香菇原生质体工程菌株的构建研究——I.香菇原生质体的分离与再生行为

本研究通过电镜观察与酶解分析确定香菇(Lentinula edodes)菌丝细胞壁组成.并进行了双核菌丝原生质体的分离和培养.研究发现:酶解过程中,菌丝原生质体一般只从菌丝顶端等新生部位释放出来.合适的渗透剂、菌龄和酶解温度是原生质体得率的重要保证;原生质体再生菌丝过程是一个间歇性的依赖于原生质团涌动流向的过程,再生菌丝的形成经历了原生质体形成突起-哑铃状-分枝状等不同形态类型的再生阶段;再生菌丝体在形态上没有观察到锁状联合,栽培后未能结实,初步判断为单核菌株.     

Greenhouse gas emissions in response to straw incorporation,water management and their interaction in a paddy field in subtropical central China

A field experiment was conducted to study the effects of combination of straw incorporation and water management on fluxes of CH₄, N₂O and soil heterotrophic respiration (Rh) in a paddy field in subtropical central China by using a static opaque chamber/gas chromatography method. Four treatments were set up: two rice straw incorporation rates at 0 (S1) and 6 (S2) t ha^?1 combined with two water managements of intermittent irrigation (W1, with mid-season drainage) and continuous flooding (W2, without mid-season drainage). The cumulative seasonal CH₄ emissions for the treatments of S1W2, S2W1 and S2W2 increased significantly by 1.84, 5.47 and 6.63 times, respectively, while seasonal N₂O emissions decreased by 0.67, 0.29 and 1.21 times, respectively, as compared to S1W1 treatment. The significant increase in the cumulative Rh for the treatments S1W1, S2W1 and S2W2 were 0.54, 1.35 and 0.52 times, respectively, in comparison with S1W2. On a seasonal basis, both the CO₂-equivalents (CO₂e) and yield-scaled CO₂e (GHGI) of CH₄ and N₂O emissions increased with straw incorporation and continuous flooding, following the order: S2W2>S2W1>S1W2>S1W1. Thus, the practices of in season straw incorporation should be discouraged, while mid-season drainage is recommended in paddy rice production from a point view of reducing greenhouse gas emissions.     

Molecular cloning and functional analysis of duck Toll-like receptor 5

Toll-like receptor 5 (TLR5) is responsible for the recognition of bacterial flagellin in vertebrates. In this study, we cloned the single-exon TLR5 gene of the Maya breed of Common Shelduck (Tadorna tadorna). The TLR5 open reading frame is 2580 bp in length and encodes an 859-amino acid protein. The putative amino acid sequence of duck TLR5 consisted of a signal peptide sequence, 11 leucine-rich repeat domains, a leucine-rich repeat C-terminal domain, a transmembrane domain, and an intracellular Toll-interleukin-1 receptor domain. The duck TLR5 gene was highly expressed in the lung, bone marrow, spleen, and liver; moderately expressed in kidney, small intestine, large intestine, and brain. A plasmid expressing duck TLR5 was constructed and transfected into HEK293T cells, and expression was confirmed by indirect immunofluorescence assay. HEK293T cells transfected with duck TLR5- and NF-κB-luciferase-containing plasmids significantly responded to flagellin from Salmonella typhimurium, indicating that it is a functional TLR5 homolog.     

猪结合珠蛋白基因的克隆、原核表达及单克隆抗体制备

本研究的目的在于利用重组猪结合珠蛋白(Haptoglobin,Hp)制备特异性单克隆抗体.以梅山猪肝脏总RNA为模板,利用RT-PCR方法扩增猪hp基因,将其克隆到原核表达栽体pGEX-KG和pET-32a上,转化至E.coli BL21中,IPTG诱导下高效表达重组GST-Hp和HIS-Hp蛋白.用纯化的重组GST-Hp免疫BALB/c小鼠,用纯化的HIS-Hp作为包被抗原进行间接ELISA检测,通过淋巴细胞杂交瘤技术筛选阳性细胞株,制备单克隆抗体.结果表明筛选到2株高效分泌抗猪Hp单克隆抗体的杂交瘤细胞株,命名为3B4和3C8.Western-blot和间接ELISA检测均表明制备的单克隆抗体具有良好的特异性.     

北京市怀柔区设施种植业废弃物处理现状及发展

针对北京市怀柔区设施种植业废弃物处理的现状,分析废弃物处理过程中存在的问题,并提出相应的解决措施。     

宁南干旱山区的节水灌溉技术

宁夏南部干旱区农业产生最突出的矛盾是干旱缺水，近年，发展节水灌溉在宁南山区农业生产与经济建设中取得了一定成效，重点推广集雨节灌（滴灌，喷灌，膜上膜下点灌及窖，井下微灌），低压管灌等节水灌溉技术，对缓解干旱地区水资源供需矛盾发挥了重要作用。