The C-terminal extension of bacterial flavodoxin-reductases: Involvement in the hydride transfer mechanism from the coenzyme

To study the role of the mobile C-terminal extension present in bacterial class of plant type NADP(H):ferredoxin reductases during catalysis, we generated a series of mutants of the Rhodobacter capsulatus enzyme (RcFPR). Deletion of the six C-terminal amino acids beyond alanine 266 was combined with the replacement A266Y, emulating the structure present in plastidic versions of this flavoenzyme. Analysis of absorbance and fluorescence spectra suggests that deletion does not modify the general geometry of FAD itself, but increases exposure of the flavin to the solvent, prevents a productive geometry of FAD:NADP(H) complex and decreases the protein thermal stability. Although the replacement A266Y partially coats the isoalloxazine from solvent and slightly restores protein stability, this single change does not allow formation of active charge-transfer complexes commonly present in the wild-type FPR, probably due to restraints of C-terminus pliability. A proton exchange process is deduced from ITC measurements during coenzyme binding. All studied RcFPR variants display higher affinity for NADP⁺ than wild-type, evidencing the contribution of the C-terminus in tempering a non-productive strong (rigid) interaction with the coenzyme. The decreased catalytic rate parameters confirm that the hydride transfer from NADPH to the flavin ring is considerably hampered in the mutants. Although the involvement of the C-terminal extension from bacterial FPRs in stabilizing overall folding and bent-FAD geometry has been stated, the most relevant contributions to catalysis are modulation of coenzyme entrance and affinity, promotion of the optimal geometry of an active complex and supply of a proton acceptor acting during coenzyme binding.     

青藏高原高寒草甸植被特征与温度、水分因子关系

以广布于青藏高原的高寒草甸为研究对象,进行模拟增温实验,探讨高寒草甸植被特征与温度、水分因子关系,并试图论证高寒草甸植被是否符合生物多样性代谢理论.结果表明:①高寒草甸植被物种多样性的对数与绝对温度的倒数呈显著线性递减关系,空气-地表-浅层土壤(0-20 cm)温度(R2 ＞0.6,P＜0.01)较深层土壤(40-100 cm)温度(R2＜0.5,P＜0.05)对物种多样性影响大;其植被新陈代谢平均活化能为0.998-1.85 eV,高于生物多样性代谢理论预测值0.6-0.7 eV,这是高寒草甸植被对长期低温环境适应进化的结果.②除趋势对应分析和冗余分析显示,温度对植被地上部分影响较大,而土壤水分对全株影响均较大,适当的增温与降水均可极显著促进高寒草甸植被生长.③逐步回归和通径分析显示,40 cm、60 cm深度土壤水分对植被地上部分产生直接影响,20 cm高度空气相对湿度和40 cm深度土壤温度对其产生间接影响;40 cm深度土壤温度和60 cm深度土壤水分对植被地下部分产生直接影响,红外地表温度对其产生间接影响.深层土壤温度和水分对高寒草甸植被具有影响作用,这可能与增温后冻土的融化有关,但其机理尚待进一步研究.     

β-葡萄糖苷酶的分离纯化和性质研究

β－葡萄糖苷酶是纤维素酶的重要组分之一,它不仅可水解纤维二糖和寡糖,更可解除纤维二糖对β－１,４－内切葡聚糖酶和外切葡聚糖酶的抑制,提高水解速率和程度．利用ＳｅｐｈａｄｅｘＧ－１５０和ＤＥＡＥ－ＳｅｐｈａｄｅｘＡ－５０层析法从黑曲霉变异株Ｌ－２２中分离提纯了β－葡萄糖苷酶,该酶是由两个分子量相同的亚基组成的二聚体,每个亚基分子量为２０３ｋＤ．该酶最适ｐＨ为４．８,ｐＨ稳定范围在３．６～６．４;最适温度是６０℃,温度稳定范围为４～６０℃;酶分子含糖量为８．３５％．它是一个酸性β－葡萄糖苷水解酶,专一性地水解β－糖苷键．而不水解α－糖苷键,对短链底物表现了相对高的活力．用动力学分析和共价化学修饰方法探讨了与该酶活力有关的必需基团．由ｐＨ对ｌｇＶｍ和ｌｇＶｍ／Ｋｍ的影响,推测出酶活性部位至少有两个可解离基团为酶活性所必需,它们在酶－底物复合物中的ｐＫｅｓ１和ｐＫｅｓ２的值分别为４．０和５．６,在游离酶中的ｐＫ值分别为４．２和５．９．由此可初步判断这两个可解离基团可能为组氨酸和含羧基的氨基酸,它们与酶的催化和底物结合可能有关．     

普通长翼蝠食性结构及其回声定位与体型特征

在普通长翼蝠(Miniopterus fuliginosus)的捕食区内用灯诱法和网捕法调查潜在食物(昆虫)种类; 用粪便分析法鉴定普通长翼蝠的食物组成,发现其主要捕食体型较大的鳞翅目和鞘翅目昆虫,体积百分比分别为55%和38%.普通长翼蝠具有相对狭长的翼,翼展比为6.94 ± 0.13;翼载为(9.85 ± 0.83)N/m2,相对较大.飞行状态下普通长翼蝠的回声定位叫声为调频下扫型,声脉冲时程为(1.45 ± 0.06)ms,脉冲间隔为(63.08 ± 21.55)ms,主频较低,为(44.50 ± 2.26)kHz.研究表明,普通长翼蝠的形态特征和回声定位特征与其捕食行为有着密切的联系.     

人工微生物混菌系统机制解析中的组学应用及进展

人工微生物混菌系统的生物工程应用价值日益受到重视,使得对于混菌系统中成员菌间的相互作用机制研究也成为近年来的一个热点.其研究结果一方面可以为现有人工混菌系统的进一步优化提供理论依据,另一方面也为全新混菌系统的人工构建提供新的思路和策略,进而促进人工微生物混菌系统未来规模化应用.基因组学、转录组学、蛋白质组学和代谢组学等...     

淀山湖基于初级生产力的鲢鳙富营养化控制

于2008年10月—2009年9月,采用黑白瓶测溶氧法对淀山湖湖南区和围隔区初级生产力进行逐月测定,依据测定的湖南区初级生产力估算鲢、鳙渔产潜力及合理放养量,讨论温度、透明度、营养盐、叶绿素a等理化因子与初级生产力的相关性,分析鲢、鳙放养对淀山湖水体营养物质的定量去除效果。结果表明:1)湖南区平均水柱日毛产量(PG)为4.02gO2.m-2,8月最高、1月最低,净产量(PN)为1.99gO2.m-2,年PN/PG=52%;2)0～0.5m水层对水柱初级生产力贡献最大,湖南区占32.3%、围隔区占32.2%;3)初级生产力与透明度、水温呈明显正相关,与其他理化因子相关性不明显;4)淀山湖浮游植物年生产量为28.18×104t,按鲢鳙3:1比例放养,鲢渔产潜力为1621.58t,鳙为1216.18t,鲢合理放养量为16.94t.km-2,鳙合理放养量为6.35t.km-2;5)以淀山湖每年渔产2837.76t计算,可以固定氮、磷分别为85.50t、7.63t,由此可使淀山湖水体中氮、磷含量分别降低0.67和0.06mg.L-1。     

葡萄基因组中KUP蛋白的生物信息学分析

植物钾(K+)转运体蛋白在K+的跨膜运输中起重要作用,进而维持植物体正常生长和代谢活动.本研究中,通过隐马尔科夫模型(HMM)和葡萄蛋白质库搜索,共找到18个葡萄钾转运体蛋白(VvKUPs).利用生物信息学方法,我们对葡萄家族12条KUP蛋白序列的系统发生和KUP基因组定位进行分析,然后对其氨基酸组成成分、理化性质以及二级结构进行预测和分析,同时还分析了葡萄与拟南芥、水稻和杨树的KUP基因家族之间的联系.基因组定位结果发现其分布在至少9条染色体上.二级结构预测结果发现不同成员间氨基酸数目、氨基酸序列间的疏水性存在一定的差异;α-螺旋和无规则卷曲为主要二级结构组成部分.基因结构分析表明,KUP基因家族成员分别含有7～10个内含子.葡萄KUP蛋白的亚细胞定位分析表明VvKUP主要定位于膜结构上.电子表达图谱分析结果表明:12条KUP基因有对应的EST序列,其中的11条KUP有相应的电子表达谱,并主要在花、果实、花序和花蕾等组织部位表达.     

The effect of the armored scale,Rhizaspidiotus donacis (Hemiptera: Diaspididae), on shoot growth of the invasive plant Arundo donax (Poaceae: Arundinoideae)

The effect of feeding by the armored scale, Rhizaspidiotus donacis (Leonardi, 1920) (Hemiptera: Diaspididae) on the growth of the plant Arundo donax L. (Poaceae) was evaluated under field conditions in its native range. The study was designed to evaluate the impact of R. donacis, a candidate agent for biological control of A. donax which is invasive in arid riparian ecosystems of the Southwestern USA and Mexico. The study was carried out at five A. donax sites in the Province of Alicante, Spain, differing in altitude and climate. At each site, 30 infested lateral shoots were selected and 15 were randomly treated monthly with imidacloprid insecticide. Shoot lengths were measured monthly over a 1-year period in a comparative growth analysis. Shoots infested with R. donacis had an over 2-fold reduced growth rate as compared to treated shoots. Growth of shoots varied by site, and the effect of R. donacis on growth was most pronounced in the late spring, when mature females produced first instar scale crawlers. The impact of R. donacis on A. donax growth under field conditions in the native range, combined with its narrow host specificity, indicate that R. donacis is a promising candidate for biological control of A. donax in North America and other areas invaded by this weed.     

谷子窄颖花突变体sins1的表型分析和突变基因定位

利用甲基磺酸乙酯(EMS,ethyl methyl sulfonate)诱变剂处理野生型Yugu1(豫谷1号),在后代中发现了一个可以稳定遗传的颖花明显变窄的突变体,将其命名为sins1。与Yugu1相比,突变体sins1的株高显著降低了3.89%,穗长和穗粗分别显著降低了17.42%和21.62%,旗叶叶长和叶宽分别显著降低了15.09%和25.78%,千粒重显著降低了40.96%,谷码数显著降低了25%,均达到显著水平(P0.05)。利用突变体sins1为母本、SSR41为父本构建F_2定位群体,F_2正常颖花与窄颖花植株数目的分离比例为3∶1,表明该突变性状由隐性单基因控制。利用F_2群体隐性单株,最终将突变基因定位在3号染色体上SSR标记3-2658与CAAS3031间约7.709 Mb的距离内,为下一步精细定位提供了基础,同时也为促进禾本科作物颖花的研究提供了方向。     

Analysis of growth during phototropic curvature of cress hypocotyls

Abstract. Growth rates throughout an organ curving phototropically under continuous, unilateral while light were monitored by lime-lapse photography of cress hypocotyls marked into 1 mm sections by two rows of ion-exchange beads. Curvature resulted from an integrated sequence of changes in growth rate on each side of the organ, but the actual patterns of change and, therefore rales of curvature, differed within even this one species, depending upon the immediate pretreatment of the seedlings. Transference of seedlings from darkness to unilateral irradiation gave immediate growth inhibition on both sides of the organ. Curvature resulted from differential recovery of growth rate, being seen first on the shaded side, most prominently in the apical regions; only 2h after initial exposure to light did growth recover on the lit (lower) side. On the other hand, transfer of seedlings from omnilateral to unilateral irradiation of the same intensity resulted in simultaneous growth inhibition on the irradiated side and stimulated growth on the shaded side: this growth stimulation of the shaded side was greater than occurred in totally darkened control plants.