BackgroundTeak (Tectona grandis L.f.) is currently the preferred choice of the timber trade for fabrication of woody products due to its extraordinary qualities and is widely grown around the world. Gene expression studies are essential to explore wood formation of vascular plants, and quantitative real-time reverse transcription PCR (qRT-PCR) is a sensitive technique employed for quantifying gene expression levels. One or more appropriate reference genes are crucial to accurately compare mRNA transcripts through different tissues/organs and experimental conditions. Despite being the focus of some genetic studies, a lack of molecular information has hindered genetic exploration of teak. To date, qRT-PCR reference genes have not been identified and validated for teak.ResultsIdentification and cloning of nine commonly used qRT-PCR reference genes from teak, including ribosomal protein 60s (rp60s), clathrin adaptor complexes medium subunit family (Cac), actin (Act), histone 3 (His3), sand family (Sand), β-Tubulin (Β-Tub), ubiquitin (Ubq), elongation factor 1-α (Ef-1α), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Expression profiles of these genes were evaluated by qRT-PCR in six tissue and organ samples (leaf, flower, seedling, root, stem and branch secondary xylem) of teak. Appropriate gene cloning and sequencing, primer specificity and amplification efficiency was verified for each gene. Their stability as reference genes was validated by NormFinder, BestKeeper, geNorm and Delta Ct programs. Results obtained from all programs showed that TgUbq and TgEf-1α are the most stable genes to use as qRT-PCR reference genes and TgAct is the most unstable gene in teak. The relative expression of the teak cinnamyl alcohol dehydrogenase (TgCAD) gene in lignified tissues at different ages was assessed by qRT-PCR, using TgUbq and TgEf-1α as internal controls. These analyses exposed a consistent expression pattern with both reference genes.ConclusionThis study proposes a first broad collection of teak tissue and organ mRNA expression data for nine selected candidate qRT-PCR reference genes. NormFinder, Bestkeeper, geNorm and Delta Ct analyses suggested that TgUbq and TgEf-1α have the highest expression stability and provided similar results when evaluating TgCAD gene expression, while the commonly used Act should be avoided.

Drought stress is an increasingly common and worrying phenomenon because it causes a loss of production in both agriculture and forestry. Teak is a tropical tree which needs alternating rainy and dry seasons to produce high-quality wood. However, a robust understanding about the physiological characteristics and genes related to drought stress in this species is lacking. Consequently, after applying moderate and severe drought stress to teak seedlings, an infrared gas analyzer (IRGA) was used to measure different parameters in the leaves. Additionally, using the root transcriptome allowed finding and analyzing the expression of several drought-related genes. As a result, in both water deficit treatments a reduction in photosynthesis, transpiration, stomatal conductance and leaf relative water content was found. As well, an increase in free proline levels and intrinsic water use efficiency was found when compared to the control treatment. Furthermore, 977 transcripts from the root contigs showed functional annotation related to drought stress, and of these, TgTPS1, TgDREB1, TgAREB1 and TgPIP1 were selected. The expression analysis of those genes along with TgHSP1, TgHSP2, TgHSP3 and TgBI (other stress-related genes) showed that with moderate treatment, TgTPS1, TgDREB1, TgAREB1, TgPIP1, TgHSP3 and TgBI genes had higher expression than the control treatment, but with severe treatment only TgTPS1 and TgDREB1 showed higher expression than the control treatment. At the end, a schematic model for the physiological and molecular strategies under drought stress in teak from this study is provided. In conclusion, these physiological and biochemical adjustments in leaves and genetic changes in roots under severe and prolonged water shortage situations can be a limiting factor for teak plantlets’ growth. Further studies of those genes under different biotic and abiotic stress treatments are needed.

Sourgrass(Digitaria insularis, L. Mez ex Ekman)is a weed that requires high rates of glyphosate ((N-[phosphonomethyl]-glycine) forcontrol, verylittle of the herbicide applied isabsorbed by theleaves. Morphological and histological differences in leaves of glyphosate-resistant and glyphosate-susceptible plantsshould explain the contrast of variance between herbicide susceptibility. Leaves of different growth andphenological stages were collected and submitted to histological andelectron microscopy scanning analysis. Those plants were also submitted to a glyphosate dose-response curve analysis. The results suggest thatbiotypes exhibits differences infoliar structures that can influence the uptake and translocation of glyphosate. A thinparenchyma, lesser distance amongvascular bundles, and higher phloem sizewere found in resistant biotypes (differences close to 10%). Minor stomata number and higher epicuticular waxy deposition in stomata and leaf surface were found in resistant plants when leaves came from regrowth, leading to a possiblelesser herbicide absorption. Resistant plants showed absence of trichomes in late stages of development (64% less trichomes than susceptible plants). The higher concentration of trichomes in susceptible plants can be an absorption pathfor glyphosate getthrough the cuticle.Keywords: Absorption, herbicide resistance, parenchyma,trichomes A anatomia foliar de capim-amargoso influencia na resistência ao glyphosate? ResumoO capim-amargoso (Digitaria insularis, L. Mez ex Ekman) é uma plantadaninha que requer altas dosesde glifosato ((N-[fosfonometilo] glicina) para seu controle, sendo pouco do herbicida aplicado absorvido pelas folhas. Diferenças morfológicas e histológicas em folhas de biótipos resistentes e suscetíveis ao glifosato podem explicar a susceptibilidade diferencial ao herbicida. Folhas de diferentes estádios de crescimento de plantas foram coletadas e submetidas à análise histológica e de microscopia eletrônica de varredura. Essas plantas foram também submetidas a uma curva de dose-resposta ao glifosato. Os resultados sugerem diferenças foliares que podem influenciar a absorção e translocação de glyphosate. Um parênquima menos espesso, menores distâncias entre feixes vasculares, e maior tamanho de floema no biótipo resistente em relação ao biótipo suscetível foram observados (diferenças próximo a 10%). Menor número de estômatos e maior deposição de cera epicuticular em estômatos e superfície foliar nos biótipos resistentes foram encontradas quando a folha veio do rebrote, levando a uma possível menor absorção do herbicida. Em biótipos resistentes encontrou-se aausência de tricomas nos últimos estágios do desenvolvimento (64% a menos do que os tricomas plantas suscetíveis). A maior concentração de tricomas em plantas sensíveis, podem ser uma via de absorção para o glyphosate por entre a cutícula. Palavras-chave:Resistência a herbicidas, absorção, tricomas, parênquima

BackgroundCurrently, Tectona grandis is one of the most valuable trees in the world and no transcript dataset related to secondary xylem is available. Considering how important the secondary xylem and sapwood transition from young to mature trees is, little is known about the expression differences between those successional processes and which transcription factors could regulate lignin biosynthesis in this tropical tree. Although MYB transcription factors are one of the largest superfamilies in plants related to secondary metabolism, it has not yet been characterized in teak. These results will open new perspectives for studies of diversity, ecology, breeding and genomic programs aiming to understand deeply the biology of this species.ResultsWe present a widely expressed gene catalog for T. grandis using Illumina technology and the de novo assembly. A total of 462,260 transcripts were obtained, with 1,502 and 931 genes differentially expressed for stem and branch secondary xylem, respectively, during age transition. Analysis of stem and branch secondary xylem indicates substantial similarity in gene ontologies including carbohydrate enzymes, response to stress, protein binding, and allowed us to find transcription factors and heat-shock proteins differentially expressed. TgMYB1 displays a MYB domain and a predicted coiled-coil (CC) domain, while TgMYB2, TgMYB3 and TgMYB4 showed R2R3-MYB domain and grouped with MYBs from several gymnosperms and flowering plants. TgMYB1, TgMYB4 and TgCES presented higher expression in mature secondary xylem, in contrast with TgMYB2, TgHsp1, TgHsp2, TgHsp3, and TgBi whose expression is higher in young lignified tissues. TgMYB3 is expressed at lower level in secondary xylem.ConclusionsExpression patterns of MYB transcription factors and heat-shock proteins in lignified tissues are dissimilar when tree development was evaluated, obtaining more expression of TgMYB1 and TgMYB4 in lignified tissues of 60-year-old trees, and more expression in TgHsp1, TgHsp2, TgHsp3 and TgBi in stem secondary xylem of 12-year-old trees. We are opening a door for further functional characterization by reverse genetics and marker-assisted selection with those genes. Investigation of some of the key regulators of lignin biosynthesis in teak, however, could be a valuable step towards understanding how rigidity of teak wood and extractives content are different from most other woods. The obtained transcriptome data represents new sequences of T. grandis deposited in public databases, representing an unprecedented opportunity to discover several related-genes associated with secondary xylem such as transcription factors and stress-related genes in a tropical tree.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0599-x) contains supplementary material, which is available to authorized users.

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