Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Concentração foliar de nutrientes e produtividade de tomateiro cultivado sob diferentes substratos e doses de ácidos húmicos Leaf nutrient content and yield of tomato grown in different substrates and doses of humic acids

Este trabalho foi realizado com o objetivo de avaliar o estado nutricional e a produtividade do tomateiro, híbrido Vênus, cultivado em substratos, com aplicação de ácidos húmicos (AH) e fertirrigação. Foram avaliadas quatro doses de AH (0, 20, 40 e 80 L ha-1) e quatro substratos: S1 (fibra de coco (FC)), S2 (FC + casca de café carbonizada (CC) na proporção 1:3), S3 (FC + CC na proporção 2:3) e S4 (CC). O delineamento experimental foi em blocos casualizados distribuídos em esquema fatorial 4x4. As mudas foram transplantadas com 35 dias para sacolas plásticas com capacidade de 7,0 L. Os AH foram aplicados quatro vezes em um intervalo de oito dias, sendo a primeira aplicação oito dias após o transplantio. Não houve efeito significativo dos tratamentos nos teores de N, K e S nas folhas e na produção de frutos não comerciais. No entanto, houve efeito significativo dos substratos nos teores de P, Ca, Mg e sobre a produção de frutos total e comercial. Doses crescentes de AH induziram redução de forma linear dos teores de B e Cu (em S1); porém, induziram aumento dos teores de Zn. Em doses de 31 a 49 L ha-1 de AH, ocorreu o máximo teor foliar de Fe, Cu e o mínimo de Mn. As maiores produtividades comerciais foram obtidas com a fibra de coco (S1) com média de 5,6 kg planta-1, apresentando incremento médio de produtividade de 22,9%, 38,7% e 49,7% em relação a S2, S3 e S4, respectivamente.<br>We evaluated the nutritional status and yield of tomato hybrid "Vênus", grown on substrates, with application of humic acids (AH) and fertigation. Four doses of AH (0, 20, 40 and 80 L-1 ha) and four substrates: S1 (coconut fiber (CF)), S2 (FC + carbonized coffee husk (CC) in the ratio 1:3), S3 (CF + CC - in the ratio 2:3) and S4 (CC) were evaluated following the randomized blocks design in factorial 4x4 scheme with four replications. The 35-day old seedlings were transplanted into plastic bags of 7 L. The humic acids were applied four times in eight-day intervals, and the first application was carried out eight days after transplanting. There was no significant effect of the treatments on the content of N, K and S in leaves and on non-marketable fruits yield. However, there was a significant effect of the substrates on the content of P, Ca, Mg and on the total and marketable fruit yield. Increasing doses of AH reduced linearly the content of B and Cu (on S1), but increased the content of Zn in leaves. Humic acid doses of 31 to 49 L ha-1 provided the highest contents of Fe, Cu, and the lowest content of Mn. The highest marketable yields were obtained using coconut fiber, with an average of 5.6 kg/plant, showing an average yield increase of 22.9%, 38.7% and 49.7% in comparison to S2, S3 and S4, respectively