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.     

镇江长江豚类省级自然保护区水域生态修复效果的监测与评价

自2006年以来,长江豚类极度濒危的生存状况得到广泛关注,在其自然保护区实施一系列生态修复工程,为镇江长江豚类省级自然保护区内的江豚提供良好的生存环境。于2015—2018年在和畅洲水域建设生态浮岛、人工鱼巢和放流底栖动物进行水域生态修复,对生态修复工程进行跟踪监测和效果评价。结果表明：生态浮岛的建设可提高区域内浮游植物和浮游动物的生物多样性和群落结构复杂程度;人工鱼巢对产黏性卵鱼类具有一定增殖效果,增殖种类主要为鲤、鲫、䱗和黄尾鲴;生态浮岛和人工鱼巢的建设对鱼类均有较好的聚集效果;底栖动物增殖放流显著增加了放流物种的丰度、生物量及保护区底栖动物群落生物量,增殖放流效果评价为中等。该自然保护区水域生态修复效果明显,为工程施工影响下水生生物保护区生态修复提供科学依据。     

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。     

5种生物制剂对马铃薯种薯萌芽、极端天气下植株生长和产量性状的影响

旨在满足马铃薯生产中茬口衔接、机械化生产技术应用、不利气候下稳产等对马铃薯出苗早、齐、壮的需求,以‘费乌瑞它’为供试品种,用基于有益活菌或工程菌提取物的5种生物制剂进行种薯处理,对多重性状进行了对比分析。5种生物制剂较常规化学制剂,均能够不同程度地促进种薯萌芽和芽根同生,出苗期提前2~7天,播种后49天的出苗率提高3.33%~17.78%。其中,表现最好的为酵母核苷酸衍生物和VDAL,种薯萌发和生根均显著高于对照。霜冻后,生物剂拌种处理在恢复前期促进植株生长,由此促进恢复后期的块茎发育,较常规化学处理增产8.39%~24.03%,体现了不同程度的保产效果。多马道黑、酵母核苷酸衍生物、根肽和VDAL体现出较好的保产效果,可作为种薯处理剂投入马铃薯生产。     

Assessing climate change adaptations for community-scale water resources using a low-frequency weather generator

Paddy and Water Environment - Water adaptation strategies are necessary for decreasing climate change impacts on local societies. This study models a community water supply system as a distributed...     

Impact of grassland contract policy on soil organic carbon losses from alpine grassland on the Qinghai–Tibetan Plateau

Carbon storage in the soils on the Qinghai–Tibetan Plateau plays a very important role in the global carbon budget. In the 1990s, a policy of contracting collective grasslands to smaller units was implemented, resulting in a change from the traditional collective grassland management to two new management patterns: a multi‐household management pattern (MMP: grassland shared by several households without enclosures) and a single‐household management pattern (SMP: grassland enclosed and used by only one household). In 2016, 50 MMP and 54 SMP winter pastures on the Qinghai–Tibetan Plateau were sampled to assess the differences in soil organic carbon (SOC) between the two management patterns. Results showed that average SOC was significantly greater under MMP than under SMP, with an estimated 0.41 Mg C/ha/yr lost due to SMP following the new grassland contract. Based on the government's grassland policy, four grassland utilization scenarios were developed for both summer and winter pastures. We found that if the grassland were managed under SMP, likely C losses ranged between 0.31 × 10⁷ and 6.15 × 10⁷ Mg C/yr across the Qinghai–Tibetan Plateau relative to MMP, which more closely resembles pre‐1990s grassland management. Previous estimates of C losses have only considered land use change (with cover change) and ignored the impacts driven by land management pattern changes (without cover change). The new data suggest that C losses from the Qinghai–Tibetan Plateau are greater than previously estimated, and therefore that the grassland contract policy should be reviewed and SMP households should be encouraged to reunite into the MMP. These findings have potential implications for land management strategies not only on the Qinghai–Tibetan Plateau but also other grazing regions globally where such practices may exist.     

Replacing Soybean Meal with Alternative Protein Sources in Diets for Pond‐raised Hybrid Catfish, ♀ Ictalurus punctatus × ♂ Ictalurus furcatus

The present study investigated the replacement of soybean meal with combinations of two or three alternative protein sources in diets for pond‐raised hybrid catfish, ♀ Ictalurus punctatus × ♂ Ictalurus furcatus. Alternative protein sources evaluated included cottonseed meal, distillers dried grains with solubles (DDGS), peanut meal, and porcine meat and bone meal (PMBM). Hybrid catfish fingerlings with a mean initial weight of 35 g/fish were stocked into 25 earthen ponds (0.04 ha) at a density of 14,826 fish/ha. Fish were fed once daily to apparent satiation for 166 d. No significant differences were observed for total diet fed, net yield, weight gain, survival, carcass yield, fillet yield, or fillet proximate composition among dietary treatments. Results show soybean meal may be completely replaced by combinations of cottonseed meal and one or two other alternative protein sources including DDGS, peanut meal, and PMBM in the diet without markedly affecting production and processing characteristics and fillet proximate composition of pond‐raised hybrid catfish. These alternative diets may be used during foodfish production when prices are favorable.     

植棉修复镉污染土壤研究进展

近年来我国耕地面积不断减少，迫使非食用作物向重金属污染耕地转移已经是大势所趋。但到目前为止仍未发现镉耐受性好和镉富集能力优异的作物，因此对非食用作物的重金属耐受性以及土壤的生物修复研究已经迫在眉睫。综述了具有在镉污染耕地上替代种植前景的棉花的镉胁迫研究进展，为全面评估棉花在镉污染耕地替代种植的可行性提供参考。     

果蔬型甜玉米新品种京科甜191的选育

京科甜191是以超甜玉米自交系T68为母本,自交系T6302为父本,通过杂交选育而成的果蔬型甜玉米品种。果穗筒型,穗长19.0 cm,穗粗5.3 cm,穗行数16~18行,行粒数36~38粒,粒行整齐。籽粒亮黄色,外观漂亮。平均每667 m2鲜果穗产量900 kg左右,出籽率约67.4%。鲜籽粒含粗蛋白2.98%,粗脂肪1.37%,总糖6.7%,口感甜脆清爽,皮薄无渣。播种至采收平均85.4 d(天),适宜京津冀及类似生态区种植。