Co‐expression of the proteinase inhibitors oryzacystatin I and oryzacystatin II in transgenic potato alters Colorado potato beetle larval development

Colorado potato beetle (CPB; Leptinotarsa decemlineata Say, Coleoptera: Chrysomelidae) has shown a remarkable adaptability to a variety of control measures. Although oryzacystatin I and II (OCI and OCII) have potential in controlling pests that use cysteine proteinases for food digestion, expression of a single OC gene in potato exhibited a minimal or no effect on CPB fitness traits. The aim of this study was to examine the effect of coexpressed OCI and OCII in potato (Solanum tuberosum L.) cultivars Desiree, Draga?evka and Jelica on CPB larvae. Growth parameters, consumption rates and food utilization, as well as activity of proteases of CPB larvae were assayed. Second and third instar larvae fed on transformed leaves molted earlier and had higher relative growth and consumption rates than larvae fed on nontransformed leaves, while efficiency of food utilization was unaffected. In contrast, fourth instar maximum weight gain and amount of leaves consumed were about 20% lower for the larvae fed on transgenic potato. Analysis of total protease activity of third instar larvae revealed reduction in overall proteolytic activity measured by azocasein hydrolysis, accompanied with inhibition of cysteine proteinase activity 24 h after ingestion of potato leaves expressing OCI and OCII. However, after long‐term feeding on transformed leaves proteolytic activities of larvae became similar to the controls. Although feeding on OCI/OCII leaves did not affect larval survival, coexpression of OC genes reduced the development time and thus significantly decreased plant damage caused by CPB larvae.     

LPS response and endotoxin tolerance in Flt-3L-induced bone marrow-derived dendritic cells

Fms-like tyrosine kinase-3 ligand (Flt-3L) stimulates the differentiation of bone marrow cells into dendritic cells (DCs) and was used as an adjuvant therapy in the experimental model of burn wound sepsis. In this study, we describe the phenotypical characteristics of an Flt-3L-dependent DC culture (FLDC) system following LPS stimulation, which induces an inflammatory response, and after a second LPS stimulation, which induces tolerance. Priming of FLDCs with LPS via TLR4 has been shown to induce the activation of all three mitogen-activated protein kinase (MAPK) families and enhance NF-κB complex translocation into the nucleus. Stimulated FLDCs express all maturation markers and exhibit an increase in IL-12p40 production and to a lesser extent, IL-10 production. In contrast, LPS stimulation of tolerized FLDCs was not associated with TLR4 up-regulation and led to MAPK inhibition. The decrease in p38 and JNK activation was correlated with an impairment of IL-12p40 production. Endotoxin tolerance in FLDCs was associated with enhanced ERK1/2 activation, an increase in MKP-1 phosphatase expression, a decrease in NF-κB translocation to the nucleus and an increase in IL-10 production. Overall, DCs generated from bone marrow with Flt-3 ligand have similar characteristics to DC subtypes found in the steady state in vivo, which can acquire endotoxin tolerance in some circumstances.     

密植条件下种植方式对夏玉米群体根冠特性及产量的影响

选用平展大穗型品种鲁单981(Ludan981,LD981)和紧凑中穗型品种鲁单818(Ludan818,LD818),在两种种植密度(60000和90000株/hm2)和两种种植方式(单株和双株)下,研究了密植及种植方式对夏玉米冠层和根系结构与功能以及子粒产量等的影响.研究发现,随种植密度增加,冠层垂直分布呈现干重比例权重上移的趋势,根系则呈现下移的趋势.密植条件下,LD981冠层对生长空间更为敏感,其根系对生长空间的竞争强于冠层,其群体产量限制因素是子粒库容;LD818根系对生长空间更为敏感,冠层对生长空间的竞争强于根系,其群体产量限制因素是单位面积穗数.60000株/hm2下,LD981的群体结构质量和功能较优,双株种植可缓解其冠层竞争,根、冠协调,表现增产;在90000株/hm2下,LD818的群体结构质量和功能较优,双株种植可缓解其根系竞争,部分改善冠层群体结构质量和功能,根、冠协调,表现增产.     

用甘蓝苗连续饲养小菜蛾的技术

室内实验结果表明 :在 ( 2 5± 1 )℃、相对湿度 5 0 %～ 70 %、光周期 1 6L∶8D条件下 ,用甘蓝苗(BrassicanapusL .)饲养小菜蛾PlutellaxylostellaL .可取得较好的饲养结果 ,连续饲养多代种群未出现退化现象 ,能够得到大量发育整齐的供试虫源。在低温 ( 4℃ )条件下 ,卵冷藏 7d ,孵化率 77 8% ,蛹冷藏 1 5d化蛹率 86 9% ,冷藏后卵和蛹的发育等与各自的对照基本一致 ,可以较好地调控小菜蛾发育进度 ,便于取得虫龄基本一致的虫源。     

A study of the genetic variation of the aquatic fern Marsilea quadrifolia L. preserved in botanical collections in Poland and originated from natural populations in Europe

Aim of the present study was to evaluate the genetic diversity of selected European populations of Marsilea quadrifolia L. and to assess the applicability of those genetic resources of Marsilea quadrifolia L. that have been preserved in Polish botanical gardens, for the reintroduction of this species into its historical range in Poland. Three Polish populations that originated from botanical collections (Zabrze, ?arów and Pu?awy) and four natural populations (two from Slovakia (Slovakia I and Slovakia II), one from France and one from Germany) were analyzed using Amplified Fragment Length Polymorphism (AFLP) markers. A very low level of genetic variation was found both within and between the populations in the study, which likely resulted from a genetic bottleneck probably caused by human activities. Plants with the same AFLP fingerprint were found across several populations; however, singleton samples with a unique AFLP band pattern were also present within all of the populations. The presence of singletons led to relatively high values of Simpson's diversity index, which may suggest a considerable effect of mutations and some possibility of sexual reproduction as sources of the observed variation. The partitioning of molecular variance was calculated using hierarchical AMOVA, which showed that a negligible value of only 0.81% of the variation was explained by the category of population, i.e. plants originating from the botanical collections or from the natural habitats. This result indicates that M. quadrifolia populations from botanical collections resemble natural populations in terms of the level of their genetic variation and that the populations that were obtained from the Polish collections could be used for the successful reintroduction of this species into its historical range in Poland, and a similar situation may be given also in other areas of occurrence of this plant that is under threat throughout its area of occurrence in Europe.     

萌发玉米种子脱水耐性与胚根细胞超微结构的变化

玉米种子脱水试验表明,25℃下萌发24h种子脱水耐性开始丧失,丧失50％和100％的时间分别为33h、58h。萌发过程中随着吸胀时间增加,玉米种子脱水耐性逐步丧失。显微观察显示,种子吸胀过程中,胚根细胞的贮藏物质逐步减少,线粒体等细胞器的分化程度则不断提高,尤其是脂类物质的分解程度与脱水耐性变化的关系似乎更明显。     

Identification of d-Erythro-Dihydroneopterin Triphosphate as a Product of Guanosine Triphosphate Cyclohydrolase from Serratia indica

Guanosine triphosphate cyclohydrolase (EC 3.5.4.16) was previously shown to exist in two forms (GTP cyclohydrolase D-I and D-II) in Serratia indica IFO 3759, and they were homogeneously isolated. The present study deals with the characterization of their reaction products. A fluorescent product formed from guanosine triphosphate by GTP cyclohydrolase D-II was identified as 7,8-dihydroneopterin triphosphate by its absorption spectra, phosphate analysis and gas chromatography-mass spectrometry of the dephosphorylated trimethylsilyl derivative. After oxidation and dephosphorylation, the d-erythro configuration of the side chain was made clear by the elution profile on ECTEOLA-cellulose chromatography, Rf values on thin-layer chromatography and by biological activity to Crithidia fasciculata ATCC 12857. The fluorescent products from GTP cyclohydrolase D-I and D-II were indistinguishable.     

Aluminium causes nutrient imbalance and structural changes in the needles of Scots pine without inducing clear root injuries

The effects of aluminium chloride (AICI3) treatments (50 and 150 mg/l) on 3-year-old Scots pine (Pinus sylvestris L.) seedlings were studied in a sand culture during 2 growing periods in an open field experiment. Even by the end of the first growing period, a decline was observed in the concentrations of Ca, Mg and P within the needles, and of Ca and Mg in the roots. After the second growing period, increased N and K concentrations were observed in the needles of Al-treated seedlings. Both the needles and roots of Al-treated seedlings showed, after the second growing period, a decline in growth and increased concentrations of AI as the amount of AICI3 in the nutrient solution increased. Al-induced changes in needle structure were found to be symptomatic of a nutrient imbalance, particularly of Mg and P. Al-stress did not result in any observable changes in root anatomy or in the number of mycorrhizas. Scots pine proved to be rather resistant to Al-stress, indicating that direct Al-injuries are not likely in the field, though Al-stress may be a contributing factor in the formation of nutrient imbalances.     

结球甘蓝下胚轴原生质体培养再生植株体系的优化研究

以结球甘蓝‘新夏50’的无菌苗下胚轴为材料,对影响原生质体分离、纯化与培养的主要因素进行研究,建立适合结球甘蓝原生质体游离、纯化、收集、培养以至再生出完整植株的实用技术体系,为其非对称细胞融合及品种改良与创新等研究奠定基础。结果表明:2.5%纤维素酶R-10+0.05%果胶酶Y-23+9CPW+5mmol/L MES的混合酶液,从4d苗龄的下胚轴上分离出高产率的原生质体。在改良B5+0.5mg/L 2,4-D+0.2mg/L 6-BA+0.2mg/L NAA的液体培养基上,原生质体分裂旺盛。形成愈伤组织后经芽诱导和生根培养,获得了再生植株。倍性检测结果表明,不同原生质体所获得的24株再生植株中,19株为正常二倍体,4株为嵌合体,1株为四倍体。     

Vertebrate evolution reflected in the evolution of nuclear ribosomal internal transcribed spacer 2

In eukaryotes, mature rRNA sequences are produced from single large (45S) precursor (pre-rRNA) as the result of successive removal of spacers through a series of rapid and intricate actions of endo- and exonucleases. The excision of internal transcribed spacer (ITS2), a eukaryotic-specific insertion, remains the most elusive processing step. ITS2 is the element mandatory for all eukaryotic pre-rRNAs that contain at least three processing cleavage sites for precise 5.8S and 28S formation. Conserved core sequences (cis-elements) binding to trans-factors provide for precise rRNA processing, whereas rapidly diverging regions between the core sequences preserve internal complementarity, which guarantees the spatial integrity of ITS2. Characteristic differences in the formation of such insertions during evolution should reflect the relationships between taxa. The phylogeny of the reptiles and the relationships between taxa proposed by scientists are controversial. To delineate the structural and functional features preserved among reptilian ITS2s, we cloned and sequenced 58 ITS2s belonging to four reptile orders: Squamata, Crocodilians, Aves, and Testudines. We studied the subsequent alignment and folding of variable regions. The sizes and packing of the loop–stems between conserved consensus segments in reptiles vary considerably between taxa. Our phylogenetic trees constructed on the basis of the reptile ITS2s primary structural alignments revealed a split between Iguania clade and all other taxa. True lizards (suborder Scleroglossa) and snakes (suborder Serpentes) show sister relationships, as well as the two other reptilian orders, Crocodilia + Aves and Testudines. In summary, our phylogenetic trees exhibit a mix of specific features deduced or, to the contrary, rejected earlier by other authors.