聚多曲霉菌原生质体和液泡的制备及优化

原生质体的大量制备是研究原生质体转化、诱变、融合等技术的关键,液泡的制备对研究液泡中分解、转运有机物的特性具有重要意义,但目前分离技术还不够成熟.本研究从聚多曲霉菌菌丝中分离原生质体和液泡并对分离条件进行优化,以聚多曲霉菌DJ515-2菌丝为材料,探索不同因素对原生质体和液泡制备分离效果的影响.结果表明,聚多曲霉菌DJ515-2在菌龄42 h,以3％纤维素酶、1％蜗牛酶和3％的溶壁酶组成复合酶液,25℃下酶解4 h,原生质体达到最大产量,为5.167×105个/mL.同时,在此基础上裂解液泡的最优条件为pH7.5、1.0mol/L KCl、0.020％Triton X-10,其产量达2.63×105个/mL,产率为原生质体的50.8％,相比采用多元碱化合物诱导真菌原生质体裂解释放液泡的产率增加了 40％～45％.本研究为真菌和植物的各种原生质体技术及基于亚细胞层面的研究提供了材料基础.     

中国濒危鱼类、两栖爬行类和哺乳类的地理分布格局与优先保护区域——基于《中国濒危动物红皮书》的分析

《中国濒危动物红皮书》依据我国动物所面临的濒危,对我国濒危动物的濒危等级划分,种群现状,致危因素和保护措施等进行了描述说明,首批收录了535种濒危动物。本文以《中国濒危动物红皮书》中所收录的352种濒危脊椎动物（不含鸟类（以下简记为“濒危植物”）为研究对象,整理统计了现有濒危物种的分布资料,在GIS支持下,对中国濒危动物的地理分布进行了研究。结果表明,中国濒危动物物种呈明显的集聚分布,最密集的地区是横断山区,海南岛,西双版纳和云贵高原;而在华北平原,内蒙古东部,黄土高原和东北平原等地区出现大片空白区。影响濒危动物分布的主要因素有热量和水分条件,地形条件等自然条件以及历史开发,人为破坏等人文条件。山地因地形屏障作用而保留了较多的古老物种,其较为复杂的环境因子也有益物种的生存,因而物种丰富度较高。大多数动物对于水分和热量的依赖性较强,因此水热条件对其分布的限制作用十分明显。人为破坏较为严重的地区,濒危物种稀少;污染,开荒等引起的环境问题对于现存动物的威胁很。运用Dobson排除算法得到云南勐腊县等9个县分布有168种濒危动物,占全国总数（海生种类及仅分布于台湾和香港的特有种除外）的51．5％,而其土地面积之和仅为全国陆地总面积的0．9％。而云南勐腊县等94个县市就分布有中国所有的濒危动物。这些地区是我国生物多样性保护应该优先考虑的地方。     

亚硝基胍诱变选育林肯霉素高产菌株

以林肯链霉菌９４７－８（Ｓｔｒｅｐｔｏｍｙｃｅｓｌｉｎｃｏｌｎｅｎｓｉｓ９４７－８）为出发菌株（产林肯霉素９４０γ／ｍｌ）。采用孢子热处理方法处理出发菌株孢子,得到变异株９４７－８ｓ,产林肯霉素１０８０γ／ｍｌ。对９４７－８ｓ菌株进行ＮＴＧ诱变处理,得变异株９４７－８ｘ,产林肯霉素为１２１８γ／ｍｌ,且生产能力稳定。     

1.42A crystal structure of mini-IGF-1(2): an analysis of the disulfide isomerization property and receptor binding property of IGF-1 based on the three-dimensional structure

Insulin and insulin-like growth factor 1 (IGF-1) share a homologous sequence, a similar three-dimensional structure and weakly overlapping biological activity, but IGF-1 folds into two thermodynamically stable disulfide isomers, while insulin folds into one unique stable tertiary structure. This is a very interesting phenomenon in which one amino acid sequence encodes two three-dimensional structures, and its molecular mechanism has remained unclear for a long time. In this study, the crystal structure of mini-IGF-1(2), a disulfide isomer of an artificial analog of IGF-1, was solved by the SAD/SIRAS method using our in-house X-ray source. Evidence was found in the structure showing that the intra-A-chain/domain disulfide bond of some molecules was broken; thus, it was proposed that disulfide isomerization begins with the breakdown of this disulfide bond. Furthermore, based on the structural comparison of IGF-1 and insulin, a new assumption was made that in insulin the several hydrogen bonds formed between the N-terminal region of the B-chain and the intra-A-chain disulfide region of the A-chain are the main reason for the stability of the intra-A-chain disulfide bond and for the prevention of disulfide isomerization, while Phe B1 and His B5 are very important for the formation of these hydrogen bonds. Moreover, the receptor binding property of IGF-1 was analyzed in detail based on the structural comparison of mini-IGF-1(2), native IGF-1, and small mini-IGF-1.     

毕赤酵母表达体系中重组蛋白的分离纯化

随着基因重组技术的快速发展,基因工程产品的利用越来越广泛,但其分离纯化的成本约占总成本的60%～70%.因此,探索一些简单有效的分离纯化方法尤为必要.简单介绍了目前较为流行的毕赤酵母表达体系,着重概述了重组蛋白分离纯化技术方法的应用情况.     

菜用和观赏甘薯种质资源遗传多样性分析

为了发掘菜用和观赏甘薯优异种质资源,通过对国家种质徐州甘薯试管苗库中1000余份资源材料进行鉴定,筛选了96份优异种质。利用30对SSR引物对入选材料进行了遗传多样性和群体结构分析,明确了这些材料遗传差异;并对入选材料12个表型质量性状进行主成分和聚类分析。结果表明:扩增的总条带数为275条,其中多态性条带为269条,多态率97.8%;利用DPS软件计算入选材料间的Nei72遗传距离为0.15~0.76,平均遗传距离0.66;群体结构分为3个组群,与分子标记聚类结果相似,表明入选材料有较大的遗传差异性;表型质量性状主成分分析得到5个主要成分,其累计贡献率达到80.50%;利用表型质量性状,可聚为8个组群。本研究通过分子标记与表型质量性状分析为下一步杂交选育菜用和观赏甘薯新品种提供了亲本选择信息。     

Primulina maciejewskii sp. nov. (Gesneriaceae) from Guangdong,China

Primulina maciejewskii F. Wen, R.L. Zhang & A.Q. Dong, a new species of Primulina (Gesneriaceae) from Guangdong, China, is described and illustrated. It is similar to P. lobulata and P. danxiaensis in leaf and flower shape, but can be distinguished by bract shape, peduncle, pedicel, and bract size, filament and anther indumentum, and number of bracts, staminodes and stigma.     

培养基质、储藏方式和盐度对三种海滨植物种子萌发的影响

研究了培养基质、储藏方式和盐度对3种海滨植物互花米草(Spartina alterniflora)、盐地碱蓬(Suaeda salsa)和芦苇(Phragmites australis)种子萌发的影响,探索在潮间带环境下海滨植物种子萌发适应策略。结果表明:3种植物的干藏种子和湿藏芦苇种子随着盐度的升高,萌发率和萌发速率均显著下降,湿藏互花米草和盐地碱蓬种子在各盐度下萌发率和萌发速率差异不显著。各盐度-土培-干藏互花米草,中、高盐度-土培-干藏盐地碱蓬,土培各处理,中、高盐度-水培-干藏,高盐度-水培-湿藏芦苇种子萌发失败。湿藏提高了各盐度处理下土培互花米草,中、高盐度-水培和土培盐地碱蓬,淡水、中盐度-水培芦苇种子的萌发率和萌发速率。干藏互花米草种子在中、高盐度和土埋条件下种子的萌发受到抑制,限制了互花米草向高潮带与潮上带的扩展;而经常受潮水浸淹保持湿润的种子能抵抗高盐和泥沙沉积,导致互花米草种群逐步向低潮带方向发展;湿藏芦苇种子在淡水中萌发率和萌发速率最高,当潮上带盐度降低时,芦苇具有很强的竞争优势,但是对盐度和土埋敏感,限制了其向海的拓展;盐地碱蓬在中、高盐度和土培条件下萌发速率最高,快速萌发的适应策略和广适应性在盐地碱蓬占据高潮带和中潮带广大区域的过程中起到了重要的作用。     

Bioassays on selection and feeding responses of Holotrichia oblita adults to different plant species

Bioassays on selection and feeding responses of Holotrichia oblita adults to different plant species were conducted both in laboratory and under field conditions. The selection response bioassay showed that the adults were significantly attracted to Ricinus communis. In paired tests of the field cages, more than 60% adults were attracted to R. communis plants, while only 35.55%, 34.45%, 33.35%, and 26.65% were attracted to plants of Glycine max, Arachis hypogaea, Ipomoea batatas, and Gossypium hirsutum, respectively. In the laboratory, the selection response percentage of the adults to R. communis leaves reached up to 93% in the presence of other host plant leaves and still arrived at 70% in the absence of other host plant leaves. Feeding deterrence bioassay showed that the adults preferred to I. batatas leaves with 229.90 ± 4.25 mg leaf consumption and 41.30 ± 5.19 mg faeces excretion after 12 h, followed by the leaves of G. hirsutum and A. hypogaea. The adult did not prefer leaves of R. communi with only 7.10 ± 0.23 mg leaf consumption and corresponding 4.00 ± 0.84 mg fecal excretion. In summary, all experimental results suggested that the attraction of H. oblita adults to R. communi is not triggered for feeding.