Nuclear division in the red algae Chondria nidifica and Chondria tenuissima

Cytological investigations are reported for two Chondria species, the Pacific species Chondria nidifica Harvey and Chondria tenuissima (Goodenough et Woodward) C. A. Agardh from the shore of the Marmara Sea in Istanbul. Nuclear division during mitosis and meiosis has been followed in somatic cells and in tetrasporangial mother cells respectively of diploid tetrasporic plants. The spherical interphase nucleus stains densely, showing many chromatin granules. Mitotic nuclei in the apical groove show a large number of chromosomes at metaphase; the chromosome number has been estimated at diakinesis to be 40 in both C. nidifica and C. tenuissima. The meiotic nuclei of tetraspore mother cells in prophase contain several relatively large nucleolar-derivatives in both species. The nucleolar derivatives disappear completely before the chromosomes begin to differentiate. In meiotic prophase the tetraspore mother cell enlarges from its original diameter. The period of the second meiotic anaphase seems to be extremely short in comparison with other nuclear phases. When the chromosomes reach the poles, they spread and subsequently form a relatively compact mass at telophase. The spindle has not been observed in C. tenuissima. Photographs are presented of nucleoli and nucleolar-derivatives in mitotic and meiotic divisions.     

O-Methylated mannogalactan from the microalga Coccomyxa mucigena, symbiotic partner of the lichenized fungus Peltigera aphthosa

A structural study of the carbohydrates from Coccomyxa mucigena, the symbiotic algal partner of the lichenized fungus Peltigera aphthosa, was carried out. It produced an O-methylated mannogalactan, with a (1 → 6)-linked β-galactopyranose main-chain partially substituted at O-3 by β-Galp, 3-OMe-α-Manp or α-Manp units. There were no similarities with polysaccharides previously found in the lichen thallus of P. aphthosa. Moreover, the influence of lichenization in polysaccharide production by symbiotic microalgae and the nature of the photobiont in carbohydrate production in lichen symbiosis are also discussed.     

铜离子胁迫对两种地衣细胞结构的影响

采用光学显微镜徒手切片技术和透射电子显微镜超薄切片制备技术,以两种叶状地衣即中国树花(Ramalina sinensis)和地卷(Peltigera rufescens)为实验材料,研究了不同浓度CuSO4(0、1、2、3、4mmol/L)处理24h后地衣体细胞的存活率以及Cu2+胁迫对细胞超微结构的影响。结果显示:(1)光学显微镜下可初步确定,Cu2+浓度越大中国树花共生藻细胞存活率越小,而同样处理条件下地卷共生藻细胞的存活率则基本保持不变。(2)低浓度Cu2+(1mmol/L)对中国树花细胞结构基本无影响,细胞壁、细胞膜及细胞内的线粒体、叶绿体完整;随着Cu2+浓度增加,当处理Cu2+浓度为2mmol/L时,细胞壁无损,但细胞膜开始破坏形成小空泡,线粒体嵴变凌乱,叶绿体也出现皱缩,类囊体膨胀,基粒排列紊乱;当Cu2+处理浓度大于2mmol/L时,共生藻的细胞膜和细胞器出现不同程度的损伤;当Cu2+浓度为3mmol/L时,细胞壁变薄,细胞膜形成的空泡变大,细胞内部结构变松散,蛋白核消失,叶绿体与细胞质混在一起,基粒片层扭曲,分布混乱,线粒体变形;当Cu2+浓度达4mmol/L时,细胞结构完全受到破坏。(3)不同浓度Cu2+处理对地卷共生藻细胞结构无明显的影响,在所有处理条件下地卷共生藻细胞壁、细胞膜都完整,且大多数共生藻细胞处于分裂状态。研究认为,中国树花对Cu2+胁迫较敏感,Cu2+耐受在1~2mmol/L之间,Cu2+浓度与中国树花细胞结构的损伤程度存在着明显的剂量效应关系,Cu2+浓度越高,其属于共球藻的真核共生藻细胞受损程度越大;地卷对Cu2+胁迫具有一定的耐性,Cu2+胁迫下地卷属于蓝藻的原核共生藻细胞仍能繁殖分裂产生子代细胞。     

Population size and fire intensity determine post‐fire abundance in grassland lichens

Questions: What is the variability in abundance of lichens on grassland soil between and within fields after prescribed fire? Is post‐fire lichen abundance an effect of pre‐fire population size? Location: Cedar Creek Natural History Area, Minnesota, USA. Methods: Lichen abundance, estimated as ground cover and dominated by Cladonia spp., was mapped in plots in two fields before prescribed burning on 06.10.2003 and 15.10.2003 for the first time since abandonment in the 1950s. The plots were resurveyed one year post‐fire. Results: Post‐fire cover of Cladonia spp. varied strongly between the fields, most likely due to different weather conditions between the burn events, which resulted in different fire intensities, one of low and one of high intensity. In the field that experienced the low intensity fire, post‐fire cover of Cladonia spp. was still relatively high, and showed a positive relationship with pre‐fire cover, while no such relationship was found after the high intensity fire. In that field Cladonia spp. experienced high mortality rates irrespective of pre‐fire cover. Conclusions: This study provides an example of how species response to disturbance can be a function of population size, but that this relationship can be non‐linear; lichens in grassland can survive a low intensity fire proportionally to pre‐fire population size, but experience high mortality rates above a fire intensity threshold. The applications of these results are that fire intensity matters to species response to prescribed fire, and that the persistence of climax lichen communities and biodiversity in the study system needs a broad range of fire intervals.     

Spatial patterns of photobiont diversity in some Nostoc-containing lichens

Patterns of photobiont diversity were examined in some Nostoc -containing lichens using the nucleotide sequence of the cyanobacterial tRNA^Leu (UAA) intron. Lichen specimens collected in northwestern USA were analysed and the sequence data were compared with tRNA^Leu(UAA) intron sequences previously obtained from lichens in northern Europe. Generally, it is the species identity of a lichen rather than the geographical origin of the specimen that determines the identity of the cyanobiont. Identical intron sequences were found in Peltigera membranacea specimens collected in Oregon (USA) and in Sweden, and very similar sequences were also found in Nephroma resupinatum thalli collected in Oregon and Finland. Furthermore, in mixed assemblages where two Peltigera species grew in physical contact with each other, the different lichen species housed different photobiont strains. There is however not a one-to-one relation between mycobiont and photobiont as some intron sequences were found in more than one lichen species, and different intron sequences were found in different samples of some lichen taxa. Peltigera venosa exhibited a higher level of photobiont diversity than any other lichen species studied, and several intron sequences could for the first time be obtained from a single thallus. It is not clear whether this is evidence of lower cyanobiont specificity, or reflects an ability to exhibit different degrees of lichenization with different Nostoc strains. In one specimen of P. venosa , which contained bipartite cyanosymbiodemes and tripartite, cephalodiate thalli, both thallus types contained the same intron sequence.     

Production of several isoforms of β-1,4-glucanase by the cyanolichen Peltigera canina

Lichen cellulase may participate in the degradation of the external substrata and/or modification of the photobiont cell wall. To promote a better understanding of the roles of cellulases in lichens, a cyanolichen was chosen because of the absence of cellulose in its symbionts. Freshly-collected thalli of Peltigera canina (L.) Wild, produce β-1,4-glucanase (EC 3.2.1.4, β-1,4-D-glucanohydrolase). This enzyme's activity was detected in the soluble and cell wall fractions and it was found to be secreted to the incubation medium when thalli were floated on water or on cellobiose. Several forms of the enzyme were detected by isoelectrofocusing. In preparative isoelectrofocusing, a single peak was obtained in each fraction, characterized by pls of 5.05, 5.25 and 4.75 in the soluble, cell wall and medium fractions, respectively. These differences were in agreement with the different pattern of bands obtained in slab-isoelectrofocusing, where the most acidic band (pl of 4.45) was present only in the soluble fraction and the band with higher pl (6.17) was more intense in the cell wall fraction. Since both symbionts in a cyanolichen lack cellulose, cellulases cannot participate in the modification of their cell wall; the presence of cellulase in Peltigera canina must therefore be related to the degradation of the tissues of the moss substratum.     

Artificial cultures of the cyanobacterial lichenPeltigera didactyla (Peltigeraceae) in the natural environment

Soredia of the cyanobacterial lichenPeltigera didactyla were cultivated on soil substratum in the natural environment. Various developmental stages (germination, formation of a soredia field, differentiation of thallus lobes with soralia and fruiting bodies) were attained during a cultivation period of 6–7 months. Our field experiments revealed that the ephemeral pioneer lichenPeltigera didactyla has a short life cycle. Thus this lichen is a well-suited object for further investigations on thallus morphogenesis of lichens with cyanobionts.Dedicated to Prof. DrLothar Geitler on the occasion of his 90th birthday.     

Co-occurrence of the multicopper oxidases tyrosinase and laccase in lichens in sub-order peltigerineae

BACKGROUND AND AIMS: Following previous findings of high extracellular redox activity in lichens and the presence of laccases in lichen cell walls, the work presented here additionally demonstrates the presence of tyrosinases. Tests were made for the presence of tyrosinases in 40 species of lichens, and from selected species their cellular location and molecular weights were determined. The effects of stress and inhibitors on enzyme activity were also studied. METHODS: Tyrosinase and laccase activities were assayed spectrophotometrically using a variety of substrates. The molecular mass of the enzymes was estimated using polyacrylamide gel electrophoresis. KEY RESULTS: Extracellular tyrosinase and laccase activity was measured in 40 species of lichens from different taxonomic groupings and contrasting habitats. Out of 20 species tested from the sub-order Peltigerineae, all displayed significant tyrosinase and laccase activity, while activity was low or absent in other species tested. Representatives from both groups of lichens displayed low peroxidase activities. Identification of the enzymes as tyrosinases was confirmed by the ability of lichen thalli or leachates derived by shaking lichens in distilled water to metabolize substrates such as L-dihydroxyphenylalanine (DOPA), tyrosine and epinephrine readily in the absence of hydrogen peroxide, the sensitivity of the enzymes to the inhibitors cyanide, azide and hexylresorcinol, activation by SDS and having typical tyrosinase molecular masses of approx. 60 kDa. Comparing different species within the Peltigerineae showed that the activities of tyrosinases and laccase were correlated to each other. Desiccation and wounding stimulated laccase activity, while only wounding stimulated tyrosinase activity. CONCLUSIONS: Cell walls of lichens in sub-order Peltigerineae have much higher activities and a greater diversity of cell wall redox enzymes compared with other lichens. Possible roles of tyrosinases include melanization, removal of toxic phenols or quinones, and production of herbivore deterrents.