Wood anatomy of major Bignoniaceae clades

The circumscription of Bignoniaceae genera and tribes has undergone major changes following an increased understanding of phylogenetic relationships within the family. While DNA sequence data have repeatedly reconstructed major clades within the family, some of the clades recovered still lack diagnostic morpho-anatomical features, complicating their recognition. In this study we investigated the wood anatomy of all major lineages of Bignoniaceae (except Tourrettieae) in search for anatomical synapomorphies for clades. We sampled 158 species of Bignoniaceae, representing 67 out of the 82 genera currently recognized. Detailed descriptions of quantitative and qualitative wood anatomical features are presented for each clade and interpreted in the light of a molecular phylogeny for the family. Jacarandae are characterized by a paratracheal winged-aliform parenchyma, with the traditional subdivision of Jacaranda into sections Monolobos and Dilobos supported by the uniseriate and homocellular rays of Monolobos versus the wide and heterocellular rays of Dilobos. Tecomeae s.s. are characterized by scanty paratracheal parenchyma, septate fibers, and heterocellular rays, traits also found in Delostoma, a genus previously included in Tecomeae s.l., but recently shown to represent a separate lineage. Crescentiina includes two sub-clades, the Tabebuia alliance and the Paleotropical clade, which share abundant aliform parenchyma, short and mainly homocellular rays, less commonly with heterocellular rays with body procumbent and one row of marginal square cells. Members of the Tabebuia alliance and the Paleotropical clade can be distinguished from each other by the narrow vessels with a widespread storied structure found in members of the Tabebuia alliance, versus the vessels with medium to wide width and a non-storied structure found in members of the Paleotropical clade. Oroxyleae are characterized by a combination of simple and foraminate perforation plates and homocellular rays, while Catalpeae are characterized by scanty paratracheal parenchyma, abundant tyloses and vessel-ray pits simple to semi-bordered. Bignonieae differ from all other clades by a variant secondary growth and a typically lianoid wood anatomy. Overall, wood anatomical characters are not very labile within the family, being distributed across clades in a very predictive manner. Several anatomical characters represent good anatomical synapomorphies and provide further support to clades identified in molecular phylogenetic studies.The authors are indebted to Alex C. Wiedenhoeft, Mike Wiemann and Regis Miller from the Forest Products Laboratory for warmly hosting us and allowing the study of the entire MADw collection (Madison, Wisconsin); Maria José Miranda and Raphael Pigozzo from the Institute of Technological Research (IPT, BCTw, São Paulo) for slides of Sparattosperma, Handroanthus and Tabebuia; Alexandre Zuntini, Anselmo Nogueira, Diana Sampaio, Mariane Souza-Baena, Milton Groppo from the Universidade de São Paulo (Brazil), Márdel Lopes from the Universidade Federal de Viçosa (Brazil), and Luzmilla Arroyo and Daniel Villaboel of the Museo Noel Kempff Mercado (Bolivia) for collecting samples or for assistance in field collections, Harri Lorenzi for allowing us to collect samples at the Instituto Plantarum/Jardim Botânico Plantarum; Antonio Carlos Franco Barbosa sectioning Stereospermum; Julio C. Majcher for sectioning samples of Oroxyleae used in Scanning Electron Microscopy; Pieter Baas, Michelle Zjhra, Guillermo Angeles, and Giuliano Locosselli for invaluable suggestions in early versions of the manuscript; Gabriella Pace, Lizana Rezende and Carolina L. Bastos for valuable support, the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, grants 481034/2007-2; 486971/2012-0), and the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, grant 2012/01099-8; 2013/10679-0) for financial support. RGO is supported by US National Science Foundation (DEB-0309065, DEB-1353761). LGL and VA by CNPq productivity 307781/2013-5 and 308441/2012-5, respectively

Evolution of the cambial variant and the secondary phloem in Bignonieae (Bignoniaceae)

Lianas de Bignoniaceae são reconhecidas por apresentarem uma variação cambial em seus caules, que promove a formação de cunhas de floema que interrompem o xilema. Uma grande diversidade de formas anatômicas foram descritas para esta variação, assim como o floema resultante dela também foi descrito como sendo distinto do floema normal presente concomitantemente nestes caules. Entretanto, nada se sabe sobre a origem, evolução e diversificação das características anatômicas neste grupo. Por essa razão, o presente estudo teve como objetivo uma análise anatômica dos caules de Bignonieae num contexto filogenético, com o intuito de lançar hipóteses para a evolução da diversidade anatômica na tribo. Para tanto, foi realizada uma análise anatômica caulinar de 54 espécies de Bignonieae, representantes dos 21 gêneros atualmente reconhecidos. Nossos resultados apontam que, não obstante a grande diversidade anatômica presente nos caules de Bignonieae, todas compartilham estágios comuns de desenvolvimento, seguidos de adições terminais que promoveram o aumento da complexidade em seus caules. Além disso, vimos que as diferenças entre o floema normal e o variante tem aumentado ao longo da evolução e está presente em todos os tipos celulares do floema. Vimos ainda que o floema secundário em Bignonieae evolui em direções opostas em diferentes linhagens da tribo, evidenciando que a evolução do floema não segue uma única direção, mas várias. Por fim, este estudo demonstra que análises anatômicas dentro de um contexto filogenético são primordiais por permitirem um maior entendimento dos processos que promoveram a evolução e diversificação dos grupos.Lianas in Bignoniaceae are well known for presenting a cambial variant in their stems, which develops into phloem wedges that deep furrows the xylem. An enormous diversity of anatomical forms were described as resulting from this cambial variant, as well as the phloem produced by the cambial variant was described as being distinct from the regular phloem concurrently present in these stems. However, nothing is known about the origin, evolution, and diversification of the anatomical traits in this group. Therefore, the present study aimed to provide an anatomical analysis of the stems of Bignonieae (Bignoniaceae) within a phylogenetic framework, in order to address questions on the evolution of anatomical diversity in this tribe. For that reason, here we analyzed the stems of 54 species of Bignonieae, representative of the 21 genera currently known for the tribe. Our results show that, despite the great anatomical diversity present in the stems of Bignonieae, all of them share common developmental stages, which are then followed by subsequent terminal additions that are though to have promoted an augment in the complexity of these stems. Furthermore, our results indicate that the differences found between regular and variant phloem is increasing along time and is present in all cell types of the phloem. Moreover, we found that the secondary phloem in Bignonieae is evolving in opposite directions in distinct lineages of the tribe, evidencing that the evolution of the phloem is not constraint to a single line of specialization. In conclusion, this study demonstrates the importance of anatomical analyses within a phylogenetic framework, allowing for the detection of the processes that have been involved in the evolution and diversification of plant groups

Evolution of the vascular system in lineages that contain lianas

O sistema vascular das lianas, em especial o xilema, mostrou−se repetidas vezes distinto nas lianas, com aspectos compartilhados mesmo dentre linhagens distantemente relacionadas, tais como a presença de variações cambiais, vasos mais largos e longos, parênquima axial mais abundante − frequentemente não−lignificado − raios mais altos e largos − geralmente heterocelulares. Não obstante todo esse conhecimento, poucos trabalhos investigaram o impacto da evolução do hábito lianescente no sistema vascular em linhagens cujos ancestrais não são lianas e sim plantas auto−suportantes. Portanto, nesta tese exploramos o lenho, o floema e a anatomia caulinar como um todo em linhagens que contêm lianas e plantas auto−suportantes, utilizando filogenias bem sustentadas e investigações anatômicas detalhadas. Em Bignoniaceae (Lamiales), investigamos em detalhe a anatomia do lenho, delimitando caracteres e estados de caráter e mapeando-os na filogenia mais recente do grupo, encontrando que modificações eco−fisiológicas e transições de hábito tiveram grande impacto na evolução do lenho na família. Anéis porosos e semi−porosos, bem como espessamento espiralado foram encontrados em plantas crescendo em latitudes mais altas ou em regimes hídricos fortemente sazonais, ao passo que fibras septadas apareceram correlacionadas com a presença de parênquima axial escasso. A evolução de lianas, por sua vez, parece ter levado a um aumento no diâmetro dos vasos, contudo dimórficos, células perfuradas de raio, parênquima axial mais escasso e surgimento de variações cambiais. Apesar da enorme diversidade dentro de Bignoniaceae, os grandes clados possuem uma anatomia bastante preditiva e 9 possíveis sinapomorfias morfológicas são sugeridas para clados delimitados somente com base em dados moleculares. Dentro das traqueófitas, investigamos 26 pares filogeneticamente controlados de lianas espécies auto−suportantes relacionadas pertencentes a todas as principais linhagens de traqueófitas (exceto licófitas), a fim de buscar caracteres que tenham evoluído em correlação com o hábito lianescente. Encontramos que os elementos crivados e os poros das placas crivadas têm sempre maior calibre nas lianas, e que os raios são mais altos e heterocelulares. Contudo, as principais características do floema das lianas se mantêm conservadas em relação às espécies auto−suportantes relacionadas, evidenciando que as lianas teriam evoluído um sistema de condução de fotossintetatos mais eficiente, porém preservando um alto sinal filogenético. Em Malpighiaceae, lianas são abundantes, tal como as variações cambiais. Contudo, pouco se sabe sobre o número de variações presentes na família ou como elas estariam distribuídas. Aqui delimitamos 6 diferentes tipos de variação cambial, que teriam evoluído independentemente 8 vezes na família, cujo ancestral é reconstruído como tendo caule simples. Muitas dessas variações compartilham estágios de desenvolvimento, ao passo que variações anatomicamente muito similares derivam de trajetórias ontogenéticas distintas. Dentro dos gêneros as variações se mostraram conservadas e mesmo dentre grupos irmãos do novo e velho mundo, evidenciando que as variações cambiais seriam bons indicadores de relações na família. De maneira geral, podemos concluir que lianas impactam significativamente o sistema vascular nas linhagens onde ocorrem e que tais modificações em geral resultam em um sistema de condução hídrico e de fotossintetatos mais eficiente e também mais flexível para a escalada.The vascular system of lianas, especially the xylem, has been repeatedly been shown to be different, with lianas having a set of features shared among even distantly related lineages, such as the presence of cambial variants, wide and long vessels, more abundant axial parenchyma, frequently non−lignified, taller and wider rays, which are generally heterocellular. In spite of this amount of knowledge, few works have investigated the impact for the vascular system of the evolution of this habit within lineages whose ancestors are not lianas, but self-supporting plants. Therefore, in this dissertation we explored wood, phloem and overall stem−anatomy evolution in lineages that contain lianas and self−supporting plants, using well−supported phylogenies and detailed anatomical investigations. Within Bignoniaceae (Lamiales), we thoroughly investigated the wood anatomy, delimiting character states and mapping them onto the last phylogeny for the group, encountering that eco−physiological and habit transition were the main drivers of modifications in the wood anatomy in the family. Ring−porous and semi−ring porous woods and helical thickening was found in plants either growing in higher latitudes or with marked seasonal water regimes, and septate fibres correlated with scanty axial parenchyma, which are eco−physiological drivers. Evolution of lianas, in turn, drove an increase in vessel diameter, wide vessels accompanied by very narrow ones, presence of perforated ray cells, scanty axial parenchyma and cambial variants. Despite the great wood anatomical diversity within the family, major clades have quite predictive wood anatomy and 9 possible anatomical synapomorphies were raised in this work to clades previously delimitated exclusively by molecular characters. Within the tracheophytes, we investigated 26 phylogenetically controlled pairs of lianas and their self−supporting relatives within all major lineages of tracheophytes (except lycophytes), in order to seek characters evolving in correlation with the lianescent habit. We found that the sieve elements and sieve pores were always wider in the lianas, and that the rays were always taller. However, all the main characters of the phloem of the lianas remained conserved with that of their self−supporting relatives. This evidenced that although a more efficient photosynthetic conductive system evolved in the phloem of lianas, overall anatomy conserved a high phylogenetic signal. Within Malpighiaceae, lianas are abundant and many cambial variants are present. However, nothing was known regarding how many types of cambial variants there were in the family and how they were distributed. We were able to delimit 6 different types of cambial variants that evolved at least 8 times independently in the family, which ancestrally lacks a cambial variant. Many of these types share common stages of development and some variants that are anatomically very similar derive from different ontogenetic trajectories. Within the genera, the variants are conserved, and even between sister groups in the new and old world, evidencing that cambial variants may be a good indicator of relationships within the family. Overall, we conclude that lianas greatly impact the evolution of the vascular system in the lineages where they have evolved, and these modifications normally result in a more efficient water and photosynthates conduction system and an increased flexibility for climbing

Diversity of metaxylem vessel elements in three Syagrus palms (Arecaceae) of different habits

Vessel elements in Monocotyledons present morphological differences according to the organ where they occur. Usually such diff erences have been explained from an evolutionary perspective, with few data on how the vessel elements could be infl uenced by the diff erent functions of organs and the growth habits of the plants. To address this question, three vegetative organs of palms of the same genus, Syagrus, growing in similar environments, but with different habits, were analyzed. Accordingly, we aimed to detect whether the vessel elements would present similar features in all species or whether the vessel elements would change according to their different habits. We found that the width and type of perforation plates varied in the same way among all species, while the lengths varied in an unusual form. First, all species presented very long elements in the roots, either as long, or longer than those of the stems and leaves. Second, the vessel elements of the stems varied considerably among the species. Specifi cally, in Syagrus romanzoffi ana, vessel elements of the stem were equal in size to those of the other organs, while in both Syagrus fl exuosa and Syagrus petraea, shorter vessel elements were found in the stems. We surmise that vessel elements in palm roots may be related to the high pressure-potential required to avoid stem embolism. The dimorphism of the vessel elements in the stems most likely reflects the distinct habits of these species. Large stems, such as of those of the arboreal palms, presented much longer vessels than those of subterranean stems. Based on these anatomical findings, we suggest that the diff erences found among the vessel elements of roots, stem, and leaves may have evolved in response not only to phylogenetic and ecological constraints, but also to specificities derived from the different plant habits

Physiological changes associated with antioxidant enzymes in response to sugarcane tolerance to water deficit and rehydration

Water deficit is the main limiting factor for sugarcane yield worldwide. Under this stress, the crop develops adaptive mechanisms that participate in maintaining the plants’ water status. A better understanding of these mechanisms may contribute to differentiating tolerant cultivars to be used in genetic improvement programs. Therefore, this study aimed to assess the physiological responses of sugarcane cultivars subjected to water deficit and rehydration. Cultivars RB92579, SP81-3250, SP83-2847 and IAC91-5155 were grown in pots in a greenhouse. These plants were physiologically and biochemically evaluated at three intervals: 0 (before stress), 15 (water deficit) and 27 days (rehydration) after the onset of treatment (DAT) at 85 days after planting. Cultivars RB92579 and SP81-3250 had greater reductions in stem height, number of green leaves, leaf area, relative water content, leaf water potential (Ψw), leaf temperature, stomatal conductance (gs), maximum photochemical efficiency of photosystem II (Fv/Fm), SPAD index and photosynthetic pigments, and inhibited catalase enzyme activity when subjected to water stress. Furthermore, SP81-3250 showed no recovery after rehydration. Water deficit also increased the levels of proline and the enzymes superoxide dismutase and ascorbate peroxidase in all cultivars. We also found increased carbohydrates under stress, except in the cultivar SP81-3250, which had a reduced content. Due to the minor damage caused by drought in the physiology and biochemistry of cultivars SP83-2847 and IAC91-5155, they have higher potential for tolerance and can acclimate to drought. Consequently, oxidative stress can be largely avoided in the chloroplasts of these cultivars by maintaining the balance of antioxidant enzyme activity, photosynthetic efficiency, stomatal control and water status.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Evaluation of a quality improvement intervention to reduce anastomotic leak following right colectomy (EAGLE): pragmatic, batched stepped-wedge, cluster-randomized trial in 64 countries

Background Anastomotic leak affects 8 per cent of patients after right colectomy with a 10-fold increased risk of postoperative death. The EAGLE study aimed to develop and test whether an international, standardized quality improvement intervention could reduce anastomotic leaks. Methods The internationally intended protocol, iteratively co-developed by a multistage Delphi process, comprised an online educational module introducing risk stratification, an intraoperative checklist, and harmonized surgical techniques. Clusters (hospital teams) were randomized to one of three arms with varied sequences of intervention/data collection by a derived stepped-wedge batch design (at least 18 hospital teams per batch). Patients were blinded to the study allocation. Low- and middle-income country enrolment was encouraged. The primary outcome (assessed by intention to treat) was anastomotic leak rate, and subgroup analyses by module completion (at least 80 per cent of surgeons, high engagement; less than 50 per cent, low engagement) were preplanned. Results A total 355 hospital teams registered, with 332 from 64 countries (39.2 per cent low and middle income) included in the final analysis. The online modules were completed by half of the surgeons (2143 of 4411). The primary analysis included 3039 of the 3268 patients recruited (206 patients had no anastomosis and 23 were lost to follow-up), with anastomotic leaks arising before and after the intervention in 10.1 and 9.6 per cent respectively (adjusted OR 0.87, 95 per cent c.i. 0.59 to 1.30; P = 0.498). The proportion of surgeons completing the educational modules was an influence: the leak rate decreased from 12.2 per cent (61 of 500) before intervention to 5.1 per cent (24 of 473) after intervention in high-engagement centres (adjusted OR 0.36, 0.20 to 0.64; P < 0.001), but this was not observed in low-engagement hospitals (8.3 per cent (59 of 714) and 13.8 per cent (61 of 443) respectively; adjusted OR 2.09, 1.31 to 3.31). Conclusion Completion of globally available digital training by engaged teams can alter anastomotic leak rates. Registration number: NCT04270721 (http://www.clinicaltrials.gov)