sites around Florida. At Miami Beach, after one growing season, the shoot mass of plants inoculated with the native isolate was approximately twice that of plants inoculated with the foreign isolate. At Katherine Abbey Hanna Park and Eglin Air Force Base there were no significant inoculum source effects on shoot mass or root length after one growing season. However, the native isolate produced a greater colonized root length than the foreign isolate in all plantings. The soil hyphal density was measured at Eglin Air Force Base, and the results showed that plants inoculated with the native isolate had more soil hyphae (4.33 mg^-1) than plants inoculated with the foreign isolate (3.65 mg^-1) or the non-inoculated plants (2.12 mg^-1). Even where there were no obvious shoot growth responses, mycorrhizal inoculation may have an important effect on dune stabilization, as soil hyphae are known to bind sand grains and improve dune stability.Publishedas Florida Agriculture Experiment Station Journal SeriesPublishedas Florida Agriculture Experiment Station Journal Series     

Contribution of arbuscular mycorrhizal fungi and soil amendments to remediation of a heavy metal-contaminated soil using sweet sorghum

Owing to their potential advantages such as waste reduction,recycling,and economic attributes,fast-growing bioenergy crops have the capacity to e?ectively phytoremediate heavy metal-contaminated soils.However,little is known about the role of microbial and chemical amendments in phytoremediation using bioenergy crops.Here,we studied the contributions of inoculation with the arbuscular mycorrhizal fungus (AMF) Acaulospora mellea ZZ and three soil amendments,i.e.,hydroxyapatite (HAP),manure,and bi...     

Changes of Bacterial Community Structure in Copper Mine Tailings After Colonization of Reed (Phragmites communis)

Soil samples were collected from both bare and vegetated mine tailings to study the changes in bacterial communities and soil chemical properties of copper mine tailings due to reed (Phragmites communis) colonization. The structures of bacterial communities were investigated using culture-independent 16S rRNA gene sequencing method. The bacterial diversity in the bare mine tailing was lower than that of the vegetated mine tailing. The former was dominated by sulfur metabolizing bacteria, whereas the latter was by nitrogen fixing bacteria. The bare mine tailing was acidic (pH = 3.78), whereas the vegetated mine tailing was near neutral (pH = 7.28). The contents of organic matter, total nitrogen, and ammonium acetate-extractable potassium in vegetated mine tailings were significantly higher than those in the bare mine tailings (P< 0.01), whereas available phosphorus and electrical conductivity were significantly lower than those in the bare mine tailings (P< 0.01). The results demonstrated that 16S rRNA gene sequencing could be successfully used to study the bacterial diversity in mine tailings. The colonization of the mine tailings by reed significantly changed the bacterial community and the chemical properties of tailings. The complex interactions between bacteria and plants deserve further investigation.     

Effects of microplastic polystyrene, simulated acid rain and arbuscular mycorrhizal fungi on Trifolium repens growth and soil microbial community composition

Microplastic pollution is a global and ubiquitous environmental problem in the oceans as well as in the terrestrial environment. We examined the fate of microplastic polystyrene (MPS) beads in experimental soil in the presence and absence of symbiotic arbuscular mycorrhizal fungi (AMF) and simulated acid rain (SAR) to determine whether the combinations of these three factors altered the growth of white clover Trifolium repens. We found that MPS, SAR, or AMF added singly to soil did not alter T. repens growth or yields. In contrast, MPS and AMF together significantly reduced shoot biomass, while SAR and MPS together significantly reduced soil available phosphorus independent of AMF presence. Microplastic polystyrene, AMF, and SAR together significantly reduced soil NO^-₃-N. Arbuscular mycorrhizal fungi added singly also enriched the beneficial soil bacteria (genus Solirubrobacter), while MPS combined with AMF significantly enriched the potential plant pathogenic fungus Spiromastix. Arbuscular mycorrhizal fungi inoculation with MPS increased the abundance of soil hydrocarbon degraders independent of the presence of SAR. In addition, the abundance of soil nitrate reducers was increased by MPS, especially in the presence of AMF and SAR. Moreover, SAR alone increased the abundance of soil pathogens within the fungal community including antibiotic producers. These findings indicate that the coexistence of MPS, SAR, and AMF may exacerbate the adverse effects of MPS on soil and plant health.     

Effects of hexachlorocyclohexane on rhizosphere fungal propagules and root colonization by arbuscular mycorrhizal fungi in Plantago lanceolata

Lindane ( γ‐hexachlorocyclohexane or γ‐HCH) is an organochlorine insecticide previously used extensively for the control of agricultural pests. We studied the effects of soil HCH contamination on vegetation and its associated arbuscular mycorrhizas (AM). The polluted and unpolluted plots had similar plant cover, with the same species richness and abundance. Plantago lanceolata plants were selected for mycorrhizal analysis because of their presence in both plots and known mycotrophy. The presence of HCH appeared to have no significant effect on the extent of colonization of Plantago roots by AM, suggesting a similar functionality of the fungal symbionts. However, infective AM propagules, the density of AM spores and viable AM hyphae in the rhizosphere were much less in the HCH‐polluted soil than in the unpolluted plot. Pre‐inoculation of four plant species with an isolate of Glomus deserticola obtained from the HCH‐contaminated soil resulted in increased growth and fungal colonization of roots compared with plants pre‐inoculated with the introduced fungus G. macrocarpum or colonized by the consortium of indigenous AM fungal species, when those plants were transplanted to an HCH‐contaminated soil. This suggests that the fungus increases the tolerance of plants to the toxic soil environment. We conclude that herbaceous and woody plants can grow in soil with little P contaminated with <100 mg HCH kg^?1 with the help of tolerant AM, despite the detrimental effect of HCH on AM fungal propagules in soil. The effects of AM fungi on plant growth and soil microbial community structure in HCH‐polluted sites could be important for remediation of the pollutant through the microbial activity in the rhizosphere.     

Characterization of Burkholderia sp. XTB‐5 for Phenol Degradation and Plant Growth Promotion and Its Application in Bioremediation of Contaminated Soil

A key issue when researching land degradation is the pollution of soils. For bioremediation of contaminated soil, Burkholderia sp. XTB‐5 cells were obtained from soil and grown on mineral salt medium with initial phenol concentrations of 650 mg L⁻¹ and 850 mg L⁻¹, which were found to degrade more than 98% of phenol content in less than 4 days. About 90% of phenol content (with initial concentration of 250 mg kg⁻¹ soil) was removed from soil inoculated with XTB‐5 cells in 6 days. More than 90% of phenol content was removed within 20 days after co‐introduction of XTB‐5 cells and plants to sterilized soil in a greenhouse or to natural soil in field trials. But under the same conditions, individual introduction of plants to sterilized soil in the greenhouse reduced phenol content by about 50% and introduction to natural soil in field trials reduced phenol content by about 38%, suggesting that phytoremediation of phenol is often inefficient and microorganisms can efficiently degrade this pollutant. In addition, strain XTB‐5 was found to solubilize phosphate and produce 1‐Aminocyclopropane‐1‐Carboxylate (ACC) deaminase and siderophore. Strain XTB‐5 promoted plant growth in both phenol‐absent and phenol‐spiked soil under greenhouse and field conditions. Considering that ACC deaminase is beneficial to plant growth under adverse environmental conditions, plant growth promotion by XTB‐5 in phenol‐contaminated soil is not only due to XTB‐5 cell‐degradation of phenol and reduced phytotoxicity but also to production of ACC deaminase. Hence, Burkholderia sp. XTB‐5 presents an attractive microorganism for phytoremediation of contaminated soil and agronomic application. Copyright © 2016 John Wiley & Sons, Ltd.     

Enhanced dissipation of chrysene in planted soil: the impact of a rhizobial inoculum

Results from an innovative approach to improve remediation in the rhizosphere by encouraging healthy plant growth and thus enhancing microbial activity are reported. Mixed grass-legume systems, together with microbial inoculants, were used to remediate a polycyclic aromatic hydrocarbon (chrysene) spiked agricultural soil. Inoculants were symbiotic rhizobia, which may play an important role in rhizoremediation by increasing plant and root growth. An inoculum of an isolate of Rhizobium leguminosarum bv. trifolii, selected for PAH tolerance, was produced using a peat carrier. The inoculum and white clover (Trifolium repens L.), were planted into soils with ryegrass (Lolium perenne L.). The soils spiked with chrysene (500 mg kg⁻¹) then aged for 4 weeks. Shoot- and root-biomass of plants, and the amount of root nodulation, were determined. Rhizobial populations, soil pH and soil nitrogen were also monitored throughout the trial. In addition, the ability of the inoculated rhizobial strain to utilise chrysene as a sole carbon source was assessed. Direct uptake and/or degradation of chrysene by the clover and ryegrass did not occur to a significant degree. Enhanced losses of chrysene were seen in planted, non-sterile soils that contained a rhizobial inoculum. No direct degradation of chrysene by R. leguminosarum bv. trifolii was observed and no enhanced losses of PAHs were detected in sterile soils after inoculation with rhizobia. Results suggest that the enhanced dissipation of chrysene, observed in the non-sterile planted inoculated pots, was not a result of degradation of chrysene by R. leguminosarum bv. trifolii. The symbiotic association with R. leguminosarum bv. trifolii improved plant vigour and growth in inoculated planted treatments. This may have stimulated the rhizospheric microflora to degrade chrysene.     

Transport and survival of GFP-tagged root-colonizing microbes: Implications for rhizodegradation

Many recalcitrant organic contaminants pose unique and vast environmental challenges, potentially addressed through phytoremediation. Using molecular engineering to enhance enzymatic capabilities of root-colonizing microbes, additional contaminants can be targeted and contaminant fate can be altered to promote rhizosphere degradation of contaminants, which is desired among phytoremediation mechanisms. In this paper, rhizosphere bacteria were tagged with the green fluorescent protein genes (gfp) in order to monitor colonization, survival, and transport within the root zone and to evaluate the effectiveness of visual identification using GFP. Transport of the gfp-tagged root colonizers was observed to a one meter depth against a hydraulic gradient in less than 180 days revealing that plant's roots clearly enhanced movement of the recombinant strains through the rhizosphere although proliferation of the recombinant bacteria was not substantial. Over a 49-day plant growth period survival and colonization by the recombinant bacteria was monitored in soil and on roots, revealing a decreasing trend. Overall, this study showed that enhanced rhizosphere degradation is mechanistically promising, but that the specific plant-microbe pairing studied herein was not ideal. Using GFP for visual identification is not 100% efficient but provides a quick and simple marker to detect tagged microorganisms. Selection of root colonizing organisms to be engineered in enhanced rhizoremediation is a critical step in advancing the technology.