Larviculture of the painted river prawn Macrobrachium carcinus in different culture systems

The objective of this study was to evaluate different hatchery systems used for the larviculture of the Macrobrachium carcinus based on survival, larval development and production of post-larvae. The experimental culture was carried out in three phases designated as Phase I (Zoea VI to VIII – Z_{VI – VIII}), Phase II (Zoea VIII to X – Z_{VIII – X}), and Phase III (Zoea X to PL – Z_{X – PL}), with densities of 30, 27.5 and 25 larvae / L, respectively. The M. carcinus larvae (Z_VI) were reared in four culture systems, two being open (Greenwater – GW and Clearwater – CW) and two being closed (Biofloc – BFT and Bio-filter – RAS), distributed in twelve 10 L plastic containers, filled with 20 ppt brackish water, equipped with constant aeration, and water circulated by air lift and heated with thermostat (∼30 °C). The GW treatment was maintained with Chlorophyceae algae in the density of 3–5 × 10⁵ cells/mL. In the CW, the water was previously filtered through a 5 μm mesh screen, sterilized with 10 ppm active chlorine and, dechlorinated with vitamin C and subjected to aeration for 24 h. The BFT received water rich in bioflocs that was matured prior to the experiment and used molasses as a source of organic carbon. In the RAS, the culture water circulated through an external “Dry-Wet” biological filter. The feeding was carried out ad libitum four times daily, alternating a wet diet formula with a commercial diet, which was supplemented with newly hatched Artemia nauplii at a rate of 40–50 per larvae/day. Temperature, dissolved oxygen and pH were monitored daily and the salinity two times per week. Total ammonia, nitrite, nitrate, orthophosphate, alkalinity, total suspended solids, chlorophyll-a, COD and BOD were also analyzed. The best water quality (P < 0.05) was obtained in the RAS, with 0.49 (±0.38), 0.23 (±0.22), and 9.0 (±1.5) mg/L of TAN, NO₂-N and NO₃-N, respectively. In the GW, the nitrogen species showed high fluctuations and higher concentrations at 2.32 (±1.68), 3.53 (±3.53) and 18.2 (±12.9) mg / L of TAN, NO₂-N and NO₃-N, respectively. Considering the three phases (Z_{VI – PL}), the overall survival was 0.03, 1.97, 2.23 and 17.32 % for the BFT, CW, GW and RAS, respectively. When considering the phases separately, the survival in Phase I (Z_{VI – VIII}) was highest in the GW system at 58.7 % while the RAS was the highest in Phases II (Z_{VIII – X}) and III (Z_{X – PL}) at 70.6 % and 60.3 %, respectively. The BFT showed 8.4 (±3.5) PL/L, which was higher (P < 0.05) than that obtained in the RAS (2.8 ± 1.2 PL/L) and the GW (1.3 ± 1.1 PL/L) and similar to that obtained in the CW (5.6 ± 2.0 PL/L). Thus, the larviculture for the M. carcinus may be optimized by adopting a multiphase management strategy, which the intermediate larval stages (Z_{VI – IX}) are reared in the GW system and the final stages (Z_{X – PL}) are reared in the BFT system.     

食用菌多糖抗运动疲劳研究进展

采用文献资料法对食用菌多糖抗运动疲劳的研究进展情况进行了研究。结果表明:食用菌多糖提取方法不同而食用菌多糖的提取率也不相同,超声波法+复合酶法及微波法提取食用菌多糖提取率较高,2种方法科学合理的结合提取食用菌多糖以及不同产地、品种和不同部位提取率是未来学者们需要研究的领域;目前,只是对极少部分食用菌多糖抗运动疲劳进行了研究,研究对象(动物)、模型、指标、评价单一而没有确定最佳剂量的时间而没确定的量。为此,提出了未来应加强和重视食用菌多糖抗疲劳运动多食用菌、多对象(尤其是人)、多模型、多指标、多评价、最佳剂量的研究以及量-效、构-效关系的研究建议。     

绿色富硒生物猪饲料中AFB1达标生产工艺优化研究

饲料中黄曲霉毒素(AFB₁)容易超标,检出率达80%~100%,毒性大,具强致癌性,可抑制生猪免疫机能,降低动物生产性能,引起动物继发感染,还会在动物产品中残留而威胁人类健康,给生猪养殖带来经济损失,降低猪肉食品安全性。本文通过使用先进固体发酵系统设备和益生菌发酵技术,采用单因素试验和响应面中试优化,获得发酵降解猪饲料AFB₁的最佳工艺参数为硒浓度0.3 mg/kg,发酵时间12 h,量子波强度30 Hz,益生菌菌种组合CGMCC NO.17328混合CGMCC NO.15611。该工艺将猪饲料AFB₁量从63.41μg/kg降解到2.98μg/kg,降解率达到95.30%,AFB₁含量达到国家饲料安全标准。生产工艺适合养猪场低成本快速生产AFB₁达标猪饲料。     

内蒙古高原紫花苜蓿土壤氮素丰缺指标和推荐施氮量

为了给内蒙古高原紫花苜蓿（Medicago sativa L.）测土施氮奠定科学基础，本研究采用“零散实验数据整合法”和“养分平衡-地力差减法”新应用公式，开展了该自然区域紫花苜蓿土壤氮素丰缺指标和推荐施氮量研究。结果表明：内蒙古高原生长第1年紫花苜蓿土壤碱解氮第1~6级丰缺指标为≥48，20~48，8~20，4~8，2~4和<2 mg·kg^-1，土壤全氮第1~5级丰缺指标为≥1.4，0.8~1.4，0.4~0.8，0.2~0.4和<0.2 g·kg^-1，土壤有机质第1~6级丰缺指标为≥17，10~17，6~10，3~6，2~3和<2 g·kg^-1。当紫花苜蓿目标产量9~18 t·hm^-2、氮肥利用率40%时，内蒙古高原紫花苜蓿第1~6级土壤推荐施氮量分别为0，68~135，135~270，203~405，270~540和338~675 kg·hm^-2。     

光照对箭筈豌豆及其子代生长和繁殖特性的影响

光照不仅会影响植物自身的生长,也可能会对其子代的生长具有潜在作用。以箭筈豌豆品种兰箭1号和兰箭3号为材料,分析遮荫和自然光照下箭筈豌豆及其子代的生长特性。结果表明:光照对箭筈豌豆生长和繁殖特性的影响因基因型而异。与光照条件相比,遮荫显著降低兰箭1号生物量及茎叶比,但对繁殖分配的比例影响不显著。与此相反,遮荫对兰箭3号的生长与繁殖无显著影响;母本光照环境对箭筈豌豆子代生长特性具有一定程度的影响,且这种影响因子代植株生长的光照环境而异。当子代在遮荫环境下,母本环境显著影响兰箭1号分枝数、地上生物量、总生物量、地上/地下生物量以及兰箭3号的分枝数。但当子代在光照环境下生长时,除兰箭3号分枝数外,其他生长特性均不受母本环境影响。     

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.     

水钾耦合对苜蓿根颈抗氧化特性及越冬率的影响

为探究不同水钾耦合处理对科尔沁沙地生境下种植苜蓿（Medicago sativa）抗寒生理特性的影响，以‘骑士T’和‘公农1号’苜蓿品种为试验材料，采用两因素裂区随机区组设计，水分处理为主区：灌水时间间隔分别为4 d，8 d和12 d（用W₁，W₂，W₃表示）；钾肥施用量水平为副区，具体施用量分别为50 kg·hm^-2 K₂O，100 kg·hm^-2 K₂O和150 kg·hm^-2K₂O（用K₁，K₂，K₃表示），并设置不施钾肥为对照（CK），于越冬前期挖取苜蓿根颈材料测定丙二醛（Malondialdehyde，MDA）、可溶性蛋白含量和抗氧化酶活性，翌年测定越冬率。结果表明：在W₃灌水时间间隔处理下，‘骑士T’和‘公农1号’品种苜蓿根颈的过氧化氢酶、过氧化物酶和超氧化物歧化酶活性、越冬率以及可溶性蛋白含量均明显高于W₁和W₂处理，其中以K₂施用量处理下的数值与CK差异最显著（P<0.05），且此施肥量下的MDA含量也显著低于CK（P<0.05）。因此在科尔沁沙地生境下生产苜蓿，灌水时间间隔为12 d、钾肥施用量在100 kg·hm^-2K₂O有利于提高苜蓿抗寒性。     

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。