The plant-pathogenic fungus Mycosphaerella graminicola (asexual stage: Septoria tritici) causes septoria tritici blotch, a disease that greatly reduces the yield and quality of wheat. This disease is economically important in most wheat-growing areas worldwide and threatens global food production. Control of the disease has been hampered by a limited understanding of the genetic and biochemical bases of pathogenicity, including mechanisms of infection and of resistance in the host. Unlike most other plant pathogens, M. graminicola has a long latent period during which it evades host defenses. Although this type of stealth pathogenicity occurs commonly in Mycosphaerella and other Dothideomycetes, the largest class of plant-pathogenic fungi, its genetic basis is not known. To address this problem, the genome of M. graminicola was sequenced completely. The finished genome contains 21 chromosomes, eight of which could be lost with no visible effect on the fungus and thus are dispensable. This eight-chromosome dispensome is dynamic in field and progeny isolates, is different from the core genome in gene and repeat content, and appears to have originated by ancient horizontal transfer from an unknown donor. Synteny plots of the M. graminicola chromosomes versus those of the only other sequenced Dothideomycete, Stagonospora nodorum, revealed conservation of gene content but not order or orientation, suggesting a high rate of intra-chromosomal rearrangement in one or both species. This observed “mesosynteny” is very different from synteny seen between other organisms. A surprising feature of the M. graminicola genome compared to other sequenced plant pathogens was that it contained very few genes for enzymes that break down plant cell walls, which was more similar to endophytes than to pathogens. The stealth pathogenesis of M. graminicola probably involves degradation of proteins rather than carbohydrates to evade host defenses during the biotrophic stage of infection and may have evolved from endophytic ancestors.

Summary Feruloylation of arabinoxylan (AX) in grass cell walls is a key determinant of recalcitrance to enzyme attack, making it a target for improvement of grass crops, and of interest in grass evolution. Definitive evidence on the genes responsible is lacking so we studied a candidate gene that we identified within the BAHD acyl‐CoA transferase family.We used RNA interference (RNAi) silencing of orthologs in the model grasses Setaria viridis (SvBAHD01) and Brachypodium distachyon (BdBAHD01) and determined effects on AX feruloylation.Silencing of SvBAHD01 in Setaria resulted in a c. 60% decrease in AX feruloylation in stems consistently across four generations. Silencing of BdBAHD01 in Brachypodium stems decreased feruloylation much less, possibly due to higher expression of functionally redundant genes. Setaria SvBAHD01 RNAi plants showed: no decrease in total lignin, approximately doubled arabinose acylated by p‐coumarate, changes in two‐dimensional NMR spectra of unfractionated cell walls consistent with biochemical estimates, no effect on total biomass production and an increase in biomass saccharification efficiency of 40–60%.We provide the first strong evidence for a key role of the BAHD01 gene in AX feruloylation and demonstrate that it is a promising target for improvement of grass crops for biofuel, biorefining and animal nutrition applications.

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-The objective of this work was to estimate the genetic parameters and variability among accessions (half-sib families) of physic nut (Jatropha curcas) during the early stages of development. For this study, 110 accessions in the first year of development of the physic nut germplasm bank, maintained at Embrapa Cerrados, DF, Brazil, were evaluated in situ. The experiment was established in a randomized complete block design, with two replicates and five plants per plot arranged in rows at 4x2 m spacing. Grain yield, total number of branches per plant, plant height, stem diameter, canopy projection on the row, canopy projection between rows, canopy volume, number of days until first flowering and height of the first inflorescence were evaluated. Estimates of vegetative genetic parameters showed the existence of genetic variability in the physic nut germplasm bank. Physic nut accessions of the germplasm bank were grouped into five similarity groups based on character divergence. Although preliminary, the obtained results are promising for showing potential for Jatropha improvement with selective efficiency.Index terms: Jatropha curcas, germplasm bank, plant breeding. Parâmetros genéticos e variabilidade em acessos de pinhão-manso no estágio inicial de desenvolvimentoResumo -O objetivo deste trabalho foi estimar os parâmetros genéticos e a variabilidade entre acessos (famílias de meios-irmãos) de pinhão-manso (Jatropha curcas) no estágio inicial de desenvolvimento. Para este estudo, foram avaliados in situ 110 acessos, no primeiro ano de desenvolvimento, do banco de germoplasma mantido na Embrapa Cerrados. O experimento foi implantado em delineamento de blocos completos ao acaso, com duas repetições e cinco plantas por parcela, dispostas em linha no espaçamento de 4x2 m. Foram avaliados os caracteres: produção de grãos, número total de ramos por planta, altura de plantas, diâmetro de caule, projeção da copa na linha, projeção da copa na entrelinha, volume da copa, numero de dias para o primeiro florescimento e altura da primeira inflorescência. As estimativas dos parâmetros genéticos de caracteres vegetativos mostraram a existência de variabilidade genética no banco de germoplasma de pinhão-manso. Os acessos do banco de germoplasma de pinhão-manso foram agrupados em cinco grupos de similaridade, com base na divergência dos caracteres avaliados. Embora preliminares, os resultados obtidos são promissores por mostrar o potencial para o melhoramento de Jatropha com eficiência seletiva.Termos para indexação: Jatropha curcas, banco de germoplasma, melhoramento vegetal.

Real-time PCR (RT-qPCR) expression analysis is a powerful analytical technique, but reliable results depend on the use of stable reference genes for proper normalization. This study proposed to test the expression stability of 13 candidate reference genes in Setaria viridis, a monocot species recently proposed as a new C4 model plant. Gene expression stability of these genes was assayed across different tissues and developmental stages of Setaria and under drought or aluminum stress. In general, our results showed Protein Kinase, RNA Binding Protein and SDH as the most stable genes. Moreover, pairwise analysis showed that two reference genes were sufficient to normalize the gene expression data under each condition. By contrast, GAPDH and ACT were the least stably expressed genes tested. Validation of suitable reference genes was carried out to profile the expression of P5CS and GolS during abiotic stress. In addition, normalization of gene expression of SuSy, involved in sugar metabolism, was assayed in the developmental dataset. This study provides a list of reliable reference genes for transcript normalization in S. viridis in different tissues and stages of development and under abiotic stresses, which will facilitate genetic studies in this monocot model plant.

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Summary Micro RNA s (mi RNA s) modulate the abundance and spatial–temporal accumulation of target mRNA s and indirectly regulate several plant processes. Transcriptional regulation of the genes encoding mi RNA s ( MIR genes) can be activated by numerous transcription factors, which themselves are regulated by other mi RNA s. Fine‐tuning of MIR genes or mi RNA s is a powerful biotechnological strategy to improve tolerance to abiotic or biotic stresses in crops of economic importance. Current approaches for mi RNA fine‐tuning are based on the down‐ or up‐regulation of MIR gene transcription and the use of genetic engineering tools to manipulate the final concentration of these mi RNA s in the cytoplasm. Transgenesis, cisgenesis, intragenesis, artificial MIR genes, endogenous and artificial target mimicry, MIR genes editing using Meganucleases, ZNF proteins, TALEN s and CRISPR /Cas9 or CRISPR /Cpf1, CRISPR / dC as9 or dC pf1, CRISPR 13a, topical delivery of mi RNA s and epigenetic memory have been successfully explored to MIR gene or mi RNA modulation and improve agronomic traits in several model or crop plants. However, advantages and drawbacks of each of these new biotechnological tools ( NBT s) are still not well understood. In this review, we provide a brief overview of the biogenesis and role of mi RNA s in response to abiotic or biotic stresses, we present critically the main NBT s used for the manipulation of MIR genes and mi RNA s, we show current efforts and findings with the MIR genes and mi RNA s modulation in plants, and we summarize the advantages and drawbacks of these NBT s and provide some alternatives to overcome. Finally, challenges and future perspectives to mi RNA modulating in important crops are also discussed.

Setaria viridis was recently described as a new monocotyledonous model species for C4 photosynthesis research and genetic transformation. It has biological attributes (rapid life cycle, small genome, diploid, short stature and simple growth requirements) that make it suitable for use as a model plant. We report an alternative method of S. viridis transformation using floral dip to circumvent the necessity of tissue culture phase for transgenic plant regeneration. S. viridis spikes at boot stage were selected to be immersed in Agrobacterium suspension. T1 seeds could be identified in 1.5–2 months after floral dipping. We demonstrated through molecular analysis and RFP expression that seeds and resulting plants from dipped inflorescences were transformed. Our results suggest the feasibility of S. viridis floral dip transformation as a time-saving and cost-effective compared with traditional methods. To our knowledge, this is the first report using floral dip in S. viridis as an Agrobacterium-mediated transformation method.