鸡Ii结合Rab5a和Rab7b分子的分析

本研究旨在探明鸡恒定链（invariant chain，Ii）与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先，用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii_{（Cyt-Tra）}]、Ii CLIP （class Ⅱ-associated invariant chain peptide）-三聚体区[Ii_{（CLIP-TRIM）}]和Ii突变体[Ii_{（M81-87aa）}、Ii_{（M91-99aa）}和Ii_{（M81-99aa）}]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次，将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T，观察它们的共定位。将构建的原核重组质粒进行表达和纯化，最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明，成功构建Ii结构域及Ii突变体的重组质粒。Ii_{（Cyt-Tra）}及Ii突变体均能与Rab5a和Rab7b在细胞内共定位，而Ii_{（CLIP-TRIM）}与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域，而不是CLIP与三聚体区。综上所述，鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区，而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制，为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。     

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.     

Comparison of the microbial communities of alpacas and sheep fed diets with three different ratios of corn stalk to concentrate

The objective of this study was to investigate the characteristics of ruminal microbial communities of alpacas (Lama pacos) and sheep (Ovis aries) fed three diets with varying ratios of roughage (corn stalk) to concentrate, 3:7 (LS), 5:5 (MS) and 7:3 (HS). Six alpacas (one-year-old and weighing 29.5 ± 7.1 kg) and six sheep (one-year-old and weighing 27.9 ± 2.7 kg) were used in this study, in a replicated 3 × 3 Latin square experiment. Total protozoa concentration was determined under the microscope; total fungi and methanogens were assessed using quantitative polymerase chain reaction and expressed as a percentage of total bacterial 16S rRNA gene copies; bacterial communities were investigated by targeted 16S rRNA gene (V3–V4 region) sequencing. The percentage of fungi was significantly higher in alpacas than in sheep under the LS diet, while the concentration of protozoa was significantly lower in alpacas under HS, MS and LS diets. The alpha diversity including Shannon, Chao l and ACE indices of bacterial communities was higher in alpacas than in sheep, under the LS diet. A total of 299 genera belonging to 22 phyla were observed in the forestomach of alpaca and sheep, with Bacteroidetes and Firmicutes dominating both animal species. Phyla Armatimonadetes and Fusobacteria, as well as 64 genera, were detected only in alpacas, whereas phyla Acidobacteria and Nitrospira, as well as 44 genera, were found only in sheep. The abundance of cellulolytic bacteria, including Butyrivibrio and Pseudobutyrivibrio, was higher in alpacas than in sheep under all three diets. These differences in the forestomach microbial communities partly explained why alpacas displayed a higher poor-quality roughage digestibility, and a lower methane production. Results also revealed that the adverse effects of high-concentrate diets (70%) were lesser in alpacas than in sheep.     

同源四倍体及其原二倍体黑皮冬瓜光合光响应模型筛选

为了筛选出最适合黑皮冬瓜Benincasa hispida (Thunb.) Cogn.的光合光响应模型,为其育种提供理论依据,以同源四倍体及其原二倍体黑皮冬瓜为试验材料,对8种经典的光合光响应模型适用性进行了比较分析。结果表明：二次多项式能够表现出光抑制情况,但在拟合过程中出现暗呼吸速率为正值、光补偿点为负值及无法解释当光强达到饱和后光合速率快速下降的问题;直角双曲线、非直角双曲线及指数函数Ⅰ、指数函数Ⅱ无法直接求取光饱和点、光补偿点,结合常用的光饱和点的计算方法得到的光饱和点与实测值均存在较大的偏差,且指数函数Ⅱ在计算光饱和点时表现出明显的人为性,也无法拟合光抑制情况,但4种模型拟合得到的光补偿点均与实测值相差不大;指数修正模型因系数β为负值,无法求取四倍体黑皮冬瓜材料的光饱和点和最大净光合速率,且拟合得到的四倍体黑皮冬瓜的光补偿点明显低于实测值;直角双曲线修正模型计算得到的暗呼吸速率及二倍体黑皮冬瓜的光饱和点明显低于实测值,但获得的四倍体及其二倍体的最大净光合速率与实测值最接近,说明其在拟合最大净光合速率上有优势;整体上分段函数计算得到的黑皮冬瓜的各光合参数与实测值最为接近,与实测值的平均相对误差最小,也能很好的拟合发生光抑制部分的光响应曲线。分段函数拟合同源四倍体及其原二倍体黑皮冬瓜光合光响应曲线效果较其他模型效果好,分段函数模型为黑皮冬瓜最适合的光合光响应模型。     

中国高粱光温类型的初步研究

1、在短光照或温室条件件下高粱品种出苗～抽穗期日数显著缩短,供试品种平均比对照分别缩短12.5天和13.635天,缩短百分率分别为18.8和20.5;伴随苗穗期日数的缩短,叶片数在短光或温室条件下分别减少3.9和4.275片,减少百分数分别为22.24和24.22;株高分别降低60.15和47.65厘米,降低百分数为25.88和20.5,表明光、温直接影响高粱的生育进程和器官建成的数量与结构。 2、高粱品种生育日数变化主要反映在出苗～抽穗期。出苗～抽穗日数在短光或温室条件下分别减少18.8％和20.5％.而抽穗～成熟日数仅减少5.1％和8.3％,表明高粱出苗～抽穗期是光温反应的最大效应期。 3、不同品种在短光或温室条件下性状变化的趋势一致,但程度明显不同;同一品种同一性状在上述条件下变化程度也不同。根据供试品种对光温敏感的程度划为四大类群,其数量与地理分布如下: Ⅰ、光温敏感型,占试验品种总数的20％.分布在长江以北,辽宁以南地区。Ⅱ、温敏感型,包括温敏感光中度敏感型和温敏感光迟钝型,占25%,分布在黄河以北,北纬45℃以南的山西、内蒙、辽宁、河北北部和吉林南部。Ⅲ、光敏感型、包括光敏感沮中度敏感型和光敏感温迟钝型,占12.5％,分布在黄河(北纬35°)以南,南岭(北纬25°)以北地区。Ⅳ、中度敏感型占42.5％,分布广且没有明显的地带性。试验未发现对光、温都迟钝的类型。北方温敏感型品种较多,短日性较弱,南方光敏感型品种较多,短日性强、感温性中等或较弱。不同光温类型的形成、分布与其所处的生态环境即光温条件和栽培制度密切相关。 4、温敏感型和光敏感型中早熟品种较多,占86％,光温敏感型品种既有早熟的,中熟的、也有晚熟的,所以光温敏感的品种不一定全是早熟品种,而晚熟品种也并非对光温反应不敏感。中度敏感型品种也有早熟、中熟和晚熟之分。在不同光温条件和栽培制度下,不同光温类型熟性变化的主导因素不同。 5、光、温在形成高粱不同类型过程中的效应是相互联系而又有区别的,光、温效应对不同环境下的不同类型有主次之分,大小之别;光、温效应在高粱不同发育阶段既是同步性的,又是顺序性的。     

湟源县草地资源基本特点及开发利用

湟源县草地面积有 85 1 85 .1 3hm2 ,占全县土地总面积的 5 6.45 % ,草地资源丰富 ,牧草种类较多 ,农业生产秸秆来源广泛 ,但由于利用不当 ,造成了草地严重破坏和秸秆的浪费 ,如何合理利用草地和秸秆资源 ,扼制草地退化 ,是促进该县畜牧业发展的重要因素     

中西药结合预防肉鸡腹水综合征的临床试及防治机理探讨

将一日龄艾维因肉仔鸡(雌雄各半)180只在同一舍内饲养,14日龄时将其随机分成4组,其中Ⅰ、Ⅱ组饲料按1％分别添加我们研制的方剂一和方剂二,连续用药15d,为药物预防组;Ⅲ组在饲料中按0.05％添加肾肿腹水消,连用7d(药品要求的预防剂量),为药物对照组;Ⅳ组饲料中不添加任何药物,为空白对照组。Ⅰ-Ⅳ组发病率分别为0、10％、3％和20％,42d时Ⅰ、Ⅳ组体重高于其它组,Ⅰ平均体重为1972.92g,Ⅳ组平均体重为1989.91g。试验表明方剂一为该试验条件下预防肉鸡腹水综合征最有效的方剂,其保护率可达100％,且不影响出栏重。对方剂一预防肉鸡腹水综合征的机理进行探讨。     

分泌抗鸡Ig单抗杂交瘤细胞系的建立与单抗生物学特性的鉴定

从传染性法氏囊病病毒（ＩＢＤＶ）单克隆抗体（ＭｃＡｂ）杂交瘤细胞系１Ｄ１０、１Ｇ１中得到了３株能稳定分泌抗鸡免疫球蛋白（Ｉｇ）ＭｃＡｂ的杂交瘤细胞系１Ｂ７、１Ｄ７、３Ｇ６。其抗体类和亚类均属ＩｇＧ２ｂ，腹水的ＥＬＩＳＡ效价≥１∶２５６００，琼扩效价为１∶８～１∶１６。３株ＭｃＡｂ能与鸡血清中的Ｉｇ出现沉淀反应，琼扩在２４ｈ以内出现清晰的沉淀线，而不能和鸭、鸽、鹌鹑等禽类及异种动物血清出现沉淀线。纯化的鸡ＩｇＧ、ＩｇＭ经ＳＤＳ－ＰＡＧＥ后，分别用纯化并经辣根过氧化物酶（ＨＰＲ）标记的１Ｂ７、１Ｄ７、３Ｇ６单抗酶结合物进行免疫印迹试验证明，３株单抗识别的均为ＩｇＧ、ＩｇＭ分子的轻链     

二甲硝咪唑盐的合成研究

本文报道以简便易行的方法合成水溶性二甲硝咪唑盐，收率高达95％以上，具工业化应用价值。     

大豆室内耐冷筛选及其在田间的应用研究

本试验把生长箱大豆耐低温萌发筛选与田间早春播研究相结合,验证耐低温发芽的可靠性。在实验室,萌发期表现抗冷的大豆,田间出昔期仍表现抗冷或较抗冷。实验表明,大豆出苗的生物学最低温度平均为7.1℃,所需有效积温136℃。地温的高低直接影响大豆早春出苗的多寡,两者呈密切的正相关。黄、青、黑豆中,以黑豆抵抗早春不良气候条件的能力最强。