小分子RNA及其在化学品毒理学中应用的展望

小分子RNA,包括siRNA、miRNA、piRNA等,在基因表达调控过程中扮演了至关重要的角色.对小分子RNA生物发生和功能的认知将有助于促进基因沉默的机制研究和基因治疗.本文归纳了近些年关于小分子RNA的重大研究成果以及应用于毒理学研究的案例,并对毒理学未来研究小分子RNA的重点和方向作出展望.     

RNAi及其在肿瘤生物学研究中的应用

siRNA(short interference RNA)是指21～25nt的短双链RNA,能特异性地抑制同源基因的表达,使内源性的mRNA降解,最终致使基因沉默,这种现象称为RNA干扰(RNA interference,RNAi).RNAi具有高效、稳定、特异性强等特点,已成为目前生物医学研究领域最为活跃的热点之一.RNAi技术为肿瘤的基因治疗提供了新思路,是肿瘤治疗的策略之一.本文就RNAi技术及其在肿瘤生物学研究中的应用进行综述.     

好氧反硝化菌种DF2的分离鉴定及生理生化特性分析

分离了新型的高效反硝化细菌并研究其生物脱氮效率。通过观察形态学、生理生化鉴定和16SrRNA基因同源分析来鉴定菌种,致力于研究微生物污水脱氮处理的实际效果和克服反硝化过程中有害中间产物的积累。利用BTB培养基从水平潜流人工湿地基质中初步筛选出了9株平板阳性菌,革兰阴阳性均有。经试验研究发现该9种菌株在好氧条件下均具有一定的脱氮能力,其中以DF2和DF3 2种菌株脱氮能力最为显著,在3 d时间内NOX--N去除率达到了95%以上,NO2--N仅有微量积累;而其它7种菌株对NO3--N的去除率可以达到98%以上,但是NO2--N积累比较严重,积累量达到了NO3--N去除的50%~70%左右。经测序分析鉴定DF2和DF3分别属于德克斯氏菌属（Derxia）和假单胞菌属（Pseudomonadaceae）。德克斯氏菌属作为除氮的反硝化菌属在以往的研究中鲜有报道,该菌在好氧条件下可以合成周质NAR的亚基基因（napA）。     

不同沉水植物净水能力与植株体细菌群落组成相关性

选取苦草(Vallisneria natans)、密刺苦草(Vallisneria denseserrulata)、黑藻(Hydrilla verticillata)、伊乐藻(Elodea canadensis)、金鱼藻(Ceratophyllum demersum)、竹叶眼子菜(Potamogeton malaianus)、篦齿眼子菜(Potamogeton pectinatus)和微齿眼子菜(Potamogeton maackianus)这8种亚热带地区常见的沉水植物为研究对象,在室内静水条件下对其氮、磷吸收和水质净化能力进行对比试验,并结合16S rRNA基因测序对沉水植物关联细菌群落组成进行测定,研究沉水植物对水体的净化能力与植株体菌群之间的相关性.结果表明,8种沉水植物对水体中的氮磷的去除主要通过植物增效作用,植物吸收富集作用去除率较低.其中密刺苦草对水体中TN、TP的去除率最高,达到了91. 58%和96. 81%.伊乐藻、金鱼藻对水中氮磷自身吸收能力高于其他组,密刺苦草和苦草的植物增效的净化能力最强.经分析,8种沉水植物对水体中氮磷的净化能力较强可能是因为植物关联的细菌大多具有降解作用.根瘤菌目(Rhizobiales)、伯克霍尔德氏菌目(Burkholderiales)、黄杆菌目(Flavobacteriales)、产碱杆菌科(Alcaligenaceae)、贪铜菌属(Cupriavidus)和芽孢杆菌属(Bacillales)的细菌可能是引起密刺苦草增效的净化能力较强的优势菌群,异常球菌纲(Deinococci)、丛毛单胞菌科(Comamonadaceae)、腐螺旋菌科(Saprospiraceae)和生丝微菌属(Hyphomicrobium)的细菌可能是引起苦草增效的净化能力较强的优势菌群.     

纳米银和银离子对土壤中硝化微生物及其氨氧化速率的影响

为研究纳米银对土壤硝化微生物及其氮转化的影响,采用土壤培养方式,对不同剂量纳米银(10、50、100 mg·kg~(-1))和银离子(1、5、10 mg·kg~(-1))暴露下黄棕壤和水稻土硝化细菌数量、土壤酶活性、amoA基因丰度、NH_4~+-N与NO_3~--N含量变化以及土壤潜在氨氧化速率进行研究.结果表明,纳米银和银离子暴露后,2种土壤亚硝酸细菌和硝酸细菌数量显著减少;土壤酶活性受到抑制,对脲酶的影响大于过氧化氢酶;土壤氨氧化细菌(AOB)和氨氧化古菌(AOA)的amoA基因丰度均降低,对AOB基因丰度的影响大于AOA.在2种土壤中外源添加(NH_4)_2SO_4时,随着纳米银和银离子暴露剂量增加,土壤NH_4~+-N含量累积,NO_3~--N含量减少,氨氧化速率降低,铵态氮向硝态氮的转化受阻.综上所述,纳米银和银离子对土壤硝化微生物产生毒害作用并影响铵态氮转化,且影响程度与土壤理化性质有关.     

四环素抗生素对污泥中四环素抗性基因丰度和表达水平的作用影响

为研究四环素(TC)抗生素对其抗性基因(TC-ARGs)转录表达的作用影响,以从活性污泥中筛选获得的四环素抗性细菌(TRB)作为研究对象,采用荧光定量PCR和逆转录PCR方法检测了7种TC-ARGs,包括tet A、tet C、tet G、tet M、tet O、tet W和tet X基因的丰度和表达水平,并探讨了TC与TC-ARGs丰度及其表达水平之间的相关关系.结果表明,在整个培养周期内,tet A、tet G和tet W基因丰度随TC暴露浓度的增大总体呈现上升趋势,但其余TC-ARGs基因丰度整体波动较大. TC胁迫对不同TC-ARGs的转录表达水平作用影响差别较大,其中,tet A基因表达水平相对稳定,且随TC浓度的升高而上调,在TC浓度为100 mg·L~(-1)时,其上调倍数高达5. 3倍.在短期(1 d) TC胁迫下,TC-ARGs转录表达水平随TC浓度的升高整体呈现上调趋势.由相关性分析可知,tet A和tet W基因丰度与其转录表达水平之间存在显著的线性相关性,表明其基因丰度可在一定程度上衡量和评价其抗性表达水平,进而反映其功能活性和环境风险.     

Different effects of foliar application of silica sol on arsenic translocation in rice under low and high arsenite stress

Foliar application of Si can generally reduce As translocation from roots to shoots in rice; however, it does not always work, particularly under high As stress. Here, the effects of foliar application of nanoscale silica sol on As accumulation in rice were investigated under low (2 μmol/L) and high (8 μmol/L) arsenite stress. The results revealed that foliar Si application significantly decreased the As concentration in shoots under low arsenite stress, but showed different effects under high arsenite stress after 7 days of incubation. The reduction in root-to-shoot As translocation under the 2As+Si treatment was related to the down-regulation of OsLsi1 and OsLsi2 expression and up-regulation of OsABCC1 expression in roots. In the 8As+Si treatment, the expressions of OsLsi1, OsLsi2, and OsABCC1 were significantly promoted, which resulted in substantially higher As accumulation in both the roots and shoots. In the roots, As predominantly accumulated in the symplasts (90.6%–98.3%), in which the majority of As was sequestered in vacuoles (79.0%–94.0%) under both levels of arsenite stress. Compared with that of the 8As treatment, the 8As+Si treatment significantly increased the As concentration in cell walls, but showed no difference in the vacuolar As concentration, which remained constant at approximately 69.1–71.7 mg/kg during days 4–7. It appeared that the capacity of root cells to sequester As in the vacuoles had a threshold, and the excess As tended to accumulate in the cell walls and transfer to the shoots via apoplasts under high arsenite stress. This study provides a better understanding of the different effects of foliar Si application on As accumulation in rice from the view of arsenite-related gene expression and As subcellular distribution in roots.     

拟南芥VTE1过量表达可以增加维生素E含量和提高烟草植株耐盐行

维生素E在动物细胞内具有抗氧化等重要作用，但在植物体内的功能却鲜为人知．本实验利用CaMV 35S启动子与来源于拟南芥的编码生育酚环化酶（TC）的cDNA（VTE1）构建的嵌合表达载体，以根癌农杆菌介导的叶盘法转化烟草W38．实验结果表明，具有卡那霉素抗性的再生植株经RT-PCR检测，得到了与阳性对照一致的495bp的目标片段；转基因植株的VE含量比对照植株高2倍左右，个别株系高达11倍．实验还发现，在耐盐性实验中转基因植株对盐的抗性明显高于野生型烟草；同时，在不同盐浓度（150、250mmoL／L）胁迫下转基因植株VE含量比未转化植株增加了1．3～1．8倍，首次证明VTE1与植物耐盐性之间的关系．     

Evaluating unintended consequences of intentional species introductions and eradications for improved conservation management

Increasingly intensive strategies to maintain biodiversity and ecosystem function are being deployed in response to global anthropogenic threats, including intentionally introducing and eradicating species via assisted migration, rewilding, biological control, invasive species eradications, and gene drives. These actions are highly contentious because of their potential for unintended consequences. We conducted a global literature review of these conservation actions to quantify how often unintended outcomes occur and to elucidate their underlying causes. To evaluate conservation outcomes, we developed a community assessment framework for systematically mapping the range of possible interaction types for 111 case studies. Applying this tool, we quantified the number of interaction types considered in each study and documented the nature and strength of intended and unintended outcomes. Intended outcomes were reported in 51% of cases, a combination of intended outcomes and unintended outcomes in 26%, and strictly unintended outcomes in 10%. Hence, unintended outcomes were reported in 36% of all cases evaluated. In evaluating overall conservations outcomes (weighing intended vs. unintended effects), some unintended effects were fairly innocuous relative to the conservation objective, whereas others resulted in serious unintended consequences in recipient communities. Studies that assessed a greater number of community interactions with the target species reported unintended outcomes more often, suggesting that unintended consequences may be underreported due to insufficient vetting. Most reported unintended outcomes arose from direct effects (68%) or simple density-mediated or indirect effects (25%) linked to the target species. Only a few documented cases arose from more complex interaction pathways (7%). Therefore, most unintended outcomes involved simple interactions that could be predicted and mitigated through more formal vetting. Our community assessment framework provides a tool for screening future conservation actions by mapping the recipient community interaction web to identify and mitigate unintended outcomes from intentional species introductions and eradications for conservation.     

稻田落干过程砷甲基化效率变化与关键影响因素分析

研究稻田落干过程砷甲基化效率变化规律,分析关键环境和生物因素的影响,为今后水稻直穗病防控提供科学依据.开展室内培养模拟稻田落干过程,以采集自贵州兴仁（XR）和广西南丹（ND）的两种砷污染水稻土壤为供试土壤,各土壤设置添加（RS）和不添加（CK）水稻秸秆处理,分析自然落干0、24、36、48和60 h过程中Eh、pH、孔隙水总有机碳（TOC）、砷形态、砷甲基化功能基因（arsM）、硫酸盐还原菌（dsrA,砷甲基化相关微生物）、产甲烷菌（mcrA,砷去甲基化相关微生物）丰度和arsM功能微生物多样性变化.稻田落干过程土壤Eh由完全淹水状态下的-300~-200 mV向落干后的-150~-50 mV变迁,而pH值变化规律不明显;孔隙水无机砷（iAs）和二甲基砷（DMAs）浓度随落干过程变化更为显著,总体呈现增加趋势,且RS处理DMAs浓度显著高于CK,ND土壤孔隙水比XR土壤孔隙水DMAs浓度更高;随落干时间延长,XR-CK和XR-RS处理土壤砷甲基化效率有一定提升,但变化不显著,而ND-CK和ND-RS处理土壤砷甲基化效率显著增加.当培养为60 h时,ND-CK和ND-RS处理砷甲基化效率相比培养初期分别提高约61.8%和23.2%;随落干时间延长arsM和dsrA基因拷贝数明显增加,而mcrA基因拷贝数显著下降.秸秆添加后显著提高全细菌和arsM、dsrA和mcrA基因丰度;进一步基于多因素方差分析和冗余分析发现,供试土壤、秸秆添加、落干时间和其交互作用对于各砷形态、砷甲基化效率和关键基因丰度变化影响显著,TOC、Eh和砷甲基化相关基因与甲基态砷呈正向关联,而与无机砷iAs呈负向关联;基于arsM微生物测序发现,伴随落干过程还发生着砷甲基化功能微生物群落的更替.研究结果有助于提升稻田落干过程中砷甲基化变化的理论认知,为今后水稻直穗病科学防控提供指导.