我国食用菌出口遭遇贸易壁垒的原因、类型及对策

贸易壁垒正在成为我国农产品出口的瓶颈。本文分析了我国食用菌产品出口遭遇的贸易壁垒类型及其原因，并提出了相关建议。     

Effect of EGFP gene transfection on the cell cycle distribution of primary cultured human chondrocytes

AIM: To investigate the effect of enhanced green fluorescence protein (EGFP) gene transfection on the cell cycle distribution of primary cultured human chondrocytes in order to establish a tracking method of cultured human nasoseptal chondrocytes. METHODS: pEGFP-N1 plasmid was amplified in E.coli, and purified by high purity kit. Primary cultured human chondrocytes,which were initially obtained from the nasoseptal cartilage, were cultured in vitro and transferred with pEGFP-N1 by means of electroporation with Amaxa nucleofector device. Transfering process and transient expression were evaluated by laser scanning confocal microscope (LSCM), the transfer efficiency and the cell cycle distribution were evaluated by flow cytometry. RESULTS: There was significant expression of EGFP at 24 h after transferring. The transfection efficiency of pEGFP-N1 into primary cultured human chondrocytes reached 35.37％ at 48 h. It didn't affect the process of cell adherance and had no effect on the cell cycle distribution. CONCLUSION: Primary cultured human chondrocytes, which were transfected with pEGFP, are alive in vitro, and the transferring process doesn't affect the cell cycle distribution. These results suggest that pEGFP-N1 is an ideal transient expression vector for primary cultured human chondrocytes and it might be a well tracer in construction tissue engineered cartilage.     

Roles of the Kv1.2, Kv1.5, Kv2.1 potassium channels in hypoxia pulmonary vasoconstriction

AIM: To determine the role of Kv1.2, Kv1.5, Kv2.1 in the hypoxia pulmonary vasoconstriction (HPV). METHODS: Male Wistar rats were divided into two groups: normoxic group and hypoxic group. The single smooth muscle cell was obtained from pulmonary artery of Wistar rats with acute enzymatic digestion method. The conventional whole-cell patch clamp technique was used to record the resting membrane potential (Em) and the potassium currents of voltage-gated potassium channel (IKv) in rat pulmonary arterial smooth muscle cells (PASMC). Intracellular application of Kv1.2/Kv1.5/Kv2.1 antibodies (1∶125) was conducted through the whole-cell patch clamp system. RESULTS: ① Em of PASMC was depolarized after 24 h hypoxia compared with that of control cells . IKv of PASMC was decreased after 24 h hypoxia, . ② The mixture of Kv1.2/Kv1.5/Kv2.1 antibodies depolarized Em and inhibited IKv in PASMC from normoxic rat, whereas the mixture of Kir2.1/Kir2.3/Kir4.1 antibodies had no effects on them. ③ The mixture of Kv1.2/Kv1.5/Kv2.1 antibodies and the mixture of Kir2.1/Kir2.3/Kir4.1 antibodies had no effects on IKv and Em from rats hypoxic for 24 h. CONCLUSION: Kv1.2, Kv1.5, Kv2.1 might be oxygen sensitive potassium channels which mediated HPV.     

家鸡Leptin成熟肽cDNA的克隆、重组蛋白表达及纯化

从 18周龄鸡卵巢组织中抽提总 RNA,使用六聚体随机引物反转录后 ,用鸡 L eptin(瘦素 )特异性引物扩增出鸡L eptin编码区第 5 2～ 4 6 0 bp的长度为 4 0 9bp的 c DNA片段。根据鸡 L eptin的 3′端第 4 6 0～ 4 92 bp序列设计了 3条部分相互重叠并且顺序串联延伸的下游反义引物 ,并在最后 1条引物 3′端连接上 Eco R 切点 ;在以上用于反转录扩增的 5′端引物的 5′端连接一 Bam H 切点。用该 5′端引物分别与 3个 3′端引物配对 ,利用反转录扩增出的 4 0 9bp的L eptin c DNA片段作为第 1模板进行扩增 ,扩增产物再作为模板与下一引物对再次扩增。经 3次扩增得到编码鸡 L ep-tin成熟肽的全长 4 5 1bp的 c DNA序列。将该 4 5 1bp的 L eptin c DNA序列经 Bam H 和 Eco R 双酶切后 ,克隆入表达质粒 p RSET A的 Bam H 和 Eco R 两酶切位点之间 ,构建成表达质粒 p L ep- SCAU。转化有重组表达质粒 p L ep-SCAU的大肠杆菌 BL 2 1(DE3)在 L B培养基中培养后 ,经 IPTG诱导表达出相对分子质量为 2 0 10 0的鸡 L eptin融合蛋白和少量 4 0 2 0 0的 L eptin融合蛋白。L eptin融合蛋白的表达在 IPTG浓度为 0 .0 5 mmol/ L 时达到最高 ,占总菌体蛋白的 32 .6 %。用 Ni- NTA凝胶从 7L 发酵培养菌裂解液中纯化出 180 m g左右     

重庆市近郊区县鸭传染性浆膜炎病病原分离鉴定及防治

通过对重庆近郊的沙坪坝区、北碚区、九龙坡区、永川市、璧山县、铜梁县的7个养鸭场病死鸭的病原分离,分离到三株细菌,经染色镜检、生化鉴定、动物回归实验,确诊为鸭里氏杆菌。药敏试验结果显示对庆大霉素、青霉素、链霉素、土霉素等高度敏感。在临床上交替使用这些药物,收到较好治疗效果。将分离菌制成灭活苗免疫1周龄的健康鸭,20日龄时用分离菌进行攻毒,平均保护率达83.3%。     

野外放归大熊猫肠道菌群变化的研究

对四川卧龙中国保护大熊猫研究中心的一只野外放归亚成体大熊猫肠道菌群的组成和季节变化规律进行了研究,同时与其圈养双胞胎兄弟的肠道菌群进行了比较。从放归大熊猫粪便中分离出17种肠道菌,优势菌群为肠杆菌、肠球菌和乳杆菌。与圈养大熊猫相比,放归大熊猫肠道菌群中优势菌群的种类未发生改变,但是肠球菌数量增加,肠杆菌和乳杆菌的数量减少。研究发现放归大熊猫肠道菌群中的肠杆菌和肠球菌的数量随季节变化有较大波动,乳杆菌的数量随季节变化波动不大;而圈养大熊猫三种优势菌的数量随季节变化波动都不大。     

旱、盐胁迫下黄芪种子萌发及其对水杨酸的响应

通过测定不同浓度聚乙二醇(PEG)和NaCl胁迫下膜荚黄芪(Astragalus membranaceus)和蒙古黄芪(A.membranaceus var.mongholicus)种子的最终发芽率、发芽势、简化活力指数、苗长、根长、苗干重、根干重等指标,以及不同施用方式和浓度的水杨酸(SA)对重度PEG和NaCl胁迫下两种黄芪种子最终发芽率的影响,旨在研究两种黄芪种子对PEG和NaCl胁迫响应方式的异同及SA对两种黄芪种子在重度PEG和NaCl胁迫下保护效果的差异。结果表明,低浓度PEG和NaCl只能促进膜荚黄芪种子萌发。在-0.5 MPa PEG和-0.7 MPa NaCl处理下,蒙古黄芪种子的活力指数显著优于膜荚黄芪种子(P0.05)。SA浸种能促进重度PEG和NaCl胁迫下两种黄芪种子的最终发芽率。蒙古黄芪种子的耐胁迫性高于膜荚黄芪种子。两种黄芪种子对SA的响应方式有区别。SA浸种处理优于拌种处理。     

黄花苜蓿花药培养再生体系的建立与单倍体鉴定

花药组织培养再生体系的构建是单倍体育种的重要途径之一。本研究对呼伦贝尔黄花苜蓿(Medicago falcate‘Hulunbeier’)进行了花药组织培养研究并建立了花药组培再生体系。结果显示,液体悬浮培养基比固体培养基更适合呼伦贝尔黄花苜蓿花药愈伤组织的培养,其愈伤形成的培养条件为B5培养基+0.5 mg·L~(-1) 2,4-D+0.25 mg·L~(-1) 6-BA+0.4mg·L~(-1) NAA+3.0mg·L~(-1) KT;分化培养基为MS+1.0mg·L~(-1) 2,4-D+0.5mg·L~(-1) 6-BA+2%蔗糖+0.7%琼脂;生根培养基为1/2MS+0.1mg·L~(-1) NAA+2%蔗糖+0.7%琼脂;获得的再生植株经流式细胞仪鉴定,其单倍体比例高达27%。     

极细链格孢激活蛋白对大豆光合特性、叶绿素荧光参数及产量的影响

为探讨激活蛋白对大豆生长、产量的影响,以大豆冀豆17为研究对象,利用光合仪与称重法测量了不同浓度极细链格孢激活蛋白(1,2,3,4,5,6μg/m L)处理后大豆的光合特性、叶绿素荧光参数及产量。结果表明,随着施用的极细链格孢激活蛋白浓度的增加,大豆叶绿素含量、净光合速率、气孔导度、蒸腾速率、最大光化学量子效率(Fv/Fm)、有效量子效率(ΦPSⅡ)和光化学猝灭系数(q P)呈先上升后下降的趋势。施用1,2,3,4μg/m L浓度的极细链格孢激活蛋白不仅提高了大豆叶绿素含量,而且增加了大豆叶片的净光合速率、气孔导度、蒸腾速率,同时还能提高叶绿素荧光的最大光化学量子效率(Fv/Fm)、有效量子效率(ΦPSⅡ)和光化学猝灭系数(q P)。其中以3,4μg/m L浓度的处理效果最好。施用更高浓度的激活蛋白(5,6μg/m L),上述光合特性与叶绿素荧光促进效果下降或者消失。施用中浓度激活蛋白(3,4μg/m L)大豆产量增加了11.21%~14.76%。结果表明,喷洒合适浓度的极细链格孢激活蛋白是一种促进大豆增产的有效措施。