Nitrogen economy of alpine plants on the north Tibetan Plateau: Nitrogen conservation by resorption rather than open sources through biological symbiotic fixation

Nitrogen (N) is one of the most important factors limiting plant productivity, and N fixation by legume species is an important source of N input into ecosystems. Meanwhile, N resorption from senescent plant tissues conserves nutrients taken up in the current season, which may alleviate ecosystem N limitation. N fixation was assessed by the ¹⁵N dilution technique in four types of alpine grasslands along the precipitation and soil nutrient gradients. The N resorption efficiency (NRE) was also measured in these alpine grasslands. The aboveground biomass in the alpine meadow was 4–6 times higher than in the alpine meadow steppe, alpine steppe, and alpine desert steppe. However, the proportion of legume species to community biomass in the alpine steppe and the alpine desert steppe was significantly higher than the proportion in the alpine meadow. N fixation by the legume plants in the alpine meadow was 0.236 g N/m², which was significantly higher than N fixation in other alpine grasslands (0.041 to 0.089 g N/m²). The NRE in the alpine meadows was lower than in the other three alpine grasslands. Both the aboveground biomass and N fixation of the legume plants showed decreasing trends with the decline of precipitation and soil N gradients from east to west, while the NRE of alpine plants showed increasing trends along the gradients, which indicates that alpine plants enhance the NRE to adapt to the increasing droughts and nutrient‐poor environments. The opposite trends of N fixation and NRE along the precipitation and soil nutrient gradients indicate that alpine plants adapt to precipitation and soil nutrient limitation by promoting NRE (conservative nutrient use by alpine plants) rather than biological N fixation (open sources by legume plants) on the north Tibetan Plateau.     

大鼠脂肪间充质干细胞诱导为类肝细胞及其细胞片的制备

成体多能干细胞,如来自骨髓和脂肪组织的间充质干细胞等具有多向分化的潜能。虽然自体干细胞移植已经发展成为器官移植的有效代替疗法之一,但是由于移植位点细胞的流失和分化条件的限制等问题使得这种疗法的效率大大降低。本研究目的是将由脂肪干细胞分化而来的类肝细胞制备成具有稳定细胞性状的可移植的肝细胞片。首先在体外分离扩增脂肪干细胞,并通过控制严格地分化条件获得类肝细胞。然后将此细胞接种到聚N-异丙基丙烯酰胺(PNIPAAm)结合的细胞培养皿表面,通过调节培养温度到20oC,使细胞成片脱离培养皿形成细胞片。对细胞片进行了常规HE染色和免疫组化观察,结果显示:这类细胞片中平均含有2～3层细胞,并且保持了细胞外基质的完整。同传统的胰酶消化收集移植用细胞相比,细胞片方法极大地减少了对移植用细胞的细胞膜和细胞外基质的损伤,这将大大促进细胞片和原位组织的相互作用,增加细胞利用效率,从而有望提高治疗效果。     

稻田生态系统中3种蜘蛛对白背飞虱的捕食效应研究

应用 二次正交旋转组合设计方法,对稻田生态系统中３种捕食性天敌（三突花蛛Ｍｉｓｕｍｅｎｏｐｓ ｔｒｉｃｕｓｐｉｄａｔａ（Ｔａｂｒｉｃｉｕｓ）、草间钻头蛛Ｈｙｌｙｐｈａｎｔｅｓ ｇｒａｍｉｎｉｃｏｒａ（Ｓｕｎｄｅｖａｌｌ）、真水狼蛛Ｐｉｒａｔａ ｐｉｒａｔｉｃｕｓ（Ｃｌｅｒｋ）和１种害虫（白背飞虱Ｓｏｇａｔｅｌｌａ ｆｕｒｃｉｆｅｒａ（Ｈｏｒｖａｔｈ）的共存系统进行研究,获得了天敌对害虫控制作用的数     

Transformation of Saussurea involucrata by Agrobacterium rhizogenes: Hairy root induction and syringin production

Agrobacterium rhizogenes-mediated genetic transformation of Saussurea involucrata was investigated. Four bacterial strains, A4, LBA 9402, R1000 and R1601 and three explant types, leaf blade, petiole and root, were examined. Over 100 hairy root lines were successfully established with strains R1601, R1000 and LBA9402, but none with A4. The highest transformation efficiency of 67% was achieved by using strain R1601 with root explants. One hairy root line isolated from this combination, HR1601-1, produced up to 43.5 ± 1.13 mg syringin g⁻¹ dw, which is about 50-fold higher than that in the wild type plants.Two other lines, HR1000-1 and HRLBA9402-1, isolated from R1000- and LBA9402-transformed roots, respectively, also displayed high capacity of syringin production, being 32.5 ± 3.08 and 39.7 ± 1.37 mg syringin g⁻¹ dw. These three lines were characterized in detail. Polymerase chain reaction analyses confirmed these root lines were of A. rhizogenes origin.     

三种不同方法固定的石蜡切片中RNA的分析

研究在 10 %中性福尔马林、丙酮、甲醇 氯仿 冰醋酸 3种方法固定的石蜡切片中提取RNA的质量和数量 .取 2 5 0g体重的Wistar大鼠的肾脏 ,分别采用 10 %中性福尔马林、丙酮、甲醇 氯仿 冰醋酸 3种方法固定 ,石蜡包埋 ,H E染色 ;采用RNA裂解液、TRIZOL试剂 2种方法提取切片RNA ,逆转录为cDNA ,采用普通PCR和SYBRGREEN 1定量PCR分析RNA质量和数量 .结果表明 ,3种固定方法都可保持组织良好的结构和形态 ;采用 2种提取方法 ,均可经RT PCR扩增出 180bp大鼠磷酸甘油醛脱氢酶 (G3PDH)、5 6 5bpβ肌动蛋白 (β actin)、10 0bp纤溶酶系活化剂抑制物 1(PAI 1) ;但采用RNA裂解液时 ,比TRIZOL试剂可提取更多的RNA .     

Optimization of sulfated modification conditions of tremella polysaccharide and effects of modifiers on cellular infectivity of NDV

Based on our previous research, sulfated modification conditions of Tremella polysaccharide (TPS), the chlorosulfonic acid to pyridine (CSA-Pry) ratio, reaction temperature and time, were optimized by L₉ (3⁴) orthogonal design taking the yield and degree of sulfation (DS) of modifiers as indexes. Two TPSs, TPS_tp and TPS_70c, were modified under optimized conditions. The effects of two modifiers, sTPS_tp and sTPS_70c, on cellular infectivity of NDV were determined by MTT method taking the non-modified TPS_tp, TPS_tc and TPS_70c as controls. The results showed that the optimized modification conditions were reaction temperature of 80 °C, CSA-Pry ratio of 1:6 and reaction time of 1.5 h. Five polysaccharides at proper concentrations could significantly inhibit the infectivity of NDV to CEF. The virus inhibitory rates of sTPS_tp at 1.563 μg mL⁻¹ group were the highest and significantly higher than those of other three non-modified polysaccharide groups in three sample-adding modes. This indicated that sulfated modification could significantly improve the antiviral activity of TPS. sTPS_tp possessed the best efficacy and would be as a component of antiviral polysaccharide drug.     

含甲肝病毒基因的重组痘苗病毒在动物体内产生针对甲肝病毒的中和抗体

用DNA重组技术得到的含甲肝病毒基因的重组痘苗病毒,可在家兔体内产生ELISA竞争抑制与中和抗体。基础免疫后,动物体内竞争抑制抗体滴度为10,加强免疫后达到80。由重组病毒产生的抗体中和指数比甲肝病毒产生者略低。     

应用纤维素结合域标签构建谷氨酸棒状杆菌表达系统

为优化谷氨酸棒状杆菌表达系统的纯化工艺,合成里氏木霉的CBD基因,将其与谷氨酸棒状杆菌分泌表达载体pXMJ19-sp连接,构建以CBD为纯化标签的重组载体pXMJ 19-sp-CBD.在该载体中插入GFP基因并转化至谷氨酸棒状杆菌,可获得分泌表达融合蛋白GFP-CBD的重组菌.该菌经IPTG诱导后的发酵液在紫外灯下显示强烈的绿色荧光,重组蛋白的分泌表达量达200 mg/L.利用CBD标签对纤维素柱的可逆性吸附,可直接对谷氨酸棒状杆菌分泌到培养基中的重组蛋白进行纯化,从而简化工艺和降低成本,为工业化大生产奠定基础.     

Comparative proteomics analysis of silkworm hemolymph during the stages of metamorphosis via liquid chromatography and mass spectrometry

The silkworm is a lepidopteran insect that has an open circulatory system with hemolymph consisting of blood and lymph fluid. Hemolymph is not only considered as a depository of nutrients and energy, but it also plays a key role in substance transportation, immunity response, and proteolysis. In this study, we used LC‐MS/MS to analyze the hemolymph proteins of four developmental stages during metamorphosis. A total of 728 proteins were identified from the hemolymph of the second day of wandering stage, first day of pupation, ninth day of pupation, and first day as an adult moth. GO annotations and categories showed that silkworm hemolymph proteins were enriched in carbohydrate metabolism, proteolysis, protein binding, and antibacterial humoral response. The levels of nutrient, immunity‐related, and structural proteins changed significantly during development and metamorphosis. Some, such as cuticle, odorant‐binding, and chemosensory proteins, showed stage‐specific expression in the hemolymph. In addition, the expression of several antimicrobial peptides exhibited their highest level of abundance in the hemolymph of the early pupal stage. These findings provide a comprehensive proteomic insight of the silkworm hemolymph and suggest additional molecular targets for studying insect metamorphosis.