Loss of S-nitrosoglutathione reductase disturbs phytohormone homeostasis and regulates shoot side branching and fruit growth in tomato

S-Nitrosoglutathione plays a central role in nitric oxide (NO) homeostasis, and S-nitrosoglutathione reductase (GSNOR) regulates the cellular levels of S-nitrosoglutathione across kingdoms. Here, we investigated the role of endogenous NO in shaping shoot architecture and controlling fruit set and growth in tomato (Solanum lycopersicum). SlGSNOR silencing promoted shoot side branching and led to reduced fruit size, negatively impacting fruit yield. Greatly intensified in slgsnor knockout plants, these phenotypical changes were virtually unaffected by SlGSNOR overexpression. Silencing or knocking out of SlGSNOR intensified protein tyrosine nitration and S-nitrosation and led to aberrant auxin production and signaling in leaf primordia and fruit-setting ovaries, besides restricting the shoot basipetal polar auxin transport stream. SlGSNOR deficiency triggered extensive transcriptional reprogramming at early fruit development, reducing pericarp cell proliferation due to restrictions on auxin, gibberellin, and cytokinin production and signaling. Abnormal chloroplast development and carbon metabolism were also detected in early-developing NO-overaccumulating fruits, possibly limiting energy supply and building blocks for fruit growth. These findings provide new insights into the mechanisms by which endogenous NO fine-tunes the delicate hormonal network controlling shoot architecture, fruit set, and post-anthesis fruit development, emphasizing the relevance of NO-auxin interaction for plant development and productivity.This work was supported by the São Paulo Research Foundation (FAPESP) (grants 16/02033-1, 16/01128-9 17/17935-3, 18/16389-8, 18/06436-9, 20/03720-8, 21/05714-8), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (grants 422287/2018-0, 305012/2018-5), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES) – Finance Code 00

Improving resource productivity at a crop sequence level

The challenge to increase agricultural production with a minimum environmental impact requires to reach themaximum effiency in the capture and use of resources such as photosynthetically active radiation (PAR), water,and nitrogen (N). Such requisites are encompassed in the ecological intensifiation (EI) concept. The aims of this work were to evaluate at a crop sequence level: i) crops yields, ii) water and radiation productivity and itscomponents, i.e. resource capture and resource use effiency, and iii) partial factor productivity of applied N(PFPN), partial nutrient balance for N (PNB), N uptake and N utilization effiency of a two-year, three-cropsequence (wheat [Triticum aestivum L.]/soybean [Glycine max (L.) Merr.] double crop – maize [Zea mays L.])carried-out under EI principles in comparison with the same crop sequence under current farmer practices (FP) intwo contrasting locations of the Argentinean Pampas, i.e. Paraná (-31°50′; -60°31′) at the northern Pampas andBalcarce (-37°45′, -58°18′) at the southern Pampas. Experiments were carried-out during four consecutive years,covering two complete cycles of the crop sequence. For the accumulated grain production of the crop sequence, EI management outyielded FP from 13 to 42%, depending on environmental conditions. Maize yield accounted formost of the variation (41–64%) of the accumulated grain yield of crop sequence, whether in EI as in FP. Averagegrain yield diffrences between EI and FP treatments were 274 g m−2 for maize, 69 g m−2 for wheat and -2 g m−2for soybean. Water and radiation productivities of the sequence were higher in EI than in FP (26% for water and17% for radiation; P < 0.0001), mainly because of increases in resource use effiencies. EI reduced partial factorproductivity of applied N, but improved partial nutrient balance for N as compared with FP. These reductions inpartial factor productivity of applied N were less than proportional than the increases in N rate. Moreover, in spiteof the higher N rate in EI respect to FP, N utilization effiency (NutE), i.e. grain per unit N uptake, was higher across all situations in EI. Our results showed that the challenge to obtain high grain yields by increasing N rate in amedium-input system could be achieved even with an increase in NutE. Grain yields improvements, and increasesin radiation and water productivity were reached by applying a set of agronomic practices that included improvedgenetics, crop and fertilizer N management englobed under EI concept

Produtividade de raízes de mandioca consorciada com milho e caupi em sistema orgânico.

Foram avaliados sistemas orgânicos de produção de mandioca “de mesa”, em Seropédica (RJ). O experimento constou dos seguintes tratamentos: monocultivo de mandioca (cv. IAC 576-70) e consórcios com milho experimental (cv. Eldorado), caupi (cv. Mauá) e milho+caupi. O manejo orgânico foi padronizado e toda a área experimental irrigada durante o período de permanência do milho no sistema. Do milho, foram colhidas espigas verdes (imaturas) e a parte aérea acamada na superfície do solo. O caupi foi incluído como adubo verde e cortado na floração, sendo mantidos os resíduos na superfície do solo. Ambos os consortes ocuparam as entrelinhas da mandioca, de modo alternado, semeados após a primeira capina da cultura principal. A cultivar IAC 576-70 mostrou-se adaptada ao manejo orgânico, com produtividade de raízes de padrão comercial próxima a 31 Mg ha-¹. Não houve diferenças significativas entre o monocultivo e os três tipos de consórcios testados. A inclusão do milho representou potencial de renda adicional ao produtor, colhendo-se,em média, 18.125 espigas ha-¹, o que correspondeu a 5,1 Mg ha-¹. Os resíduos provenientes da roçada do caupi proporcionaram um aporte de biomassa fresca de 12 Mg ha-¹, com uma expressiva contribuição em nitrogênio (cerca de 44 kg de N ha-1). A fabácea leguminosa cobriu por completo as entrelinhas da mandioca,demonstrando seu potencial de controle à erosão e a ervas espontâneas. O consórcio triplo mostrou-se vantajoso tendo em vista que a receita obtida com a venda do milho verde justificaria os custos da irrigação, além dos benefícios da inclusão do caupi e da não interferência dos consortes na produtividade da mandioca

Efficacy of the dietary histone deacetylase inhibitor butyrate alone or in combination with vitamin A against proliferation of MCF-7 human breast cancer cells

The combined treatment with histone deacetylase inhibitors (HDACi) and retinoids has been suggested as a potential epigenetic strategy for the control of cancer. In the present study, we investigated the effects of treatment with butyrate, a dietary HDACi, combined with vitamin A on MCF-7 human breast cancer cells. Cell proliferation was evaluated by the crystal violet staining method. MCF-7 cells were plated at 5 x 10(4) cells/mL and treated with butyrate (1 mM) alone or combined with vitamin A (10 µM) for 24 to 120 h. Cell proliferation inhibition was 34, 10 and 46% following treatment with butyrate, vitamin A and their combination, respectively, suggesting that vitamin A potentiated the inhibitory activities of butyrate. Furthermore, exposure to this short-chain fatty acid increased the level of histone H3K9 acetylation by 9.5-fold (Western blot), but not of H4K16, and increased the expression levels of p21WAF1 by 2.7-fold (Western blot) and of RARβ by 2.0-fold (quantitative real-time PCR). Our data show that RARβ may represent a molecular target for butyrate in breast cancer cells. Due to its effectiveness as a dietary HDACi, butyrate should be considered for use in combinatorial strategies with more active retinoids, especially in breast cancers in which RARβ is epigenetically altered

Phosphorothioate antisense oligonucleotides induce the formation of nuclear bodies

Antisense oligonucleotides are powerful tools for the in vivo regulation of gene expression. We have characterized the intracellular distribution of fluorescently tagged phosphorothioate oligodeoxynucleotides (PS-ONs) at high resolution under conditions in which PS-ONs have the potential to display antisense activity. Under these conditions PS-ONs predominantly localized to the cell nucleus where they accumulated in 20-30 bright spherical foci designated phosphorothioate bodies (PS bodies), which were set against a diffuse nucleoplasmic population excluding nucleoli. PS bodies are nuclear structures that formed in cells after PS-ON delivery by transfection agents or microinjection but were observed irrespectively of antisense activity or sequence. Ultrastructurally, PS bodies corresponded to electron-dense structures of 150-300 nm diameter and resembled nuclear bodies that were found with lower frequency in cells lacking PS-ONs. The environment of a living cell was required for the de novo formation of PS bodies, which occurred within minutes after the introduction of PS-ONs. PS bodies were stable entities that underwent noticeable reorganization only during mitosis. Upon exit from mitosis, PS bodies were assembled de novo from diffuse PS-ON pools in the daughter nuclei. In situ fractionation demonstrated an association of PS-ONs with the nuclear matrix. Taken together, our data provide evidence for the formation of a nuclear body in cells after introduction of phosphorothioate oligodeoxynucleotides