Overexpression of Rice Genes OsNRT1.1A and OsNRT1.1B Restores Chlorate Uptake and NRT2.1/NAR2.1 Expression in Arabidopsi
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Overexpression of Rice Genes OsNRT1.1A and OsNRT1.1B Restores Chlorate Uptake and NRT2.1/NAR2.1 Expression in Arabidopsis thaliana chl1‑5 Mutant Juan Sebastian Vera1 · Marcus Vinícius Loss Sperandio2 · Manlio Silvestre Fernandes1 · Leandro Azevedo Santos1 Received: 17 January 2020 / Accepted: 8 September 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Nitrogen uptake by plants is a key step for efficient nitrogen use, which affects plant growth and yield. Arabidopsis thaliana gene NRT1.1 was identified as a transporter related to nitrate (NO3−) signaling and uptake. In rice, three orthologs of NRT1.1, named OsNRT1.1A, OsNRT1.1B, and OsNRT1.1C, have been identified. This study evaluated the potential of OsNRT1.1A, OsNRT1.1B, and OsNRT1.1C in NO3− signaling and uptake through overexpression in the Arabidopsis chl1-5 mutant. The expression of OsNRT1.1A, OsNRT1.1B, and OsNRT1.1C was evaluated in the roots and shoots of rice cultivated with NO3− or NH4+. OsNRT1.1A was expressed in the roots and shoots cultivated with NO3− and NH4+. OsNRT1.1B was expressed predominantly in roots of rice cultivated with N O3−, while the expression of OsNRT1.1C was low in roots and shoots. Arabidopsis chl1-5 plants were transformed by the floral dip method using Agrobacterium tumefaciens to overexpress OsNRT1.1A and the alternative splicing product named OsNRT1.1As, OsNRT1.1B, and OsNRT1.1C. The chlorate test showed the ability of OsNRT1.1A, OsNRT1.1B or OsNRT1.1C to take up chlorate, as evidenced by the decrease in fresh weight. The OsNRT1.1B lineages presented higher toxicity to chlorate. Gene expression analyses showed that the insertion of OsNRT1.1A and OsNRT1.1B into Arabidopsis chl1-5 induced the expression of NRT2.1 and NAR2.1. OsNRT1.1As overexpression did not significantly affect the expression of NRT2.1 and NAR2.1. The results show the differential ability of NRT1.1 orthologs in rice to take up chlorate and signal the expression of other nitrate transporters, which may affect the efficiency of nitrogen utilization and its uptake. Keywords Nitrate signaling · Gene expression · Oryza sativa · Gateway cloning · Chlorate toxicity · Nitrate transport
Introduction Rice is one of the most consumed cereals worldwide and is part of the daily and staple diet of more than half the world’s population; it is one of the main crops for the maintenance Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00344-020-10219-9) contains supplementary material, which is available to authorized users. * Juan Sebastian Vera [email protected] 1
Federal Rural University of Rio de Janeiro, BR 465, Km 7.0, Seropédica, Rio de Janeiro, Brazil
Department of Biology, Federal Rural University of Pernambuco, R. Dom Manuel de Medeiros, Dois Irmãos, Recife, Pernambuco, Brazil
2
of food security of various countries. The majority of crops remove significant amounts of N from the soil, and significant applications of nitrogen fertilizers are necessar
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