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。     

铅(Pb2+)和镉(Cd2+)对火炬树种子萌发及幼苗生长的影响

【目的】研究不同浓度重金属铅(Pb^2+)和镉(Cd^2+)对火炬树Rhus typhina种子萌发及幼苗生长的影响,探索火炬树对重金属铅、镉胁迫的耐受程度及机理。【方法】采用培养皿滤纸发芽法,分别用不同浓度Pb^2+(300、600、900、1200、1500 mmol/L)、Cd^2+(140、280、420、560、700 mmol/L)溶液处理火炬树种子,测定种子发芽率、发芽指数、幼苗胚根长和茎长、硫代巴比妥酸(TBARS)含量、可溶性蛋白含量、超氧化物歧化酶(SOD)活性及过氧化物酶(POD)活性。【结果】与对照相比,不同浓度Pb^2+、Cd^2+均对火炬树种子的发芽率、发芽指数和幼苗生长有抑制作用,但低浓度Pb^2+可促进根长和芽长的伸长。随着Pb^2+和Cd^2+溶液浓度的增加,幼苗叶片中SOD活性和可溶性蛋白含量表现为先增加后降低,POD活性和TBARS含量均逐渐升高。其中,Cd^2+浓度为700 mmol/L时,发芽率、发芽指数、芽长和SOD活性达到最低值,而TBARS含量最高;Pb^2+浓度为1500 mmol/L时,根长达到最低值,POD活性达到最高值;Pb^2+浓度为600 mmol/L时,可溶性蛋白含量最高。【结论】Pb^2+浓度为300 mmol/L时会抑制火炬树种子萌发,但会促进幼苗生长,其余浓度Pb^2+、Cd^2+胁迫下,火炬树种子与幼苗受到一定影响,但是均较好完成了萌发与幼苗生长,并表现出多种积极的适应性特征,未出现无根苗现象,火炬树较其他木本植物对铅、镉2种重金属表现出更强的耐性。本试验结果可为今后选用火炬树作为重金属污染土壤的植物修复材料提供理论参考。     

Sorghum extrusion process combined with biofortified sweet potato contributed for high iron bioavailability in Wistar rats

This study aimed to evaluate the effect of sorghum and sweet potato on the bioavailability of iron, gene expression of proteins involved in iron metabolism and the plasma antioxidant capacity in animals fed with whole sorghum grains processed by dry heat or extrusion, combined or not with sweet potato flour with high content of carotenoids. Five experimental groups were tested (n = 7): dry heat sorghum flour (DS); extruded sorghum flour (ES); whole sorghum flour + sweet potato flour (DS + SP); extruded sorghum flour + sweet potato flour (ES + SP) and positive control (FS). The evaluations included: hemoglobin gain, hemoglobin regeneration efficiency, gene expression of divalente metal transporter 1 (DMT-1), duodenal citochroma B (DcytB), ferroportin, hephaestin, transferrin and ferritin and total plasma antioxidant capacity (TAC). The ES + SP group showed higher (p < 0.05) expression of DcytB, ferroportin and hephaestin when compared to the control group. The DS group showed high (p < 0.05) expression of DMT-1 and the ES showed high mRNA expression of transferrin and ferritin. The changes in the sorghum physicochemical properties from extrusion process reduced the iron and phytate content, and increased the gene expression of proteins involved in iron metabolism, improving iron bioavailability. The combination of sweet potato and sorghum flour (dry or extruded) improved the iron capture and total antioxidant capacity, probably due to the presence of β-carotene and antioxidant compounds.     

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.     

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。     

基于镧系金属有机框架材料检测水体中酰胺类除草剂残留的可视化技术研究

研发水体中酰胺类农药残留简便快捷的检测技术，对评估农药造成的水体污染和提出相应的治理对策具有重要意义。以共掺杂镧系金属离子 (Eu3+ 和Tb3+) 作为配位金属离子，分别利用水热法和微波法制备了镧系金属有机框架材料 (lanthanide metal-organic frameworks, Ln-MOF)，对其结构和性质进行了表征，并采用所制备的Ln-MOF对水体中残留的5种常见酰胺类除草剂 (乙草胺、异丙草胺、丙草胺、敌草胺和异丙甲草胺) 进行了定性和定量分析。结果表明:与水热法相比，采用微波法制备的Ln-MOF样品呈蓬松的簇状、结晶度好，且具有更高的发光效率，故选择微波法制备Ln-MOF。农药残留检测试验结果表明，不同除草剂对Tb3+和Eu3+的发光会产生差别性猝灭，并使整体发射光发生变化，进而实现水体中农药的可视化检测。进一步研究发现，荧光强度比值I (Tb3+)/I (Eu3+) 与丙草胺浓度在0.1~1 mmoL/L范围时呈线性相关，线性相关系数R2 = 0.998，检测限 (LOD) 为0.08 mmoL/L。另外，该材料对于用湖水配制的除草剂溶液也具有一定的定性检测能力。本研究结果展现了Ln-MOF用于水体中酰胺类除草剂的定性、定量分析具有良好应用前景。     

重金属污染土壤几种生物修复方式比较

土壤重金属污染是全球普遍存在的问题,生物修复因其环境友好且成本效益高而得到广泛关注。但不同生物修复技术有其优势和局限性,充分了解每种修复技术的特点,才能更经济、有效地对污染土壤进行修复。本研究阐述对比了目前的土壤重金属生物修复方法,包括植物修复（植物挥发、植物固定和植物提取）、转基因植物提取、螯合辅助植物修复、微生物辅助植物修复等技术的机制、优势、局限性和适用性等方面的差异。综述提出有效的生物修复技术需要土壤化学、植物生物学、遗传学、微生物学和环境工程等多学科的有机结合。根据污染土壤的特点,结合具有相应改良特性的转基因植物,是实现污染土壤大面积修复的有效方法。同时,农艺措施对天然超级积累植物的生物量和重金属提取能力的刺激作用还需要进一步挖掘。植物修复可以与其他几种传统修复技术有效结合,利用转基因技术建立土壤+植物+微生物的组合是未来修复技术发展的最佳途径。     

Community structure of arbuscular mycorrhizal fungi associated with Robinia pseudoacacia in uncontaminated and heavy metal contaminated soils

The significance of arbuscular mycorrhizal fungi (AMF) in soil remediation has been widely recognized because of their ability to promote plant growth and increase phytoremediation efficiency in heavy metal (HM) polluted soils by improving plant nutrient absorption and by influencing the fate of the metals in the plant and soil. However, the symbiotic functions of AMF in remediation of polluted soils depend on plant–fungus–soil combinations and are greatly influenced by environmental conditions. To better understand the adaptation of plants and the related mycorrhizae to extreme environmental conditions, AMF colonization, spore density and community structure were analyzed in roots or rhizosphere soils of Robinia pseudoacacia. Mycorrhization was compared between uncontaminated soil and heavy metal contaminated soil from a lead–zinc mining region of northwest China. Samples were analyzed by restriction fragment length polymorphism (RFLP) screening with AMF-specific primers (NS31 and AM1), and sequencing of rRNA small subunit (SSU). The phylogenetic analysis revealed 28 AMF group types, including six AMF families: Glomeraceae, Claroideoglomeraceae, Diversisporaceae, Acaulosporaceae, Pacisporaceae, and Gigasporaceae. Of all AMF group types, six (21%) were detected based on spore samples alone, four (14%) based on root samples alone, and five (18%) based on samples from root, soil and spore. Glo9 (Rhizophagus intraradices), Glo17 (Funneliformis mosseae) and Acau3 (Acaulospora sp.) were the three most abundant AMF group types in the current study. Soil Pb and Zn concentrations, pH, organic matter content, and phosphorus levels all showed significant correlations with the AMF species compositions in root and soil samples. Overall, the uncontaminated sites had higher species diversity than sites with heavy metal contamination. The study highlights the effects of different soil chemical parameters on AMF colonization, spore density and community structure in contaminated and uncontaminated sites. The tolerant AMF species isolated and identified from this study have potential for application in phytoremediation of heavy metal contaminated areas.