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JXB Advance Access originally published online on August 1, 2005
Journal of Experimental Botany 2005 56(419):2443-2452; doi:10.1093/jxb/eri237
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org

RESEARCH PAPER

Soluble methionine enhances accumulation of a 15 kDa zein, a methionine-rich storage protein, in transgenic alfalfa but not in transgenic tobacco plants

Golan Amira1, Matityahu Ifat1, Avraham Tal1, Badani Hana2, Galili Shmuel2 and Amir Rachel1,3,*

1Plant Science Laboratory, Migal–Galilee Technology Center, PO Box 831, Kiryat Shmona, 11016, Israel
2Department of Agronomy and Natural Resources Department, Agricultural Research Organization, The Volcani Center, Bet Dagan 50250, Israel
3Tel-Hai Academic College, Upper Galilee 10120, Israel

* To whom correspondence should be addressed. Fax: +972 4 6944980. E-mail: rachel{at}migal.org.il

With the general aim of elevating the content of the essential amino acid methionine in vegetative tissues of plants, alfalfa (Medicago sativa L.) and tobacco plants, as well as BY2 tobacco suspension cells, were transformed with a ß-zein::3HA gene under the 35S promoter of cauliflower mosaic virus encoding a rumen-stable methionine-rich storage protein of 15 kDa zein. To examine whether soluble methionine content limited the accumulation of the 15 kDa zein::3HA, methionine was first added to the growth medium of the different transgenic plants and the level of the alien protein was determined. Results demonstrated that the added methionine enhanced the accumulation of the 15 kDa zein::3HA in transgenic alfalfa and tobacco BY2 cells, but not in whole transgenic tobacco plants. Next, the endogenous levels of methionine were elevated in the transgenic tobacco and alfalfa plants by crossing them with plants expressing the Arabidopsis cystathionine {gamma}-synthase (AtCGS) having significantly higher levels of soluble methionine in their leaves. Compared with plants expressing only the 15 kDa zein::3HA, transgenic alfalfa co-expressing both alien genes showed significantly enhanced levels of this protein concurrently with a reduction in the soluble methionine content, thus implying that soluble methionine was incorporated into the 15 kDa zein::3HA. Similar phenomena also occurred in tobacco, but were considerably less pronounced. The results demonstrate that the accumulation of the 15 kDa zein::3HA is regulated in a species-specific manner and that soluble methionine plays a major role in the accumulation of the 15 kDa zein in some plant species but less so in others.

Key words: Amino acids, cystathionine {gamma}-synthase, forage plants, 15 kDa zein, methionine, methionine-rich storage protein, nutritional quality, transgenic plants


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