Macroscopic and Microscopic Morphological Features of Stromatolites Related To Activity of Eukaryote-Dominated Biofilms in an Acid Mine Drainage Environment: Biosignatures and Understanding Preservation of Stromatolites as Trace Fossils

Euglena-, diatom-, and algae-dominated biofilms are the principal producers of iron-rich biolaminates that result in biosedimentary structures, or stromatolites, in an acid mine drainage (AMD) environment in Indiana. These structures are considered trace fossils because they are produced by organism-sediment interactions and record physicochemical conditions of the environment. Our purpose was to link the biofilm types to specific micro- and micromorphological features and the physicochemical conditions under which they were formed. Analyses revealed that Euglena-dominated biofilm produced thin, porous microlaminae by trapping, binding, and relocating AMD precipitates as the biofilm kept pace with chemical sedimentation. More massive microlaminae were produced by high rates of chemical sedimentation brought on by increased discharge and dilution of acidity. Diatom- and algae-dominated biofilms produced thick, mm–cm-scale, porous, spongelike micro- to macrolaminae through oxygenic photosynthesis and/or metal uptake in extracellular polymeric substances, which promoted mineral precipitation on cell walls to create a rigid, porous structure. The variations in biolaminate textures within the stromatolites record seasonal changes in the microbial populations and physicochemical conditions of the AMD environment. These iron-rich stromatolites represent trace fossils that record morphological biosignatures of eukaryote-dominated microbial biofilms and may serve as appropriate proxies in the search for similar evidence of eukaryotic life in other iron-rich paleoenvironments, such as those on early Earth and Mars.     

Morphometrics and SEM analysis of the species pair Lithobius mutabilis L. Koch, 1862 and L. glacialis Verhoeff, 1937 (Chilopoda: Lithobiomorpha)

The sets of morphological characters of two nominal species in the genus Lithobius Leach, 1814, Lithobius mutabilis L. Koch, 1862 and Lithobius glacialis Verhoeff, 1937, were studied in order to test specific dissimilarity. Morphometrics were examined and analysed statistically. In addition, a comparative SEM analysis of external characters was made. Morphometrically significant differences between the species were found concerning body length, the width ratio of the head to the 5th tergite, the number of antennal articles, and the length/width proportions of certain articles (e.g. the femur) of the 15th legs of males. Because of these and several additional differences, such as in tergite surface structure, modifications to the 15th legs of males, the structure of the female gonopod claws, and between the ecological profiles, L. glacialis, which is found in the high Alps at altitudes above timberline (1300 m), can be distinguished clearly from L. mutabilis. This study demonstrates another example for a cryptic species pair, a common phenomenon between lowland and alpine populations of small invertebrates. The combination of morphometrics and SEM offers a powerful methodology for resolving previously uncertain questions in the species-level taxonomy of centipedes.     

Investigating the potential of Andean lupin as a lignocellulosic feedstock for Europe: First genome-wide association study on Lupinus mutabilis biomass quality

The development of multipurpose crops will drive the transformation of agricultural “waste” into added-value products, helping to meet biomass demands without competing with food production or increasing environmental pressure. Lupinus mutabilis, has been proposed not only as a valid source of protein and oil for Europe but also as a possible source of lignocellulosic feedstock for the biorefinery industry. In this study, the quality of L. mutabilis lignocellulosic biomass and its genetic architecture are investigated for the first time, using a panel of 223 accessions planted across three locations in two different European cropping conditions. Biomass quality was evaluated based on the estimation of neutral detergent fiber, cellulose, hemicellulose and acid detergent lignin fractions, and on the basis of the monosaccharide composition of cell wall polysaccharides. The broad variation in yield and composition of biomass encountered in the panel confirms the potential of L. mutabilis as lignocellulosic feedstock and points out the value of this panel as a breeding tool for the improvement of biomass quality. A genome-wide association study was conducted to identify single-nucleotide polymorphisms (SNPs) associated with biomass quality, both across locations and per specific location. Scanning of 16,781 SNPs across the whole genome identified 46 unique quantitative trait loci for biomass quality, 4 of which were detected as common either among traits or GWAS models. For each of the traits analyzed, between 3 and 10 SNPs were detected, explaining 2.7%–15.9% of the phenotypic variation. Underlying these loci, 28 genes were proposed as candidate genes for biomass quality. Important genes involved in cellulose and sucrose synthesis (CESA4, SPP1,WRKY33, GONST2), monolignol biosynthesis (SKIP31, WAT1, CCR-SNL6) and pectin degradation (RAV1, PE) were identified and will require validation to confirm their value for application in L. mutabilis breeding.     

Cultural manipulations affecting callus formation from seedling explants of the pearl lupin (Lupinus mutabilis Sweet)

The influences of several cultural factors were investigated in relation to callus formation from seedling explants of L. mutabilis Sweet. The interaction of low culture temperature (18°C), light differences, rapid callus transfer, and media additions led to the successful abolition of media browning which usually accompanied callus senescence. The supplement of activated charcoal slowed down the growth rate of the callus produced, and made it appear friable. Low levels of picloram and dicamba were found to produce large quantities of friable callus especially when combined with kinetin. The availability of nitrogen, as well as type of explant, markedly influenced callus induction and growth, whilst the addition of glutamine enhanced callus formation.     

木芙蓉三个品种及近缘种的叶绿体基因组比较分析

木芙蓉(Hibiscus mutabilis)栽培历史悠久,是原产中国的古老园林树种和药用植物。为了探讨木芙蓉品种及近缘种的进化特征,厘清木芙蓉品种间及其与近缘种间的亲缘关系,以及探究木芙蓉叶绿体基因组(chloroplast DNA, cpDNA)的遗传方式,该文选择了一个杂交组合中的3个木芙蓉栽培品种(‘单瓣白''‘金秋颂''‘牡丹粉''),用高通量测序平台Illumina NovaSeq对其cpDNA进行首次测序。经组装注释后得到3条完整的cpDNA序列,结合该团队已经完成的近缘种台湾芙蓉(H. taiwanensis)和来自基因库的木槿、朱槿的cpDNA,对木槿属4种及木芙蓉种下的3个品种进行了cpDNA组成和结构特征的比较分析,并完成了其系统发育树重建。结果表明:(1)‘单瓣白''‘金秋颂''‘牡丹粉''的cpDNA序列长度分别为160 880、160 879、160 920 bp,基因数目均为130个,其中蛋白编码基因85个、核糖体RNA 8个和转运RNA 37个。(2)比较分析结果显示,木芙蓉的种下3个品种及其近缘种台湾芙蓉在cpDNA上高度保守,反向重复区(IR)均为26 300 bp; 木槿和朱槿在IR区发生了收缩,分别为25 745、25 598 bp。(3)系统发育分析结果显示,种下3个品种先聚成一个单系支,再与台湾芙蓉聚成一个高支持率分支,表明木芙蓉和台湾芙蓉的亲缘关系最近; 相较于木槿和朱槿,木芙蓉、台湾芙蓉2种与海滨木槿、黄槿、大麻槿在亲缘关系上更近。(4)木芙蓉3个品种之间能通过cpDNA序列区分开,在大/小单拷贝区(LSC/SSC)长度上,‘单瓣白''‘金秋颂''‘牡丹粉''分别为89 355 bp/18 925 bp、89 353 bp/18 926 bp、89 400 bp/18 920 bp,并且从重复序列和核苷酸多样性分析中开发出了候选分子标记和DNA条形码,可以作为品种鉴定的分子条码。(5)木芙蓉品种‘单瓣白''与‘金秋颂''cpDNA差异最小,亲缘关系最近,根据两者母本与子代的关系,证明了木芙蓉cpDNA的母系遗传特征。该研究结果有助于更好地了解3个木芙蓉品种及台湾芙蓉cpDNA的进化特征和物种间的系统发育关系,为木芙蓉品种的准确鉴定和优良品种选育提供了cpDNA方面的基础资料。     

Characterization of a non-thermophilic strain of the red algal genus Galdieria isolated from Soos (Czech Republic)

Volcanic areas with highly acidic solfatara soils and temperatures of up to 56?°C are inhabited by the red algal genus Galdieria. We examined three highly acidic but non-volcanic habitats in the western part of the Czech Republic for the occurrence of this red alga. In soil samples from the National Nature Reserve of Soos we found, together with Euglena mutabilis, Pseudococcomyxa simplex and species of Chlorella, a new strain of Galdieria. In contrast to all other Galdieria strains described so far, the strain from Soos exhibited a low temperature optimum for growth of about 30?°C. Other properties, such as the substrate spectrum for heterotrophic growth, ultrastructure, fatty acid composition, thermostability of enzymes and the nitrogen source, showed no obvious differences from other strains of Galdieria. Within a phylogenetic tree based on 18S rRNA sequence data, the strain from Soos occupied a position at the base of the ‘Galdieria’-branch. Our findings indicate that the genus Galdieria is not restricted to volcanic and mining areas and that strains of Galdieria are able to compete successfully with green algae in habitats like Soos.     

Evaluation of responses of five cowpea (Vigna unguiculata (L.) Walp.) varieties to field infestation by major insect pests of cowpea

The study was conducted to evaluate yield response of five cowpea (Vigna unguiculata (L.) Walp.) varieties to field infestation and damage by major insect pests of cowpea. Three local varieties of cowpea, namely, Sokoto, Oloyin and Drum, and two International Institute of Tropical Agriculture (IITA) varieties, IT 90K-277-2 and IT90K-76, were planted on the Teaching and Research Farm of University of Agriculture, Abeokuta (UNAAB), Ogun State, Nigeria in September, 2006 and repeated in September, 2007. The experiment was laid out in Randomised Complete Block Design in split plots arrangement. Insecticidal spray constitutes the main plot while variety was the sub-plot. The main plot size was 19.4 m × 19 m and each of the sub-plot was 2.4 m × 3 m. The space between sub-plots was 1 m. Data from the sprayed plots were significantly (P < 0.05) higher than the unsprayed ones in the two seasons. Irrespective of season, the unsprayed plot of IT 90K-277-2 has a significantly (P < 0.05) high incidence of Ootheca mutabilis. The local variety, “Drum” has the highest incidence of flower thrip – Megalurothrips sjostedti in the two planting seasons. The highest mean number of peduncles/plant and pods/plant was in “IT 90K-277-2”. In both seasons, “IT90K-76” has a significantly (P < 0.05) lower number of damaged seeds and pods and “Oloyin” a significantly (P < 0.05) higher mean number of total pod sucking bugs. The grain yield was in the order IT90K-76 > Oloyin > IT 90K-277-2 > Sokoto > Drum.     

Purification of Pseudomonas sp. proteases through aqueous biphasic systems as an alternative source to obtain bioactive protein hydrolysates

Aqueous biphasic systems (ABSs) are an interesting alternative for separating industrial enzymes due to easy scale-up and low operational cost. The proteases of Pseudomonas sp. M211 were purified through ABS platforms formed by polyethylene glycol (PEG) and citrate buffer salt. Two experimental designs 2³ + 4 were performed to evaluate the following parameters: molar mass of PEG (M_PEG), concentration of PEG (C_PEG), concentration of citrate buffer (C_Cit), and pH. The partition coefficient (K), activity yield (Y), and purification factor (PF) were the responses analyzed. The best purification performance was obtained with the system composed of M_PEG = 10,000 g/mol, C_PEG = 22 wt%, C_Cit = 12 wt%, pH = 8.0; the responses obtained were K = 4.9, Y = 84.5%, PF = 15.1, and tie-line length = 52.74%. The purified proteases of Pseudomonas sp. (PPP) were used to obtain hydrolysates of Lupinus mutabilis (Peruvian lupin cultivar) seed protein in comparison with the commercial protease Alcalase® 2.4L. A strong correlation between hydrolysis degree and radical scavenging activity was observed, and the highest antioxidant activity was obtained with Alcalase® (1.40 and 3.47 μmol Trolox equivalent/mg protein, for 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) and oxygen radical absorbance capacity, respectively) compared with PPP (0.55 and 1.03 μmol Trolox/mg protein). Nevertheless, the IC₅₀ values were lower than those often observed for antioxidant hydrolysates from plant proteins. PEG/citrate buffer system is valuable to purify Pseudomonas proteases from the fermented broth, and the purified protease could be promising to produce antioxidant protein hydrolysates.