Evaluation of embryonic age and the effects of different proteases on the isolation and primary culture of chicken intestinal epithelial cells in vitro

The present study evaluates the effects of embryonic age and proteolytic enzymes on the isolation and primary culture of chicken enterocyte and to establish an effective technique for chicken intestinal epithelial cell (IEC) cultivation. Fourteen‐day‐old, 16‐day‐old and 18‐day‐old embryos (average weight: 52.23 ± 0.76 g, 50.86 ± 0.99 g, 48.98 ± 1.03 g) were the source for preparation of enterocyte culture, and trypsin‐ethylene diamine tetraacetic acid, collagenase, thermolysin and combination of collagenase and thermolysin were used for digestion medium. Optimal culture protocols were determined by qualitative assays of proliferation. Cells isolated by using 14‐day‐old embryo and collagenase obtain the best attachment and growth in culture, and the production of continuously growing IEC cultures. Thus, we conclude that the use of collagenase as a dissociating enzyme and 14‐day‐old embryo as a source can be advantageously applied to the isolation of chicken IEC and this method may be useful for various applications and basic studies of the intestinal tract concerning such objects as physiology, immunology and toxicology.     

猪流行性腹泻病毒灭活疫苗的制备及攻毒保护研究

为研究现用猪流行性腹泻病毒(PEDV)CV777疫苗毒株对流行毒株的免疫保护效果,以RTPCR进行PEDV CV777疫苗株、JS12流行株S基因克隆测序,进行PEDV S基因核酸序列分析与氨基酸序列分析。分别以转瓶工艺与细胞悬浮培养工艺培养Vero E6细胞,制备PEDV CV777株转瓶灭活苗与悬浮灭活苗。分别以PEDV转瓶灭活苗、悬浮灭活苗、PEDV流行株组织灭活苗(JS12株)产前30d免疫经产母猪;母猪产仔后14d进行仔猪攻毒保护试验。核酸序列分析显示,PEDV CV777株S基因与流行毒株的相似性在96.4%~99.7%之间,氨基酸序列相似性在96.7%~99.5%之间。转瓶工艺制备PEDV抗原效价为107.0 TCID50/mL,悬浮培养制备PEDV抗原效价为108.5 TCID50/mL。攻毒保护试验显示,悬浮灭活苗免疫母猪后,仔猪有2/10发病,保护率为8/10,转瓶灭活苗组发病率为8/10,保护率仅为2/10;组织灭活苗组9/10发病;对照组10头全部发病。该试验证明,提高PEDV CV777株的抗原含量,能够有效提高疫苗对PEDV流行毒株的保护效果。     

Effect of water extract from spent mushroom substrate after Ganoderma balabacense cultivation by using JUNCAO technique on production performance and hematology parameters of dairy cows

The spent mushroom substrate of Ganoderma balabacense cultivation (SMSGB) contains a large amount of bioactive substances. However, the potentials of SMSGB for improving milk production in dairy cows have not been thoroughly studied. In this study, the effects of hot water extract (HWE) from spent mushroom substrate after G. balabacense cultivated with JUNCAO, the herbaceous plants long‐known to be suitable for cultivating edible and medicinal fungi, on production performance and hematology parameters of dairy cows, were determined. Holstein dairy cows were fed different doses of HWE. After a 60‐day administration period with 100 g/day of HWE, milk yield, milk protein and triglyceride (TG) levels increased by 4.02% (P < 0.01), 4.49% (P < 0.05) and 32.65% (P < 0.05), respectively; somatic cell count (SCC) and alanine aminotransferase (ALT) were significantly decreased (P < 0.05). The production performance of dairy cows suggests that HWE with SMSGB treatment is effective in improving milk yield (P < 0.01) and hematology parameters of dairy cows, and may be useful as a functional feed additive.     

斑马水塔花的组织培养和快速繁殖

试验筛选出斑马水塔花的诱导愈伤组织培养基MS+6-BA1.0mg·L-1+NAA0.2mg·L-1，愈伤组织诱导率达96.7%；继代增殖培养基MS+6-BA2.0mg·L-1+NAA0.2mg·L-1，增殖倍数达8～9倍；生根培养基MS+KT 0.5mg·L-1+NAA0.5mg·L-1，生根率达100%。幼苗经炼苗后，移栽到经菌肥处理的甘蔗渣∶泥炭土∶珍珠岩＝6∶3∶1的基质上，成活率达95%以上。     

植物组织培养快速繁殖类型综述

应用植物组织培养方法进行快速繁殖,具有繁殖速度快、能保持植株的优良性状等优点。其繁殖类型有无菌扦插、器官发生、器官直接转变型、胚状体发生和原球茎等类型。指出了组织培养快速繁殖技术目前存在的主要问题。     

芳樟工厂化育苗技术研究

通过对芳樟工业原料林良种茎尖的离体培养和繁殖 ,探讨芳樟茎尖诱导、分化、不定芽生长分化、生根的适宜的培养基和适宜的培养条件。结果表明 :适宜芳樟茎尖诱导培养基为MS +6 -BAa～ 2amg .L-1+NAAbmg .L-1,诱导率达 70 %以上 ,适宜分化丛生芽的理想培养基为改良MS +6 -BAa′～ 2a′mg .L-1+NAA0～ 2bmg .L-1+GA0～cmg .L-1芽年增殖系数达4 5 12 以上 ,有效芽苗率达 80 %以上 ,适合生根培养基为 1/2改良MS +NAAb′mg .L-1+IBA0～b′mg .L-1或 1/2改良MS +IBAb′mg .L-1+NAA0～b′mg .L-1,或R +IBAc2 mg .L-1+NAAc1 mg .L-1生根率达 98%以上。移栽土壤基质以黄心土加沙(10∶1)理想 ,移栽成活率可达 85 %以上     

杉木优良无性系组培繁育试验

通过对杉木优良无性系的组培繁育试验，筛选出一组广谱、高效的培养基和组培苗最佳移栽期及移栽后的培育管理方法；营造了组培苗与实生苗及各种源无性系不同繁殖方式的对比试验林，进一步研究其造林效果的差异。结果表明，组培苗造林在树高、地径等性状上均高于实生苗及常规无性繁殖苗。     

胡杨组织培养研究概述与问题讨论

胡杨插条繁殖生根极难，因而进行胡杨试管繁殖研究具有重要意义。对胡杨试管苗再生的途径及影响因子进行了综述，对胡杨组织培养中存在的黄萎、玻璃化等主要问题进行了讨论。试管苗黄萎问题没有解决，有必要进一步开展黄萎发生机理和高效再生体系的研究。     

观赏植物组培与辐射结合育种研究进展

综合分析了组培与辐射结合育种方式的优点,对这种复合育种方式在观赏植物上的应用进行了分析归纳。总结出组培与辐射结合育种体系的基本技术路线,指出不同植物、不同材料的辐射效果的差异,并列出一些供育种者参考辐射剂量。最后分析了以后的研究方向。