Potential function of CbuSPL and gene encoding its interacting protein during flowering in Catalpa bungei.

BACKGROUND: "Bairihua", a variety of the Catalpa bungei, has a large amount of flowers and a long flowering period which make it an excellent material for flowering researches in trees. SPL is one of the hub genes that regulate both flowering transition and development. RESULTS: SPL homologues CbuSPL9 was cloned using degenerate primers with RACE. Expression studies during flowering transition in "Bairihua" and ectopic expression in Arabidopsis showed that CbuSPL9 was functional similarly with its Arabidopsis homologues. In the next step, we used Y2H to identify the proteins that could interact with CbuSPL9. HMGA, an architectural transcriptional factor, was identified and cloned for further research. BiFC and BLI showed that CbuSPL9 could form a heterodimer with CbuHMGA in the nucleus. The expression analysis showed that CbuHMGA had a similar expression trend to that of CbuSPL9 during flowering in "Bairihua". Intriguingly, ectopic expression of CbuHMGA in Arabidopsis would lead to aberrant flowers, but did not effect flowering time. CONCLUSIONS: Our results implied a novel pathway that CbuSPL9 regulated flowering development, but not flowering transition, with the participation of CbuHMGA. Further investments need to be done to verify the details of this pathway.

Identification and Analysis of a Candidate WRKY Transcription Factor Gene Affecting Adventitious Root Formation Using Association Mapping in Catalpa Scop.

The capacity to develop adventitious roots (AR) from cuttings is a key factor for the mass deployment of superior genotypes in the woody plant, including Catalpa Scop. Catalpa Scop. was characterized as having excellent woody qualities in China. However, the knowledge of the molecular mechanisms of AR formation is limited in Catalpa Scop. In this study, for the first time, association mapping for AR formation was performed on a selected sample of 108 Catalpa accessions. Genetic diversity and population structure was estimated on the basis of 54 gene-derived simple sequence repeat markers. Genetic diversity analysis revealed that four accessions belonging to Catalpa duclouxii and eight belonging to Catalpa fargesii formed one clade, providing molecular evidence for C. duclouxii belonging to C. fargesii. Marker-trait association analysis revealed four genes associated with three rooting traits, namely AR rating, adventitious root numbers (ARN), and maximal AR length, with phenotypic variation explained for these traits of 10.77-18.49% in experiments in 2 years. Among the four genes, a WRKY transcription factor gene CbNN1 was the only gene that showed association with the ARN in both years, and expression of this gene (determined by analysis by real-time quantitative polymerase chain reaction) increased with increasing rooting ability. These results indicated that the gene CbNN1 might play a positive role in AR formation. The findings from this study will not only be beneficial to the research of AR formation, but also contribute to the phylogeny of interspecies in Catalpa Scop.

Construction of a high-density genetic map and QTL mapping of leaf traits and plant growth in an interspecific F1 population of Catalpa bungei × Catalpa duclouxii Dode.

BACKGROUND: Catalpa bungei is an important tree species used for timber in China and widely cultivated for economic and ornamental purposes. A high-density linkage map of C. bungei would be an efficient tool not only for identifying key quantitative trait loci (QTLs) that affect important traits, such as plant growth and leaf traits, but also for other genetic studies. RESULTS: Restriction site-associated DNA sequencing (RAD-seq) was used to identify molecular markers and construct a genetic map. Approximately 280.77 Gb of clean data were obtained after sequencing, and in total, 25,614,295 single nucleotide polymorphisms (SNPs) and 2,871,647 insertions-deletions (InDels) were initially identified in the genomes of 200 individuals of a C. bungei (7080) × Catalpa duclouxii (16-PJ-3) F1 population and their parents. Finally, 9072 SNP and 521 InDel markers that satisfied the requirements for constructing a genetic map were obtained. The integrated genetic map contained 9593 pleomorphic markers in 20 linkage groups and spanned 3151.63 cM, with an average distance between adjacent markers of 0.32 cM. Twenty QTLs for seven leaf traits and 13 QTLs for plant height at five successive time points were identified using our genetic map by inclusive composite interval mapping (ICIM). Q16-60 was identified as a QTL for five leaf traits, and three significant QTLs (Q9-1, Q18-66 and Q18-73) associated with plant growth were detected at least twice. Genome annotation suggested that a cyclin gene participates in leaf trait development, while the growth of C. bungei may be influenced by CDC48C and genes associated with phytohormone synthesis. CONCLUSIONS: This is the first genetic map constructed in C. bungei and will be a useful tool for further genetic study, molecular marker-assisted breeding and genome assembly.

Genome-wide analysis of long non-coding RNAs in Catalpa bungei and their potential function in floral transition using high-throughput sequencing.

BACKGROUND: Long non-coding RNAs (lncRNAs) have crucial roles in various biological regulatory processes. However, the study of lncRNAs is limited in woody plants. Catalpa bungei is a valuable ornamental tree with a long cultivation history in China, and a deeper understanding of the floral transition mechanism in C. bungei would be interesting from both economic and scientific perspectives. RESULTS: In this study, we categorized C. bungei buds from early flowering (EF) and normal flowering (NF) varieties into three consecutive developmental stages. These buds were used to systematically study lncRNAs during floral transition using high-throughput sequencing to identify molecular regulatory networks. Quantitative real-time PCR was performed to study RNA expression changes in different stages. In total, 12,532 lncRNAs and 26,936 messenger RNAs (mRNAs) were detected. Moreover, 680 differentially expressed genes and 817 differentially expressed lncRNAs were detected during the initiation of floral transition. The results highlight the mRNAs and lncRNAs that may be involved in floral transition, as well as the many lncRNAs serving as microRNA precursors. We predicted the functions of lncRNAs by analysing the relationships between lncRNAs and mRNAs. Seven lncRNA-mRNA interaction pairs may participate in floral transition. CONCLUSIONS: This study is the first to identify lncRNAs and their potential functions in floral transition, providing a starting point for detailed determination of the functions of lncRNAs in C. bungei.

Germination and seed conservation of a pioneer species, Tecoma stans (Bignoniaceae), from tropical dry forest of Colombia / Germinación y conservación de semillas de la especie pionera, Tecoma stans (Bignoniaceae), del bosque seco tropical de Colombia

Abstract Seed germination and seed longevity under different environmental conditions are fundamental to understand the ecological dynamics of a species, since they are decisive for its success within the ecosystem. Taking this into account, seed germination and seed storage behavior of a pioneer species of tropical dry forest (Tecoma stans) were studied in the laboratory, to establish the effect of different environmental conditions on a local tree population. Two seed lots collected in July 2011, from Cali (Colombia), were evaluated under three alternating temperatures (20/30, 20/25, 25/30 ºC; 16/8 h) and four light qualities (12-hour white light photoperiod, darkness, and 15 minutes of red light -R and far red light -FR). Final germination was recorded for all treatments; for white light treatment the daily germination was recorded to calculate mean germination rate, mean germination time, and two synchronization indices. To assess the effect of light quality on physiological variables, a destructive germination test was carried out. For this test, another seed lot was evaluated under the same light conditions using an alternating temperature of 20/30 °C - 16/8 h, recording germination during six days for every treatment. In addition, seeds were stored at two different moisture contents (7.7, 4.1 %) and three storage temperatures (20, 5, -20 ºC), during two time periods (one and three months); a seed germination test was conducted for each treatment. Four replicates of 35 seeds per treatment were used for all experiments. Germination was high (GP > 90 %) with all alternating temperatures under white light, whereas under R, FR, and darkness germination was evenly successful at low temperatures, but at higher temperature, half of the seeds entered into secondary dormancy (GP= 45-65 %). However, mean germination rate and synchronization under R and FR decreased significantly in comparison to white light treatment and, consequently, mean germination time increased. Seed storage behavior of this species is orthodox due to the high germination (GP > 90 %) obtained under all treatments. In conclusion, T. stans seeds have a negative germination response at high incubation temperatures in the absence of white light, entering into a secondary dormancy. In contrast, an environment with a lower temperature and without white light delays the germination, but at the end seeds are able to reach the same germination values. This seed dependence on incident light in limiting conditions suggests a physiological mechanism on the seed tissues of this species, probably mediated by phytochromes. Finally, the orthodox seed storage behavior of T. stans is a reason to include this species in ex situ seed conservation programs for restoration and recovery of the tropical dry forest; however, long-term studies should be conducted in order to evaluate the maintenance of this characteristic throughout longer periods of time. Rev. Biol. Trop. 66(2): 918936. Epub 2018 June 01.

Resumen La germinación y la longevidad de las semillas de una especie bajo diferentes condiciones ambientales son fundamentales para las dinámicas ecológicas de una especie, debido a que son decisivas en el éxito de la misma en un ecosistema. Teniendo en cuenta esto, se estudió la germinación y el comportamiento en el almacenamiento de las semillas de una especie pionera de bosque seco tropical (Tecoma stans) a nivel de laboratorio, para establecer el efecto de diferentes condiciones ambientales en una población local de árboles. Dos lotes de semillas recolectados en julio 2011, de Cali (Colombia), se evaluaron a tres temperaturas alternadas (20 / 30, 20 / 25, 25 / 30 °C; 16 / 8 h) y cuatro calidades de luz (fotoperiodo de 12 horas de luz blanca, oscuridad, y 15 minutos de luz roja -R y roja lejana -RL). Se registró la germinación final para todos los tratamientos; para el tratamiento de luz blanca se registró la germinación diaria para calcular la tasa media de germinación, el tiempo medio de germinación y dos índices de sincronización. Para evaluar el efecto de la calidad de luz sobre las variables fisiológicas, se realizó una prueba de germinación destructiva. Para esta prueba, otro lote de semillas fue puesto a las mismas condiciones de luz usando una temperatura alternada de 20 / 30 °C - 16 / 8 h, registrando la germinación durante seis días para cada tratamiento. Además, se almacenaron semillas a dos contenidos de humedad (7.7, 4.1 %) y a tres temperaturas de almacenamiento (20, 5, -20 °C), durante dos periodos de tiempo (uno y tres meses); se realizó una prueba de germinación a cada tratamiento. Cuatro repeticiones de 35 semillas por cada tratamiento se usaron en cada experimento. La germinación fue alta (PG > 90 %) en todas las temperaturas alternadas con luz blanca, mientras que en los tratamientos de luz R, RL y en oscuridad, la germinación fue igualmente exitosa a bajas temperaturas, pero a temperaturas más altas la mitad de las semillas entraron en latencia secundaria (PG= 45-65 %). Sin embargo, la tasa media de germinación y la sincronización en R y RL disminuyeron significativamente en comparación con el tratamiento de luz blanca y consecuentemente el tiempo medio de germinación aumentó. El comportamiento de las semillas de T. stans en el almacenamiento es ortodoxo debido a la alta germinación obtenida (PG > 90 %) en todos los tratamientos. En conclusión, las semillas de T. stans tienen una respuesta germinativa negativa a temperaturas de incubación alta en ausencia de luz blanca, donde entran a latencia secundaria. En contraste, un ambiente con baja temperatura y sin luz blanca retrasa la germinación, pero al final las semillas son capaces de alcanzar los mismos valores de germinación. Esta dependencia de las semillas a la luz incidente en condiciones limitantes sugiere la presencia de un mecanismo fisiológico en los tejidos de esta especie, probablemente mediado por fitocromos. Finalmente, el comportamiento ortodoxo de las semillas de T. stans en el almacenamiento abre la posibilidad de incluirla en programas de conservación ex situ para la restauración y recuperación del bosque seco tropical; no obstante, se deben llevarse a cabo pruebas más largas para evaluar el mantenimiento de esta característica por periodos de tiempo más largos.

Physiological and transcriptional responses of Catalpa bungei to drought stress under sufficient- and deficient-nitrogen conditions.

Many semi-arid ecosystems are simultaneously limited by soil water and nitrogen (N). We conducted a greenhouse experiment to address how N availability impacts drought-resistant traits of Catalpa bungei C. A. Mey at the physiological and molecular level. A factorial design was used, consisting of sufficient-N and deficient-N combined with moderate drought and well-watered conditions. Seedling biomass and major root parameters were significantly suppressed by drought under the deficient-N condition, whereas N application mitigated the inhibiting effects of drought on root growth, particularly that of fine roots with a diameter <0.2 mm. Intrinsic water-use efficiency was promoted by N addition under both water conditions, whereas stable carbon isotope compositions (δ13C) was promoted by N addition only under the well-watered condition. Nitrogen application positively impacted drought adaptive responses including osmotic adjustment and homeostasis of reactive oxygen species, the content of free proline, soluble sugar and superoxide dismutase activity: all were increased upon drought under sufficient-N conditions but not under deficient-N conditions. The extent of abscisic acid (ABA) inducement upon drought was elevated by N application. Furthermore, an N-dependent crosstalk between ABA, jasmonic acid and indole acetic acid at the biosynthesis level contributed to better drought acclimation. Moreover, the transcriptional level of most genes responsible for the ABA signal transduction pathway, and genes encoding the antioxidant enzymes and plasma membrane intrinsic proteins, are elevated upon drought only under sufficient-N addition. These observations confirmed at the molecular level that major adaptive responses to drought are dependent on sufficient N nutrition. Although N uptake was decreased under drought, N-use efficiency and transcription of most genes encoding N metabolism enzymes were elevated, demonstrating that active N metabolism positively contributed drought resistance and growth of C. bungei under sufficient-N conditions.

Mixed pollen load and late-acting self-incompatibility flexibility in Adenocalymma peregrinum (Miers) L.G. Lohmann (Bignonieae: Bignoniaceae).

Mixed cross and self-pollen load on the stigma (mixed pollination) of species with late-acting self-incompatibility system (LSI) can lead to self-fertilized seed production. This "cryptic self-fertility" may allow selfed seedling development in species otherwise largely self-sterile. Our aims were to check if mixed pollinations would lead to fruit set in LSI Adenocalymma peregrinum, and test for evidence of early-acting inbreeding depression in putative selfed seeds from mixed pollinations. Experimental pollinations were carried out in a natural population. Fruit and seed set from self-, cross and mixed pollinations were analysed. Further germination tests were carried out for the seeds obtained from treatments. Our results confirm self-incompatibility, and fruit set from cross-pollinations was three-fold that from mixed pollinations. This low fruit set in mixed pollinations is most likely due to a greater number of self- than cross-fertilized ovules, which promotes LSI action and pistil abortion. Likewise, higher percentage of empty seeds in surviving fruits from mixed pollinations compared with cross-pollinations is probably due to ovule discounting caused by self-fertilization. Moreover, germinability of seeds with developed embryos was lower in fruits from mixed than from cross-pollinations, and the non-viable seeds from mixed pollinations showed one-third of the mass of those from cross-pollinations. The great number of empty seeds, lower germinability, lower mass of non-viable seeds, and higher variation in seed mass distribution in mixed pollinations, strongly suggests early-acing inbreeding depression in putative selfed seeds. In this sense, LSI and inbreeding depression acting together probably constrain self-fertilized seedling establishment in A. peregrinum.

Seedling survival of Handroanthus impetiginosus (Mart ex DC) Mattos in a semi-arid environment through modified germination speed and post-germination desiccation tolerance.

Uniform rapid seed germination generally forms a great risk for the plant population if subsequent intermittent precipitation causes desiccation and seedling death. Handroanthus impetiginosus can be found commonly in a wide range of biomes within Brazil including those that are semi-arid. Germination and early growth was studied to understand how germinated seeds survive under these stringent conditions. Accessions were sampled from four seasonally dry biomes in Brazil. Precipitation at the start of the rainy season in the Caatinga, a semi-arid biome, is less predictable and the number of successive dry days per dry interval in the first four months of the rainy season was higher than in the other studied biomes. Plants from the Caatinga produced thicker seeds and this trait concurred with slow germination and stronger osmotic inhibition of germination across the accessions, forming a stress avoidance mechanism in the Caatinga. Post-germination desiccation tolerance was high in the Caatinga accession, could be re-induced in accessions from biomes with more regular precipitation (Cerrado and transition zone), but remained poor in the Cerradão accession; thus forming a stress tolerance mechanism. Production of adventitious roots ascertained survival of all tested individuals from all four locations, even if protruded radicles did not survive desiccation, forming an additional stress tolerance mechanism. A sequence of stress avoidance and stress tolerance mechanisms in seeds and germinated seeds was associated with precipitation patterns in different biomes. These mechanisms purportedly allow rapid seedling establishment when conditions are suitable and enable survival of the young seedling when conditions are adverse.

Seedling survival of Handroanthus impetiginosus (Mart ex DC) Mattos in a semi-arid environment through modified germination speed and post-germination desiccation tolerance / Sobrevivência de plântulas de Handroanthus impetiginosus (Mart ex DC) Mattos em ambiente semi-árido ocorre por modificação da velocidade de germinação e da tolerância à dessecação pós germinação

Abstract Uniform rapid seed germination generally forms a great risk for the plant population if subsequent intermittent precipitation causes desiccation and seedling death. Handroanthus impetiginosus can be found commonly in a wide range of biomes within Brazil including those that are semi-arid. Germination and early growth was studied to understand how germinated seeds survive under these stringent conditions. Accessions were sampled from four seasonally dry biomes in Brazil. Precipitation at the start of the rainy season in the Caatinga, a semi-arid biome, is less predictable and the number of successive dry days per dry interval in the first four months of the rainy season was higher than in the other studied biomes. Plants from the Caatinga produced thicker seeds and this trait concurred with slow germination and stronger osmotic inhibition of germination across the accessions, forming a stress avoidance mechanism in the Caatinga. Post-germination desiccation tolerance was high in the Caatinga accession, could be re-induced in accessions from biomes with more regular precipitation (Cerrado and transition zone), but remained poor in the Cerradão accession; thus forming a stress tolerance mechanism. Production of adventitious roots ascertained survival of all tested individuals from all four locations, even if protruded radicles did not survive desiccation, forming an additional stress tolerance mechanism. A sequence of stress avoidance and stress tolerance mechanisms in seeds and germinated seeds was associated with precipitation patterns in different biomes. These mechanisms purportedly allow rapid seedling establishment when conditions are suitable and enable survival of the young seedling when conditions are adverse.

Resumo A germinação rápida e uniforme geralmente pode apresentar riscos para a população de plantas caso a precipitação ocorra de maneira intermitente, provocando seca e morte das plântulas. Handroanthus impetiginosus pode ser encontrado em uma grande variedade de biomas no Brasil. A germinação e o crescimento inicial das plântulas de Handroanthus impetiginosus foram estudadas para compreender como as sementes germinadas sobrevivem nestas condições. Acessos de quatro biomas que apresentam secas sazonais foram amostrados. A precipitação, no início da estação chuvosa na Caatinga, um bioma semi-árido, é menos previsível e o número de dias secos consecutivos por intervalo de seca, nos primeiros quatro meses da estação chuvosa, foi maior do que nos outros biomas estudados. Plantas do bioma Caatinga produzem sementes mais espessas e essa característica está associada com uma germinação lenta e com a inibição osmótica maior da germinação através dos acessos, formando um mecanismo para evitar o estresse. Tolerância à dessecação em sementes germinadas foi superior no bioma Caatinga e que podem ser reinduzida em acesso com uma precipitação mais regular (Cerrado e na zona de transição), mas manteve-se baixa no acesso Cerradão, formando assim um mecanismo para evitar o estresse. Produção de raízes adventícias foi observada nos indivíduos estudados de todos os quatro biomas, mesmo quando as raízes primárias não sobreviveram à dessecação, formando um mecanismo adicional de tolerância ao estresse. Assim, foi observado estratégias para evitar o estresse e mecanismos de tolerância ao estresse em sementes e sementes germinadas associada com padrões de precipitação nos diferentes biomas estudados. Estes mecanismos podem favorecer o estabelecimento das plântulas de forma rápida quando as condições são adequadas e permitirem a sobrevivência de plântulas quando as condições são adversas.

Secondary phloem diversity and evolution in Bignonieae (Bignoniaceae).

BACKGROUND AND AIMS: Phloem evolution has been explored in the literature across very broad scales, either for vascular plants as a whole or for major plant groups, such as the monocotyledons or the former dicotyledons. However, it has never been examined in a way that would elucidate evolutionary shifts leading to the diversification of phloem in single lineages. Therefore, the present study explores in detail the patterns of phloem evolution in the tribe Bignonieae (Bignoniaceae). This group represents a particularly good model for phloem studies since it is known to have a very conspicuous and diverse phloem. METHODS: A total of 19 phloem characters were coded in 56 species from all 21 genera currently recognized in the tribe Bignonieae, accounting for phloem wedge growth and for all the anatomical cell diversity encountered in the phloem. Phloem evolution was explored by reconstructing ancestral character states using maximum-likelihood assumptions with a time-calibrated molecular phylogeny for the group. Directionality and the effect of phylogenetic transformations in the current variation of quantitative traits and evolutionary correlations of selected discrete phloem traits were also tested under a maximum-likelihood approach. KEY RESULTS: Individual phloem features are quite diverse in the tribe, but generally conserved within smaller clades. Contrasting phloem patterns were found when comparing major groups, with certain lineages having the phloem marked by a background of phloem fibres where all other cells are embedded, tangentially arranged sieve tubes and sieve-tubecentric parenchyma. In contrast, other lineages exhibited a scarcely fibrous phloem, regularly stratified phloem, sieve tube elements in radial or diffuse arrangement, and diffuse parenchyma. We found signals of directional evolution in fibre abundance and number of sieve areas, which increased in the 'Fridericia and allies extended clade' and decreased in the 'Multiples of four extended clade', resulting in no signal of directionality when the whole Bignonieae was considered. In contrast, no indication of directional evolution was found for the axial parenchyma, either in single clades within Bignonieae or in the entire tribe. Positive correlation was found between sieve element length and both sieve plate type and the presence of a storied structure. Correlated evolution was also found between fibre abundance and several traits, such as sieve tube arrangement, sieve plate type, parenchyma arrangement, ray lignification and number of companion cells. CONCLUSIONS: The secondary phloem of Bignonieae is extremely diverse, with sister lineages exhibiting distinct phloem anatomies derived from contrasting patterns of evolution in fibre abundance. Fibre abundance in the tribe has diversified in correlation with sieve tube arrangement, sieve tube morphology, number of companion cells and parenchyma type. The results challenge long-standing hypotheses regarding general trends in cell abundance and morphological cell evolution within the phloem, and demonstrate the need to expand studies in phloem anatomy both at a narrow taxonomic scale and at a broad one, such as to families and orders.

[Seed germination of four tree species from the tropical dry forest of Valle del Cauca, Colombia]. / Germinación de semillas de cuatro especies arbóreas del bosque seco tropical del Valle del Cauca, Colombia.

The ecological restoration strategies for highly threatened ecosystems such as the tropical dry forest, depend on the knowledge of limiting factors of biological processes for the different species. Some of these include aspects such as germination and seed longevity of typical species present in those forests. In this study, we evaluated the effect of light and temperature on seed germination of two Fabaceae (Samanea saman and Jacaranda caucana) and two Bignoniaceae (Pithecellobium dulce and Tabebuia rosea) species having potential use in restoration, and we analyzed the seed storage behavior of these species for a three months period. To study the light effect, four levels of light quality on seeds were used (photoperiod of 12 hours of white light, darkness and light enriched in red and far-red, both for an hour each day), and we combined them with three levels of alternated temperatures (20/25, 20/30 and 25/30*C-16/8h). For the storage behavior, two levels of seed moisture content particular for each species were used (low: 3.5-6.1% and high: 8.3-13.8%), with three storage temperatures (20, 5 and -20 degrees C) and two storage times (one and three months). The criterion for germination was radicle emergence which was measured in four replicates per treatment, and was expressed as percentage of germination (PG). There were significant differences in germination of Samanea saman and Jacaranda caucana among light and temperature treatments, with the lowest value in darkness treatments, whereas germination of Pithecellobium dulce and Tabebuia rosea did not differ between treatments (PG>90%). The most suitable temperature regime to promote germination in all species was 25/30 degrees C. These four species showed an orthodox seed storage behavior. We concluded that seeds of R dulce, J. caucana and T. rosea did not have an apparent influence of all light conditions tested in their germination response, which might confer advantages in colonization and establishment processes, while S. saman did not germinate well in darkness. We suggest the use of seeds of P dulce, J. caucana and T rosea in ecological restoration processes, due to their tolerance and germination under a wide range of temperature and light conditions. Futhermore, seeds of S. saman might be used in open areas such as forest gaps.

Exploring 1,4-dihydroxyanthraquinone as long-range emissive ratiometric fluorescent probe for signaling Zn(2+)/PO4(3-): Ensemble utilization for live cell imaging.

Fluorescent 1,4-dihydroxyanthraquinone 1 was found to demonstrate its ratiometric signaling property upon interaction with divalent zinc (Zn(2+)). While the probe itself exhibited fluorescence emission in the yellow region (λem=544 nm and 567 nm), binding with Zn(2+) induced strong emission in the orange region (λem=600 nm) which was mainly due to a combination of CHEF and ICT mechanism. The probe was found to be highly sensitive toward the detection of zinc and the limit of detection (LOD) was calculated to be 9×10(-7) M. The possibility of using this probe for real-time analysis was strongly supported by the striking stability of fluorescence signal for more than five days with similar fluorescence intensity as observed during instant signaling. The present probe works within physiological pH range and is devoid of any interference caused by the same group elements such as Cd(2+)/Hg(2+). The probe possesses excellent excitation/emission wavelength profile and can penetrate cell membrane to image low concentration of zing inside living system. The in situ formed zinc-probe ensemble was further explored as ratiometric sensing platform for detecting another bio-relevant analyte phosphate anion through a zinc-displacement approach.

Germinación de semillas de cuatro especies arbóreas del bosque seco tropical del Valle del Cauca, Colombia / Seed germination of four tree species from the tropical dry forest of Valle del Cauca, Colombia

The ecological restoration strategies for highly threatened ecosystems such as the tropical dry forest, depend on the knowledge of limiting factors of biological processes for the different species. Some of these include aspects such as germination and seed longevity of typical species present in those forests. In this study, we evaluated the effect of light and temperature on seed germination of two Fabaceae (Samanea saman and Jacaranda caucana) and two Bignoniaceae (Pithecellobium dulce and Tabebuia rosea) species having potential use in restoration, and we analyzed the seed storage behavior of these species for a three months period. To study the light effect, four levels of light quality on seeds were used (photoperiod of 12 hours of white light, darkness and light enriched in red and far-red, both for an hour each day), and we combined them with three levels of alternated temperatures (20/25, 20/30 and 25/30°C-16/8h). For the storage behavior, two levels of seed moisture content particular for each species were used (low: 3.5-6.1% and high: 8.3-13.8%), with three storage temperatures (20, 5 and -20°C) and two storage times (one and three months). The criterion for germination was radicle emergence which was measured in four replicates per treatment, and was expressed as percentage of germination (PG). There were significant differences in germination of Samanea saman and Jacaranda caucana among light and temperature treatments, with the lowest value in darkness treatments, whereas germination of Pithecellobium dulce and Tabebuia rosea did not differ between treatments (PG>90%). The most suitable temperature regime to promote germination in all species was 25/30°C. These four species showed an orthodox seed storage behavior. We concluded that seeds of P. dulce, J. caucana and T. rosea did not have an apparent influence of all light conditions tested in their germination response, which might confer advantages in colonization and establishment processes, while S. saman did not germinate well in darkness. We suggest the use of seeds of P. dulce, J. caucana and T. rosea in ecological restoration processes, due to their tolerance and germination under a wide range of temperature and light conditions. Futhermore, seeds of S. saman might be used in open areas such as forest gaps.

Las estrategias de restauración ecológica en ecosistemas altamente amenazados como el bosque seco tropical, dependen del conocimiento de factores limitantes de procesos biológicos, como la germinación y la longevidad de semillas de especies típicas de los mismos. En este estudio se evaluó el efecto de la luz y la temperatura en la germinación de semillas de dos especies de Fabaceae y dos de Bignoniaceae potencialmente útiles para restauración, y se determinó su comportamiento en el almacenamiento. Para el primer objetivo, se utilizaron cuatro niveles de calidad lumínica (fotoperiodo con 12 horas de luz, oscuridad y estímulos de luz enriquecida en rojo y luz enriquecida en rojo lejano, ambas por una hora), combinados con tres niveles de temperaturas alternadas (20/25, 20/30 y 25/30°C - 16/8h). Para el segundo objetivo, se utilizaron dos niveles de contenido de humedad de las semillas, con tres niveles de temperatura de almacenamiento (20, 5 y -20°C) y dos tiempos de almacenamiento (1 y 3 meses). La variable de respuesta para ambos experimentos fue el número de semillas germinadas, expresado en porcentaje (PG). Samanea saman y Jacaranda caucana presentaron un PG significativamente diferente entre los tratamientos lumínicos, con el valor menor en la oscuridad, mientras Pithecellobium dulce y Tabebuia rosea no presentaron diferencias significativas entre tratamientos (PG>90%). La respuesta a la temperatura mostró que 25/30°C es el régimen más adecuado para promover la germinación de la mayoría de las especies. Las semillas de las cuatro especies evaluadas presentaron un comportamiento ortodoxo en el almacenamiento. Se concluye que las semillas de P. dulce, J. caucana y T. rosea son indiferentes a las condiciones de luz y temperatura probadas, confiriéndoles ciertas ventajas en procesos de colonización y establecimiento frente a S. saman, cuya germinación no se ve favorecida en la oscuridad. Se sugiere el uso de semillas de P. dulce, J. caucana y T. rosea en proyectos de restauración ecológica del bosque seco Tropical, debido a su tolerancia y germinación en condiciones ambientales con intervalos amplios de temperatura y luminosidad. Mientras que, semillas de S. saman podrían ser usadas en sitios abiertos como claros de bosques.

Histological features, starch accumulation and sprouting in the early root development of Jacaranda ulei (Bignoniaceae).

The plant species occurring in the savanna region of the Cerrado biome in Brazil present typical morphological and physiological adaptations to a dry climate with seasonal occurrence of wildfires. In this study, the histological features of the root system, the main sites of synthesis and storage of starch and the initial phases of the bud development were characterized in Jacaranda ulei. The anatomical features observed in the root system of J. ulei are related to the needs of the species to survive in the Cerrado. The histochemical analyses demonstrated high synthesis of glucose and glycoprotein after the third day of in vitro culture, in the proximal cells of the cortical parenchyma of the exoderm. Meristematic primordia were observed in the ninth day and the beginning of the meristem formation was observed after 21 days of in vitro culture. Jacaranda ulei displays morphological, anatomical and storage features typical from resprouter species. However, it may be vulnerable to unsustainable exploitation. Considering the importance of this species for local people, more studies regarding its therapeutic properties should be performed, including the planning of appropriate programs for the species management and the production of selected clones through in vitro micropropagation.

Influência da densidade de plantio e da cama de frango na produção da carobinha (Jacaranda decurrens Cham. ssp. symmetrifoliolata Farias & Proença) / Effects of plant density and the use of broiler litter on the Jacaranda decurrens Cham. ssp. symmetrifoliolata Farias & Proença yield

Carobinha (Jacaranda decurrens Cham. ssp. symmetrifoliolata Farias & Proença) é uma planta medicinal que ocorre no Cerrado de Mato Grosso do Sul. A espécie vem sendo amplamente utilizada pela medicina popular como depurativa do sangue e cicatrizante de feridas uterinas e dos ovários, resultando na sua exploração predatória. Por ser uma espécie identificada recentemente, os estudos sobre adaptação ex situ são ainda escassos. O objetivo deste trabalho foi avaliar o crescimento, o desenvolvimento, e a produção da carobinha cultivada ex situ em cinco espaçamentos entre plantas dentro das fileiras, em parcelas adubadas com e sem cama semidecomposta de frango de corte. O experimento foi desenvolvido no Horto de Plantas Medicinais (HPM) da Universidade Federal da Grande Dourados - UFGD, em Dourados-MS, de abril de 2006 a outubro de 2007. Os espaçamentos entre plantas foram: 0,30; 0,35; 0,40; 0,45 e 0,50 m, sendo a cama de frango aplicada na dose de 10 t ha-1 nas parcelas com este tratamento. O ensaio fatorial 5x2, foi conduzido no delineamento experimental de blocos casualizados, sendo a parcela constituída de 4,5 m², com quatro repetições. As mudas foram obtidas a partir de sementes coletadas de uma população de 40 plantas nativas, distribuídas ao acaso, em área de cerca de 100 m² com vegetação característica de Cerrado. Durante o cultivo avaliou-se a altura das plantas a cada 30 dias a partir de 150 até 480 dias após o transplante (DAT). As plantas foram colhidas aos 480 DAT, quando avaliou-se as massas fresca e seca da parte aérea, raízes, área foliar, e o comprimento e o diâmetro da maior raiz. A altura máxima (1,58 m) foi alcançada aos 471 DAT no espaçamento de 0,50 m entre plantas, tratadas com a cama de frango. Os dados de produtividade foram influenciados significativamente pelos espaçamentos, mas não pela adição da cama. O rendimento de massa seca das folhas por hectare não foi influenciado pelo espaçamento. A máxima área foliar foi obtida no espaçamento 0,50 m. Os maiores valores de massa seca de caules por hectare ocorreram no espaçamento de 0,30 m. Os maiores diâmetros de raiz, de xilopódio, e de caule, ocorreram no espaçamento 0,30 m, sendo os máximos valores de comprimento de raiz e de xilopódio também observados neste espaçamento. Os maiores rendimentos de massa seca de raiz (4,564 t ha-1) e de xilopódio (2,102 t ha-1) foram obtidos no espaçamento 0,30 m. Pelos resultados obtidos, concluiu-se que para se obter maiores produções de carobinha ela deve ser cultivada no espaçamento de 0,30 m, independente da cama de frango.

Jacaranda decurrens Cham. ssp. symmetrifoliolata Farias & Proenca, commonly known as carobinha, is a medicinal plant which occurs in the Cerrado of Mato Grosso do Sul. The species is widely used in folk medicine as a blood purifying and for healing wounds of uterine and ovarian cancers, which leads to its extensive and predatory exploitation. As it is a newly identified species, studies on its adaptation ex situ are still rare. Thus, the objective of this experiment was to evaluate the early development and yield of the species cultivated ex situ using five different plant spacing within the ranks and applying semicomposed broiler litter in the plots. The experiment took place at the Garden of Medicinal Plants from the Federal University of Grande Dourados ( UFGD), between April 2006 and October 2007. Plant spacings were: 0.30, 0.35, 0.40, 0.45 and 0.50 m, with broiler litter applied at 10 ton ha-1 in plots. The 5x2 factorial trial was designed as randomized blocks, with 4.5 m² plot area and four replications each. The seedlings were obtained from seeds collected in a natural germoplasm bank of 40 plants, randomly distributed in an area of about 100 m² with typical Cerrado vegetation. During cultivation, the height of the plants was evaluated every 30 days from 150 to 480 days after transplanting (DAT). Plants were harvested at 480 DAT, when fresh and dry matter of shoots, roots, leaf area and length and diameter of the largest root were assessed. The maximum height (1.58 m) was reached at 471 DAT at 0.50 m spacing between plants when poultry litter was applied. The yield data were influenced significantly by the spacing, but not by the addition of the broiler litter. The yield per hectare of dry mass of leaves was not influenced by plant spacing, however maximum leaf area was also obtained at 0.50 m spacing. The highest values of dry mass of stems per hectare were obtained at 0.30 m between plants, as well as the largest diameters of root, xylopodium and stem and the maximum length of root and xylopodium was also observed under this plant spacing. Someway, the highest yields of dry mass of root (4.564 t ha-1) and xylopodium (2.102 t ha-1) occurred when used 0.30 m plant spacing. Based on these results, we concluded the species achieves higher yields when grown in 0.30 m plant spacing, independent of the addition of poultry litter.

Longevity of the Brazilian underground tree Jacaranda decurrens Cham.

Underground trees are a rare clonal growth form. In this survey we describe the branching pattern and estimate the age of the underground tree Jacaranda decurrens Cham. (Bignoniaceae), an endangered species from the Brazilian Cerrado, with a crown diameter of 22 meters. The mean age calculated for the individual was 3,801 years, making it one of the oldest known living Neotropical plants.

Anti-inflammatory activity of Markhamia tomentosa (Benth.) K. Schum. Ex Engl. ethanolic leaf extract.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Markhamia tomentosa (Benth.) K. Schum (Bignoniaceae) are used traditionally for the treatment of oedema and rheumatoid arthritis in Nigeria. AIM OF THE STUDY: The aim of the work was to investigate the anti-inflammatory activity of the ethanolic leaf extract of Markhamia tomentosa. MATERIALS AND METHODS: The extract was screened using the carrageenan-induced paw oedema in rats, xylene-induced oedema in mice and the formalin-induced oedema in mice at 50, 100, 200mg/kg doses p.o respectively. The mechanism by which the extract mediated the anti-inflammatory activity was assessed using the histamine-induced rat paw oedema and serotonin-induced rat paw oedema at the highest dose (200mg/kg). RESULTS: The results showed that the extract produced a significant dose-dependent inhibition in carrageenan-induced, xylene-induced and the formalin tests. The extract exerted a significant inhibition of 54.55% (P<0.0001) and 42.11% (P<0.01) at 90 min in the histamine-induced and serotonin-induced rat paw oedema models respectively. CONCLUSIONS: These findings suggest that the ethanolic leaf extract of Markhamia tomentosa possesses anti-inflammatory activity possibly mediated by histamine. The results justify the use of the plant in the preparation of ethno medicines used in the treatment of ailments associated with inflammation.

Influence of arbuscular mycorrhizal fungi (AMF) on zinc biogeochemistry in the rhizosphere of Lindenbergia philippensis growing in zinc-contaminated sediment.

The association of arbuscular mycorrhizal fungi (AMF) with the roots of Lindenbergia philippensis (Cham.) Benth., sampled from a Zn-contaminated settling pond at a zinc smelter, significantly enhanced Zn accumulation (72,540 ± 5,092 mg kg⁻¹ dry weight) in rhizosphere sediment amended with 1,000 mg L⁻¹ of Zn sulfate solution compared to fungicide-treatments that suppressed AMF colonization. This can be explained by a significant proportion of Zn being found in rectangular crystals that were associated with the root mucilaginous sheath. Despite this, all treatments maintained the same Zn coordination geometry in both Zn oxidation state and the coordinated neighbouring atoms. X-ray absorption spectroscopy (XAS) showed a Zn(II) oxidation state as a core atom and associated with six oxygen atoms symmetrically arranged in an octahedral coordination and coordinated with sulfur. The results may indicate a role for AMF in enhancing Zn immobilization in the rhizosphere of indigenous plants that successfully colonize Zn mining and smelting disposal sites.

The complex leaves of the monkey's comb (Amphilophium crucigerum, Bignoniaceae): a climbing strategy without glue.

PREMISE OF THE STUDY: Monkey's comb (Amphilophium crucigerum) is a widely spread neotropical leaf climber that develops attachment pads for anchorage. A single complex leaf of the species comprises a basal pair of foliate, assimilating leaflets and apical, attaching leaflet tendrils. This study aims to analyze these leaves and their ontogenetic development for a better understanding of the attachment process, the form-structure-function relationships involved, and the overall maturation of the leaves. METHODS: Thorough morphometrical, morphological, and anatomical analyses incorporated high-resolution microscopy, various staining techniques, SEM, and photographic recordings over the entire ontogenetic course of leaf development. KEY RESULTS: The foliate, assimilating leaflets and the anchorage of the more apical leaflet tendrils acted independently of each other. Attachment was achieved by coiling of the leaflet tendrils and/or development of attachment pads at the tendril apices that grow opportunistically into gaps and fissures of the substrate. In contact zones with the substrate, the cells of the pads differentiate into a vessel element-like tissue. During the entire attachment process of the plant, no glue was excreted. CONCLUSION: The complex leaves of monkey's comb are highly differentiated organs with specialized leaf parts whose functions-photosynthesis or attachment-work independently of each other. The function of attachment includes coiling and maturation process of the leaflet tendrils and the formation of attachment pads, resulting in a biomechanically sound and persistent anchorage of the plant without the need of glue excretion. This kind of glue-less attachment is not only of interest in the framework of analyzing the functional variety of attachment structures evolved in climbing plants, but also for the development of innovative biomimetic attachment structures for manifold technical applications.