On the propensity of lignin to associate: a size exclusion chromatography study with lignin derivatives isolated from different plant species

Despite evidence that lignin associates under both aqueous and organic media, the magnitude and nature of the underlying driving forces are still a matter of discussion. The present paper addresses this issue by examining both solution properties and size exclusion behaviour of lignins isolated from five different species of softwoods, as well as from the angiosperms Eucalyptus globulus and wheat straw. This investigation has used the recently described protocol for isolating enzymatic mild acidolysis lignin (EMAL), which offers lignin samples highly representative of the overall lignin present in the wood cell wall. The molecular weight distributions of these EMALs were found to be dependent upon the wood species from which they were isolated and upon the incubation conditions used prior to size exclusion chromatography. While the chromatograms of EMALs isolated from softwoods displayed a bimodal behaviour, the elution profiles of EMAL from E. globulus and straw were nearly unimodal. A marked tendency to dissociate prevailed under incubation at room temperature for all examined species with the exception of the straw lignin preparation; furthermore, lignin solutions incubated at 4 degrees C showed an associative behaviour manifested by an increase in the weight and number average molecular weights for some species. The extent of such association/dissociation, as well as the time needed for the process to reach completion, was also found to depend upon the wood species, i.e. lignins from softwoods were found to associate/dissociate to a greater extent than lignins from E. globulus and straw. The origin of such effects within the lignin structure is also discussed.     

三种桉叶油化学成分研究

通过水蒸汽蒸油试验,发现已贮藏半年多的风干桉叶,仍有较高的含油量。云南省弥勒产的直杆桉(Eucalyptus maidenii)得油率为4.14％,蓝桉(E.globulus)为3.5％,福建省惠安产的窿缘桉(E.exserta)为1.2％,桉叶油生产厂也可以通过贮藏的风干桉叶生产桉叶油。通过气相色谱和质谱分析,从直杆桉叶油中鉴定出44个组份,蓝桉叶油中鉴定出35个组份,窿缘桉叶油中鉴定出26个组份。三种桉叶油单萜部分的组份基本一致,只是相对含量各有不同。桉油素含量,以直杆桉最高为68.02％,其次是蓝桉为67.54％,窿缘桉最低为34.33％。直杆桉、蓝桉可以怍材、油两用树种,窿缘桉不宜作油用树种。     

The numerical intraspecific taxonomy of Eucalyptus globulus Labill. (Myrtaceae)

The history of the taxonomic perception of Eucalyptus globulus Labill. is described. Morphological-geographical clusters within E. globulus are identified through the use of a polythetic, agglomerative classificatory strategy. The results of the numerical analysis and unpublished data on geographic variation strongly suggest that four subspecies should be recognized. Thus three taxa formerly regarded as species are reduced to subspecies of E. globulus.     

Antimicrobial activities of eucalyptus leaf extracts and flavonoids from Eucalyptus maculata

AIMS: We investigated the antimicrobial activities of eucalyptus leaf extracts to find effective antibacterial agents. METHODS AND RESULTS: The antimicrobial activities of leaf extracts from 26 species of eucalyptus were measured. Extracts of Eucalyptus globulus, E. maculata and E. viminalis significantly inhibited the growth of six Gram-positive bacteria (Staphylococcus aureus, MRSA, Bacillus cereus, Enterococcus faecalis, Alicyclobacillus acidoterrestris, Propionibacterium acnes), and of a fungus (Trichophyton mentagrophytes), but they did not show strong antibacterial activity against Gram-negative bacteria (Escherichia coli, Pseudomonas putida). 2',6'-dihydroxy-3'-methyl-4'-methoxy-dihydrochalcone, eucalyptin and 8-desmethyl-eucalyptin, isolated from E. maculata extracts, exhibited potent antimicrobial activities against seven micro-organisms with minimum inhibitory concentrations (MIC) ranging from 1.0 to 31 mg l(-1). CONCLUSIONS: The eucalyptus extracts and three compounds from E. maculata were found to be effective against micro-organisms that cause food poisoning, acne and athlete's foot. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows potential uses of extracts from E. globulus, E. maculata and E. viminalis, and antimicrobial compounds isolated from E. maculata.     

A perennial ryegrass CBF gene cluster is located in a region predicted by conserved synteny between Poaceae species

CBF/DREB1 proteins are the most important regulators of the cold temperature signaling pathway in many plants. CBF genes are candidates for low-temperature tolerance QTL in wheat and barley. Ten novel putative CBF cDNAs of perennial ryegrass (Lolium perenne L.) have been isolated from cold-treated leaf tissue. Their primary structures contain some conserved motifs, characteristic of the gene class. Phylogenetic analysis revealed that LpCBF genes were attributable to the HvCBF3-, and HvCBF4-subgroups following the previously proposed classification of barley CBF genes. RT-PCR analysis revealed that the expression of LpCBF genes was rapidly induced in response to low temperature and that the expression pattern under the low-temperature conditions for a long period was different between the various LpCBF genes. Five of the ten LpCBF genes were assigned to the genetic linkage map using the p150/112 reference mapping population. LpCBFIb, LpCBFII, LpCBFIIIb and LpCBFIIIc were mapped on LG5 forming a cluster within 2.2 cM, while LpCBFVb was located on LG1. Based on comparative genetic studies, conserved synteny for CBF gene family was observed between the Triticeae cereals and perennial ryegrass. Information on the perennial ryegrass CBF genes at both the molecular and genetic level obtained in this study would be useful for the further study on the role of CBF genes and low-temperature tolerance in grasses.     

Temperatures lethal for citrus blackfly parasites,Encarsia opulenta andE. Smithi [Hym.: Aphelinidae]

Seasonally acclimatized adult and immature parasites of the citrus blackfly (CBF),Aleurocanthus woglumi Ashby, were exposed to high or low temperature extremes for 3 h periods. Death of all summer adults ofEncarsia opulenta Silvestri andE. smithi Silvestri occurred between 35° and 40°C. Within CBF hosts,E. opulenta were not able to emerge when temperatures reached between 45° and 50°C. In winter experiments adults of bothEncarsia species succumbed between −5° and −10°C. In a comparison of the 2 seasonal tests, a higher percentage ofE. smithi adults were able to survive both higher and lower temperatures thanE. opulenta, but the main interspecific difference was the ability ofE. opulenta within CBF to survive −10° to − 15°C whileE. smithi did not. Limited data forAmitus hesperidum Silvestri [Hym.: Platygasteridae] indicated that the immatures survived better at low, and not as well at high, temperatures as either species ofEncarsia. Florida Agricultural Experiment Station Journal Series # 5549.     

An essential yeast protein, CBF5p, binds in vitro to centromeres and microtubules.

Yeast centromere DNA (CEN) affinity column chromatography has been used to purify several putative centromere and kinetochore proteins from yeast chromatin extracts. The single yeast gene (CBF5) specifying one of the major low-affinity centromere-binding proteins (p64'/CBF5p) has been cloned and shown to be essential for viability of Saccharomyces cerevisiae. CBF5 specifies a 55-kDa highly charged protein that contains a repeating KKD/E sequence domain near the C terminus, similar to known microtubule-binding domains in microtubule-associated proteins 1A and 1B, CBF5p, obtained by overexpression in bacterial cells, binds microtubules in vitro, whereas C-terminal deleted proteins lacking the (KKD/E)n domain do not. Dividing yeast cells containing a C-terminal truncated CBF5 gene, producing CBF5p containing only three copies of the KKD/E repeat, delay with replicated genomes at the G2/M phase of the cell cycle, while depletion of CBF5p arrests most cells in G1/S. Overproduction of CBF5p in S. cerevisiae complements a temperature sensitivity mutation in the gene (CBF2) specifying the 110-kDa subunit of the high-affinity CEN DNA-binding factor CBF3, suggesting in vivo interaction of CBF5p and CBF3. A second low-affinity centromere-binding factor has been identified as topoisomerase II.     

Biomass production and nutrient cycling in Eucalyptus short rotation energy forests in New Zealand. I: Biomass and nutrient accumulation

Accumulation of biomass and nutrients (N, P, K, Ca, Mg and Mn) was measured during the first 3-year rotation of three Eucalyptus short rotation forest species (E. botryoides, E. globulus and E. ovata) irrigated with meatworks effluent compared with no irrigation. E. globulus had the highest biomass and nutrient accumulation either irrigated with effluent or without irrigation. After 3-year growth, E. globulus stands irrigated with effluent accumulated 72 oven dry t/ha of above-ground total biomass with a total of 651 kg N, 55 kg P, 393 kg K, 251 kg Ca, 35 kg Mg and 67 kg Mn. Effluent irrigation increased the accumulation of biomass, N, P, K and Mn, but tended to reduce the leaf area index and leaf biomass, and decreased the accumulation of Ca and Mg.     

萝卜低温胁迫转录因子的克隆及遗传转化

采用RT-PCR和电子克隆技术从抗寒植物萝卜(Raphanus sativusL.)中分离了一个低温胁迫转录因子的cDNA全长序列,命名为RsICE1(GenBank登录号为HQ891287).序列分析显示,该基因全长1 375 bp,编码区为1266 bp,编码421个氨基酸.序列比对表明,该蛋白C端含有一个典型的bHLH结构域,与其他植物的ICE1蛋白具有较高的同源性.进化树分析表明,RsICE1编码的蛋白与油菜的ICE1编码蛋白亲缘关系最近,处在同一进化分枝.半定量RT-PCR分析表明,RsICE1是冷诱导条件下差异表达的基因.构建该基因的植物表达载体,利用农杆菌介导法转化粳稻品种龙粳24,经PCR和RT-PCR分子检测,证明RsICE1基因已经整合到水稻基因组中,并正常表达.与对照相比,低温处理后转基因株系存活率和脯氨酸含量明显增加,相对电导率积累速率明显下降,提高了抗低温胁迫能力.     

Characterization of a cellulose-binding, cellulase-containing complex in Clostridium thermocellum.

The isolation and biochemical characterization of the extracellular form of a cellulose-binding factor (CBF) from Clostridium thermocellum is described. The CBF was isolated from the culture supernatant by a two-step procedure which included affinity chromatography on cellulose and gel filtration on Sepharose 4B. The isolated CBF was homogeneous as determined by immunoelectrophoresis, polyacrylamide gel electrophoresis, gel filtration, and analytical ultracentrifugation analysis. The CBF was found to form a complex which exhibited a molecular weight estimated at 2.1 million. Electron microscopic analysis of negatively stained preparations of the isolated CBF revealed a particulate, multisubunit entity of complicated quaternary structure. The molecule appeared to be about 18 nm in size. Although urea failed to break the complex into its component parts, polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate resolved the CBF complex into 14 polypeptide bands. Immunoprecipitation experiments confirmed that these polypeptides indeed formed part of the same complex. Interestingly, by using the whole-cell immunization procedure described in the accompanying article (Bayer et al., J. Bacteriol., 156:818-827, 1983) only one CBF subunit (Mr = 210,000) was found to be antigenically active. By using a gel-overlay assay technique, at least eight of the remaining CBF-associated polypeptide components were shown to exhibit cellulolytic activity. The results are consistent with the contention that the CBF comprises a discrete, multisubunit complex or group of closely related complexes which exhibit separate antigenic and multiple cellulase activities in addition to the property of cellulose binding.(ABSTRACT TRUNCATED AT 250 WORDS)     

The rare silver gum, Eucalyptus cordata, is leaving its trace in the organellar gene pool of Eucalyptus globulus

The process of genetic assimilation of rare species by hybridizing congeners has been documented in a number of plant genera. This raises the possibility that some of the genetic diversity found in phylogeographical studies of widespread species has been acquired through hybridization with species that are now rare or extinct. In this fine-scale phylogeographical analysis, we show that a rare eucalypt species is leaving its trace in the chloroplast genome of a more abundant congener. The heart-leafed silver gum, Eucalyptus cordata, is a rare endemic of south-eastern Tasmania. Its populations are scattered amidst populations of more abundant related species, including the Tasmanian blue gum, Eucalyptus globulus. Using 339 samples from across the full range of both species, we compared chloroplast (cp) DNA haplotype phylogeography in E. globulus and E. cordata. The genealogy and distribution of chloroplast haplotypes suggest that E. globulus has acquired cpDNA from E. cordata in at least four different mixed populations. Shared haplotypes are highest in E. globulus sampled within 2 km of known E. cordata populations and drop to zero at a distance of 25 km from the nearest known E. cordata population. Localized haplotype sharing occurs in the absence of obvious hybrid zones or locally shared nuclear ribosomal DNA sequences. Given that the future loss of E. cordata from some mixed populations is likely, these findings indicate that phylogeographical analyses of organellar DNA should consider the possibility of introgression, even from species that have been eliminated from the sites of interest.