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III. SIMULTANEOUS BIOFORTIFICATION OF Zn AND Fe IN Triticum<br />

aestivum L. – NUTRIENTES ACCUMULATION IN THE SEEDS<br />

Fernando H. Reboredo 1 , Ana Luísa Fernando 1 , Fernanda Simões 3 , Ana Rita Costa 3 , José<br />

Matos 3 , António E. Leitão 2 , José C. Ramalho 2 , Ana Sofia Almeida 3 , Paula Scotti-Campos 3 ,<br />

Isabel P. Pais 3 , Benv<strong>in</strong>do Maçãs 3 , José P. Cout<strong>in</strong>ho 3 , Maria Manuela Silva 4 , Maria Paula<br />

Duarte 1 , Luís F. Goulão 3 , Ana I. Ribeiro 2 , Fernando C. Lidon 1<br />

1 DCTB, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Qu<strong>in</strong>ta da Torre,<br />

2829-516 Caparica, Portugal.<br />

2 Centro de Ecofisiologia, Bioquímica e Biotecnologia Vegetal, Instituto de Investigação<br />

Científica Tropical, Qu<strong>in</strong>ta do Marquês, 2784-505 Oeiras, Portugal.<br />

3 Instituto Nacional de Recursos Biológicos, Estrada de Gil Vaz, Apartado 6, 7350-591 Elvas /<br />

Estrada do Paço do Lumiar, 22 Ed. 5, 1649-038 Lisboa/ URGEMP, Qu<strong>in</strong>ta do Marquês,<br />

2784-505, Oeiras, Portugal.<br />

4 ESE Almeida Garrett, Grupo Universidade Lusófona, COFAC, Palácio de Santa Helena,<br />

Largo do Sequeira nº 7, Lisboa, Portugal.<br />

Keywords: Nutrients localization, Triticum biofortification, X-ray microanalysis<br />

Certified seeds of Triticum aestivum L. cv Nabão (0Ts) were grown <strong>in</strong> walk-<strong>in</strong> growth<br />

chamber (EHHF10000, Aralab, Portugal), under environmental controlled conditions (80%<br />

RH; 24/20C day/night temperatures; PPFD of ca 700 µmol m -2 s -1 , 12 h photoperiod), <strong>in</strong> pots<br />

irrigated <strong>with</strong> a standard solution (1Ts), dur<strong>in</strong>g 1 month follow<strong>in</strong>g germ<strong>in</strong>ation. Thereafter,<br />

until <strong>the</strong> physiological maturity, half of <strong>the</strong> pots kept <strong>the</strong> same irrigation solution (1Ts), while<br />

<strong>the</strong> o<strong>the</strong>r half was submitted to a five fold concentration of all <strong>the</strong> nutrients (1T5s). The seeds<br />

1T5s were sown and grown aga<strong>in</strong> us<strong>in</strong>g <strong>the</strong> <strong>in</strong>itial standard nutrient solution (now called 2Ts)<br />

and solutions conta<strong>in</strong><strong>in</strong>g 5, 7, 10 and 13 times <strong>the</strong> concentration of all <strong>the</strong> nutrients (2T5s,<br />

2T7s, 2T10s and 2T13s, respectively). In <strong>the</strong> seeds of 1Ts, 1T5s, 2Ts, 2T5s, 2T7s, 2T10s and<br />

2T13s, Zn concentration <strong>in</strong>creased about 7.18, 6.10, 9.92, 8.87, 4.73, 9.00 and 9.23 fold,<br />

whereas Fe augmented 1.01, 1.35, 3.91, 4.95, 5.10, 8.98 and 11.06 times, respectively,<br />

compared <strong>with</strong> <strong>the</strong> 0Ts Zn and Fe levels. The pattern of nutrient accumulation, as seen by<br />

SEM, coupled <strong>with</strong> X-ray microanalysis, throughout a l<strong>in</strong>e scan on transversal sections of <strong>the</strong><br />

gra<strong>in</strong>s, revealed a non-homogeneous distribution of <strong>the</strong> nutrients, despite <strong>the</strong> distribution of<br />

<strong>the</strong> mean values shown were: Cu 0Ts

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