Knockdown of OsNRT2.4 modulates root morphology and alters nitrogen metabolism in response to low nitrate availability in rice.

The expression patterns of the NRT2 genes have been well described; however, the role of OsNRT2.4 in root growth is not well known. In this study, we thus aimed at investigating the role of high-affinity NO3- transport OsNRT2.4 in root growth modulation. Through the amiRNA-mediated gene silencing technique, we successfully obtained osnrt2.4 knockdown lines to study the role of OsNRT2.4 on root growth under low nitrate conditions. We performed real-time PCR analysis to investigate the relative gene expression level in root and shoot, soluble metabolites, and measurement of root system. Knockdown of OsNRT2.4 decreased rice growth. The comparison with wild-type (WT) plants showed that (i) knockdown of OsNRT2.4 inhibited root formation under low NO3- supply; (ii) we demonstrated that the mutant lines had significantly increased NO3- uptake than WT plants when grown in different nitrate supplies; (iii) osnrt2.4 knockdown lines showed an alteration in nitrogen metabolism, and this affected the root growth; and (iv) the downregulation of OsNRT2.4 enhanced the expression of gene response of low external NO3- concentrations. Herein we provide new insights in OsNRT2.4 functions. Our data demonstrated that OsNRT2.4 plays a role in root growth, nitrogen metabolic pathway and probably have functions in nitrate transport from root to shoot under low nitrate availability in rice. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-021-01273-6.

Metabolic characterization and oil profile of sunflower cultivars cultivated with increasing nitrogen doses / Caracterização metabólica e perfil do óleo de cultivares de girassol cultivados sob doses crescentes de nitrogênio

Sunflower (Helianthus annuus L.) has great economic importance due to its oil yield and quality. Among the factors influencing these parameters, nitrogen fertilization stands out. In this study we evaluated the effect of different N-urea doses (0,10,30,50,90,130 kg N-urea ha-1) at planting and 30 days after emergency on soluble fractions, oil profile and yield of three sunflower cultivars (BRS324, Catissol 01and Neon). Plants were harvested at the flowering stage for metabolic study, and at the end of the cycle for oil extraction and characterization. The Catissol 01 genotype presented the highest N-NO3- accumulation capacity, mainly in the stem. The Neon genotype presented the highest achene production at 30 and 50 kg N ha-1. Oleic oil had the highest yield, with the Catissol 01 and Neon cultivar standing out. Among the cultivars, Catissol 01 is economically more suitable for biofuel production, when cultivated at 50 kg N ha-1 since higher N doses did not result in increased oil yield.

O girassol é uma espécie com importante valor econômico pela qualidade e rendimento de óleo. Entre os fatores que influenciam esses parâmetros se destaca a fertilização nitrogenada. Este trabalho avaliou o efeito de doses crescentes de N-uréia (0, 10, 30, 50, 90, 130 kg de N-Uréia ha-1 aplicadas no plantio e aos 30 dias após a emergência) sobre as frações solúveis, rendimento e perfil do óleo de três genótipos de girassol (BRS324, Neon e Catissol 01). As plantas foram coletadas no estádio de floração para o estudo metabólico e ao final do ciclo para a extração e caracterização de óleo. O genótipo Catissol 01 apresentou maior capacidade de acúmulo de N-NO3-, preferencialmente no caule. A maior produção de óleo por planta foi observada no BRS324, nas doses entre 30 e 50 kg N ha-1, embora o genótipo Neon tenha apresentado a maior produção de aquênios. O ácido oléico foi o de maior rendimento com destaque para os genótipos Catissol 01 e Neon. Dentre os genótipos avaliados, o Catissol 01 é economicamente mais indicado para a produção de óleo destinada ao biocombustível, quando cultivado com a dose de 50 kg N ha-1, pois doses superiores de N não resultam em aumento na produção de óleo.

Silencing the Oryza sativa plasma membrane H+-ATPase isoform OsA2 affects grain yield and shoot growth and decreases nitrogen concentration.

The plasma membrane (PM) H+-ATPase (EC 3.6.1.3.) is a key component involved in nutrient uptake. There are 10 PM H+-ATPase isoforms in the rice genome (OsA1-OsA10), and OsA2 is highly responsive to nitrate (NO3-). We investigated the role that the OsA2 isoform plays in the total N and growth of rice (Oryza sativa). By the use of artificial microRNA, mutant osa2 rice lines presented ∼70 % downregulated levels of OsA2. Three osa2 lines and control plants (transformed with an empty IRS154 vector and named IRS) were cultivated in the greenhouse to evaluate grain and shoot production. For hydroponic experiments, the same lines were grown in Hoagland solution under two different NO3- levels for 30 days - 0.2 mM NO3--N (low N) or 2.0 mM NO3--N (sufficient N) - or were grown for three days without NO3- (starvation) after 27 days under 2.0 mM NO3--N. In the greenhouse experiments, compared with the IRS plants, the osa2 lines had lower shoot fresh weights, grain yields and SPAD values. Moreover, compared with the IRS plants, the three osa2 lines grown hydroponically under low NO3- levels had lower N concentration and net flux of NO3-. PM H+-ATPase activity was lower in the osa2 mutants than in the IRS plants. The relatively low N concentration in the osa2 lines was not due to lower expression of OsNRT2.1, OsNRT2.2, or OsNAR2.1. These results indicate that the specific PM H+-ATPase isoform OsA2 affects the net flux of NO3-, N concentration, and grain yield.

Isoforms of plasma membrane H(+)-ATPase in rice root and shoot are differentially induced by starvation and resupply of NO3⁻ or NH4+.

The goal of this study was to evaluate the effects of nitrogen starvation and resupply in 10 PM H+-ATPase isoforms and the expression of NO3⁻ and NH4+ transporters in rice. The net uptake of both forms of NO3⁻-N or NH4+-N was increased with its resupply. Resupply of NO3⁻ resulted in induction of the following PM H+-ATPase isoforms, OsA1, OsA2, OsA5 and OsA7 in the shoots and OsA2, OsA5, OsA7 and OsA8 in the roots. Resupply of NH4+ resulted in the induction of the following OsA1, OsA3 and OsA7 isoforms in the roots while OsA1 was induced in the shoots. It was observed that increased PM H+-ATPase activity also resulted in increased net uptake of NO3⁻ and NH4+. In the roots, OsNRT2.1 and OsNRT2.2 were induced by NO3⁻ resupply, while OsAMT1.1 and OsAMT1.2 were induced by NH4+ deficiency. The results showed that the expression of PM H+-ATPase isoforms is related to NO3⁻ and NH4+ transporters as well as in which section of the plant it takes place. PM H+-ATPase isoforms OsA2 and OsA7 displayed the strongest induction in response to N resupply, therefore indicating that these genes could be involved in N uptake in rice.