Electrophoretic evaluation of the taxonomic status of two species of sea anemone

Puerto Rican populations of two species of sea anemones (Bunodosoma cavernata and B. granulifera) which had previously been considered one were assayed electrophoretically for enzymes encoded by 12 loci. The two species shared no common allozymes at 6 of the 12 loci. Genetic distance and identity values based on these allozymes were computed for the Puerto Rican populations and for B. cavernata from Florida and B. granulifera from Panama. The Puerto Rican populations of both species had much higher genetic identities for their geographically distant conspecifics than for each other. These results indicate that the two species are reproductively isolated and should be considered as separate valid species. Average heterozygosities are presented which are the first published for coelenterate species.     

Isoenzyme diversity and phylogenetic relationships among the American beans of the genus Vigna Savi (Fabaceae)

Electrophoretic variation of 13 isoenzyme systems encoded by 25 gene loci has been studied among the 12 American bean species of the genus Vigna, subgenera Sigmoidotropis and Lasiospron, in comparison with the two pantropical species Vigna luteola and V. vexillata of the subgenera Vigna and Plectrotropis, respectively. Of the 202 electromorphs (putative allozymes) discovered, 106 were phylogenetically informative, i.e. shared by at least two taxa, and 96 were taxon-specific. Parsimony, neighbour-joining and UPGMA analyses of the isozyme data as unordered presence–absence characters revealed the division of the American Vigna species into six clades which mostly correspond to the grouping of species into subgenera and sections in the traditional morphology-based systems, except in the placement of species V. gentryi and V. caracalla. The type species of the section Caracallae, V. caracalla, appears on the cladogram as basally paraphyletic in the subgenus Sigmoidotropis, but apart from the V. linearis group of the same section which forms a separate subclade. Vigna gentryi is not linked to other species of the section Sigmoidotropis, but appears in a distant subclade as a sister species to V. adenantha s. l. of the section Leptospron. The subgenus Lasiospron is sister to the pantropical V. vexillata of the African subgenus Plectrotropis.     

基于GC-MS代谢组学技术委陵菜与星毛委陵菜的初生代谢比较研究

为明确药用植物委陵菜与其同属植物星毛委陵菜在不同基原之间的代谢差异，更有效的利用药用植物资源。基于气相色谱与质谱联用技术对两药用植物间的小分子初生代谢物进行非靶向代谢组学分析，利用主成分分析和偏最小二乘法分析，寻找可区分两物种的差异代谢物以及在初生代谢途径上的差异。结果：利用GC-MS在两植物中共检测到134个色谱峰，通过与数据库比对，共鉴定出43种初生代谢产物。PCA分析结果显示代谢物在委陵菜和星毛委陵菜间的分布有较大的差异；采用OPLS-DA筛选获得导致2物种区分的差异代谢物共33种，两物种在半乳糖代谢、淀粉和蔗糖代谢、氨酰生物合成、氨基糖和核苷酸糖代谢和果糖和甘露糖代谢途径有显著的差异。委陵菜和星毛委陵菜的初生代谢方面存在较大差异，虽然两植物来源于同科同属植物，但是星毛委陵菜不适合代替委陵菜作为药材使用。     

The novel peptide NbPPI1 identified from Nicotiana benthamiana triggers immune responses and enhances resistance against Phytophthora pathogens

In plants, recognition of small secreted peptides, such as damage/danger‐associated molecular patterns (DAMPs), regulates diverse processes, including stress and immune responses. Here, we identified an SGPS (Ser‐Gly‐Pro‐Ser) motif‐containing peptide, Nicotiana tabacum NtPROPPI, and its two homologs in Nicotiana benthamiana, NbPROPPI1 and NbPROPPI2. Phytophthora parasitica infection and salicylic acid (SA) treatment induced NbPROPPI1/2 expression. Moreover, SignalP predicted that the 89‐amino acid NtPROPPI includes a 24‐amino acid N‐terminal signal peptide and NbPROPPI1/2‐GFP fusion proteins were mainly localized to the periplasm. Transient expression of NbPROPPI1/2 inhibited P. parasitica colonization, and NbPROPPI1/2 knockdown rendered plants more susceptible to P. parasitica. An eight‐amino‐acid segment in the NbPROPPI1 C‐terminus was essential for its immune function and a synthetic 20‐residue peptide, NbPPI1, derived from the C‐terminus of NbPROPPI1 provoked significant immune responses in N. benthamiana. These responses led to enhanced accumulation of reactive oxygen species, activation of mitogen‐activated protein kinases, and up‐regulation of the defense genes Flg22‐induced receptor‐like kinase (FRK) and WRKY DNA‐binding protein 33 (WRKY33). The NbPPI1‐induced defense responses require Brassinosteroid insensitive 1‐associated receptor kinase 1 (BAK1). These results suggest that NbPPI1 functions as a DAMP in N. benthamiana; this novel DAMP provides a potentially useful target for improving plant resistance to Pytophthora pathogens.     

Genetic detection of cryptic species in the frillfin goby Bathygobius soporator

We compared, through allozyme and cytochrome b sequence analyses, populations of Bathygobius soporator from four localities on the Brazilian coast with those from the Oceanic Islands of the Rocas Atoll (230 km off the Brazilian coast) and the Bahamas (5700 km northwest from Rocas). The population from the Rocas Atoll was genetically more similar to the geographically distant Bahamas (gene identity, I=0.92) than to the supposedly conspecific populations from the Brazilian coast (I<0.55). Five diagnostic allozyme loci and a high level of nucleotide divergence (Kimura 2-parameters=0.21) were found between the two groups, further confirming that they must belong to different biological species. Since the type locality of B. soporator is in the Caribbean, the binomial should be maintained for the Bahamas/Rocas Atoll populations, and the other Brazilian populations of Bathygobius analysed either represent the occurrence of a species of Bathygobius hitherto not cited for the Brazilian coast, or belong to a new species. The coastal populations of this species were also found to be highly structured (F_ST=0.18; p<0.05), indicating that, even along the coast, levels of gene flow of this species are limited. This could be related to their reproductive biology (adhesive benthic eggs with parental care and short length of larval life). The timings of the divergence between the Caribbean and the Southwest Atlantic species found here, estimated from allozymes and cytb sequences, are very similar (around 10 MY bp), and correlate with the start of the Amazon river outflow. The colonization of the Rocas Atoll by populations from the Caribbean seems to have taken place after the Caribbean and the Brazilian coast species had diverged.     

Structural comparison of lactate dehydrogenase homologs differing in sensitivity to hydrostatic pressure

The muscle-type (M4) lactate dehydrogenases (L-lactate: NAD+ oxidoreductase EC 1.1.1.27) of two teleost fishes, Sebastolobus alascanus and Sebastolobus altivelis , differ in the susceptibility of ligand binding to perturbation by moderate hydrostatic pressures. The enzyme homologs were purified by affinity chromatography. The amino-acid compositions of these enzymes are virtually identical. The proteins were digested with trypsin and the peptide mixtures mapped using reverse-phase HPLC. Although there was variation in elution times of some peaks, the amino-acid compositions of the fractions from the two profiles were highly similar. Only one clear difference in amino-acid composition was found and this peptide was sequenced using the manual dansyl-Edman method. The enzyme of S. alascanus , which is susceptible to pressure-perturbation, had a histidine at position 115; the S. altivelis enzyme had an asparagine. Ionization of histidine is affected by pressure and may be involved in the differences between the two lactate dehydrogenase homologs. There is no covalently bound phosphate associated with either enzyme, and thus phosphorylation cannot account for the differences between the enzyme homologs. Acquisition of pressure-tolerance appears to involve only minor changes in primary structure.     

The phylogeny and systematics of the endemic Ethiopian Lophuromys flavopunctatus species complex based upon random amplified polymorphic DNA (RAPD) analysis

Random amplified polymorphic DNA (RAPD) analysis was performed to clarify systematics and phylogenetic relationships within the Ethiopian endemic representatives of Lophuromys flavopunctatus species complex. Data were analysed by both phenetic (UPGMA) and phylogenetic (neighbor-joining (NJ) and maximum parsimony) procedures. NJ and maximum parsimony analyses yielded identical phylogenetic trees that demonstrate the basal position of L. melanonyx with L. brevicaudus splitting next and sister-group relationship for L. brunneus–L. chrysopus. This phylogenetic pattern is congruent with inferences from allozymes for the considered species suggesting early divergence of Afroalpine species and recent origin of forest taxa. Thus, the results demonstrate that RAPD-PCR might be a useful technique for phylogenetic analysis at the species levels in vertebrates. Controversial taxonomy of L. brevicaudus, L. brunneus and L. chrysopus is clarified, with their specific ranks confirmed on the basis of tree topology and genetic distances.     

Pore-forming peptides of Entamoeba dispar. Similarity and divergence to amoebapores in structure, expression and activity.

Amoebapore, a 77-residue peptide with pore-forming activity from the human pathogen Entamoeba histolytica, is implicated in the killing of phagocytosed bacteria and in the cytolytic reaction of the amoeba against host cells. Previously, we structurally and functionally characterized three amoebapore isoforms in E. histolytica but recognized only one homolog in the closely related but non-pathogenic species Entamoeba dispar. Here, we identified two novel amoebapore homologs from E. dispar by molecular cloning. Despite strong resemblance of the primary structures of the homologs, molecular modeling predicts a species-specific variance between the peptide structures. Parallel isolation from trophozoite extracts of the two species revealed a lower amount of pore-forming peptides in E. dispar and substantially higher activity of the major isoform from E. histolytica towards natural membranes than that from E. dispar. Differences in abundance and activity of the lytic polypeptides may have an impact on the pathogenicity of amoebae.     

The geographic distribution and host range of Capillaria hepatica (Bancroft) (Nematoda) in Australia.

The geographic distribution, host range and prevalence of Capillaria hepatica were recorded in 4629 house mice, Mus domesticus, 263 black rats, Rattus rattus, and 58 Norway rats, R. norvegicus. The parasite was found at five localities, all in or near large towns along the coast. The two Rattus species appeared to be the primary hosts of C. hepatica in Australia. Published and unpublished data on helminth infections of Australian native mammals from 1162 murids (26 species), 3018 marsupials (67 species) and 99 monotremes (two species) were compiled. Only seven animals from three murid species were infected with C. hepatica; all were from the same rainforest in northern Queensland. C. hepatica was distributed widely, occurring in the house mouse, black rat and Norway rat on a 10,850 ha farm but there was no infection in cattle, sheep or goats (abattoir records). Also, 52 rabbits, four cats and one fox (shot samples) and 27 marsupial mice, Sminthopsis crassicaudata (museum specimens), had no sign of C. hepatica infection. Overall, the results indicate that transmission of C. hepatica to native, domestic and feral mammals is rare, presumably because of ecological constraints on egg embryonation and survival. In the light of these findings, the potential use of C. hepatica as a biological agent to control mouse plagues in Australia is discussed.     

Trichinella zimbabwensis n.sp. (Nematoda), a new non-encapsulated species from crocodiles (Crocodylus niloticus) in Zimbabwe also infecting mammals

Since 1995, Trichinella larvae have been detected in 39.5% of farmed crocodiles (Crocodylus niloticus) in Zimbabwe. Morphological, biological, biochemical and molecular studies carried out on one isolate from a farmed crocodile in 2001 support the conclusion that this parasite belongs to a new species, which has been named Trichinella zimbabwensis n.sp. This species, whose larvae are non-encapsulated in host muscles, infects both reptiles and mammals. The morphology of adults and larvae is similar to that of Trichinella papuae. Adults of T. zimbabwensis cross in both directions with adults of T. papuae (i.e. male of T. zimbabwensis per female of T. papuae and male of T. papuae per female of T. zimbabwensis), producing F1 offspring which produce very few and less viable F2 larvae. Muscle larvae of T. zimbabwensis, like those of T. papuae, do not infect birds. Three allozymes (of a total of 10) are diagnostic between T. zimbabwensis and T. papuae, and five are diagnostic between T. zimbabwensis and Trichinella pseudospiralis, the third non-encapsulated species. The percentage of the pairwise alignment identity between T. zimbabwensis and the other Trichinella species for the cytochrome oxidase subunit I gene, the large subunit ribosomal-DNA (mt-lsrDNA) gene and the expansion segment five, shows that T. zimbabwensis is more similar to the two non-encapsulated species T. papuae (91% for cytochrome oxidase I; 96% for mt-lsrDNA; and 88% for expansion segment five) and T. pseudospiralis (88% for cytochrome oxidase I; 90% for mt-lsrDNA; and 66–73% for expansion segment five) than to any of the encapsulated species (85–86% for cytochrome oxidase I; 88–89% for mt-lsrDNA; and 71–79% for expansion segment five). This is the first non-encapsulated species discovered in Africa. The finding of a new Trichinella species that infects both reptiles and mammals suggests that the origin of Trichinella parasites dates back further than previously believed and can contribute to understanding the phylogeny and the epidemiology of the genus Trichinella.     

A comparison of hemoglobins in five species of Microtus

The composition of tryptic peptides was determined for Hb (major or fast electrophoretic component) from four additional species of Microtus; M. p. pennsylvanicus, M. abbreviatus, M. miurus, and M. oeconomus, The amino acids from the four Hb were compared with Hb from M. p. tananaensis, and, on the basis of the combination of analyses for the several Hb. Ca 95% of the overall amino acid composition was considered. The compositions of most of the homologous peptides were identical, and two deletions in the sequence of 21 amino acids β 41–61 are believed to characterize the Hb of all four species. From differences in peptide composition the following substitutions were inferred. In the β chain, M. pennsylvanicus (2 ssp) differed from other species at two positions, 8: Ser vs Thr and 12: Thr vs Ash. In the β chain M. pennsylvanicus (2 ssp) differed from other species at two positions, β45: Phe vs Leu, and β50: Ser-Glx vs Thr. M. p. pennsylvanicus differed from M. p. tananaensis at position β88: Val-Leu vs Leu. All species showed ambiguity among the amino acids Ala-Ser-Phe-Leu centred presumably in positions β129 and β130. On the basis of an incomplete examination, the slow electrophoretic component of M. abbreviatus Hb could not be seen to differ from the fast component in its peptide map or in the general composition of its and β peptides.     

Caffeine metabolism in Coffea arabica and other species of coffee

High performance liquid chromatography has been used to measure the quantities of caffeine, theobromine, and theophylline in aqueous extracts of endosperm from immature and mature fruits of Coffea arabica and six other species of Coffea. Caffeine was the alkaloid present in largest amounts and, with one exception, in concentrations that were broadly similar in immature and mature fruit. The highest concentrations of caffeine were found in C. canephora at 35.1 and 24.5 mg g⁻¹, respectively, in immature and mature endosperm. The lowest concentrations were in C. bengalensis, where caffeine was not detected in extracts from mature fruit. [8-³H]Caffeine was metabolised relatively slowly by immature endosperm of C. arabica and C. canephora. In contrast, C. dewevrei, C. eugenioides, C. stenophylla, C. salvatrix and C. bengalensis all appeared to metabolise [8-³H]caffeine much more rapidly, as the percentage recovery of the applied label was much lower and there was more extensive incorporation of radioactivity into theobromine, theophylline, 3-methylxanthine and two unidentified polar metabolites. The endogenous caffeine concentrations and the metabolism data indicate that there may be marked differences in the rate of turnover of caffeine in the various species of Coffea. Potential sources of material for the production of naturally decaffeinated coffee are discussed.     

拟南芥ACOL8基因在乙烯合成与响应中的功能分析

植物激素乙烯在多种生理生化过程中发挥重要作用,但其在特定组织器官中的合成机制尚不完全清楚。拟南芥中存在12个功能未知的ACC氧化酶类似蛋白（ACO-like homolog,ACOL）,运用基因定点编辑技术构建了ACOL8的功能丧失型突变体,发现该基因的突变削弱了经典的乙烯“三重反应”。与野生型相比,突变体黄化幼苗下胚轴及主根的长度显著增加,这与突变体对外源ACC的敏感性下降现象一致。同时还发现ACOL8基因的表达受乙烯信号的正反馈调控,EIN3过表达增强其表达水平,而etr1-3的突变则产生相反效应。再者,在正常条件下,ACOL8基因的突变并未影响拟南芥的生长;但在盐胁迫条件下,突变体的根冠比显著下降,这说明该基因参与植物的盐胁迫响应。综上,这些结果说明ACOL8可能具有ACC氧化酶的功能,参与乙烯的合成与响应。     

The Chinese bamboo leaf odorous frog (Rana (Odorrana) versabilis) and North American Rana frogs share the same families of skin antimicrobial peptides

The Chinese bamboo leaf odorous frog (Rana (Odorrana) versabilis) and the North American pickerel frog (Rana palustris) occupy different ecological niches on two different continents with no overlap in geographical distribution. R. palustris skin secretions contain a formidable array of antimicrobial peptides including homologs of brevinin-1, esculentin-1, esculentin-2, ranatuerin-2, a temporin and a family of peptides considered of unique structural attributes when isolated, palustrins 1-3. Here we describe the structures of mature peptides and precursors of eight putative antimicrobial peptides from the skin secretion of the Chinese bamboo leaf odorous frog (Rana (Odorrana) versabilis). Each peptide represents a structural homolog of respective peptide families isolated from R. palustris, including two peptides identical in primary structure to palustrin 1c and palustrin 3b. Additionally, two peptides were found to be structural homologs of ranatuerin 2B and ranatuerin 2P from the closely-related North American species, Rana berlandieri (the Rio Grande leopard frog) and Rana pipiens (the Northern leopard frog), respectively. Both palustrins and ranatuerins have hitherto been considered unique to North American ranid frogs. The use of primary structures of amphibian skin antimicrobial peptides is thus questionable as a taxonomic device or alternatively, the micro-evolution and/or ancestry of ranid frogs is more highly complex than previously thought.     

Flavonoids and the systematics of Luffa

Flavonoids were characterized from leaves and flowers of six species of Luffa. Each species had a distinct flavonoid pattern. Based on leaf flavonoids, Luffa is separable into two groups of species: L. graveolens and L. operculata contain only flavonols. L. acutangula, L. aergyptiaca, L. echinata and L. forskalii contain only flavones. Flavonoid data indicate that the New World disjunct species. L. operculata, is most closely related to L. graveolens.     

Pituitary adenylate cyclase-activating polypeptide (PACAP) type 1 receptor (PAC1R) co-localizes with activity-dependent neuroprotective protein (ADNP) in the mouse brains

The crustacean hyperglycemic hormone is the most abundant neuropeptide present in the eyestalk of Crustacea and its main role is to control the glucose level in the hemolymph. Our study was aimed at assessing the importance of C-terminal amidation for its biological activity. Two recombinant peptides were produced, Asl-rcHH-Gly with a free carboxyl terminus and Asl-rcHH-amide with an amidated C-terminus. Homologous bioassays performed on the astacid crayfish Astacus leptodactylus showed that the amidated peptide had a stronger hyperglycemic effect compared to the non-amidated peptide. To assess the relevance of amidation also in other decapods and how much the differences in the cHH amino acid sequence can affect the functionality of the peptides, we carried out heterologous bioassays on the cambarid Procambarus clarkii and palaemonid Palaemon elegans. The Asl-rcHH-amide elicited a good response in P. clarkii and in P. elegans. The injection of Asl-rcHH-Gly evoked a weak response in both species. These results prove the importance of C-terminal amidation for the biological activity of cHH in crayfish as well as the role of the peptide primary sequence for the species-specificity hormone-receptor recognition.