三角褐指藻固定化培养的初步研究

用褐藻酸钙对三角褐指藻进行固定化培养实验,测定了不同胶粒大小,不同胶粒密度及不同接种量对该藻的影响,比较了自由生长和固定细胞的生长曲线,还进行了常温组和低温组固定化细胞的活化复苏实验,三角褐指藻在褐藻酸钙胶中仍具有呼吸和光合作用的能力,球径为２．５ｍｍ时,细胞生长快,每５０ｍｌ培养液中加入１５０个胶球时细胞生长量大,接种量不能低于１０＾５个细胞／ｍｌ,在冰箱（４℃）中可保存２１个月,本藻适合用固定     

Immobilization antigen-independent protection of olive flounder (Paralichthys olivaceus) against Philasterides dicentrarchi (Ciliophora: Scuticociliatia) infection

To determine whether olive flounder (Paralichthys olivaceus) immunized with lysate of Philasterides dicentrarchi could be protected immobilization antigen (i-antigen) type independently, fish were immunized with lysate of ciliates obtained from in vitro cultures or ciliates obtained from infected fish. Fish immunized with ciliates obtained from infected fish were completely protected against artificial infection with in vitro cultured P. dicentrarchi, in spite of a weak or no serum agglutination activity against in vitro cultured ciliates. The present results indicate that i-antigen-independent protection was elicited by immunization of fish with the ciliates lysate, and the cultured ciliates would provide a good source for preparation of vaccines, which are cross-protective against various i-antigen types of P. dicentrarchi.     

Nitrogen transformations in cold and frozen agricultural soils following organic amendments

Though microbial activity is known to occur in frozen soils, little is known about the fate of animal manure N applied in the fall to agricultural soils located in areas with prolonged winter periods. Our objective was to examine transformations of soil and pig slurry N at low temperatures. Loamy and clay soils were either unamended (Control), amended with ¹⁵NH₄-labeled pig slurry, or amended with the pig slurry and wheat straw. Soils were incubated at −6, −2, 2, 6, and 10 °C. The amounts of NH₄, NO₃ and microbial biomass N (MBN), and the presence of ¹⁵N in these pools were monitored. Total mineral N, NO₃ and ¹⁵NO₃ increased at temperature down to −2 °C in the loam soil and −6 °C in the clay soil, indicating that nitrification and mineralization proceeded in frozen soils. Nitrification and mineralization rates were 1.8-4.9 times higher in the clay than in the loamy soil, especially below freezing point (3.2-4.9), possibly because more unfrozen water remained in the clay than in the loamy soil. Slurry addition increased nitrification rates by 3-14 times at all temperatures, indicating that this process was N-limited even in frozen soils. Straw incorporation caused significant net N immobilization only at temperatures ≥2 °C in both soils; the rates were 1.4-3.4 higher in the loam than in the clay soil. Nevertheless, up to 30% of the applied ¹⁵N was present in MBN at all temperatures. These findings indicate that microbial N immobilization occurred in frozen soils, but was not strong enough to induce net immobilization below the freezing point, even in the presence of straw. The Q₁₀ values for estimated mineralization and nitrification rates were one to two orders-of-magnitude larger below 2 °C than above this temperature (13-208 versus 1.5-6.9, respectively), indicating that these processes are highly sensitive to a small increase in soil temperature around the freezing point of water. This study confirms that net mineralization and nitrification can occur at potentially significant rates in frozen agricultural soils, especially in the presence of organic amendments. In contrast, net N immobilization could be detected essentially above the freezing point. Our results imply that fall-applied N could be at risk of overwinter losses, particularly in fine-textured soils.     

采用15N同位素稀释法研究不同层次土壤氮素总转化速率

采用15N同位素稀释方法,开展短期(7天)室内培养实验,估算了一水稻土0~20、20~60和60~90 cm土层土壤主要N素转化过程的总转化速率,结果表明,标记N溶液加入后2 h内各土层土壤的总矿化、硝化、固定速率显著高于其他时间段(p<0.01)。2 h后,矿化速率在小范围内起伏。0~20 cm土层土壤N素的硝化速率随培养时间延长而降低,另外两层土壤则基本保持稳定,硝化速率的变化与硝化作用底物NH4+-N浓度的变化呈显著正相关。值得注意的是,外源无机N溶液加入后2 h内,大量NH4+-N和NO3--N被固定,并认为N素的非生物固定起主导作用。2 h后,出现了N素在固定与再矿化间反复转换的现象。实验结果表明,与净转化速率相比总转化速率能更好地描述单个N素转化过程,但由于外源N加入对N素转化的影响、再矿化作用以及忽略了N素转化过程中的气体损失、DNRA(硝态氮异化还原为铵)过程等,本研究结果与真实值间存在一定差异。     

微胶囊法固定化糖化酶的研究

酶或细胞固定化技术是微生物发酵的主要手段之一,它可使生产速率和效率获得显著提高。为确定微胶囊法固定糖化酶的最佳条件,以壳聚糖和海藻酸钠为载体,采用微胶囊法固定糖化酶。试验结果表明,最佳固定化条件为:壳聚糖脱乙酰度为85%、海藻酸钠浓度为3%、戊二醛浓度为1.5%、氯化钙浓度为0.2 mol.L-1。固定化酶的最适作用温度为75℃,比游离酶提高了20℃,最适pH为4.5,比游离酶下降0.5个单位。固定化酶的活力最高达2 013.42 U.g-1干胶,相对活力为89.3%,米氏常数Km为1.28%,半衰期为223 h。采用壳聚糖和海藻酸钠为载体微胶囊法固定糖化酶的方法是可行的。     

固定化唾液链球菌的酶学性质研究

以海藻酸钙为载体包埋唾液链球菌嗜热亚种Y-2的菌体细胞,对固定化细胞催化合成γ-氨基丁酸进行了较详细的研究。研究结果表明:固定化细胞谷氨酸脱羧酶(Glutamate decarboxylase,GAD)反应的最适温度为40℃,同时具有良好的温度稳定性。固定化细胞酶活最适反应pH为3.8。细胞经固定化后pH稳定性明显增加,GAD酶活回收率普遍高于游离细胞。0.1%Triton X-100具有较强的酶活促进作用。固定化细胞的GAD在酶促反应中并不存在底物抑制现象。在上述最优条件下进行菌体生产力测试,固定化细胞转化L-谷氨酸单钠盐11 h后,转化液γ-氨基丁酸的浓度达到了2.79 g/L。     

壳聚糖固定化胰蛋白酶研究

以自制的壳聚糖为载体,以戊二醛为交联剂,制备了壳聚糖固定化胰蛋白酶。经测定该酶活性回收率达到72.7%,操作半衰期为12 d,表观米氏常数为5.88 mg/ml,最适pH值为8.0,最适温度为40℃;与游离胰蛋白酶相比,酶的耐热性和操作半衰期有明显提高,但表观米氏常数也略有增大。     

D301大孔阴离子树脂固定化β-淀粉酶的研究

［目的］对β-淀粉酶的固定化工艺进行探讨。［方法］以D301大孔阴离子树脂为载体,采用离子交换吸附法固定化β-淀粉酶,测定酶活性。［结果］加酶量为500 U/g（树脂,湿重）,pH值为5.6,在45 ℃下恒温振荡3 h,固定化酶的酶活可达180 U/g载体。［结论］制备的固定化酶,其温度稳定性、pH稳定性、储藏稳定性和操作稳定性较好。     

Temporal variation in mineral nitrogen in soil as influenced by incorporation of legume or cereal residues under submergence or well drained conditions

A laboratory study was conducted at the Indian Agricultural Research Institute, New Delhi on a sandy clay loam soil of pH 7.9 and organic C content of 0.34% to study the effect of incorporating Sesbania or Vigna legume residues or wheat straw at 15 and 30t ha^?1 on temporal variation in ammoniacal and nitrate‐N in soil under submergence and well drained conditions. Under submergence most mineral N was present as ammoniacal‐N, while under well drained conditions it was present as Nitrate‐N. The content of ammoniacal N in soil was the highest after 30 days of incubation and declined thereafter under submergence. On the other hand under well drained conditions the mineral‐N (mostly nitrate) content in soil at 30 DAI was very little and showed increases only later, reaching the highest level at 90 DAI. Application of wheat straw specially at 301 ha^?1 level resulted in immobilization of native soil‐N. These results show that rice which is grown under submergence can be transplanted soon after incorporation of legume residues, but for wheat or other crops which are grown under well drained condition a time interval of 30 days or more needs to be provided before sowing the crop.     

Litter dynamics of six multipurpose trees in a homegarden in Southern Kerala, India

Multipurpose trees, the integral components of homegardens, contribute significantly to the closed nutrient cycling processes and sustainability of the ecosystem. Although, the litter production and probable nutrient returns via litter in homegardens have been documented, quantification and characterization of the decomposition and bioelement release from the litter have received relatively little scientific attention. The objective of the present study is to explore the litter dynamics of six locally important multipurpose trees (Mangifera indica L., Artocarpus heterophyllus Lamk., Anacardium occidentale L., Ailanthus triphysa Dennst., Artocarpus hirsutus L. and Swietenia macrophylla L.), in an agroforestry homegarden in Southern Kerala, India. Litterfall and nutrient additions in the six species ranged from 383 to 868 g m⁻² yr⁻¹, nitrogen, 6.4 to 8.8, phosphorus, 0.17 to 0.42 and potassium, 1.1 to 2.8 g m⁻² yr⁻¹. The annual litter output in the homegarden was 425 kg with A. hirsutus, M. indica, A. heterophyllus and A. occidentale recording significantly higher litter and nutrient additions. Leaf litter decay studies revealed A. heterophyllus and A. occidentale to be the most labile litter species and S. macrophylla the most recalcitrant. The decay rate coefficients varied significantly among the species. Foliage decomposition rates related to the initial chemical composition of the litter revealed best correlation with lignin. NPK release was almost complete by the end of decay in all species inspite of the initial phases of accumulation observed for nitrogen and phosphorus. Two-way analysis of variance test revealed significant differences in the contents of the three elements as a function of species and time elapsed. Macronutrients were released in the order K>N/P. The higher rates of decay and nutrient turnover in M. indica, A. heterophyllus and A. occidentale foliage indicated the potential of using these species’ litter as nutrient inputs in agriculture while A. triphysa, A. hirsutus and S. macrophylla perform better as organic mulches taking a longer time for decay and hence nutrient release.