A whole-transcriptome approach to evaluate reference genes for quantitative diurnal gene expression studies under natural field conditions in Tamarix ramosissima leaves

Background: Tamarix ramosissima is a desert forest tree species that is widely distributed in the drought-stricken areas to sustain the fragile ecosystem. Owing to its wide usage in the desert restoration of Asia, it can be used as an ecophysiological model plant. To obtain reliable and accurate results, a set of reference genes should be screened before gene expression. However, up to date, systematical evaluation of reference genes has not been conducted in T. ramosissima. Results: In this study, we used eigenvalues derived from principal component analysis to identify stable expressed genes from 72,035 unigenes from diurnal transcriptomes under natural field conditions. With combined criteria of read counts above 900 and CV of FPKM below 0.3, a total of 7385 unigenes could be qualified as candidate reference genes in T. ramosissima. By using three statistical algorithm packages, geNorm, NormFinder, and BestKeeper, the stabilities of these novel reference genes were further compared with a panel of traditional reference genes. The expression patterns of three aquaporins (AQPs) suggested that at least UBQ (high expression), EIF4A2 (low expression), and GAPDH (moderate expression) could be qualified as ideal reference genes in both RT-PCR and RNA-seq analysis of T. ramosissima. Conclusions: This work will not only facilitate future studies on gene expression and functional analysis of genetic resources of desert plants but also improve our understanding of the molecular regulation of water transport in this plant, which could provide a new clue to further investigate the drought adaptation mechanism of desert plant species under harsh environments.

Regulation of aquaporins in plants under stress

Abstract Aquaporins (AQP) are channel proteins belonging to the Major Intrinsic Protein (MIP) superfamily that play an important role in plant water relations. The main role of aquaporins in plants is transport of water and other small neutral molecules across cellular biological membranes. AQPs have remarkable features to provide an efficient and often, specific water flow and enable them to transport water into and out of the cells along the water potential gradient. Plant AQPs are classified into five main subfamilies including the plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), nodulin 26 like intrinsic proteins (NIPs), small basic intrinsic proteins (SIPs) and X intrinsic proteins (XIPs). AQPs are localized in the cell membranes and are found in all living cells. However, most of the AQPs that have been described in plants are localized to the tonoplast and plasma membranes. Regulation of AQP activity and gene expression, are also considered as a part of the adaptation mechanisms to stress conditions and rely on complex processes and signaling pathways as well as complex transcriptional, translational and posttranscriptional factors. Gating of AQPs through different mechanisms, such as phosphorylation, tetramerization, pH, cations, reactive oxygen species, phytohormones and other chemical agents, may play a key role in plant responses to environmental stresses by maintaining the uptake and movement of water in the plant body.

Ajuste de modelos e difusividade efetiva na secagem de folhas de Azadirachta indica A. Juss / Ajuste de los modelos y la difusividad efectiva en el secado de las hojas de Azadirachta indica A. Juss / Adjustment of models and effective diffusivity in the drying of Azadirachta indica A. Juss. leaves

Introdução: a espécie Azadirachta indica A. Juss. conhecida como neem indiano, possui ampla utilização nas áreas da medicina, agropecuária e cosmética, proporcionada particularmente pelas folhas desidratadas. Devido à importância da espécie, tem-se a necessidade de conhecimentos específicos no seu pré-processamento. Objetivo: determinar a curva de secagem, ajustar modelos não lineares aos dados, estimar o coeficiente de difusividade efetiva e a energia de ativação durante a secagem das folhas de A. indica, em diferentes temperaturas. Métodos: as folhas da referida espécie foram submetidas à secagem em diferentes temperaturas com quatro repetições em secador de camada fixa. Utilizaram-se doze modelos para testar o comportamento da cinética de secagem, e seleciono-os que melhor desempenharam respostas funcionais considerando a magnitude do coeficiente de determinação, qui-quadrado, erro médio relativo e desvio padrão. Ainda, ajustou os dados, ao modelo matemático da difusão líquida com a solução analítica para a placa plana infinita, com aproximação de oito termos. Os gráficos foram elaborados pelo Sigma Plot. Resultados: evidenciou ajuste satisfatório para os modelos de Verma, Newton, Midilli, Logarítmico e Aproximação da Difusão, porém selecionou-se o modelo de Newton por apresentar maior simplicidade de aplicação. Constatou-se que com aumento da temperatura, ocorre um decréscimo no tempo de secagem e aumento na intensidade do fenômeno de transporte de água. Conclusões: a energia de ativação foi de 27,77 kJ mol-1. O modelo de Newton pode ser utilizado para descrever a cinética de secagem das folhas de A. indica. A energia de ativação se mostrou dentro do intervalo indicado para produtos agrícolas(Introdução: a espécie Azadirachta indica A. Juss. conhecida como neem indiano, possui ampla utilização nas áreas da medicina, agropecuária e cosmética, proporcionada particularmente pelas folhas desidratadas. Devido à importância da espécie, tem-se a necessidade de conhecimentos específicos no seu pré-processamento. Objetivo: determinar a curva de secagem, ajustar modelos não lineares aos dados, estimar o coeficiente de difusividade efetiva e a energia de ativação durante a secagem das folhas de A. indica, em diferentes temperaturas. Métodos: as folhas da referida espécie foram submetidas à secagem em diferentes temperaturas com quatro repetições em secador de camada fixa. Utilizaram-se doze modelos para testar o comportamento da cinética de secagem, e seleciono-os que melhor desempenharam respostas funcionais considerando a magnitude do coeficiente de determinação, qui-quadrado, erro médio relativo e desvio padrão. Ainda, ajustou os dados, ao modelo matemático da difusão líquida com a solução analítica para a placa plana infinita, com aproximação de oito termos. Os gráficos foram elaborados pelo Sigma Plot. Resultados: evidenciou ajuste satisfatório para os modelos de Verma, Newton, Midilli, Logarítmico e Aproximação da Difusão, porém selecionou-se o modelo de Newton por apresentar maior simplicidade de aplicação. Constatou-se que com aumento da temperatura, ocorre um decréscimo no tempo de secagem e aumento na intensidade do fenômeno de transporte de água. Conclusões: a energia de ativação foi de 27,77 kJ mol-1. O modelo de Newton pode ser utilizado para descrever a cinética de secagem das folhas de A. indica. A energia de ativação se mostrou dentro do intervalo indicado para produtos agrícolas(AU)

Introducción: la especie Azadirachta indica A. Juss. conocida como indiano, posee amplia utilización en las áreas de la medicina, agropecuaria y cosmética, proporcionada particularmente por las hojas deshidratadas. Debido a la importancia de la especie, existe la necesidad de un conocimento específico en su pre-procesamiento. Objetivo: determinar la curva del secado, ajustar los modelos no lineales con los datos y estimar el coeficiente de difusividad efectiva y de la energia de activación durante el secado de las hojas de A. indica, a diferentes temperaturas. Métodos: las hojas de dicha especie, fueron sometidas a un secado a diferentes temperaturas con cuatro repeticiones en el secador de capa fija. Se utilizaron doce modelos para probar el comportamiento de la cinética de secado, y se seleccionaron según los que mejor desempeñaron respuestas funcionales, teniendo en cuenta la magnitud del coeficiente de determinación, chi-cuadrado, error medio relativo y la desviación típica. Además, se ajustaron los datos al modelo matemático de difusión líquida con la solución analítica para la placa plana infinita, con la aproximación de 8 terminos. Los gráficos fueron confeccionados por Sigma Plot. Resultados: se mostró un ajuste satisfactório para los modelos de Verma, Newton, Midilli, Logarítmico y la aproximación de difusión, sin embargo se selecionó el modelo de Newton, por presentar mayor simplicidad de aplicación. Se constató que con el aumento de la temperatura ocurre una disminución en el tiempo de secado y aumento en la intensidad del fenomeno de transporte del agua. Conclusiones: la energia de activación fue de 27,77 kJ mol-1. El modelo de Newton puede ser utilizado para describir la cinética de secado de las hojas de A. indica. La energía de activación se mostró dentro del alcance indicado para productos agrícolas(AU)

Introduction: The species Azadirachta indica A. Juss, commonly known as Indian neen, is broadly used in medicine, agriculture and the cosmetic industry, mainly in the form of dehydrated leaves. Due to the importance of this species, specific knowledge is required for its preprocessing. Objective: Determine the drying curve, adjust non-linear models to the data, and estimate the effective diffusivity and activation energy coefficient during the drying of A. indica leaves at different temperatures. Methods: Leaves from the study species were dried at different temperatures with four replications in the fixed layer dryer. Twelve models were used to test the behavior of the drying kinetics. These were selected from among those providing better functional responses, bearing in mind the magnitude of the determination coefficient, chi-square, relative mean error and typical deviation. Additionally, data were adjusted to the liquid diffusion mathematical model with the analytical solution for the infinite plane plate with an approximation of 8 terms. The charts were developed by Sigma Plot. Results: A satisfactory adjustment was achieved for the models Verma, Newton, Midilli, logarithmic, and diffusion approximation, but the Newton model was chosen, due to the simplicity of its application. It was found that as temperature increases, there is a decrease in drying time and an increase in water transport intensity. Conclusions: Activation energy was 27.77 kJ mol-1. The Newton model may be used to describe the drying kinetics of A. indica leaves. Activation energy was within the scope established for agricultural products(AU)

Comparative Study of Effects of Spleen-strengthening Recipes on Water-Electrolyte Metabolism and Water Transport in Rats with Spleen-deficiency Syndrome / 广州中医药大学学报

Objective To compare the effects of different spleen-strengthening recipes on water-electrolyte metabolism and water transport in rat model of spleen-deficiency syndrome. Methods The male Wistar rats were randomly assigned into the normal group and spleen-deficiency model group. The rat model of spleen-deficiency was established by overstrain plus disordered diet method. On the 15th modeling day, the model rats were randomly assigned into model group , Buzhong Yiqi Tang(BYT) group, Si Junzi Tang(SJT) group, Shenling Baizhu San(SBS) group, and the rats except for the model group were treated with gastric gavage of the corresponding medicine in the dosage of 4.05 g·kg-1·d-1. Rats of normal group and model group were given equal volume of normal saline. The medication lasted for 14 days. On the 29th experimental day, we detected serum Na+ and K+, plasma antidiuretic hormone(ADH) and aldosterone(ALD), aquaporin 3(AQP3) in the small intestine and colon, and renal AQP2. Results Compared with the normal group, plasma ADH and ALD levels, serum Na+ level, renal AQP2 content in the model rats were increased significantly, serum K+ and AQP3 content of small intestine and colon mucosa was decreased significantly(P<0.01). Compared with the model group, plasma ADH and ALD levels, serum Na+ level, renal AQP2 content in the three medication groups were decreased significantly, serum K+level and AQP3 in the small intestine and descending colon mucosa was increased significantly(P<0.05 or P<0.01). Of the three medication groups, SBS group had lower ADH, ALD, Na+levels, and higher serum K+level and AQP3 in the small intestine and descending colon mucosa than BYT group and SJT group(P<0.01); SBS group and BYT group had lower renal AQP2 than SJT group(P<0.05 or P<0.01). Conclusion Rat model with spleen-deficiency syndrome has disordered water-electrolyte metabolism and abnormal water transport. The spleen-strengthening recipes, which have the actions of strengthening spleen, strengthening spleen to drain dampness, strengthening spleen to elevate yang, can improve the disorder of water metabolism to some degrees, and SBS has the best effect.

Altered Expression of Peritoneal Aquaporin-1 and Water Transport during Long-term Peritoneal Dialysis in Rats / 대한신장학회잡지

BACKGROUND: The present work was designed to examine the altered expression of peritoneal AQP-1 and water transport of peritoneal membrane during the long-term peritoneal dialysis with hypertonic glucose solution in rats. METHODS: Eighteen Sprague-Dawley male rats were randomly divided into 2 groups: control rats (n=6) with peritoneal catheter but not dialyzed; rats with peritoneal dialysis (PD) (n=12) were dialyzed with 4.25% glucose dialysate for all exchanges. Before completion of the study, 4 animals in PD group were euthanized owing to nonfunctional catheters or peritonitis, leaving 14 animals for the analysis. Dialysis exchanges were performed 3 times a day with 25 mL/each exchange for 12 weeks. Immunoperoxidase staining was performed using monoclonal anti-AGE antibody and polyclonal anti-AQP-1 antibody. The slides were read by 5 different examiners in a blind fashion. The staining intensity was graded semiquantitively from 0 to 3. The peritoneal membrane function was assessed by performing one-hour peritoneal equilibration tests every 6 week for comparing transport characteristics. Peritoneal membrane transport rate was assessed by D/P of urea nitrogen and D/Do of glucose. Water transport of peritoneal membrane was assessed by D/P of sodium at 12 week. RESULTS: The expression of peritoneal AQP-1 was increased in rats with PD, compared to control rats. Consistent with this, D/P of sodium in rats with PD was significantly decreased compared to control rats (0.58+/-0.04 vs 0.86+/-0.07, p<0.05), indicating high peritoneal water permeability in response to long-term peritoneal dialysis. Moreover, rats with PD were associated with significantly lower D/Do of glucose and higher D/P of urea nitrogen, suggesting high peritoneal membrane transport. CONCLUSION: High expression of peritoneal AQP-1 was associated with an increased peritoneal water permeability in response to long-term peritoneal dialysis with 4.25% glucose for 12 weeks. The underlying mechanisms for the increased AQP-1 expression need to be examined whether it is due to the continuous exposure to the dialysis solution containing high glucose concentration itself or compensatory effects of slowly developed concomitant ultrafiltration failure in chronic peritoneal dialysis.

Effects of Vasopressin or Prolactin on Li+ Transport through Amniotic Membrane during the Volume Reduction Perod of Amniotic Fluid in Pregnant Rabbits / 대한산부인과학회잡지

No abstract available.