Identification, expression and DNA variation analysis of high affinity nitrate transporter NRT2/3 gene family in Sorghum bicolor / 生物工程学报

Nitrate is the main form of inorganic nitrogen that crop absorbs, and nitrate transporter 2 (NRT2) is a high affinity transporter using nitrate as a specific substrate. When the available nitrate is limited, the high affinity transport systems are activated and play an important role in the process of nitrate absorption and transport. Most NRT2 cannot transport nitrates alone and require the assistance of a helper protein belonging to nitrate assimilation related family (NAR2) to complete the absorption or transport of nitrates. Crop nitrogen utilization efficiency is affected by environmental conditions, and there are differences between varieties, so it is of great significance to develop varieties with high nitrogen utilization efficiency. Sorghum bicolor has high stress tolerance and is more efficient in soil nitrogen uptake and utilization. The S. bicolor genome database was scanned to systematically analyze the gene structure, chromosomal localization, physicochemical properties, secondary structure and transmembrane domain, signal peptide and subcellular localization, promoter region cis-acting elements, phylogenetic evolution, single nucleotide polymorphism (SNP) recognition and annotation, and selection pressure of the gene family members. Through bioinformatics analysis, 5 NRT2 gene members (designated as SbNRT2-1a, SbNRT2-1b, SbNRT2-2, SbNRT2-3, and SbNRT2-4) and 2 NAR2 gene members (designated as SbNRT3-1 and SbNRT3-2) were identified, the number of which was less than that of foxtail millet. SbNRT2/3 were distributed on 3 chromosomes, and could be divided into four subfamilies. The genetic structure of the same subfamilies was highly similar. The average value of SbNRT2/3 hydrophilicity was positive, indicating that they were all hydrophobic proteins, whereas α-helix and random coil accounted for more than 70% of the total secondary structure. Subcellular localization occurred on plasma membrane, where SbNRT2 proteins did not contain signal peptides, but SbNRT3 proteins contained signal peptides. Further analysis revealed that the number of transmembrane domains of the SbNRT2s family members was greater than 10, while that of the SbNRT3s were 2. There was a close collinearity between NRT2/3s of S. bicolor and Zea mays. Protein domains analysis showed the presence of MFS_1 and NAR2 protein domains, which supported executing high affinity nitrate transport. Phylogenetic tree analysis showed that SbNRT2/3 were more closely related to those of Z. mays and Setaria italic. Analysis of gene promoter cis-acting elements indicated that the promoter region of SbNRT2/3 had several plant hormones and stress response elements, which might respond to growth and environmental cues. Gene expression heat map showed that SbNRT2-3 and SbNRT3-1 were induced by nitrate in the root and stem, respectively, and SbNRT2-4 and SbNRT2-3 were induced by low nitrogen in the root and stem. Non-synonymous SNP variants were found in SbNRT2-4 and SbNRT2-1a. Selection pressure analysis showed that the SbNRT2/3 were subject to purification and selection during evolution. The expression of SbNRT2/3 gene and the effect of aphid infection were consistent with the expression analysis results of genes in different tissues, and SbNRT2-1b and SbNRT3-1 were significantly expressed in the roots of aphid lines 5-27sug, and the expression levels of SbNRT2-3, SbNRT2-4 and SbNRT3-2 were significantly reduced in sorghum aphid infested leaves. Overall, genome-wide identification, expression and DNA variation analysis of NRT2/3 gene family of Sorghum bicolor provided a basis for elucidating the high efficiency of sorghum in nitrogen utilization.

Ethanol production from sweet sorghum by Saccharomyces cerevisiae DBKKUY-53 immobilized on alginate-loofah matrices

Abstract Ethanol production from sweet sorghum juice (SSJ) using the thermotolerant Saccharomyces cerevisiae strain DBKKUY-53 immobilized in an alginate-loofah matrix (ALM) was successfully developed. As found in this study, an ALM with dimensions of 20 × 20 × 5 mm3 is effective for cell immobilization due to its compact structure and long-term stability. The ALM-immobilized cell system exhibited greater ethanol production efficiency than the freely suspended cell system. By using a central composite design (CCD), the optimum conditions for ethanol production from SSJ by ALM-immobilized cells were determined. The maximum ethanol concentration and volumetric ethanol productivity obtained using ALM-immobilized cells under the optimal conditions were 97.54 g/L and 1.36 g/L h, respectively. The use of the ALM-immobilized cells was successful for at least six consecutive batches (360 h) without any loss of ethanol production efficiency, suggesting their potential application in industrial ethanol production.

High-temperature ethanol production using thermotolerant yeast newly isolated from Greater Mekong Subregion

Abstract The application of high-potential thermotolerant yeasts is a key factor for successful ethanol production at high temperatures. Two hundred and thirty-four yeast isolates from Greater Mekong Subregion (GMS) countries, i.e., Thailand, The Lao People's Democratic Republic (Lao PDR) and Vietnam were obtained. Five thermotolerant yeasts, designated Saccharomyces cerevisiae KKU-VN8, KKU-VN20, and KKU-VN27, Pichia kudriavzevii KKU-TH33 and P. kudriavzevii KKU-TH43, demonstrated high temperature and ethanol tolerance levels up to 45 °C and 13% (v/v), respectively. All five strains produced higher ethanol concentrations and exhibited greater productivities and yields than the industrial strain S. cerevisiae TISTR5606 during high-temperature fermentation at 40 °C and 43 °C. S. cerevisiae KKU-VN8 demonstrated the best performance for ethanol production from glucose at 37 °C with an ethanol concentration of 72.69 g/L, a productivity of 1.59 g/L/h and a theoretical ethanol yield of 86.27%. The optimal conditions for ethanol production of S. cerevisiae KKU-VN8 from sweet sorghum juice (SSJ) at 40 °C were achieved using the Box-Behnken experimental design (BBD). The maximal ethanol concentration obtained during fermentation was 89.32 g/L, with a productivity of 2.48 g/L/h and a theoretical ethanol yield of 96.32%. Thus, the newly isolated thermotolerant S. cerevisiae KKU-VN8 exhibits a great potential for commercial-scale ethanol production in the future.

Improvement of ethanol production from sweet sorghum juice under batch and fed-batch fermentations: effects of sugar levels, nitrogen supplementation, and feeding regimes

Background: Fermentation process development has been very important for efficient ethanol production. Improvement of ethanol production efficiency from sweet sorghum juice (SSJ) under normal gravity (NG, 160 g/L of sugar), high gravity (HG, 200 and 240 g/L of sugar) and very high gravity (VHG, 280 and 320 g/L of sugar) conditions by nutrient supplementation and alternative feeding regimes (batch and fed-batch systems) was investigated using a highly ethanol-tolerant strain, Saccharomyces cerevisiae NP01. Results: In the batch fermentations without yeast extract, HG fermentation at 200 g/L of sugar showed the highest ethanol concentration (PE, 90.0 g/L) and ethanol productivity (QE, 1.25 g/L·h). With yeast extract supplementation (9 g/L), the ethanol production efficiency increased at all sugar concentrations. The highest PE (112.5 g/L) and QE (1.56 g/L·h) were observed with the VHG fermentation at 280 g/L of sugar. In the fed-batch fermentations, two feeding regimes, i.e., stepwise and continuous feedings, were studied at sugar concentrations of 280 g/L. Continuous feeding gave better results with the highest PE and QE of 112.9 g/L and 2.35 g/L·h, respectively, at a feeding time of 9 h and feeding rate of 40 g sugar/h. Conclusions: In the batch fermentation, nitrogen supplementation resulted in 4 to 32 g/L increases in ethanol production, depending on the initial sugar level in the SSJ. Under the VHG condition, with sufficient nitrogen, the fed-batch fermentation with continuous feeding resulted in a similar PE and increased QP by 51% compared to those in the batch fermentation.

The use of dried spent yeast as a low-cost nitrogen supplement in ethanol fermentation from sweet sorghum juice under very high gravity conditions

Dried spent yeast (DSY) was used as a low-cost nitrogen supplement for ethanol fermentation from sweet sorghum juice under very high gravity (VHG) conditions by Saccharomyces cerevisiae NP 01. The fermentation was carried out at 30ºC in a 5-litre bioreactor. The results showed that DSY promoted ethanol production efficiencies. The ethanol concentration (P), productivity (Qp) and yield (Yp/s) of the sterile juice (total sugar of 280 g l-1) supplemented with 8 g l-1 of DSY were not different from those supplemented with yeast extract and/or peptone at the same amount. The initial yeast cell concentration of 5 x 10(7) cells ml-1 was found to be optimal for scale-up ethanol production. In addition, an increase in sugar concentration in inoculum preparation medium (from 10 to 100 g l-1) improved the ability of the inoculum to produce ethanol under the VHG conditions. When S. cerevisiae NP 01 grown in the juice containing 100 g l-1 of total sugar was used as the inoculum for ethanol fermentation, the P, Qp and Yp/s obtained were 108.98 +/- 1.16 g l-1, 2.27 +/- 0.06 g l-1 h-1 and 0.47 +/- 0.01 g g-1, respectively. Similar results were also observed when the ethanol fermentation was scaled up to a 50-litre bioreactor under the same conditions. The cost of the sweet sorghum for ethanol production was US$ 0.63 per litre of ethanol. These results clearly indicate the high potential of using sweet sorghum juice supplemented with DSY under VHG fermentation for ethanol production in industrial applications.

Níveis crescentes de parede de levedura sobre a resposta imune celular e perfil hematológico de frangos de corte / Cellular immune response and hematological parameters of broilers from different age broiler breeders, fed with a sorghum meal with different yeast wall levels

Um experimento foi conduzido para avaliar o efeito de níveis crescentes de parede de levedura e idade das matrizes reprodutoras sobre o peso dos órgãos linfóides, a resposta imune celular e o perfil hematológico de frangos de corte. Foram utilizados 3.360 pintos de corte da linhagem Cobb, distribuídos em delineamento inteiramente casualisado, em esquema fatorial 2x5, mais dois controles, sendo duas idades de matrizes (34 e 57 semanas de idade) e cinco níveis de suplementação de parede de levedura (zero, um, dois, três e quatro kg de parede de levedura/tonelada de ração). A idade das matrizes influenciou a resposta de todas as variáveis. A inclusão de 3kg de parede de levedura/tonelada de ração promoveu, na progênie de reprodutoras de 57 semanas, reação inflamatória mais intensa quando comparada a dieta controle, no entanto não houve aumento significativo no número de heterófilos e linfócitos circulantes. Conclui-se que a utilização da parede de levedura associada ao sorgo ou não em rações de frangos de corte ainda necessita de estudos complementares, que incluam, por exemplo, os componentes purificados da parede de levedura (MOS e ß-glucano).

An experiment was carried out to evaluate the effect increasing levels of yeast wall and broiler breeders' age levels on lymphoid organs weight, cellular immune response and hematological parameters in broilers. A total of 3,360 Cobb broilers were allotted, in a completely randomized design and a 2x5 factorial arrangement, and two controls, compound of two broiler breeders age (34 and 57 weeks of age) and five yeast wall levels (zero, one, two, three and four kg of yeast wall/ton of diet). Broiler breeders' age affected all studied variables. The inclusion of 3kg of yeast wall/ton of meal increased, at 57 weeks age broiler breeders, more intense inflammatory reaction when compared to control diet; however the circulated heterophils and lymphocytes numbers were not increased. In conclusion, the use of yeast wall, associated or not to sorghum on broilers diet still needs complementary studies, for example, purified components of yeast wall (MOS and ß-glucano).

Evaluación agronómica de genotipos de sorgo y millo (Sorghum vulgare) forrajeros para alimentación de bovinos en el medio Sinú / Agronomic evaluation of forages sorghum (Sorghum vulgare) and millo genotypes for feeding bovine in the Sinú valley

Objetivo. Evaluar siete genotipos de millo y cinco genotipos de sorgo, para determinar el rendimiento forrajero y su utilidad en la alimentación de bovinos en el valle del medio Sinu. Materiales y métodos. Las semillas de sorgo y millo (Sorghum vulgare), fueron materiales híbridos y variedades tipo comercial. Se utilizó un diseño de bloques completos al azar con 4 cuatro repeticiones para millo y tres para sorgo. Resultados. Respecto al vigor los genotipos de millo que mejor comportamiento presentaron fueron el redondo cuba y blanco panoja larga; en sorgo el LC 603, el P-8239 y el DR-1125. Los millos que mostraron mayor senescencia fueron el cuarentano alto, el bastón y el blanco panoja larga; en sorgo los de mayor senescencia fueron el P-8239, el LC 603 y el 74 CO. La mayor producción de forraje verde y materia seca la presentaron los genotipos de millo, el batea cuba, el redondo cuba y el blanco panoja larga, los de sorgo fueron el blanco criollo y el DR-1125. El mayor rendimiento de grano fue de los genotipos el batea cuba y bastón en millo, y en sorgo fueron DR-1125 y LC 603. Se seleccionaron como materiales de buen comportamiento dentro de todos los evaluados el batea cuba, redondo cuba, y blanco panoja larga en millo y el blanco criollo y DR-1125 en sorgo. Conclusiones. Los genotipos de millo que mejor comportamiento presentaron fueron en su orden por rendimiento (ton/ha): el batea cuba, el redondo cuba y el blanco panoja larga (ton/ha), obteniéndose una mayor capacidad de carga animal. Los genotipos de mejor comportamiento de sorgo fueron el blanco criollo y el DR-1125.

Substituição de milho por sorgo baixo tanino em dietas para suínos: digestibilidade e metabolismo / Replacement of corn by low tannin sorghum in pig diets: digestibility and metabolism

Um experimento foi realizado para avaliar os efeitos digestivos e metabólicos da substituição de milho por sorgo em dietas para suínos. Foram utilizados 12 suínos machos castrados, meio-irmãos paternos, com peso vivo médio inicial de 40,8kg, mantidos em gaiolas metabólicas. O delineamento experimental foi inteiramente casualizado com três tratamentos (T1 = dieta controle; T2 = substituição de 50 por cento do milho por sorgo e T3 = substituição de 100 por cento) e quatro repetições. As digestibilidades da proteína e da energia brutas foram 5 e 4 por cento superiores (P<0,05), respectivamente, nos suínos alimentados com a dieta que tinha 50 por cento de substituição, comparadas àquela com substituição total do milho por sorgo. A substituição total do milho por sorgo reduziu (P<0,05) as digestibilidades, em relação à dieta-controle, em 3 por cento para MS, 8 por cento para PB e 5 por cento para EB. O N fecal foi 25 por cento maior (P<0,05), o N absorvido 10 por cento menor (P<0,05) e a excreção fecal de energia 15 por cento maior (P<0,05) para a dieta com substituição total, em relação à dieta-controle. A excreção fecal de N e energia e a absorção de N são influenciadas negativamente pela substituição total de milho por sorgo nas dietas. A substituição em até 50 por cento do milho por sorgo em dietas para suínos não influi na digestibilidade das dietas e no metabolismo dos animais.

An experiment was carried out to evaluate metabolic and digestive effects replacement of corn by sorghum in pig diets. Twelve half brothers barrows weighing 40.8kg and housed in metabolic cages in a completely randomized design with three treatments (T1 = control diet; T2 = replacement of 50 percent corn by sorghum and T3 = replacement of 100 percent) with four replicates. Crude protein and gross energy the digestibilities were 5 percent and 4 percent higher (P<0.05), respectively, for pigs fed 50 percent of corn replacement diet, in comparison with the diet where corn was totally replaced by sorghum. Total replacement of corn by sorghum diets decreased (P<0.05) the digestibilities, in comparison to control diet, in 3 percent for DM, 8 percent for CP and 5 percent for GE. Fecal N was 25 percent higher (P<0.05), absorbed N was 10 percent lower (P<0.05), and the fecal excretion energy was 15 percent higher (P<0.05) for total substitution diet in comparison to control diet. Fecal excretion and absorption of nitrogen are negatively influenced by total replacement of corn by sorghum in the diets. Replacements of up to 50 percent of corn by sorghum in pig diets do not affect the digestibility diet and metabolism of animals.