Characterization of bovine ileal epithelial cell line for lectin binding,susceptibility to enteric pathogens,and TLR mediated immune responses

In this study, primary and immortalized bovine intestinal epithelial cells (BIECs) were characterized for the expression of surface carbohydrate moieties. Primary BIEC-c4 cells showed staining greater than 90 % for 16 lectins but less than 50 % staining for four lectins. Immortalized BIECs showed significantly different lectin binding profile for few lectins compared to BIEC-c4 cells. BIEC-c4 cells were studied for infectivity to E. coli, Salmonella enterica, bovine rotavirus, bovine coronavirus, and bovine viral diarrhea virus. Bovine strain E. coli B41 adhered to BIEC-c4 cells and Salmonella strains S. Dublin and S. Mbandaka showed strong cell invasion. BIEC-c4 cells were susceptible to bovine rotavirus. LPS stimulation upregulated IL-10, IL-8, and IL-6 expression and Poly I:C upregulated TLR 8 and TLR 9 expression. This study provides important knowledge on the glycoconjugate expression profile of primary and immortalized BIECs and infectivity and immune responses of primary BIECs to bacterial and viral pathogens or ligands.     

Replication of neurovirulent equine herpesvirus type 1 (EHV-1) in CD172a+ monocytic cells

Equine herpesvirus type 1 (EHV-1) is responsible for respiratory disorders, abortion and myeloencephalopathy (EHM) in horses. Two pathotypes of EHV-1 strains are circulating in the field: neurovirulent (N) and non-neurovirulent (NN). For both strains, CD172a⁺ monocytic cells are one of the main carrier cells of EHV-1 during primary infection, allowing the virus to invade the horse’s body. Recently, we showed that EHV-1 NN strains showed a restricted and delayed replication in CD172a⁺ cells. Here we characterize the in vitro replication kinetics of two EHV-1 N strains in CD172a⁺ cells and investigate if the replication of these strains is similarly silenced as shown for EHV-1 NN strains. We found that EHV-1 N replication was restricted to 7–8% in CD172a⁺ cells compared to 100% in control RK-13 cells. EHV-1 N replication was not delayed in CD172a⁺ cells but virus production was significant lower (10^3.0 TCID₅₀/10⁵ inoculated cells) than in RK-13 cells (10^8.5 TCID₅₀/10⁵ inoculated cells). Approximately 0.04% of CD172a⁺ cells produced and transmitted infectious EHV-1 to neighbour cells compared to 65% of RK-13 cells. Unlike what we observed for the NN strain, pretreatment of CD172a⁺ cells with histone deacetylases inhibitors (HDACi) did not influence the replication of EHV-1 N strains in these cells. Overall, these results show that the EHV-1 replication of N strains in CD172a⁺ cells differs from that observed for NN strains, which may contribute to their different pathogeneses in vivo.     

Compatibility of root growth and tuber production of potato cultivars with dynamic and static water‐saving irrigation managements

The important root characteristics of root length density (RLD) and root mass density (RMD) generally differ among irrigation managements and potato cultivars. The objective of this study was to investigate the RLD and RMD variations and their functional relationships with gross potato tuber yield for two commercial potato cultivars, Agria and Sante, under different irrigation strategies. Full irrigation and water‐saving irrigation strategies, deficit and partial root drying irrigations, were applied statically (S) and dynamically (D) based on daily crop evapotranspiration. Results showed that SPRD had significantly greater RLD (3.64 cm/cm³) and RMD (132.7 μg/cm³) than other irrigation treatments. Between the potato cultivars, Agria had significantly larger values of RLD (3.50 cm/cm³) and RMD (138.7 μg/cm³) than Sante. The functional relationship between the root growth characteristics and tuber yield showed that under water‐saving irrigations, Agria increased root mass at the expense of gross tuber yield but Sante increased root mass to maintain larger gross tuber yields. However, Agria produced more roots and gross tuber yield than Sante, and it is concluded that Agria is a more drought‐tolerant potato cultivar, which is recommended for tuber production in regions where water might be scarce. It was shown that larger root production in potatoes was associated with improved tolerance to water stress.     

Propagation techniques on root turion sprout browse ofPopulus davidiana Dode

The root segments selected from dominant trees ofPopulus davidiana Dode were taken as reproductive material and were buried in different depths to carry out the reproduction of root turion sprout. The affecting factors of gemination rate, survival rate, and height growth for cutting wood of root sprout were comparatively analyzed. The results showed that the best suitable substrate for burying root is pearlite, with a germination rate of 15.16%. 3–4-cm root segments has the highest rate of germination (12.4%). The mixture of sand and soil (2∶1) is the best cutting substrate, with a survival rate of 81.3%, while as to height growth of cutting wood, the mixture of turfy and soil (1∶1) is the best. The cutting woods selected from different positions of stem show difference in height growth. The cutting wood from top stem is higher obviously than those from middle or low stem. Biography: XING Ya-juan (1969-), female, assistant researcher in Forest Research Institute of Heilongjiang Province, Harbin 150040, P. R. China. Responsible editor: Song Funan     

水分胁迫对荔枝幼树根系与梢生长的影响

以盆栽沙培1年生荔枝空中压条苗为试材,研究了水分胁迫对荔枝幼树根系和梢生长的影响以及二者的相互关系。3个处理为:沙体积含水量为7.5%(严重缺水),15.0%(一般缺水),22.5%(充分供水)。结果表明:水分胁迫严重抑制荔枝幼树地上部分和各级侧根的生长,使地上部枝梢生长严重受阻,叶片数量、平均叶面积、叶片厚度均显著变小;根总干重、侧根长度、根表面积显著减少。水分胁迫下促进须根的生长,须根的长度、根表面积比水分充足时显著增加。水分胁迫时叶片淀粉含量下降,而根系积累更多的淀粉。比较不同处理的根梢比发现,干旱对根系生长的抑制作用比枝梢更显著。     

紫花苜蓿受冻害后促进根颈枝条再生方法的研究

对蔚县苜蓿根颈冻害后切除冻伤部分处理的比较研究结果表明,处理区的根颈苗数、地上高度、地上生物量、分蘖丛数是对照区的1.6～2倍;处理区苜蓿的各个数量指标的b值大于对照处理苜蓿b值,说明除冻害根颈后苜蓿的生长比不除冻害根颈部分的苜蓿生长更为合理。通过除去苜蓿冻害部位,防止根系继续腐烂,保护了未冻害部分,增加了苜蓿发芽的有效部位。同时,使根系与地面的距离变短或露出地面,增加光照强度,提高地温,促使未冻害苜蓿根系发芽,返青提前增加产量。     

牛皮肤成纤维细胞的体外培养与冻存

利用牛皮肤组织块直接培养法，得到牛皮肤细胞的原代培养物，再用酶消化法和反复贴壁法处理，能够纯化成纤维细胞。成纤维细胞的冻存是通过选用6种分别含有二甲基亚砜（DMSO）、甘油（GL）及乙二醇（EG）的保护液，以相同的冻前处理方法，对牛皮肤成纤维细胞进行缓慢冷冻，冰箱预冷平衡1－2h，逐步投入液氮（－196℃）中保存，再经37℃水浴解冻，Hanks液脱保护剂，以贴壁率评价冻存效果。结果表明，20％DMSO保护液对牛皮肤成纤维细胞表现出较好且稳定的冷冻保护效果，其平均贴壁率达87．9％。     

Feline Ubiquitin Fusion Protein Genes

Using cDNA from a CRFK cell line as a template, PCR amplification was performed with the Ub1S and poly(dT) primers to isolate feline ubiquitin genes. Sequencing of the 495 bp PCR fragment revealed that the putative amino acids induced by this fragment gave a fusion protein consisting of a ubiquitin polypeptide (76 amino acids) and an extension protein of ribosomal proteins L40 (52 amino acids). The putative amino acid sequence of ubiquitin was identical to those of humans, rats and pigs.The recombinant glutathione S-transferase (GST)–feline ubiquitin fusion proteins were produced in Escherichia coli and purified. The fusion proteins had a molecular weight of about 42 kDa and were detected by immunoblot assay with rabbit anti-ubiquitin antiserum.The mRNAs from heat-shocked and non-heat-shocked cells were subjected to RT-PCR (Ub1S and poly(dT) primers) analysis. The molecular weights of the ubiquitinated proteins in heat-shocked CFRK cells were between 18 kDa and 24 kDa by immunoblot assay.These results suggested that there were more ubiquinated proteins in the heat-shocked CRFK cells than in the pre-heat-shocked cells.     

桑悬浮细胞原生质体培养的研究

采用继代培养三个月后的桑子叶悬浮细胞为材料,进行了原生质体的分离和培养。桑悬浮细胞在纤维素酶、果胶酶、半纤维素酶的混合溶液中,酶解获得产量高、活力强的原生质体。原生质体在K8p(附加6—BA、NAA、2,4—D、LH)液体培养基中,再生细胞经多次分裂,得到肉眼可见的小愈伤组织。再通过增殖继代培养,获得浅黄色、具有明显颗粒结构的愈伤组织,转至各种激素含量的MSB固体培养基中,尚未获得绿苗分化。