O gênero Vaucheria D.C. (Vaucheriaceae, Xanthophyceae) no sul do Brasil: aspectos taxonômicos e ecológicos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chloroplast genes are expressed during intracellular symbiotic association of Vaucheria litorea plastids with the sea slug Elysia chlorotica.

The marine slug Elysia chlorotica (Gould) forms an intracellular symbiosis with photosynthetically active chloroplasts from the chromophytic alga Vaucheria litorea (C. Agardh). This symbiotic association was characterized over a period of 8 months during which E. chlorotica was deprived of V. litorea but provided with light and CO2. The fine structure of the symbiotic chloroplasts remained intact in E. chlorotica even after 8 months of starvation as revealed by electron microscopy. Southern blot analysis of total DNA from E. chlorotica indicated that algal genes, i.e., rbcL, rbcS, psaB, psbA, and 16S rRNA are present in the animal. These genes are typically localized to the plastid genome in higher plants and algae except rbcS, which is nuclear-encoded in higher plants and green (chlorophyll a/b) algae. Our analysis suggests, however, that similar to the few other chromophytes (chlorophyll a/c) examined, rbcS is chloroplast encoded in V. litorea. Levels of psbA transcripts remained constant in E. chlorotica starved for 2 and 3 months and then gradually declined over the next 5 months corresponding with senescence of the animal in culture and in nature. The RNA synthesis inhibitor 6-methylpurine reduced the accumulation of psbA transcripts confirming active transcription. In contrast to psbA, levels of 16S rRNA transcripts remained constant throughout the starvation period. The levels of the photosystem II proteins, D1 and CP43, were high at 2 and 4 months of starvation and remained constant at a lower steady-state level after 6 months. In contrast, D2 protein levels, although high at 2 and 4 months, were very low at all other periods of starvation. At 8 months, de novo synthesis of several thylakoid membrane-enriched proteins, including D1, still occurred. To our knowledge, these results represent the first molecular evidence for active transcription and translation of algal chloroplast genes in an animal host and are discussed in relation to the endosymbiotic theory of eukaryote origins.

海带蓝光受体及相关基因的研究

本研究中运用四种不用的方法提取海带孢子体RNA，通过对比分析，确定了采用CTAB提取液[2% (w/v) CTAB,100 mM Tris-HC1 (pH 8.0), 50 mM EDTA, pH 7.5, 2 M NaCl, 50 mM DTT]提取RNA的方法，该方法的主要改进之处： (1) 高浓度（50 mM）的 DTT作为防止多酚氧化的还原剂加入提取液中；(2) 1/4 体积的乙醇和 1/9 体积的3 M 醋酸钾（pH 4.8）用来沉淀去除多糖；(3) 1/10倍体积的3 mol/L醋酸钠（pH 5.0）和两倍体积的乙醇选择性沉淀RNA；(4) -80℃沉淀30 min沉淀RNA。CTAB改进法提取的海带孢子体RNA质量好（A260/280 =1.96±0.05)，产量高（68 μg/g），分离的RNA可用于RT-PCR反应，扩增相关基因片段。该方法提取海带配子体RNA效果也很好。 根据拟南芥(Arabidopsis thaliana)、 莱茵衣藻(Chlamydomonas reinhardtii) 、铁线蕨(Adiantum capillus-veneris)、转板藻(Mougeotia scalaris)和无隔藻(Vaucheria frigida)等的蓝光受体蛋白保守序列，设计一系列简并引物，进行RT-PCR扩增，筛选并克隆测序了4条序列片段，通过比对分析，所得序列与已知的蓝光受体基因序列不一致。在所获得的序列中，一条783 bp的序列与肌醇-1-磷酸合成酶基因的片段有较高的相似性，通过RACE法克隆出该基因的3’末端，5’末端克隆没有成功，全长序列有待深入分析。

Flora de comunidades de macroalgas lóticas de fragmentos florestais remanescentes da região noroeste do Estado de São Paulo, Brasil

Foram amostrados 17 riachos, em 12 fragmentos durante a estação seca (junho a agosto de 2007 e de 2008). Foram identificadas 16 espécies de macroalgas, pertencentes a 14 gêneros. Cyanophyta e Chlorophyta foram os grupos mais representativos (44 e 38%, respectivamente), seguidos por Rhodophyta (12%) e Heterokontophyta (6%). O número global de espécies foi baixo, fato atribuído à predominância de substrato areno-argiloso (65%) e de trechos sombreados (53%). A maioria das espécies (69%) ocorreu exclusivamente em um único riacho, padrão frequentemente observado para macroalgas lóticas. Vaucheria pseudogeminata foi reportada pela primeira vez para o Brasil e Trichocoleus sociatus para a região noroeste do Estado de São Paulo. Comparando a flora de macroalgas encontrada com aquelas de outras regiões/biomas do Estado de São Paulo, notou-se maior semelhança com Floresta Tropical (56% espécies em comum), conforme esperado, por ser o bioma mais próximo e composto também por Floresta Estacional Semidecidual.

Flora e distribuição ecológica de comunidades de macroalgas lóticas de fragmentos florestais da região noroeste do estado de São Paulo

Pós-graduação em Ciências Biológicas (Biologia Vegetal) - IBRC

The relationship between macroalgal morphological complexity and hydraulic conditions in stream habitats

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)