No abstract

microRNAs (miRNAs) are derived from self-complementary hairpin structures, while small-interfering RNAs (siRNAs) are derived from double-stranded RNA (dsRNA) or hairpin precursors. The core mechanism of sRNA production involves DICER-like (DCL) in processing the smallRNAs (sRNAs) and ARGONAUTE (AGO) as effectors of silencing, and siRNA biogenesis also involves action of RNA-Dependent RNA Polymerase (RDR), Pol IV and Pol V in biogenesis. Several other proteins interact with the core proteins to guide sRNA biogenesis, action, and turnover. We aimed to unravel the components and functions of the RNA-guided silencing pathway in a non-model plant species of worldwide economic relevance. The sRNA-guided silencing complex members have been identified in the Coffea canephora genome, and they have been characterized at the structural, functional, and evolutionary levels by computational analyses. Eleven AGO proteins, nine DCL proteins (which include a DCL1-like protein that was not previously annotated), and eight RDR proteins were identified. Another 48 proteins implicated in smallRNA (sRNA) pathways were also identified. Furthermore, we identified 235 miRNA precursors and 317 mature miRNAs from 113 MIR families, and we characterized ccp-MIR156, ccp-MIR172, and ccp-MIR390. Target prediction and gene ontology analyses of 2239 putative targets showed that significant pathways in coffee are targeted by miRNAs. We provide evidence of the expansion of the loci related to sRNA pathways, insights into the activities of these proteins by domain and catalytic site analyses, and gene expression analysis. The number of MIR loci and their targeted pathways highlight the importance of miRNAs in coffee. We identified several roles of sRNAs in C. canephora, which offers substantial insight into better understanding the transcriptional and post-transcriptional regulation of this major crop.

Small RNAs influence the gene expression at the post-transcriptional level by guiding messenger RNA (mRNA) cleavage, translational repression, and chromatin modifications. In addition to model plants, the microRNAs (miRNAs) have been identified in different crop species. In this work, we developed a specific pipeline to search for coffee miRNA homologs on expressed sequence tags (ESTs) and genome survey sequences (GSS) databases. As a result, 36 microRNAs were identified and a total of 616 and 362 potential targets for Coffea arabica and Coffea canephora, respectively. The evolutionary analyses of these molecules were performed by comparing the primary and secondary structures of precursors and mature miRNAs with their orthologs. Moreover, using a stem-loop RT-PCR assay, we evaluated the accumulation of mature miRNAs in genomes with different ploidy levels, detecting an increase in the miRNAs accumulation according to the ploidy raising. Finally, a 5' RACE (Rapid Amplification of cDNA Ends) assay was performed to verify the regulation of auxin responsive factor 8 (ARF8) by MIR167 in coffee plants. The great variety of target genes indicates the functional plasticity of these molecules and reinforces the importance of understanding the RNAi-dependent regulatory mechanisms. Our results expand the study of miRNAs and their target genes in this crop, providing new challenges to understand the biology of these species.

Due to the weather changes prognostic for the coming years, the understanding of water deficit and physiological responses of plants to drought becomes an important requirement in order to develop technologies such as mechanisms to assist plants to cope with longer drought periods, which will be essential to maintenance of Brazilian and worldwide production. This study aimed to evaluate ecophysiological and anatomical aspects as well as the nitrate reductase activity in Siriema coffee seedlings subjected to four treatments: Daily irrigated, non-irrigated, re-irrigated 24 hours and re-irrigated 48 hours after different stress periods. Non-irrigation promoted a reduction in leaf water potential being accented from the ninth day of evaluation onwards. Re-irrigation promoted a partial recovery of the plant water potential. Non-irrigated plants showed an increase in stomatal resistance and reduction of transpiration and nitrate reductase activity. In the roots, there was a decrease in nitrate reductase activity under water stress. Leaf anatomical modifications were significant only for the adaxial surface epidermis and palisade parenchyma thickness, this latter characteristic being higher in control plants. Stomatal density and polar and equatorial diameter ratios showed the highest values in plants under water stress. In the roots, differences only in the cortex thickness being bigger in the non-irrigated treatment could be observed. Therefore, Siriema coffee plants under water stress show physiological, biochemical and anatomical modifications that contribute to the tolerance of this genotype to these conditions. Index terms: Coffee tree, ecophysiology, water stress, nitrate reductase. rESuMOEm razão do prognóstico de mudanças climáticas nos próximos anos, a compreensão do déficit hídrico e das respostas fisiológicas de plantas ao mesmo, torna-se uma importante condição para o desenvolvimento de tecnologias, como mecanismos que auxiliem as plantas a suportar períodos mais longos de seca, o que será essencial para a manutenção da produção mundial e brasileira. Neste estudo, objetivou-se avaliar aspectos ecofisiológicos e anatômicos além da atividade da redutase do nitrato em mudas de café Siriema submetidas a quatro tratamentos: irrigadas diariamente, não irrigadas, re-irrigadas 24 horas e re-irrigadas 48 horas após os diferentes períodos de estresse. A ausência de irrigação promoveu redução no potencial hídrico foliar, sendo mais acentuada a partir do nono dia de avaliação. A re-irrigação das mudas promoveu uma recuperação parcial do estado hídrico das plantas. Nas plantas não irrigadas observou-se um aumento na resistência estomatal e diminuição da transpiração, reduzindo a atividade da redutase do nitrato nas folhas. Nas raízes, houve um declínio na redutase do nitrato sob estresse hídrico. Ocorreram modificações na anatomia foliar apenas na espessura da epiderme da face adaxial e parenquimama paliçádico, sendo maior nas plantas controle. A densidade estomatal e razão diâmetro polar e equatorial demonstra...

No abstract

Cultivated tomato, Solanum lycopersicum, is one of the most common fruits in the global food industry. Together with the wild tomato Solanum pennellii, it is widely used for developing better cultivars. MicroRNAs affect mRNA regulation, inhibiting its translation and/or promoting its degradation. Important proteins involved in these processes are ARGONAUTE and DICER. This study aimed to identify and characterize the genes involved in the miRNA processing pathway, miRNA molecules and target genes in both species. We validated the presence of pathway genes and miRNA in different NGS libraries and 6 miRNA families using quantitative RT-PCR. We identified 71 putative proteins in S. lycopersicum and 108 in S. pennellii likely involved in small RNAs processing. Of these, 29 and 32 participate in miRNA processing pathways, respectively. We identified 343 mature miRNAs, 226 pre-miRNAs in 87 families, including 192 miRNAs, which were not previously identified, belonging to 38 new families in S. lycopersicum. In S. pennellii, we found 388 mature miRNAs and 234 pre-miRNAs contained in 85 families. All miRNAs found in S. pennellii were unpublished, being identified for the first time in our study. Furthermore, we identified 2471 and 3462 different miRNA target in S. lycopersicum and S. pennellii, respectively.

ABSTRACT. The interaction between polygalacturonase-inhibiting proteins (PGIPs), produced by plants, and endopolygalacturonases (PGs), produced by fungi, limits the destructive potential of PGs and can trigger plant defense responses. This study aimed to i) investigate variation in the expression of different common bean (Phaseolus vulgaris L.) genotypes and its relationship with resistance to white mold (Sclerotinia sclerotiorum); ii) determine the expression levels of PvPGIP genes at different time points after inoculation with white mold; and iii) investigate differences in PvPGIP gene expression between two white mold isolates with different levels of aggressiveness. Four bean lines were analyzed, including two lines from a recurrent selection for white mold (50/5 and 84/6), one resistant line that was not adapted to Brazilian conditions (Cornell 605), and one susceptible line (Corujinha). Gene expression was investigated at 0, 1, 2, 3, and 5 days after inoculation. The isolate UFLA 03 caused no significant difference in the relative expression of any gene examined, and was inefficient in discriminating among the genotypes. For the isolate UFLA 116, all of the genes were differentially expressed, as they were associated with resistance to white mold, and the expressions increased until the third day after inoculation. The 50/5 line was not significantly different from the Corujinha line for all of the genes analyzed. However, this line had a resistance level that was similar to that of Cornell 605, according to the straw test. Therefore, the incorporation of PvPGIP genes can increase the resistance of lines derived from recurrent selection.

Bud necrosis (BN) is a common disorder that affects Vitis vinifera L. and reduces its potential yield. to minimize the losses caused by Bn, the double pruning management was applied in Brazilian Southeast vineyards. in this management strategy plants are pruned at the winter to promote a vegetative cycle and then, at summer, to promote the reproductive cycle at optimal environmental conditions. to investigate the relationship of Bn and the double pruning management RnA-seq libraries were sequenced from healthy and necrotic tissues at four different stages of the year. The comparison of differentially expressed genes in necrotic and non-necrotic tissues showed an enhanced expression of genes related to cell death possibly induced by endophytic microorganisms in the necrotic tissues. the de novo assembly, characterization and quantification of transcripts within the RNA-seq libraries showed that genes from the endophytic fungus Alternaria alternata, responsible for the production of toxic compounds were highly expressed under BN. Here we propose a model in which unfavorable conditions and reduced carbohydrate levels in buds can promote the switch from a biotrophic lifestyle to a necrotrophic lifestyle in the endophytic fungi, which seems to be involved in the development of Bn. Grapevine (Vitis vinifera L.) is an important horticultural crop with a large variety of derived products. Its berries can be consumed as fresh or dried fruits and also used for the production of juices, liquors and wines. One of the main factors affecting grape production worldwide is the occurrence of a disorder known as primary Bud Necrosis (BN) 1,2. This disorder is widespread and has been reported to occur in Australia 2 , United States 3 , Japan 4 , Iran 5 , Europe 6 and Brazil 7 leading to reduction of yield potential. Primary BN is characterized by the death of the primary buds, considered to be the most productive. Secondary and tertiary buds often derive shoots believed to be less fruitful and with smaller clusters 2. BN is difficult to be diagnosed once buds may appear healthy externally for a long time and at this condition, a dissection analysis of the buds under an stereomicroscope can detect it 2. Under primary BN, photoassimilates will be diverted to the less fruitful buds 8. The causes of BN in grapevines, including the necrosis of the primary, secondary and tertiary buds, are still unknown. Its incidence can vary according to the vineyard, location, year and cultivar 2. Many explanations are being proposed to be the cause of this phenomenon including high shoot vigor 2,8-11 , influence of gibberellins 4 , high temperature 1 , vigor of the rootstocks 1 , canopy shading 3,12 and reduced carbohydrate levels 1,9. Even low temperature might be attributed as the cause 13. However, it is still unknown if, and how, all these factors interact to promote BD in different environmental conditions.