白色金针菇工厂化生产技术与装备体系研究

白色金针菇工厂化生产使用规范化的硬件设施,采用标准化的生产流程和层层把关的有效管理,生产受农业环境影响小,确保了产品的产量和品质。白色金针菇工厂化生产流程为:菌种选育与培养基(料)优化(包括基料选配、搅拌、装袋合灭菌等)→规范化生产(包括接种、培养、搔菌、出菇蕾、驯养、生长及采收等环节)与病虫害综合防治技术→商品菇分级包装。     

Anatid herpesvirus 1 CH virulent strain induces syncytium and apoptosis in duck embryo fibroblast cultures

Anatid herpesvirus 1 (AHV-1) CH virulent strain was first isolated from an infected duck and it was found that this virus strain could induce cytopathic effect (CPE) in duck embryo fibroblast (DEF). Following AHV-1 infection, DEF showed morphological changes such as cell rounding, improved refractivity and detachment from the culture surface. However, its pathological characteristics were not adequately known. Related studies were performed and the results showed that syncytium formation could be observed as the other type of CPE in AHV-1 infection. Hematoxylin-eosin staining and 4’, 6-diamidino-2-phenylindole (DAPI) staining of infected DEF were each used to visualize the shape and distribution of chromatin within nuclei and nuclear fragmentation was observed. Chromatin condensation and margination, as well as formation of apoptotic bodies were observed by transmission electron microscopy (TEM). DNA ladder formation was detected in AHV-1 infected cells and apoptosis of the infected DEF was also detected by flow cytometry analysis of Annexin V-FITC/PI staining method. Therefore, it was suggested that AHV-1 virulent strain can induce syncytium and apoptosis in DEF. Syncytium formation and apoptosis observed in this study may contribute to the elucidation of AHV-1 pathogenesis.     

N2O production in the organic and mineral horizons of soil had different responses to increasing temperature

Journal of Soils and Sediments - The responses of N2O emission to increasing temperature in different soil horizons are not clearly understood yet. Here, we investigated the effects of increasing...     

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.     

水稻雄性不育突变体012S-3的遗传分析和基因定位

雄性不育是水稻杂种优势利用的重要资源,对雄性不育现象的研究具有重要理论意义和实践价值。本研究以自然突变的水稻雄性不育突变体012S-3为试验材料,对其表型特征和花粉育性等进行调查,并构建遗传群体,利用分子标记对目的基因进行初步定位,然后应用基因组重测序技术对其进行精细定位。结果表明,012S-3是一个典型的无花粉普通型雄性不育材料,其不育性状受1对隐性核基因控制。初步定位分析目的基因与SSR标记RM6081存在连锁关系,其遗传距离约为34.4 c M;进一步的精细定位分析,找到3个候选基因:LOC_Os07g35880、LOC_Os07g35920和LOC_Os07g35940,其中LOC_Os07g35880和LOC_Os07g35940编码β-淀粉酶,属于水稻中新发现的花粉致死基因。该不育基因的成功定位为其进一步的分离克隆及其在水稻分子设计育种中的应用奠定了基础。     

水稻恶苗病病原菌的研究

 1977~1987年采集103个稻恶苗病标本,分离后获8种镰刀菌。其中串珠镰孢(Fusarium moniliforme)的一个变种占总标本数的60.2%,其特征为:菌落生长直径2-4cm;大型分生孢子镰刀形,3-5隔;小型分生孢子卵形,串生和假头生;产孢细胞单、复粳并存;具厚垣孢子,球至亚球形。此变种定名为(F. moniliforme var. Zhejiangensis Wang et Chen var. nov.)。其他7种镰刀菌为木贼镰孢(F. equiseti),禾谷镰孢(F. graminearum),砖红镰孢(F. lateritium),雪腐镰孢(F. nivale),尖镰孢(F. oxysporum),半裸镰孢(F. semitectum),腐皮镰孢(F. solani)。试验证明,串珠镰孢浙江变种是水稻恶苗病的主要致病菌;其他6种(除腐皮镰孢未接种外)均有不同程度的弱致病性。串珠镰孢浙江变种也能侵染小小麦、大麦、玉米、西瓜、豌豆等作物的幼苗,并有徒长症状。     

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.