水稻不育突变株沈农05-9的形态和细胞学特征鉴定

以辽粳5号与丰锦杂交的高代系的突变株沈农05-9和其原始株系为材料,采用醋酸洋红染色压片技术及石蜡切片技术.进行了形态学特征、小孢子发生的细胞学特征观察.结果表明:该不育突变株花粉表现典败和染败,多数花粉母细胞减数分裂不形成二分体,而形成多核的花粉母细胞;花药表现干瘪且纤维化严重,绒毡层细胞在小孢子发育过程中提前解体.该不育突变株的减数分裂方式类似于萍乡型显性不育系及早稻昆植不育系.导致败育的直接原因为绒毡层细胞提前解体.     

基体改进剂对重楼中砷·汞含量测定的影响

[目的]探讨重楼中微量元素砷、汞含量的测定方法,为重楼药材毒副作用评价体系提供参考。[方法]采用石墨炉原子吸收光谱法（GFAAS）,通过对基体改进剂硝酸镍、氯化钯的用量,酸性介质,灰化温度,原子化温度等试验参数进行优化,探索了基体改进剂对重楼中砷、汞含量测定的影响。[结果]加入硝酸镍、氯化钯不仅能提高砷、汞的灰化温度和原子化温度,还能提高测定的灵敏度和精密度,且重现性好。[结论]加入基体改进剂的方法快速、可靠,为重楼中砷、汞含量测定提供了一种简便有效的方法。     

肉鸽H9亚型禽流感的诊断与病毒分离及鉴定

通过分子PCR检测,初步证明在湖北一发病肉鸽群感染H9亚型禽流感病毒。病原分离后,盲传3代,其血凝效价达到109~1011,经生物学特性进行测定,其鸡胚半数致死量为107.5,9日龄鸡胚平均致死时间为54.3 h,1日龄雏鸡脑内接种致病指数为1.8,6周龄小鸡静脉接种致病指数为2.0,证明该H9亚型禽流感为毒力较强。将该病毒回归非免疫肉鸽,能产生典型的H9亚型禽流感症状,采集其肺、喉头、泄殖腔等病变组织进行病毒分离,均可分离到H9亚型AIV。     

H9亚型禽流感病毒血凝素基因重组禽痘病毒的构建

【目的】获得表达H9亚型禽流感病毒血凝素(HA)基因的重组禽痘病毒。【方法】将含禽痘病毒启动子LP2EP2驱动的HA基因,插入到禽痘病毒转移载体pSY681中,获得重组转移载体pSY681/HA。用脂质体将其转染已感染亲本禽痘病毒S-FPV-017株的鸡胚成纤维细胞,使其在鸡胚成纤维细胞内与禽痘病毒基因组发生同源重组,产生表达HA的重组禽痘病毒rFPV-HA。在含有X-gal的营养琼脂培养基上进行蓝斑筛选后,对重组病毒进行多次蚀斑克隆,并用间接免疫荧光法对感染重组病毒鸡胚成纤维细胞中HA的表达产物进行鉴定。【结果】以重组禽痘病毒DNA为模板,利用HA基因特异引物进行PCR,扩增出1条约1.7 kb左右的带。以间接免疫荧光法证实重组禽痘病毒能表达HA。【结论】成功构建了表达H9亚型禽流感病毒HA基因的重组禽痘病毒,且构建的重组禽痘病毒能表达具有生物学活性的HA。     

Susceptibility of various ceil culture systems to pseudorabies virus

A comparative study was carried out to determine the susceptibility of five different cell lines to pseudorabies virus (PRV), a herpes virus of pigs. The cell systems tested were swine testicle (ST), mink lung (ML), equine dermal (ED), porcine kidney (PK₁₅), and bovine turbinate (BT) cells. Virus titers obtained were 10^4.88, 10^4.38, 10^3.75, 10^2.63, and 10^0.25 for ML, ST, PK₁₅, BT and ED cells, respectively indicating that ML, ST, and PK₁₅ are optimal cell lines for the growth of PRV whereas BT and ED are not very sensitive.     

利用CRISPR/Cas9系统构建可诱导型猪Sohlh1基因敲除细胞系

目的 将Tet-on系统与CRISPR/Cas9系统相结合建立诱导敲除猪Sohlh1基因的细胞系。方法 分离培养猪胎儿成纤维细胞(PFF),进行慢病毒载体PCW-eSpCas9(1.1)侵染并通过1 μg/mL嘌呤霉素筛选及强力霉素(Dox)诱导;改造pLV-sgRNA载体,进行293FT细胞转染验证;构建含Sohlh1基因目标靶点的pLV-sgRNA-2A-GFP特异性载体,进行慢病毒包装检测;利用包装病毒侵染稳转PCW-eSpCas9(1.1)的PFF细胞,3 μg/mL灭稻瘟菌素进行筛选,随后添加Dox诱导进行表达分析和敲除效率检测。结果 建立了受Dox调控可诱导eSpCas9(1.1)蛋白表达的PFF细胞系,不添加Dox,eSpCas9(1.1)蛋白不表达;293FT细胞表达绿色荧光,表明pLV-sgRNA-2A-GFP载体改造成功。3、4、6号Sohlh1基因靶点具有敲除活性,利用最优的2个靶点构建了pLV-sgRNA-2A-GFP特异性载体,荧光表达显示载体慢病毒包装成功。构建稳转PCW-eSpCas9(1.1)和pLV-sgRNA-2A-GFP的PFF,经Dox诱导72 h后,Sohlh1基因的敲除效率为85%。结论 利用Tet-on系统和CRISPR/Cas9系统成功构建了可诱导型猪Sohlh1基因敲除细胞系,为研究Sohlh1基因的功能以及制备条件性敲除Sohlh1基因去除生殖细胞的模型猪奠定了基础。     

禽流感病毒H9亚型血凝素特异性单克隆抗体的制备和鉴定

以常规方法纯化H9亚型禽流感病毒，制备了两株分泌抗禽流感病毒H9亚型血凝素特异性单克隆抗体的杂交瘤细胞11A5和1182，其上清ELISA效价均为1：10^4，腹水效价为1：10^7和1：10^6，HI效价上清分别为8、7log2，腹水效价均为15log2。采用HI方法证明其对禽流感病毒H9亚型的特异性，通过Western blot方法证明两株单抗都针对H9亚型禽流感病毒HA1蛋白。     

Negligible contribution from roots to soil-borne phospholipid fatty acid fungal biomarkers 18:2ω6,9 and 18:1ω9

The phospholipid fatty acid biomarkers 18:1ω9, 18:2ω6,9 and 18:3ω3,6,9 are commonly used as fungal biomarkers in soils. They have, however, also been found to occur in plant tissues, such as roots. Thus, the use of these PLFAs as fungal biomarkers in sieved soil, which may still contain small remains of roots, has been questioned. We used data from a recent beech tree girdling experiment to calculate the contribution of roots to these biomarkers and were able to demonstrate that not more than 0.61% of 18:1ω9 and 18:2ω6,9 in sieved soil samples originated from roots (but 4% of 18:3ω3,6,9). Additionally, the abundance of the biomarker 18:2ω6,9 in the soil was found to be highly correlated to ectomycorrhizal root colonization, which further corroborates its fungal origin. PLFA biomarkers were substantially reduced in vital roots from girdled trees compared to roots of control trees (by up to 76%), indicating that the major part of PLFAs measured in roots may actually originate from ectomycorrhizal fungi growing inside the roots. We calculated, that even a near to 50% reduction in fine root biomass - as observed in the girdling treatment - accounted for only 0.8% of the measured decrease of 18:2ω6,9. Our results demonstrate that both 18:1ω9 and 18:2ω6,9 are suitable biomarkers for detecting fungal dynamics in soils and that especially 18:2ω6,9 is a reliable biomarker to study mycorrhizal dynamics in beech forests.     

不锈钢管的焊接技术

汽轮机润滑油系统油管道焊接是汽轮机专业焊接的一个重点.为了保证汽轮杌润滑油管道系统的清洁度及汽轮机主机润滑系统、调节保安系统安全可靠地工作,其材质全部由不锈钢组成,所以油管道安装的内在质量格外重要.总结出一套焊接组装工艺措施,提高了生产效能,满足了汽轮机油管的质量要求.