Anatomia floral de espécies de Bulbophyllum sect. Micranthae (Orchidaceae, Asparagales)

Bulbophyllum, which comprises 1876 species, is considered the second largest genus of angiosperms, with a pantropical distribution. The morphological and anatomical floral studies in the genus are incipient, with data restricted to the gynostemium and lip of some species. Based on molecular data, six sections were recognized within Bulbophyllum at the Neotropics, amongst them Bulbophyllum sect. Micranthae, which comprises 12 species distributed in central South America. We aimed to study the floral anatomy of six species of Bulbophyllum sect. Micranthae, in order to determine useful characters to differentiate them and contribute to the anatomical characterization of the section as a whole. Floral anatomy was assessed through usual techniques of light microscopy. The data found here for B. adiamantinum, B. chloroglossum, B. epiphytum, B. mentosum, B. micranthum and B. rupicolum allowed to identificate the presence of glandular trichomes and the possible presence of a secretory region on the lip, which might produce substances used as a reward to pollinators. The most significant anatomical characters to the species characterization were the shape and ornamentation of the outer periclinal walls of the epidermal cells, as well as the number of vascular bundles in dorsal and lateral sepals and at the lip. The data also allowed the differentiation between B. epiphytum and B. rupicolum, species very similar in morphology and phylogenetically related. Besides that, the data also allowed the discussion regarding the maintenance of B. mentosum within the section: although its inclusion is supported by molecular studies, the anatomical data here presented shows greater differences compared to the other species, not supporting its maintenance in Bulbophyllum sect. Micranthae

Micromorfologia e anatomia floral das seções neotropicais de Bulbophyllum Thouars (Orchidaceae, Asparagales): considerações taxonômicas e evolutivas

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

Anatomia floral de espécies de Cyperaceae (Poales)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Glandular trichome density and essential oil composition in leaves and inflorescences of Lippia origanoides Kunth (Verbenaceae) in the Brazilian Cerrado

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

Structure and secretion mechanisms of floral glands in Diplopterys pubipetala (Malpighiaceae), a neotropical species

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

Comparative floral micromorphology and anatomy of species of Bulbophyllum section Napelli (Orchidaceae), a Neotropical section widely distributed in forest habitats

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

Paepalanthus sphaeroides, a new species of Eriocaulaceae from the Atlantic Forest, Brazil

We describe and illustrate Paepalanthus sphaeroides (Eriocaulaceae, Paepalanthoideae) from the Mantiqueira Range in South-eastern Brazil and compare it with the morphologically most similar species: Paepalanthus aequalis and Paepalanthus eriophaeus. Paepalanthus sphaeroides has unique membranaceous sheaths tightly adpressed to the scapes, patent involucral bracts slightly surpassing the capitula, and sepals of the staminate flowers fused from the base to the middle as distinctive characteristics. Comments on morphological variation, geographical distribution, ecology, conservation status, as well as a distribution map, line drawings, and photos are provided.

A New Species of Pelexia (Orchidaceae, Spiranthinae) from Sao Paulo, Brazil

A new species of Pelexia Poit. ex Until. (Orchidaceae, Spiranthinae) occurring in central Sao Paulo, southeastern Brazil, is described and illustrated as P. vinosa A. W. C. Ferreira, M. I. S. Lima & Pansarin. Pelexia vinosa is recognized by its leaves that are present at flowering and its dark purple leaf blades with reddish margins. Inflorescences are sparsely pubescent and reddish. The red sepals contrast with the white hyaline petals and labellum. The species is notable for its spurlike nectary that is parallel and adnate to the ovary. The new species is morphologically similar to P. laxa (Poepp. & Endl.) Lindl. In addition, the need to preserve native areas of the interior of Sao Paulo State (habitat of P. vinosa) is discussed.

Developmental and Environmental Effects on the Expression of the C3-C4 Intermediate Phenotype in Moricandia arvensis1

Cellular anatomy and expression of glycine decarboxylase (GDC) protein were studied during leaf development of the C3-C4 intermediate species Moricandia arvensis. Leaf anatomy was initially C3-like and the number and profile area of mitochondria in the bundle-sheath cells were the same as those in adjacent mesophyll cells. Between a leaf length of 6 and 12 mm there was a bundle-sheath-specific, 4-fold increase in the number of mitochondrial profiles, followed by a doubling of their individual profile areas as the leaves expanded further. Subunits of GDC were present in whole-leaf extracts before the anatomical development of bundle-sheath cells. Whereas the GDC H-protein content of leaves increased steadily throughout development, the increase in GDC P-protein was synchronous with the development of mitochondria in the bundle sheath. The P-protein was confined to bundle-sheath mitochondria throughout leaf development, and its content in individual mitochondria increased before the anatomical development of the bundle sheath. Anatomical and biochemical attributes of the C3-C4 character were present in the cotyledons and sepals but not in other photosynthetic organs/tissues. In leaves and cotyledons that developed in the dark, the expression of the P-protein and the organellar development were reduced but the bundle-sheath cell specificity was retained.

Eucalypt MADS-Box Genes Expressed in Developing Flowers

Three MADS-box genes were identified from a cDNA library derived from young flowers of Eucalyptus grandis W. Hill ex Maiden. The three egm genes are single-copy genes and are expressed almost exclusively in flowers. The egm1 and egm3 genes shared strongest homology with other plant MADS-box genes, which mediate between the floral meristem and the organ-identity genes. The egm3 gene was also expressed strongly in the receptacle or floral tube, which surrounds the carpels in the eucalypt flower and bears the sepals, petals, and numerous stamens. There appeared to be a group of genes in eucalypts with strong homology with the 3′ region of the egm1 gene. The egm2 gene was expressed in eucalypt petals and stamens and was most homologous to MADS-box genes, which belong to the globosa group of genes, which regulate organogenesis of the second and third floral whorls. The possible role of these three genes in eucalypt floral development is discussed.

Organ-specific and Agamous-regulated expression and glycosylation of a pollen tube growth-promoting protein.

Transmitting tissue-specific (TTS) protein is a pollen tube growth-promoting and attracting glycoprotein located in the stylar transmitting tissue extracellular matrix of the pistil of tobacco. The TTS protein backbones have a deduced molecular mass of about 28 kDa, whereas the glycosylated stylar TTS proteins have apparent molecular masses ranging between 50 and 100 kDa. TTS mRNAs and proteins are ectopically produced in transgenic tobacco plants that express either a cauliflower mosaic virus (CaMV) 35S promoter-TTS2 transgene or a CaMV 35S-promoter-NAG1 (NAG1 = Nicotiana tabacum Agamous gene) transgene. However, the patterns of TTS mRNA and protein accumulation and the quality of the TTS proteins produced are different in these two types of transgenic plants. In 35S-TTS transgenic plants, TTS mRNAs and proteins accumulate constitutively in vegetative and floral tissues. However, the ectopically expressed TTS proteins in these transgenic plants accumulate as underglycosylated protein species with apparent molecular masses between 30 and 50 kDa. This indicates that the capacity to produce highly glycosylated TTS proteins is restricted to the stylar transmitting tissue. In 35S-NAG transgenic plants, NAG1 mRNAs accumulate constitutively in vegetative and floral tissues, and TTS mRNAs are induced in the sepals of these plants. Moreover, highly glycosylated TTS proteins in the 50- to 100-kDa molecular mass range accumulate in the sepals of these transgenic, 35S-NAG plants. These results show that the tobacco NAGI gene, together with other yet unidentified regulatory factors, control the expression of TTS genes and the cellular capacity to glycosylate TTS proteins, which are normally expressed very late in the pistil developmental pathway and function in the final stage of floral development. The sepals in the transgenic 35S-NAG plants also support efficient pollen germination and tube growth, similar to what normally occurs in the pistil, and this ability correlates with the accumulation of the highest levels of the 50- to 100-kDa glycosylated TTS proteins.

Desenvolvimento floral e do óvulo e aspectos da reprodução em Aechmea sp. e Vriesea sp. (Bromeliaceae)

A utilização de Bromélias tem sido crescente no mercado de plantas onamentais, por outro lado, muitas espécies encontram-se ameaçadas, grande parte pelos impactos humanos no ambiente. Aechmea correia-araujoi E. Pereira & Moutinho, Aechmea gamossepala Wittm, Vriesea ensiformis (Vell.) Beer e Vriesea saundersii (Carrière) E. Morren ex Mez, espécies nativas da Mata Atlântica brasileira, têm sido alvo de extrativismo. Informações básicas sobre a espécie são essenciais para subsidiar a condução de programas de conservação e melhoramento genético, que aliados a ferramentas biotecnológicas permitem a incorporação de estratégias inovadoras aos métodos de melhoramento. Neste sentido, o objetivo do presente trabalho foi descrever essas espécies, quanto à micromorfologia floral, aspectos reprodutivos envolvidos no processo de polinização, desenvolvimento floral e deesenvolvimento gametofítico, como mecanismo de preservação e produção comercial. A caracterização morfológica e anatômica das flores das espécies de Aechmea e Vriesea contribuiu para a compreensão do processo reprodutivo. As espécies apresentam grãos de pólen com alta capacidade reprodutiva, viabilidade polínica superior a 93%, germinação in vitro maior que 80% e o estigma apresenta-se receptivo da antese ao final do dia. A ontogênese floral de A. correia-araujoi é centrípeta, os primórdios desenvolvem-se na ordem, sépala, pétala, androceu e gineceu. O apêndice petalar é formado na fase final do desenvolvimento. O primórdio de óvulo tem origem placentária e caráter trizonal, o óvulo é anátropo, bitegumentado e crassinucelado. O meristema floral de A. gamosepala se desenvolve de forma centrípeta, de forma unidirecional reversa. O estigma diferencia-se na fase inicial do desenvolvimento e os apêndices petalares, na fase final. O óvulo é anátropo, crassinucelado, bitegumentado, tétrade linear, megásporo calazal funcional, desenvolvimento tipo monospórico e Polygonum. As anteras são bitecas, tetraesporangiadas, com tapete secretor. Botões florais de 8,7 - 13,0 mm são indicados no estudo de embriogênese a partir de micrósporo. As alterações celulares e o padrão de distribuição de pectinas e AGPs foram caracterizadas por análise citoquímica com azul de toluidina, KI e DAPI e imunocitoquímica por imunofluorescência com os anticorpos para RNA, pectinas esterificadas (JIM7), não esterificadas (JIM5) e AGPs (LM2, LM6, MAC207, JIM13, JIM14) e analisadas por microscopia de fluorescência. Foram caracterizados padrões de distribuição espaço-temporal de pectinas e AGP que podem ser utilizados como marcadores de desenvolvimento gametofítico masculino. As observações feitas nesse trabalho fornecem dados sobre aspectos reprodutivos das espécies que podem ser utilizados em programas de melhoramento genético, conservação e desenvolvimento de haploides

Characterization of a novel lipoxygenase-independent senescence mechanism in Alstroemeria peruviana floral tissue

The role of lipoxygenase (lox) in senescence ofAlstroemeria peruviana flowers was investigated using a combination of in vitro assays and chemical profiling of the lipid oxidation products generated. Phospholipids and galactolipids were extensively degraded during senescence in both sepals and petals and the ratio of saturated/unsaturated fatty acids increased. Lox protein levels and enzymatic activity declined markedly after flower opening. Stereochemical analysis of lox products showed that 13-lox was the major activity present in both floral tissues and high levels of 13-keto fatty acids were also synthesized. Lipid hydroperoxides accumulated in sepals, but not in petals, and sepals also had a higher chlorophyll to carotenoid ratio that favors photooxidation of lipids. Loss of membrane semipermeability was coincident for both tissue types and was chronologically separated from lox activity that had declined by over 80% at the onset of electrolyte leakage. Thus, loss of membrane function was not related to lox activity or accumulation of lipid hydroperoxides per se and differs in these respects from other ethylene-insensitive floral tissues representing a novel pattern of flower senescence.

Chemotaxonomic studies on Schwenckia americana LINN.

This study investigated the chemotaxonomic studies on Schwenckia americana Linn., a member of the family Solanaceae predominantly found mostly in low grass fields, Nigeria. The habit is annual herbaceous weed with slender stem characterized with free branching and growing up to 45cm in height.They are used mainly as medicine. The leaves are simple, entire, elliptic to ovate in shape, smooth, variable, petiolate and larger at the lower region of stem and narrowing to smaller almost sessile and oblanceolate towards the apical regions which are alternately arranged and acrescently structured from the top to the base upto 3.7 ± 1.5cm long and 2.4±0.6cm wide. The inflorescence is a panicle of 15 or more flowers occurring at stem terminal.The flowers are whitish tubular structures measuring up to 1.0±0.2cm in length and 0.1cm in diameter. The petals are whitish up to 0.9 ±0.2cm in length and sepals are greenish up to 0.1cm in length. The berry fruit is greenish when unripe and pale yellowish when ripe up to 0.3±0.1cm in diameter. The seeds are blackish and spherical or triangular shaped with rough edges measuring up to 0.1cm in diameter. The epidermal studies reveal anomocytic stomata whereas the trichomes are simple uniseriate forms wth bulge heads. The anatomy of mid-ribs and petioles showed bicollateral vascular systems. There are three vascular traces and the node is unilacunar. The petioles have 2 rib traces at primary growth phase. At secondary growth phase, the mid-rib and petiole revealed vascular arcs and the stem has a ring of open vascular system. The cytological studies showed a diploid chromosome number of 2n = 24 and n = 12 for the haploids. Alkaloids, saponins, tannins, phlobatannins, flavonoids, combined anthraquinones, free anthraquinones and cardiac glycosides are present.

Targeted gene modification in Fragaria vesca mediated by CRISPR/Cas9 system

Genome editing is becoming an important biotechnological tool for gene function analysis and crop improvement, being the CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeat-CRISPR associated protein 9) system the most widely used. The natural CRISPR/Cas9 system has been reduced to two components: a single-guide RNA (sgRNA) for target recognition via RNA-DNA base pairing, which is commonly expressed using a promoter for small-RNAs (U6 promoter), and the Cas9 endonuclease for DNA cleavage (1). To validate the CRISPR/Cas9 system in strawberry plants, we designed two sgRNAs directed against the floral homeotic gene APETALA3 (sgRNA-AP3#1 and sgRNA-AP3#2). This gene was selected because ap3 mutations induce clear developmental phenotypes in which petals and stamens are missing or partially converted to sepals and carpels respectively (2). In this work, we used two different U6 promoters to drive the sgRNA-AP3s expression: AtU6-26 from Arabidopsis (4), and a U6 promoter from Fragaria vesca (FvU6) (this work). We also tested two different coding sequences of Cas9: a human- (hSpCas9) (3) and a plant-codon optimized (pSpCas9) (this work). Transient expression experiments using both CRISPR/Cas9 systems (AtU6-26:sgRNA-AP3#1_35S:hSpCas9_AtU6-26:sgRNA-AP3#2 and FvU6:sgRNA-AP3#1_35S:pSpCas9_FvU6:sgRNA-AP3#2) were performed infiltrating Agrobacterium tumefaciens into F. vesca fruits. PCR amplification and sequencing analyses across the target sites showed a deletion of 188-189 bp corresponding to the region comprised between the two cutting sites of Cas9, confirming that the CRISPR/Cas9 system is functional in F. vesca. Remarkably, the two systems showed different mutagenic efficiency that could be related to differences in expression of the U6 promoters as well as differences in the Cas9 transcripts stability and translation. Stable transformants for both F. vesca (2n) and Fragaria X anannassa (8n) are currently being established to test whether is possible to obtain heritable homozygous mutants derived from CRISPR/Cas9 strategies in strawberry. Thus, our work offers a promising tool for genome editing and gene functional analysis in strawberry. This tool might represent a more efficient alternative to the sometimes inefficient RNAi silencing methods commonly used in this species.