诱导Paracin1.7生成的刺激因子的分离与纯化

旨在分离纯化枯草芽孢杆菌(Bacillus subtilis)中能够诱导细菌素Paracin1.7分泌的刺激因子,并探明其对副干酪乳杆菌(Lactobacillus paracasei) HD1.7细菌素生成量及群体感应相关基因luxS表达的影响。本研究通过60%硫酸铵沉淀、CM Sepharose Fast Flow弱阳离子交换柱层析中度纯化及Superdex 75凝胶层析精度纯化,将获得的层析分离物与L. paracasei HD1.7共培养,检测共培养发酵液的抑菌活性及luxS基因的转录水平,并用SDS-PAGE与Native-PAGE检测分离物质。结果表明L. paracasei HD1.7抑菌活性为126.68%;luxS基因上调表达,为对照菌株的2.43倍;刺激因子的表观分子量约为30 kDa。本研究利用三步法初步分离纯化出诱导Paracin1.7生成的刺激因子,明确该刺激因子可以启动种间群体感应相关基因luxS,为L. paracasei HD1.7的群体感应研究奠定了理论和技术基础。     

共培养对土壤重金属污染植物修复的调控作用

A co-culture of two plant materials, Astragalus sinicus L., a leguminous plant with concomitant nodules, and Elsholtzia splendens Naki-a Cu accumulator, along with treatments of a chelating agent (EDTA), root excretions (citric acid), and a control with E. splendens only were used to compare the mobility of heavy metals in chelating agents with a co-culture and to determine the potential for co-culture phytoremediation in heavy metal contaminated soils. The root uptake for Cu, Zn, and Pb in all treatments was significantly greater (P < 0.05) than that of the control treatment. However with translocation in the shoots, only Cu, Zn, and Pb in plants grown with the EDTA treatment and Zn in plants co-cropped with the A. sinicus treatment increased significantly (P < 0.05). In addition, when a co-culture in soils with heavy and moderate contamination was compared, for roots in moderately contaminated soils only Zn concentration was significantly less (P < 0.05) than that of heavily contaminated soils, however, Cu, Zn, and Pb concentrations of shoots were all significantly lower (P < 0.05). Overall, this "co-culture engineering" could be as effective as or even more effective than chelating agents, thereby preventing plant metal toxicity and metal leaching in soils as was usually observed in chelate-enhanced phytoremediation.     

茭白黑粉菌对紫红曲霉部分代谢产物的影响

茭白黑粉菌(Ustilago esculentaHenn)与紫红曲霉(Monascus purpureusWent)共同培养,测定培养物中洛伐他汀、橘霉素、色价的含量。结果表明,在第8 d添加茭白黑粉菌的效果较好。当接种量为2 mL,孢子浓度3.42×108个/mL,洛伐他汀、橘霉素的含量比对照提高203%、274%,但色价却下降24%。形态观察发现,HF菌抑制红曲霉菌丝分支和子囊孢子的形成并导致菌丝的空泡增多。     

Transwell contact co-culture promotes growth and differentiation of single-dissociated iPSCs

AIM：To investigate the promoting role of Transwell contact co-culture system in the growth and differentiation of single-dissociated induced pluripotent stem cells (iPSCs). METHODS：Bovine corneal endothelial cells (CECs) at passage 1~2 (P1~2) were seeded on the underside of Transwell inserts placed into culture plates and were cultured in 37 ℃ and 5% CO2 for 8 h. Accutase digestion and 40 μm filter process disaggregated colony-aggregated iPSCs into single-dissociated iPSCs, and the cells were seeded on the inside of Transwell inserts with CECs in medium of mTeSR1 for 3 d and then in low-glucose DMEM supplemented with 10% FBS for 2 weeks. The characteristics and differentiation markers were evaluated by real-time fluorescence quantitative polymerase chain reaction (qPCR), immunofluorescence staining, live & dead cell staining and alkaline phosphatase (ALP) staining. The group of iPSCs cultured in conventional medium was used as control group 1. The group of single-dissociated iPSCs co-cultured with CECs was set as experimental group, while single-dissociated iPSCs without co-culture were as control group 2. RESULTS：The bovine CECs showed typical hexagonal cobblestone shape. iPSCs showed colony-like growth, while became single-dissociated cells after Transwell contact co-culture with bovine CECs for 3 d. The single-dissociated iPSCs positively expressed the undifferentiated markers, Nanog and Oct4. The mRNA expression levels of Nanog, Oct4 and Sox2 between experimental group and control group 1 were both positive and had no statistical significance difference (P>0.05). The dead cells in experimental group decreased significantly, and there was statistically significant difference compared to control group 2 (P<0.01). After 14 d of induced differentiation co-culture, the single-dissociated iPSCs showed rather uniform polygonal morphology, increased dimension and no obvious colony existence. Negative ALP staining, positive immunofluorescence staining for ZO-1, AQP1 and CD31, and negative for CD34 and CD133 were also observed. The results of qPCR showed that the mRNA expression of Oct4, Nanog and Sox2 significantly decreased, and had statistically significant difference compared with control group 1 (P<0.01). CONCLUSION：When co-cultured with bovine CECs, iPSCs morphologically changed to endothelial-like cells and expressed some markers of CECs. Transwell contact co-culture system not only enhances the growth of single-dissociated iPSCs, but also promotes their differentiation.     

饲料蛋白含量对稻-黄颡鱼共作模式气态氮排放、黄颡鱼生长和氮利用率的影响

【目的】明确饲料蛋白含量对稻-黄颡鱼共作模式下N₂O和NH₃挥发特征及黄颡鱼生长的影响,有利于稻-黄颡鱼综合种养的高效、绿色发展。【方法】采用盆栽模拟试验,以常规单养黄颡鱼模式为对照,系统研究了不同饲料蛋白含量对稻-黄颡鱼共作模式下N₂O和NH₃的排放特征、水体和底泥的氮含量以及黄颡鱼生长的影响。【结果】在相同蛋白含量下,稻-黄颡鱼共作模式的N₂O和NH₃挥发分别比单养黄颡鱼模式降低18.3%和76.20%,水体总氮和无机氮含量降低41.30%和48.85%,黄颡鱼蛋白累积量增加20.00%,氮利用率增加171.50%。在稻-黄颡鱼共作模式下,提高饲料蛋白含量会显著增加N₂O排放量和水体残留氮含量,但对NH₃挥发无显著影响;黄颡鱼特定生长率和蛋白累积量与饲料蛋白含量呈二次曲线关系。饲料氮利用效率随饲料蛋白含量增加呈线性降低趋势。【结论】综合考虑黄颡鱼生长和N₂O排放以及养殖水体氮残留等因素,确定稻-黄颡鱼共作模式饲料蛋白含量不宜超过43.04%。     

序贯共培养体系对小鼠早期胚胎体外发育效果的影响

将体外分离培养经超排处理的新西兰大白兔输卵管上皮及子宫内膜细胞接种于4孔板培养。处理1以CZB为培养液;处理2以CZB加输卵管上皮细胞为培养体系;处理3以CZB加子宫内膜细胞为培养体系;处理4以CZB为培养液,胚胎培养的前24h在输卵管上皮细胞共培养体系中,之后则转入子宫内膜细胞共培养体系中培养(处理4为序贯共培养体系)。将2-细胞期小鼠胚胎随机分配给4个处理,统计各处理组小鼠胚胎的各期发育率及发育质量。结果表明,序贯共培养体系胚胎的囊胚体外发育率显著高于其他共培养组(P<0.05);囊胚孵化率及孵化囊胚的贴附率显著高于其他共培养组(P<0.05);囊胚细胞总数各共培养组显著高于单独培养液组(P<0.05)。可见序贯共培养体系较单一体细胞共培养体系能更好地模拟体内环境,提高早期胚胎的发育率和发育质量。     

Effects of Rice-Fish Co-culture on Oxygen Consumption in Intensive Aquaculture Pond

Rice-fish co-culture has gained increasing attention to remediate the negative environmental impacts induced by intensive aquaculture. However, the effect of rice-fish co-culture on oxygen depletion has rarely been investigated. We constructed a rice-fish co-culture system in yellow catfish (Pelteobagrus fulvidraco) and freshwater shrimp (Macrobrachium nipponense) ponds using a new high-stalk rice variety, and conducted a field experiment to investigate the effect of rice-fish co-culture on water parameters and oxygen consumption. The results showed that rice-fish co-culture reduced the nutrients (total nitrogen, ammonia-N, total phosphorous and potassium) and the dissolved oxygen content in fish and shrimp ponds. However, they showed similar seasonal change of dissolved oxygen in the water of fish and shrimp ponds. Rice-fish co-culture reduced the total amount of oxygen consumption and optimized the oxygen consumption structure in pond. The respiration rates in water and sediment were significantly reduced by 66.1% and 31.7% in the catfish pond, and 64.4% and 38.7% in the shrimp pond, respectively, by additional rice cultivation. Rice-fish co-culture decreased the proportions of respiration in sediment and water, and increased the proportion of fish respiration. These results suggest that rice-fish co-culture is an efficient way to reduce hypoxia in intensive culture pond.     

“花菜－蚯蚓”种养模式对杂草发生的影响及除草剂对蚯蚓的急性毒性评价

为了摸清蚯蚓活动对杂草发生的影响以及除草剂对蚯蚓的急性毒性,2017年－2019年连续3年调查了“花菜－蚯蚓”种养模式下田间杂草物种数、总草密度和生物量,同时测定了17种除草剂对威廉环毛蚓的急性毒性。结果表明,与花菜单一种植模式相比,“花菜－蚯蚓”种养模式显著降低了田间杂草物种数、总草密度和生物量,减轻了杂草危害。不同除草剂对威廉环毛蚓的急性毒性存在较大的差异。采用滤纸法处理48 h,丙草胺、高效氟吡甲禾灵和噁唑酰草胺对威廉环毛蚓的急性毒性最高,LD50分别为1.1、1.8 μg/cm2和7.1 μg/cm2。其次为灭草松,LD50为70.3 μg/cm2。乙羧氟草醚、吡嘧磺隆、氰氟草酯、五氟磺草胺、硝磺草酮、嘧啶肟草醚和烟嘧磺隆对威廉环毛蚓的急性毒性相对较低,LD50均大于1 200 μg/cm2。采用人工土壤法处理14 d,灭草松对威廉环毛蚓的急性毒性最高,LD50为50.3 mg/kg。其次为二甲戊灵、草铵膦和丙草胺,LD50分别为111.9、137.4 mg/kg和169.5 mg/kg。双草醚、氰氟草酯、硝磺草酮和苯唑草酮对威廉环毛蚓的急性毒性相对较低,LD50均大于500 mg/kg。基于滤纸法和人工土壤法的测定结果,丙草胺、灭草松、噁唑酰草胺、高效氟吡甲禾灵、草铵膦和二甲戊灵对威廉环毛蚓具有相对较高的急性毒性,不建议用于“作物－蚯蚓”种养农田防除杂草。     

NaCl筛选法获得无标记耐盐转基因水稻的高效转化技术研究

NaCl筛选法是培养无标记耐盐转基因水稻的有效方法。以中华11、秀水11、春江06这3个粳稻品种成熟胚愈伤组织为材料,以SKC1为目标基因,研究NaCl筛选条件下,热激与液体共培养对农杆菌介导耐盐基因转化水稻效率的影响。结果显示,热激处理和液体共培养都能提高耐盐基因对水稻的转化率,其中,热激处理最高转化提高率达到46.2%,液体共培养后最高转化提高率达到60.7%。     

稻虾共作模式下秸秆还田与投食对稻米品质的影响

为提高稻虾共作模式的稻米品质和效益,通过设置秸秆还田投食（SF）、秸秆还田不投食（SNF）、秸秆不还田投食（NSF）和秸秆不还田不投食（NSNF）处理,并以水稻单作秸秆还田（CK-S）和水稻单作秸秆不还田（CK-NS）为对照,对秸秆还田和投食对稻米品质的影响进行研究。结果显示：投食（F）处理能显著提高水稻、小龙虾产量和稻米营养品质,在秸秆还田（S）与秸秆不还田（NS）的条件下,F处理比不投食（NF）处理的蛋白质含量分别增加了27.41%和36.16%,差异显著。S处理与F处理的交互作用可显著影响稻米加工品质;整精米率和精米率在NSNF处理下最高,显著高于SF、SNF、NSF;秸秆还田、投食及其交互作用对蒸煮、食味品质无显著影响,稻米RVA黏滞性谱差异性较小;稻虾模式可提高稻米外观品质,稻虾共作模式下各处理垩白粒率和垩白度都低于稻田单作,与CK-S相比,SF处理垩白粒率和垩白度分别降低了15.09%、15.65%。结果表明,稻虾共作秸秆还田与投食2种措施可以改善稻米品质。