ZINC ADSORPTION AND TRANSPORT BY CHLAMYDOMONAS VARUIABILIS AND SCENEDESMUS SUBSPICATUS (CHLOROPHYCEAE) GROWN IN SEMICONTINUOUS CULTURE1

The amount of zinc adsorbed onto the cell surface of the unicellular green algae Scenedesmus subspicatus Hodat and Chlamydomonas variabilis Dangeard was operationally defined by extraction with EDTA; it was a function of the concentration of free ionic zinc remaining in the growth medium, rather than that of the total (free plus complexed) zinc concentration, and could be described by Langmuir isotherms. Conditional adsorption equilibrium constants for zinc were 0.123 and 0.039 L ·μmol^?1 for S. subspicatus and C. variabilis, respectively. A portion of the zinc adsorbed onto C. variabilis was released into solution after 1 h of contact with the metal, providing a possible tolerance mechanism for this alga; the division rate of C. variabilis was not altered by up to 12 μmol Zn²⁺· L^?1, although the cell yield obtained during the stationary phase was significantly decreased. The amount of transported or cellular zinc, for both algal species, was operationally defined as the zinc remaining with the cell after EDTA-extraction; it was a linear function of the free ionic zinc concentration remaining in solution, suggesting that the zinc transported into the cell was not derived from the total adsorbed fraction, although the latter may contain some zinc originating from specific sites leading to zinc transport.     

Synthesis of nitrogenase by isolated heterocysts

Heterocysts isolated from Anabaena variabilis incorporate [¹⁴C]leucine and [³⁵S]methionine into trichloroacetic acid-precipitable material in the light. Analysis by polyacrylamide gel electrophoresis shows that the radioactivity is present in polypeptides of discrete sizes. However, the relative proportions of different proteins synthesized by isolated heterocysts differ from the relative proportions of those proteins incorporated by the heterocysts in intact filaments. The two components of nitrogenase are among the proteins synthesized by the isolated heterocysts.     

Distribution and abundance of the hedgehog (Erinaceus europaeus) L. in New Zealand, 1869–1973

An account is given of early liberations and the spread of hedgehogs throughout New Zealand between 1869 and 1973. Evidence, gathered largely from questionnaires and personal observations, is summarised in tables and maps to show the present range and abundance of the species. Hedgehogs dispersed from many points of liberation; their numbers increased dramatically in the South Island between 1910 and 1940, and in the North Island between 1920 and the 1950s. By the 1940s they had colonised most of the lowland areas, and their range had extended to the foot of the bushed mountainous regions; a few were found at altitudes of 2000 m. Since 1948 they have extended their range a little to include parts of the central volcanic plateau of the North Island, areas of inland Nelson, and northern Westland. Their numbers appear to have stabilised over the past 25 years. Today, hedgehogs are most abundant in intensively farmed lowland districts, towns, and suburbs. They become less numerous with increasing altitude, and few are encountered above 800 m or in wet, bushed areas. They are absent from areas where rainfall exceeds 250 cm/year or where more than 250 frosts occur annually. Factors limiting the abundance of hedgehogs in New Zealand are discussed.     

Production of D-amino acid oxidase (DAO) of Trigonopsis variabilis in Schizosaccharomyces pombe and the characterization of biocatalysts prepared with recombinant cells

The cDNA of D-amino acid oxidase (DAO) gene isolated from Trigonopsis variabilis was expressed in Schizosaccharomyces pombe. A clone, ASP327-10, transformed with plasmid vector, pTL2M5DAO, expressed catalytically active DAO in the presence of G418, and converted Cephalosprin C to alpha-ketoadipyl-7-cephalosporanic acid (KA-7-ACA) and glutaryl-7-aminocephalosporanic acid (GL-7-ACA). Biocatalysts were prepared using ASP327-10 and T. variabilis, and evaluated to demonstrate the feasibility of recombinant S. pombe for industrial application. The cells were immobilized by crosslinking polyethylene imine after glutardialdehyde (GDA) fixation and permeabilization by alkaline treatment. Although the biocatalyst prepared from ASP327-10 exhibited DAO activity, catalase activity still remained fully even after permeabilization, under which condition, the catalase activity of T. variabilis decreased to 20-30%. Heat treatment was required before cell fixation by GDA to inactivate the catalase in S. pombe. This improved the efficiency of bioconversion to GL-7-ACA, but caused poor mechanical strength in the biocatalyst of S. pombe. To overcome this weakness, a catalase-deficient host strain was obtained by ethylmethansulfate mutagenesis. Moreover, taking economics into consideration, the integrative vector, pTL2M5DAO-8XL, with multi-copies of expression cassette was constructed to express DAO in S. pombe even in the absence of G418. The newly established integrant, ASP417-7, did not exhibit any catalase activity so that heat treatment was not required. The obtained integrant and its biocatalyst were significantly improved in GL-7ACA conversion ability and mechanical strength. This study demonstrates that the established integrant is a potential candidate as an alternative source of DAO enzyme.     

云居山栓皮栎群落特征及多样性研究

栓皮栎群落是我国暖温带和亚热带落叶阔叶林的重要森林群落类型之一,也是赣北珍稀森林群落之一,在森林演替和植物资源利用中占有重要的地位。通过对江西云居山栓皮栎(Quercus variabilis)群落进行实地调查,采用区系分析、生活型谱分析、物种多样性和双向聚类等方法对其群落特征进行了研究,发现云居山自然保护区栓皮栎群落结构简单,层次明显,多样性程度不高。物种调查及区系研究结果显示,云居山栓皮栎群落维管束植物共计43科56属70种,植物区系主要以泛热带、东亚及北美间断、北温带分布为主,表现出从温带区系向热带区系过渡的特征;生活型谱以高位芽植物为主(占70%),其他生活型相对较少,反映出中亚热带森林以高位芽植物为主的特点。群落物种丰富度、多样性指数均为乔木层小于灌木层和草本层,均匀度指数分析表明,乔木层为聚集分布、灌木层和草本层为均匀分布。双向聚类分析表明,调查的5个样地均为栓皮栎群落,以乔木层物种多度可将5个样地分为3类,组成群落的24个主要物种可分为10类。     

Tracking the role of alternative prey in soybean aphid predation by Orius insidiosus: a molecular approach

The soybean aphid, Aphis glycines (Hemiptera: Aphididae), is a pest of soybeans in Asia, and in recent years has caused extensive damage to soybeans in North America. Within these agroecosystems, generalist predators form an important component of the assemblage of natural enemies, and can exert significant pressure on prey populations. These food webs are complex and molecular gut-content analyses offer nondisruptive approaches for examining trophic linkages in the field. We describe the development of a molecular detection system to examine the feeding behaviour of Orius insidiosus (Hemiptera: Anthocoridae) upon soybean aphids, an alternative prey item, Neohydatothrips variabilis (Thysanoptera: Thripidae), and an intraguild prey species, Harmonia axyridis (Coleoptera: Coccinellidae). Specific primer pairs were designed to target prey and were used to examine key trophic connections within this soybean food web. In total, 32% of O. insidiosus were found to have preyed upon A. glycines, but disproportionately high consumption occurred early in the season, when aphid densities were low. The intensity of early season predation indicates that O. insidiosus are important biological control agents of A. glycines, although data suggest that N. variabilis constitute a significant proportion of the diet of these generalist predators. No Orius were found to contain DNA of H. axyridis, suggesting intraguild predation upon these important late-season predators during 2005 was low. In their entirety, these results implicate O. insidiosus as a valuable natural enemy of A. glycines in this soybean agroecosystem.     

鱼腥藻苯丙氨酸脱氨酶的基因克隆、表达及最适反应pH改造

【目的】表达鱼腥藻苯丙氨酸脱氨酶(AvPAL),并经分子改造降低其最适反应pH。【方法】PCR克隆AvPAL编码基因,并在大肠杆菌中表达,用Ni2+亲和层析柱和凝胶柱纯化重组蛋白。利用GETAREA软件筛选与催化残基距离较近的暴露于酶分子表面的氨基酸位点,将其突变为带电性质不同的氨基酸,并对突变体进行酶学性质研究。【结果】在大肠杆菌中成功表达了AvPAL,纯化后得到电泳纯的重组酶。突变体E75Q和E75R的最适反应pH从8.5分别偏移到7.5和7.0。E75Q在pH 7.5时的比酶活较原酶提高了25%,在pH 6.5–9.5之间酶的稳定性良好,其最适反应温度为50 °C,在此温度下保温1 h酶活无显著变化。在最适反应条件下,E75Q的kcat/Km值较原酶提高了26.6%。【结论】改变AvPAL酶分子中起路易斯碱作用的关键氨基酸残基(质子受体)附近与之有相互作用的氨基酸的带电性质,降低了AvPAL的最适反应pH,提升了其在医疗领域的应用前景。     

秦岭北坡不同生境栓皮栎种子雨和土壤种子库动态

为了阐明栓皮栎种子雨和种子库动态,以秦岭北坡林下、林窗、林缘3种生境栓皮栎天然林为对象,采用离地和地面收集种子的方法,连续定位观测了栓皮栎种子雨、种子库的数量、质量动态,以及幼苗发育过程.结果表明:栓皮栎种子雨从8月中旬开始,9月中旬到10月上旬达到高峰期,11月上旬结束;3种生境种子雨降落历程、发生时间和组成不同;林下种子雨强度最大,为(39.55±5.56)粒·m-2,林窗种子雨降落时间最早,持续时间最长,种子活力最高,而林缘成熟种子占其种子雨的比例最大,达58.7％.从种子雨降落结束到翌年8月,土壤种子库总储量均以林下最大,林缘最小;各生境的土壤种子库中成熟、未成熟、被啃食种子数量和种子活力均随时间变化呈递减趋势,而霉烂种子数量则相反.各生境的土壤种子库中的种子均主要集中在枯落物层,其次为0～2 cm土层,2～5 cm土层中只有极少量种子存在.3种生境中栓皮栎幼苗的密度差异显著,林窗幼苗最多,林缘次之,林下最少.说明林窗更适合栓皮栎种子的萌发和幼苗生长.在栓皮栎林的经营中,通过适当间伐、增加林窗数量,可以促进栓皮栎林的天然更新.     

太行山南麓低山丘陵区栓皮栎直径变化及其与环境因子的关系

使用周长传感器(Circumference DC2)研究了太行山南麓低山丘陵上栓皮栎人工林树干直径的日变化及其影响因子.结果表明:在季节性干旱期间,栓皮栎树干的直径变化周期性明显,直径收缩与液流启动时间基本一致,直径最小值滞后于液流速率最大值3 ～4 h;栓皮栎直径日最大收缩量(MDS)呈现低-高-低的变化趋势,与累积液流通量和叶片水势日差值极显著相关,与土壤含水量呈显著的二次方程关系.MDS值受气象因子变化的影响,与温度日差值、蒸汽压亏缺日差值和相对湿度日差值显著相关,而与太阳辐射日差值的相关性不显著.连续降水后,土壤水分不再是栓皮栎直径变化的限制因子,MDS值与累积液流通量、叶片水势、土壤含水量和气象因子差值的相关性均不显著.季节性干旱期和雨季的土壤含水量和温度是影响树干直径日变化的主要因子.     

李肖叶甲成虫数量及三维空间格局动态

对合肥地区李肖叶甲成虫数量动态及三维空间格局进行研究,结果表明该成虫5月下旬初见,6月4日至7月16日为发生高峰期,9月上旬仍有零星虫口。6月4日至9月10日种群增长模型为y=1692.2258e-0.0545t,6月4日至8月27日树冠上部部位增长模型为y=205.60e-0.0523t,树冠西部部位的增长模型为y=257.13e-0.0505t。样地中的李肖叶甲5月21日至8月13日半变异函数方程是y=0.0709x3-10.479x2+391.67x-300.71、y=-0.0122x3+1.1201x2-19.781x+317.84、y=-0.0013x3+0.1613x2-4.4862x+67.363、y=-0.0016x3+0.9177x2-11.495x+551.94、y=-0.0029x3+0.3034x2-7.5906x+103.37和y=-0.0002x3+0.0172x2-0.4975x+13.691,变程在20.3938—65.0289之间,均为聚集格局,聚集强度指标表明也均为聚集格局;树冠东、西、南、北方位的水平分布5月21日至8月13日的C值均大于1,I值均大于0,CA值均大于0,Iw值均大于1,为聚集格局;树冠上、中、下方位的垂直分布6月4日至8月13日的C值均大于1,I值均大于0,CA值均大于0,Iw值均大于1,为聚集格局;5月21日至8月13日的树冠东、南、西、北、上、中、下部位的C值均大于1,I值均大于0,CA值均大于0,Iw值均大于1,三维分布均为聚集格局。用Iwao公式计算的结果与聚集强度指标分析的结果一致。6月4日—8月13日林间李肖叶甲的种群聚集均数λ值均大于2,聚集是李肖叶甲本身的行为所致。