小鼠精原细胞体外存活、增殖特性及冷冻保存研究

为研究小鼠精原细胞体外存活、增殖特性及其以单细胞或处于功能性结构中(曲细精管或完整睾丸)耐受冷冻-解冻程序的能力。本研究收集生后7d昆白系小鼠睾丸，一步酶法分离获得生精上皮单细胞悬液(主要含精原细胞和支持细胞)，分别接种于D-MEM、D-MEM/F12、KSOM（均含10%的FBS）后系统观察了精原细胞的存活和增殖情况；对生精上皮单细胞进行了-70℃冷冻保存和液氮冷冻保存，对曲细精管及完整睾丸进行了-70℃冷冻保存。结果显示，D-MEM和D-MEM/F12均能使小鼠精原细胞存活和增殖，精原细胞分别在培养的第1~9d或第1~8d 呈逐渐减少趋势，第9~13d或第8~12d呈增殖趋势，而第17d或第16d以后增殖减缓，KSOM只能使小鼠精原细胞短暂存活而不能使其增殖。精原细胞单独培养时增殖不明显而逐渐呈现出分化迹象。生精上皮单细胞、曲细精管及完整睾丸-70℃短期(1wk和2wk)冷冻保存或单细胞悬液液氮短期(2wk)和中长期(1个月和3个月)冷冻保存后均可以获得较理想的细胞存活率(均大于85%)。     

BRL饲细胞及血清浓度对体外小鼠精原干细胞的影响

分别以大鼠肝细胞(BRL)或小鼠胚胎成纤维细胞(STO)为饲养层．采用含5％、10％及20％胎牛血清的培养液，培养6日龄小鼠生精上皮细胞，研究饲养层和血清浓度对精原干细胞生物学行为的影响。结果表明：在培养的第1周内，2个及4个连成串的精原细胞合胞体数量明显增加，同时大多数精原细胞逐渐退化。培养1周后，大多数精原细胞经短暂的存活及分裂活动后退化消失，培养体系中仅保留下少量单个及由细胞间桥相连的双个及4个成串或成团的A型精原细胞。这些细胞在随后的培养过程中，不表现明显的分裂活动，呈圆球形，表达碱性磷酸酶。不同条件培养体系中的精原细胞及精原干细胞的生物学行为无明显不同，培养20～30d时均有少量精原干细胞存活。这说明BRL细胞能用作饲养层促进精原干细胞存活，但对其更新性增殖没有明显作用；培养液中高浓度胎牛血清对精原干细胞存活和增殖无明显影响，但能促进睾丸体细胞增殖。     

小鼠生后不同发育阶段睾丸组织中Crumbs-3(Crb3)的表达

为确定极性调控蛋白Crumbs-3(Crb3)在小鼠(Mus musculus)睾丸组织的定位及其在曲精小管精子发生及间质细胞发育过程中的分布规律,本研究运用常规石蜡切片-HE染色方法和免疫荧光组织化学技术,分别对小鼠生后不同发育阶段睾丸组织结构及Crb3在不同发育时期睾丸组织中的分布进行了研究。结果显示,随着个体发育,曲精小管的直径及生精上皮的厚度不断增大,管腔则由小增大、变小再增大;睾丸曲精小管内生精细胞处于动态的发育变化中,2周龄的生精小管内仅有增殖分裂的数层精原细胞和支持细胞,4周龄时出现各级生精细胞、但无精子形成;之后,继续增殖分裂,至8周管腔内有大量精子形成;并随日龄增加生精细胞密度不断加大;支持细胞的形态随着个体发育逐渐变得清晰而易于辨认;间质细胞则随个体发育体积不断增大、核质比例逐渐下降,而胞质嗜酸性逐渐增强。免疫荧光染色结果显示,Crb3蛋白主要分布在各周龄睾丸曲精小管生精细胞胞膜以及睾丸间质细胞胞膜,且间质细胞上的阳性信号明显强于生精细胞。研究结果提示,Crb3不仅与生精上皮细胞的极性建立和维持有关,而且可能参与睾丸类固醇激素的合成和分泌。     

仔猪睾丸支持细胞的增殖与发育

为了阐明支持细胞增殖和分化能力对精子生成的影响,本实验以7、21和35d仔猪为材料,利用细胞直接计数、免疫组织化学、流式细胞术和免疫印迹分析了仔猪睾丸支持细胞、生精细胞数量和增殖能力变化以及支持细胞的成熟分化状况。结果:(1)从7～21d,曲细精管中支持细胞与其它细胞之间的界线更加明显;从7～35d,每克睾丸中间质细胞和支持细胞的数量在21d时达到高峰(P<0.05),而生精细胞的数量则一直增加(P<0.05),但并没有在曲细精管中发现圆形精子;(2)从7～35d,支持细胞和间质细胞都能表达GATA-4,支持细胞中GATA-4的累积光密度(IOD)在21d时最高(P<0.05),而间质细胞中的IOD在21d和35d之间没有明显差异(P>0.05);(3)从7～35d,整个睾丸细胞都有增殖能力,睾丸细胞的细胞增殖指数(PI)和S期细胞指数(SPF)在21d时最高(P<0.05);三种细胞在这一时期都能表达PCNA,三种细胞PCNA的IOD值和睾丸中PCNA的表达水平在21d时最高(P<0.05);(4)从7～35d,Connexin 43(Cx43)主要表达于支持细胞的细胞质,支持细胞和间质细胞中的IODs在2...     

仔猪睾丸组织块生精细胞体外培养及鉴定初报

目的：建立仔猪睾丸组织块生精细胞爬片体外培养模型。方法：应用溴脱氧核苷尿嘧啶(5＇-Bromo-2＇-deoxyuridine, BrdU)免疫荧光技术检测生精细胞体外增殖分化情况；采用HE染色鉴定体外培养生精细胞分化程度。结果：结果表明体外培养第3天支持细胞开始游出睾丸组织块并贴壁；体外培养第4～8天睾丸组织块周围可见生精细胞，但未见明显增殖分化情况；体外培养第9～20天可明显观察到生精细胞增殖分化，间桥连接(gap junction)的生精细胞集落呈葡萄串或串珠状，并有精子细胞形成。结论：本培养体系能够提供睾丸组织块生精细胞增殖分化所需要的条件，在没有添加睾酮的情况下能够使非精子细胞分化为精子细胞。     

GnRH主动免疫对雄性大鼠垂体-睾丸轴功能的影响

本研究用GnRH并列体二聚物(GnRH-TDK)主动免疫SD雄性大鼠观察其对大鼠垂体-睾丸轴功能的影响,以探讨GnRH主动免疫去势的分子机理.24只性成熟SD雄鼠随机分为免疫组和对照组,免疫组雄鼠于12周龄时初免,8周后加免,对照组雄鼠不做任何注射.每两周采集血液放免法检测血清抗体滴度及激素含量变化.加免后4周脱颈处死所有雄鼠,收集垂体、睾丸实时荧光定量PCR分析相关生殖基因mRNA的变化.结果显示,GnRH主动免疫后12只免疫雄鼠中11只血清GnRH抗体滴度显著上升,血清LH、FSH及睾酮(T)均极显著下降到检测限附近或以下(p＜0.01),同时睾丸严重萎缩,其重量及体积下降到对照组睾丸的20％(p＜0.01),组织切片显示,曲细精管上皮组织严重受损,管内仅有少数退化的精原细胞.与对照组相比,GnRH主动免疫极显著下调雄鼠垂体GnRH受体、LH-β、FSH-β和睾丸LH受体及FSH受体mRNA表达水平(p＜0.01).由此可见,GnRH主动免疫可通过下调垂体GnRH受体、促性腺激素亚基及睾丸LH、FSH受体基因表达,影响垂体-睾丸轴的功能.     

哲罗鲑精子冷冻保存

哲罗鲑(Hucho taimen)是分布于额尔齐斯河和黑龙江等水域的冷水性珍稀鱼类,对其精子冷冻保存技术进行研究并建立精子冷冻库,对于哲罗鲑种质保存、人工繁殖发育、苗种培育及种群恢复都具有重要的意义.本研究利用生理盐水、葡萄糖、胎牛血清(fetal bovine serum,FBS)等配制了9种精子稀释液(MPRS,RS,Hanks,ELS,EG1,EG2,Stein,SS-2和D1S),对哲罗鲑精子在稀释液中活力、加入抗冻剂后活力和液氮中冷冻后活力进行检测,发现精子在以上稀释液中均具较高活力(73.00±2.65)％～(96.67±2.08)％,加入二甲基亚砜(dimethyl sulfoxide,DMSO)后活力下降,冷冻后仅在ELS和D15中具有极低活力(1.21±0.00)％和(1.1±0.00)％.进而利用ELS对甲醇、二甲基亚砜浓度进行筛选,发现精子在4％～12％甲醇中平衡20 min后活力仍为(79.00±6.52)％～(88.80±7.89)％,冷冻后仅在6％甲醇中具有相对较高活力(18.33± 10.61)％,但与鲜精活力相比显著下降(P＜0.05).精子在4％～12％ DMSO中平衡5 min后精子活力为(43.75± 11.09)％～(81.67±2.89)％,平衡20 min后精子活力极大下降(3.25±0.30)％～(45.00±5.77)％,冷冻后精子活力极低(1.00±0.00)％～(2.75±1.50)％(P＜0.05).以D15为基础液对葡萄糖浓度进行筛选,发现当葡萄糖浓度为30％(D30)时,冷冻后精子活力相对较高(17.80±2.59)％,但与鲜精相比活力显著下降(P＜0.05).另外对精子在D15、D30、Stein和ELs4种稀释液中短期保存的效果进行比较表明,Stein中精子存活在时间最长(45 h),D30次之(15h).综上所述,分别在ELS和D30中加入6％甲醇冷冻保存哲罗鲑精子具有一定活力,本研究结果为哲罗鲑精子大量冷冻保存和精子库建立等方面研究奠定了基础.     

牛体外受精胚胎冷冻保存的研究

本研究在国内条件下对牛体外受精胚胎的冷冻保存进行了系统研究。胚胎在含 4 .0mol/ L乙二醇的 PBS中停留 15～ 2 0 min,对其随后孵化无明显影响 ( 72 .2 %比 82 .4 % ,P>0 .0 5) ;当乙二醇的浓度升高到 5.0 mol/ L和 6.0 mol/ L时 ,胚胎的孵化率明显下降 ( 2 5.0 %和3.2 % ,P<0 .0 1)。在胚胎的常规冷冻保存试验中 ,当冷冻液为 1.8mol/ L乙二醇加 0 .2 0 mol/ L蔗糖时 ,- 7℃植冰后缓慢降温到 - 30℃投液氮保存的胚胎冻后存活率 ( 86.3% )和孵化率 ( 72 .5% )明显高于 ( P<0 .0 5)缓慢降温到 - 2 5℃投液氮保存的胚胎冻后存活率 ( 57.1% )和孵化率( 35.7% ) ;当冷冻液不加蔗糖时 ,胚胎的冻后存活率明显下降 ( 60 .9% ,P<0 .0 5) ,且胚胎冻后无一孵化。在胚胎的超速冷冻保存试验中 (在 - 2 3℃平衡 30 min直接置液氮保存 ) ,冷冻在冷冻液为 3.0 mol/ L乙二醇加 0 .30 mol/ L蔗糖中的胚胎冻后存活率 ( 84 .0 % )和孵化率 ( 60 .6% )明显高于冷冻液为 2 .5mol/ L乙二醇加 0 .30 mol/ L蔗糖、 2 .0 mol/ L乙二醇加 0 .30 mol/ L蔗糖和2 .0 mol/ L乙二醇加 0 .2 3mol/ L蔗糖的胚胎冻后存活率 (分别为 59.0 % ,4 3.5%和 2 5.0 % ,P<0 .0 5)和孵化率 (分别为 4 5.5% ,30 .4 %和 0 % ,P<0 .0 5) ;7日龄胚胎的冻     

氟对小鼠生精细胞凋亡的影响及锌的保护作用

为观察氟对成年雄性小鼠生精细胞凋亡的影响及锌的保护作用,通过腹腔注射氟化钠建立氟中毒动物模型及锌保护实验。采用石蜡切片、苏木精-伊红染色(HE染色)、末端原位标记(TUNEL)及琼脂糖凝胶电泳的方法检测小鼠生精细胞的凋亡情况。结果显示:(1)氟感染组无论是低剂量组(NaF10 mg/kg)或是高剂量组(NaF 20 mg/kg)生精细胞都发生了明显的凋亡。生精细胞凋亡主要发生在精母细胞和精原细胞,凋亡指数与对照组差异极显著(P<0.05)。(2)无论是高剂量锌(ZnSO430 mg/kg)和低剂量锌(ZnSO415 mg/kg)都可以使凋亡指数明显降低,高剂量锌组的凋亡指数与对照组差异不显著(P>0.05)。     

奶牛X性控冻精应用研究

利用SX-MOFLO精子分离设备分离奶牛XY精子,将分离后的奶牛X精子冷冻保存。探讨奶牛X性控冻精解冻后存活时间、不同方法进行人工授精的受胎率,为奶牛X性控冻精的大规模推广应用提供依据。方法与结果:(1)取不同种公牛、不同批次的X性控冻精及普通冻精解冻后倒入5 mL试管内摇匀,至37°C水浴锅中,每2 h检测一次精子活力,4 h后每小时检测一次精子活力,直至无运动精子。结果奶牛X性控冻精存活时间为5~12 h,比同种普通精子的存活时间短2~10 h。(2)应用奶牛X性控冻精人工授精的母牛的情期受胎率(59.4%=342/576)与普通冻精的情期受胎率(60.6%=129/213)相比,无显著差异(P>0.05)。奶牛X性控冻精母犊出生准确率93.3%(319/342),极显著地高于普通冻精48.8%(63/129)的母犊率(P<0.001)。且应用奶牛X性控冻精人工授精后的青年牛的情期受胎率(6 5.6%=2 4 0/3 6 6)极显著的高于经产牛的情期受胎率(4 8.6%=1 0 2/2 1 0)(P<0.001)。(3)在母牛发情结束后6~8 h左右进行人工授精,其情期受胎率为67.1%(116/173),显著地高于在母牛发情开始后8~1 2 h进行人工授精5 6.1%(2 2 6/4 0 3)的情期受胎率(P<0.0 1)。(4)在准确判断母牛卵巢上卵泡发育状态的前提下,将精液全部输至发育侧子宫角大弯处,其情期受胎率62.7%(289/461),高于母牛每侧子宫角输1/2支X性控冻精46.1%(53/115)的情期受胎率,但二者之间差异不显著(P<0.05)。研究结果表明应用奶牛X性控冻精进行人工授精,其受胎率与奶牛的经产与否、授精时间和输精部位有关。     

低氮和干旱胁迫对富士和秦冠生长及氮素利用的影响

【目的】以富士（Fuji）、 秦冠（Qinguan）嫁接在平邑甜茶（Malus hupehensis Rehd.）上的当年生盆栽苗为试验材料,采用砂培方法,研究了缺氮胁迫和干旱对富士和秦冠生长情况、 光合参数、 植株各部位氮磷钾含量及氮素利用效率的影响,分析比较了低氮干旱条件下富士和秦冠生长及氮素利用的差异,以期为果树生产高效肥水利用提供理论指导。【方法】试验共设四个处理: 正常氮正常水（ZZ）、 低氮正常水（DZ）、 正常氮干旱（ZG）、 低氮干旱（DG）。氮素和水分均设置两个水平,分别为正常氮（6 mmol/L NO-3-N）、 低氮（0.3 mmol/LNO-3-N）、 正常供水（保持盆中砂子相对含水量为饱和含水量的80%~85%）、 干旱处理（保持盆中砂子相对含水量为饱和含水量的60%~65%）。【结果】富士和秦冠的生物量（茎和叶）、 株高茎粗等生长指标以及光合速率、 气孔导度、 蒸腾速率均为正常氮正常水（ZZ）＞低氮正常水（DZ）＞正常氮干旱（ZG）＞低氮干旱（DG）,并且相对应处理下秦冠的以上指标均高于富士;正常供水下,缺氮处理使富士、 秦冠的根冠比比正常氮处理均有所增加,富士提高了2.05%,秦冠提高了22.40%。富士和秦冠的氮、 磷、 钾含量均表现出正常氮正常水（ZZ）＞低氮正常水（DZ）＞正常氮干旱（ZG）＞低氮干旱（DG）; 氮、 钾元素含量在植株各部位的分布顺序依次是叶＞根＞茎,磷元素则是根＞叶＞茎;光合氮素利用效率（PNUE）和氮素利用效率表现为秦冠处理之间差异极显著,富士处理之间差异不显著;秦冠的PNUE和NUE明显高于富士,在低氮正常水（DZ）处理下,秦冠氮肥利用率比富士高42.07%,在低氮干旱（DG）处理下高64.14%;低氮胁迫下富士和秦冠的NUE显著提高,并且秦冠提高的幅度高于富士。【结论】施用氮肥能够显著提高富士与秦冠的干物质量,同等水肥条件下,秦冠生长优于富士;水分亏缺会减少叶片对氮的吸收,干旱条件下适度增施氮肥,可提高果树的抗旱能力;低氮干旱胁迫下秦冠的生长指标、 光合指标及氮素利用效率指标均优于富士,表现出较强的抗低氮干旱胁迫的能力。     

Pasture Vegetation Elemental Analysis by Laser-Induced Breakdown Spectroscopy

Laser-induced breakdown spectroscopy (LIBS) is a new technique for the analysis of plant material. This study investigates the application of LIBS to pasture-based plant samples. The LIBS measurements were obtained from pelletized pasture samples (100 samples) that had been also analyzed by inductively coupled plasma–optical emission spectroscopy (ICP-OES) following microwave digestion for calibration and comparison purposes. Comparisons for elements sodium (Na), potassium (K), magnesium (Mg), calcium (Ca), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), boron (B), phosphorus (P), and sulfur (S) showed that LIBS could be used for almost all the standard profile total elements with concentrations down to low mg/kg levels (observed error of Na: 0.024 percent, K: 0.18 percent, Mg: 0.016 percent, Ca: 0.073 percent, P: 0.017 percent, Mn: 31 mg/kg, Fe: 150 mg/kg, Zn: 6.6 mg/kg, and B: 1.1 mg/kg). Elemental analysis at less than mg/kg levels was not possible using LIBS. The elements S and Cu were particularly difficult to analyze with reliability using LIBS at the concentration levels found in the plant samples. Replacing microwave digestion and subsequent ICP analysis with a direct analysis of dried plant samples using LIBS has the potential to improve the productivity and reduce the cost of testing.     

Faculty positions:Center for Agricultural Resources Research,Chinese Academy of Sciences

正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,     

Macroinvertebrates in North American tallgrass prairie soils: effects of fire, mowing, and fertilization on density and biomass

The responses of tallgrass prairie plant communities and ecosystem processes to fire and grazing are well characterized. However, responses of invertebrate consumer groups, and particularly soil-dwelling organisms, to these disturbances are not well known. At Konza Prairie Biological Station, we sampled soil macroinvertebrates in 1994 and 1999 as part of a long-term experiment designed to examine the effects and interactions of annual fire, mowing, and fertilization (N and P) on prairie soil communities and processes. For nearly all taxa, in both years, responses were characterized by significant treatment interactions, but some general patterns were evident. Introduced European earthworms (Aporrectodea spp. and Octolasion spp.) were most abundant in plots where fire was excluded, and the proportion of the total earthworm community consisting of introduced earthworms was greater in unburned, unmowed, and fertilized plots. Nymphs of two Cicada genera were collected (Cicadetta spp. and Tibicen spp.). Cicadetta nymphs were more abundant in burned plots, but mowing reduced their abundance. Tibicen nymphs were collected almost exclusively from unburned plots. Treatment effects on herbivorous beetle larvae (Scarabaeidae, Elateridae, and Curculionidae) were variable, but nutrient additions (N or P) usually resulted in greater densities, whereas mowing usually resulted in lower densities. Our results suggest that departures from historical disturbance regimes (i.e. frequent fire and grazing) may render soils more susceptible to increased numbers of European earthworms, and that interactions between fire, aboveground biomass removal, and vegetation responses affect the structure and composition of invertebrate communities in tallgrass prairie soils.     

Variation in quality parameters between and within 14 Nordic tree fruit and berry species

Abstract

A 3-year study was carried out to investigate quality parameters in 14 tree fruit and berry species grown in southern Norway. The species were blueberry, apple, aronia, sour cherry, sweet cherry, red raspberry, strawberry, blackcurrant, gooseberry, red currant and elderberry, harvested along with wild bilberry, cloudberry and lingonberry. Significant differences between species were identified for all quality parameters. The coefficient of variation between species was lowest for pH (12.5%), dry matter (18.9%) and soluble solids (25.3%), followed by titratable acids (59.3%), total phenolics (83.8%), antioxidant capacity FRAP (85.7%) and antiradical power by the DPPH-assay (97.8%), total monomeric anthocyanins (132%) and ascorbic acid (137%). Average coefficient of variation within species were lower and ranged from 4 (pH) to 62% (ascorbic acid). Only the FRAP values were significantly affected by harvesting year with lower levels in 2004 than in 2005 and 2006. There were significant interactions between species and harvesting year for dry matter, soluble solids, pH, ascorbic acid and FRAP. The results indicate generic ranges in composition within species independent upon growing location and climate, and the composition of the tree fruits and berries is not likely to deviate from these ranges. It is concluded that desirable composition of tree fruits and berries and their products should primarily be achieved by selection among species rather than searching fors broadened variation within individual species.     

Potassium Fixation and Release Characteristics in Rhizosphere and Nonrhizosphere Soils for a Rapeseed–Rice Cropping Sequence

Potassium (K) fixation and release in soil are important factors in the long-term sustainability of a cropping system. Changes in K concentration and characteristics of K fixation and release in rhizosphere and nonrhizosphere soils in the rapeseed (Brassica napus L.)–rice (Oryza sativa L.) rotation were investigated using a rhizobox system. The concentrations of different forms of K in both rhizosphere and nonrhizosphere soils decreased with plants compared to without plants, regardless of K fertilizer application. Potassium uptake by crops mainly came from the rhizosphere soil. In the treatment without K fertilizer (–K), the main form of K supplied by the soil to the crops was 1.0 mol L^?1 nitric acid (HNO₃) nonextractable K, followed by nonexchangeable K, and then exchangeable K. In the treatment with K fertilizer (+K), the main K forms supplied by the soil to the crops were exchangeable K and nonexchangeable K. The amount and rate of K fixation after one cycle of the rapeseed–rice rotation was greater in rhizosphere soil than in nonrhizosphere soil. The amount and rate of K fixation of soil in the +K treatment were significantly less than in the –K treatment. The cumulative amounts of K released with 1.0 mol L^?1 ammonium acetate (NH₄OAc) and 1.0 mol L^?1 HNO₃ extraction increased with the increasing numbers of extractions, but the K-releasing power of soil by successive extraction decreased gradually and finally became almost constant. The release of K was less in rhizosphere soil than in nonrhizosphere soil. The release of K in the +K treatment was similar to that in the –K treatment in rhizosphere soil, but the K release in nonrhizosphere soil was greater with the +K than the –K treatment. Overall, the information obtained in this study will be helpful in formulating more precise K fertilizer recommendations for certain soils.     

Electron donor and pH relationships for biologically enhanced dissolution of chlorinated solvent DNAPL in groundwater

Biologically enhanced dissolution offers a method to speed removal of chlorinated solvent dense non-aqueous-phase liquid (DNAPL) sources such as tetrachloroethene (PCE) and trichoroethene (TCE) from aquifers. Bioremediation is accomplished by adding an electron donor to the source zone where fermentation to intermediates leading to acetic acid and hydrogen results. The hydrogen and possibly acetic acid are used by dehalogenating bacteria to convert PCE and TCE to ethene and hydrochloric acid. Reductive dehalogenation is thus an acid forming process, and sufficient alkalinity must be present to maintain a near neutral pH. The bicarbonate alkalinity required to maintain pH above 6.5 is a function of the electron donor: 800 mg/L of bicarbonate alkalinity is sufficient to achieve about 1.2 mM TCE dechlorination with glucose, 1.7 mM with lactate, and a much higher 3.3 mM with formate. Laboratory studies indicate that in mixed culture, formate can be used as an electron donor for complete conversion to ethene, contrary to pure cultures studies indicating it cannot. Various strategies can be used to add electron donor to an aquifer for DNAPL dehalogenation while minimizing pH problems and excessive electron donor usage, including use of injection-extraction wells, dual recirculation wells, and nested injection-extraction wells.     

Biodiversity and ecogeography of wild Lactuca spp. in some European countries

Staff members of the Department of Botany of Palacký University in Olomouc and Gene Bank Department – Workplace Olomouc, Research Institute of Crop Production in Prague, Czech Republic, conducted an expedition in seven European countries (Austria, Czech Republic, France, Germany, Italy, the Netherlands, Switzerland) in August/September 1999 to collect wild Lactuca spp. germplasm and study its geographic distribution, ecology and biodiversity. During the mission, more than 600 locations were visited resulting in the collection of 602 seed samples (accessions) of wild Lactuca species and 13 seed samples of related genera (Chondrilla and Mycelis). Lactuca serriola f. serriola, L. serriola f. integrifolia, L. saligna and L. viminea subsp. chondrilliflora were prevalent in southern Europe (Italy, France), however, only L. serriola was common in central and western Europe (Austria, Czech Republic, Germany, Netherlands, Switzerland). The greatest diversity of Lactuca species was found in France, where also the most seed samples (165) were collected. The most characteristic habitats with a high density of Lactuca spp. populations were observed along roads and highways, grassy ditches, ruderal communities, and dust-heaps. Natural infections by powdery mildew (Erysiphe cichoracearum) and downy mildew (Bremia lactucae) on some wild Lactuca spp. were observed. Recent observations concerning the geographic distribution, population structure, habitats, and natural occurrence of diseases of Lactuca spp. are discussed. This assemblage of genetic resources of Lactuca spp. can serve as the basis of future studies of species diversification, spatial population structure, plant microevolution, domestication processes, and genetic variability of host-parasite interactions.     

Increase in auxiliary photoprotective photosynthetic pigments in wheat seedlings induced by Azospirillum brasilense

Inoculation of wheat seedlings with the plant growth-promoting bacterium Azospirillum brasilense Cd was immobilized in alginate microbeads and, without applying any stress, significantly increased the quantity of several photosynthetic pigments, such as chlorophyll a, chlorophyll b, and the auxiliary photoprotective pigments violaxanthin, zeaxanthin, antheroxanthin, lutein, neoxanthin, and β-carotene. This resulted in greener plants with no apparent visible stress. After monitoring the quantity of photosynthetic pigments for 4 weeks, we observed that inoculated plants had higher quantities of pigments in shoot and stem. The greatest difference in the quantity of all pigments between inoculated and noninoculated plants occurred in the first week of growth. Regardless of treatment, the quantity of pigments in stems was three to four times less than the quantity of these pigments in shoots. Application of Azospirillum, either as liquid inoculant or as alginate microbeads, did not alter the positive effect of the bacteria on pigment production or the positive response of the plants towards A. brasilense Cd inoculation.     

In-situ enrichment and analysis of atrazine-degrading microbial communities using atrazine-containing porous beads

We examined the community composition of microbes that colonized atrazine-containing beads buried in agricultural soils that differed in atrazine treatment history. Bacterial abundance was 5-40-fold greater in atrazine-fortified beads. In beads containing 20 mg atrazine kg⁻¹ buried in soil with a history of atrazine application (conditioned soil), the abundance of Actinobacteria increased approximately 80-fold whereas in control soil, Actinobacteria were enriched only 10-fold and the gamma-Proteobacteria and Planctomycetes increased by 60- and 25-fold, respectively. The gamma-Proteobacteria were enriched by 120- and 230-fold in beads containing 200 mg atrazine kg⁻¹ in conditioned and control soil, respectively. The results demonstrate that BioSep^® beads are a suitable matrix for recruiting a diverse subset of the bacterial community involved in atrazine degradation.