Nitrogen uptake and expressions of nitrate transporter genes in two tropical forage grasses

Cristiane P. Silveira * 1, Joni E. Lima2, Jos Lavres Jr.1, Jos A. Bendassoli3,

Antonio V.O. Figueira2, Adibe L. Abdalla4
* Post Doctoral, USP/CENA; 303 Centenario Ave.; Piracicaba, SP 13400-970 Brazil;
1
Plant Nutrition Lab. USP/CENA; 2Plant Breeding Lab. USP/CENA,
3
Stable Isotopes Lab. USP/CENA, 4Animal Nutrition Lab. USP/CENA
* crispsad@gmail.com
The degradation of pastures coupled with the intensive system of grassland management has led
to the inevitable use of nitrogen (N) fertilizer in grasses. N is one of the most expensive nutrients
to supply and commercial fertilizers represent the major cost in plant production. An efficient
plant development relies on the ability of roots for N uptake and translocation at rate that
matches the demand of the plant. Therefore, a coordinated N transport system is fundamental for
nitrogen use efficiency (NUE). Thus, understand the molecular feature that regulates N
acquisition is relevant to improve NUE in Signal grass (Brachiaria decumbens) and Massai grass
(Panicum maximum x Panicum infestum). Herein, we report the physiological and molecular
characterization N uptake in roots of two tropical forage grasses. Grass plants grown under
nutrient solution displayed differences for NUE features in which the Signal grass showed high
NUE values compared to Massai grass. To characterize the N uptake, plants were grown under N
sufficient conditions, with external ammonium-nitrate concentration of 2.0 mM NH4NO3 (+N) or
without nitrogen supply (-N) for 3 days. After that, roots of intact plants were subjected to short
term 15N-influx studies by transferring the plants to nutrient solution labeled with 0.2 mM or 2.0
mM of 15NH415NO3. Under N sufficient growth conditions (+N), the N uptake rates at high
affinity concentration levels (0.2 mM 15N-labeled) in Signal grass roots was 62 % lower than that
of Massai grass. Contrary, during N deficiency (-N) no significant differences were observed
between the genotypes. Furthermore, the uptake studies performed under low-affinity range, at
2.0 mM of 15NH415NO3-influxes showed similar response for the Signal grass. These results
indicate that the N inefficient Massai grass has increased high and low-affinity systems that
operate during N acquisition under optimal N supply compared to Signal grass. To investigate
the molecular features of this response, the NITRATE TRANSPORTERS (NRTs) were identified
in grass EST genome. Expression levels for nitrate transporter gene NRT1.1 responsible for
nitrate uptake under low-affinity range was higher in Signal grass versus Massai grass. Similar
response was found for the main nitrate transporters active during high-affinity concentrations
with 17-fold higher expression levels for NRT2.1 and 8-fold more transcripts accumulation for
NRT2.2 gene in the roots of Signal grass compared to Massai grass under N-replete growth
condition. The increased expression levels found in Signal grass roots indicates that nitrate
uptake is higher in this species in comparison to Massai grass despite that the Massai grass have
shown a higher uptake levels of double-labeled ammonium nitrate. Altogether, these results
suggest that ammonium is the main N source taken up by Massai grass roots responsible for the
higher N-labeled uptake rates. In addition, the N efficient species Signal grass has enhanced
nitrate uptake capacity.
Keywords: nitrate transporter, regulation, root, influx, nitrogen use efficiency, 15N-labeled
Acknowledgments: We are grateful to The Coordenao de Aperfeioamento de Pessoal de
Nvel Superior (CAPES) for providing scholarships to the first author and for the research
financial support.