Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity

Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.     

青海省祁连圆柏天然林单木生物量模型构建

利用祁连圆柏整株生物量与生长指标数据，为估算祁连圆柏林的生物量估算提供参考。通过野外调查，共获取了63株祁连圆柏天然林样木生物量与生长指标实测数据。用其中50株样木数据进行回归模拟，用其余的13株样木数据对模型可靠性进行检验，构建器官生物量与生长指标间的回归模型。结果表明，祁连圆柏单木水平下，树干生物量模型的R²_adj为0.96，均方根、模型有效性和残差系数分别为0.50、0.85和0.05；枝条生物量模型的R²_adj为0.897，均方根、模型有效性和残差系数分别为0.69、0.80和-0.66；叶生物量模型的R²_adj为0.61，均方根、模型有效性和残差系数分别为0.54、0.86和0.15；根生物量模型的R²_adj为0.93，均方根、模型有效性和残差系数分别为0.12、0.997 和-0.01。在调查数据范围内构建的模型较好地反映了祁连圆柏生物量与生长指标间的关系，形式简单、使用方便；与实测值相比，树干与叶生物量模拟值偏小，枝和根偏大。     

过表达ZmIBH1-1提高玉米苗期抗旱性

【目的】干旱是严重影响玉米生长发育进程的一个重要因素。挖掘玉米抗旱相关基因,通过转基因功能验证和转录组分析,解析关键基因在响应干旱胁迫过程中的分子调控机制,为抗旱分子育种和遗传改良提供理论依据。【方法】以玉米自交系B104（WT）为背景材料,利用农杆菌介导方法构建过表达ZmIBH1-1转基因株系（ZmIBH1-1-OE）;通过对转基因植株进行草铵膦抗性筛选、标记基因和目的基因PCR检测,以及运用实时荧光定量PCR检测目的基因的表达情况,鉴定阳性植株和株系;以WT和ZmIBH1-1-OE转基因株系为材料,通过干旱处理（20% PEG6000）,进行表型鉴定和耐旱生理生化指标测定,验证ZmIBH1-1的抗旱功能;通过对干旱胁迫下玉米4叶期转录组的比较分析,鉴定出差异表达的基因（differentially expressed genes,DEGs）;结合DAP-seq（DNA affinity purification sequencing）分析,初步确定ZmIBH1-1蛋白直接调控与抗旱相关的下游靶基因,利用基因组可视化软件IGV（integrative genomics viewer）分析ZmIBH1-1蛋白结合候选靶基因的位置,然后通过Dual-Luciferase试验验证ZmIBH1-1蛋白与靶基因的调控关系。【结果】通过玉米遗传转化获得12个转化事件;T₃代中,能同时检测到标记基因Bar和目的基因ZmIBH1-1的植株有458个,实时荧光定量PCR检测结果表明,ZmIBH1-1-OE中ZmIBH1-1的表达量显著高于WT,株系3和株系8表达量最高,将其自交获得T₄代转基因株系用于后续试验。在干旱胁迫条件下,ZmIBH1-1-OE株系存活率、叶片相对含水量、叶绿素含量、可溶性蛋白含量及其生理生化指标（超氧化物歧化酶、过氧化物酶、过氧化氢酶活性）均显著高于WT,说明玉米中过量表达ZmIBH1-1赋予玉米更高的耐旱性。转录组分析结果表明,WT与ZmIBH1-1-OE株系在干旱胁迫下有1 214个差异表达基因;Gene Ontology（GO）功能富集分析结果表明,差异表达基因主要涉及生物过程、细胞组分和分子功能,如在生物过程中主要涉及到光合作用、应激响应、脱水响应等;KEGG富集分析表明,差异表达基因主要参与植物激素信号传导、新陈代谢等过程。结合转录组显著差异表达基因和DAP-Seq分析所得到ZmIBH1-1蛋白的靶基因,初步确定ZmIBH1-1蛋白直接调控与抗旱相关的11个候选靶基因,包括2个钙信号相关基因、3个半胱氨酸代谢相关基因、1个bHLH转录因子、1个应激响应蛋白、1个谷胱甘肽转移酶、1个氧化还原过程蛋白和2个乙烯响应因子;基因组可视化结果显示ZmIBH1-1蛋白可以结合靶基因启动子区;随后通过Dual-Luciferase试验进一步表明,ZmIBH1-1蛋白可以直接作用于11个候选靶基因,其中,ZmIBH1-1蛋白可以促进ZmCa-M、ZmSYCO、ZmbHLH54、ZmGlu-r1、ZmCLPB3和ZmP450-99A2的表达,抑制ZmAGD12、ZmCYS、ZmCYSB、ZmERF-107和ZmEIN3的表达。此外,在干旱胁迫下NAC、WRKY、MYB等转录因子在ZmIBH1-1-OE和WT株系中也存在差异表达。【结论】ZmIBH1-1的过表达可以增强玉米苗期的耐旱性;ZmIBH1-1蛋白通过直接调控乙烯信号通路中的ZmERF-107和ZmEIN3的表达提高玉米的耐旱性;ZmIBH1-1蛋白通过直接调控钙信号相关基因ZmCa-M和ZmAGD12增强玉米的耐旱性;ZmIBH1-1蛋白可能通过间接调控NAC、WRKY、MYB等转录因子响应干旱胁迫。     

Mapping the spatial distribution of shade provision of street trees in Boston using Google Street View panoramas

Street trees provide shade and increase human thermal comfort during hot summer. In this study, we investigated the spatial distribution of shade provision of street trees in Boston, Massachusetts. The sky view factor (SVF), which influences the solar radiation to the ground and affects human thermal comfort, was used to indicate the contribution of street trees on shade provision. Google Street View (GSV) panoramas were used to calculate the photographic method based SVF (SVF_P), with the consideration of all kinds of obstructions within street canyons. A building height model was used to calculate the simulation based SVF (SVFs), with consideration of obstruction of building blocks only. Considering the fact that street trees and building blocks are the two major obstructions of radiation within street canyons, therefore, the difference between the two SVF estimation results can be considered as the shade provision of street trees. The results show that street trees help to decrease the SVF by 24.61% in Boston, Massachusetts. The shading level varies spatially in the study area. Generally, the southwestern area has much higher shading level than the north and the east. We further explored the shading variation among different socioeconomic groups in the study area. Result shows that Hispanics tend to live in neighborhoods with lower shading level. This study can help to provide a reference for future urban greening projects for global climate change adaption.     

The economic benefits and costs of trees in urban forest stewardship: A systematic review

Understanding the benefits provided by urban trees is important to justify investment and improve stewardship. Many studies have attempted to quantify the benefits of trees in monetary terms, though fewer have quantified the associated costs of planting and maintaining them. This systematic review examines the methods used to jointly analyse the costs and benefits of trees in the urban landscape, assesses the relative balance of benefits and costs, and attempts to understand the wide variation in economic values assigned in different studies. The benefits most frequently studied are those related to environmental regulation and property values, and the available data show that these usually outweigh the costs. Aesthetic, amenity, and shading benefits have also been shown to provide significant economic benefits, while benefits in terms of water regulation, carbon reduction and air quality are usually more modest. Variation in benefits and costs among studies is attributed largely to differences in the species composition and age structure of urban tree populations, though methodological differences also play a role. Comparison between studies is made difficult owing to differences in spatiotemporal scope, and in the way urban forest composition and demographic structure were reported. The overwhelming majority of studies concern deciduous trees in Northern America, and much less is known about urban forests in other regions, especially in the tropics. Future work should thus seek to fill these knowledge gaps, and standardise research protocols across cities. In light of ambitious goals in many cities to increase tree cover, ongoing advances in valuation methods need to provide a more comprehensive accounting of benefits and costs, and to better integrate economic assessment into the decision-making process.     

欧洲葡萄‘粉红亚都蜜’NAC基因DRL1负向调节植物抗旱性

以欧洲葡萄‘粉红亚都蜜’（Vitis vinifera‘Yatomo Rose’）为材料，利用荧光定量PCR技术和转基因技术研究葡萄NAC转录因子DRL1基因对逆境的响应。欧洲葡萄‘粉红亚都蜜’在激素和逆境胁迫下，DRL1表达呈下降趋势，其中以ABA和干旱胁迫处理最为显著。在ABA处理下DRL1转基因烟草株系种子萌发率和根长均高于野生型。干旱处理下，转基因植株对干旱的耐受性降低，同时胁迫相关基因NtLEA5、NtP5CR1、NtPSCS1、NtERD10C和NtDREB3的表达水平比野生型显著下降。此外，DRL1转基因烟草茎中柱发育受到抑制，尤其是导管横切面积仅为野生型的58%。以上结果表明，DRL1基因可能作为1个负向调节子参与植物的干旱胁迫。     

MYB转录因子参与植物非生物胁迫响应与植物激素应答的研究进展

基因的时空表达受转录因子的精确调控。植物在面对不利环境因素比如高温、低温、干旱、盐碱等胁迫时其细胞生理生化会迅速地从“舒适”状态转变进入“胁迫响应”状态。这种快速响应的状态转变依赖于植物对胁迫信号的感知及传递、激素通路(脱落酸、茉莉酸等)的激发、转录因子的活化等复杂的过程;最终植物通过胁迫相关基因的表达、次生代谢转变、抗氧化物质的积累等实现胁迫条件下细胞内环境的再平衡从而获得生存。植物MYB(v-MYB avian myeloblastosis viral oncogene homolog)转录因子就是上述转变中的重要参与者。本文介绍了植物MYB转录因子的结构特征、分类,综述了近些年来MYB转录因子与非生物胁迫,以及植物激素应答过程相关的研究进展。     

铯－134在幼龄茶树叶片中的分布特点

茶树上喷洒￣（１３４）Ｃｓ后，老叶上的沉积量和新叶上的吸收量均按指数回归形式下降，下降的速率以老叶上最快，春茶叶和夏茶叶上次之，秋茶叶上最慢；灌溉和土壤中拌入￣（１３４）Ｃｓ后，茶叶对￣（１３４）Ｃｓ的吸收分两个阶段进行，前期随着时间延长而增加。后期则减少。吸收量有个最大值；茶叶中￣（１３４）Ｃｓ的含量在喷洒情况下比灌溉和拌土的高。     

阔叶树种的研究现状与江西研究的发展方向

通过对阔叶树种在我国及江西省的研究状况的论述，初步总结了阔叶树种的研究水平及研究方向，指出了江西省阔叶树种研究存在的问题，探讨了江西省今后阔叶树种研究的方向。