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Microtubules play important roles during growth and morphogenesis of plant cells. Multiple isoforms of alpha- and beta-tubulin accumulate in higher plant cells and originate either by transcription of different genes or by post-translational modifications. The use of different tubulin isoforms involves the binding of microtubules to different associated proteins and therefore generates microtubules with different organizations and functions. Tubulin isoforms are differentially expressed in vegetative and reproductive structures according to the developmental program of plants. In grapevine (Vitis vinifera L.), vegetative and reproductive structures appear on the same stem, making this plant species an excellent model to study the accumulation of tubulin isoforms. Proteins were extracted from grapevine samples (buds, leaves, flowers and tendrils) using an optimized extraction protocol, separated by two-dimensional electrophoresis and analyzed by immunoblot with anti-tubulin antibodies. We identified eight alpha-tubulin and seven beta-tubulin isoforms with pI around 4.8-5 that group into separate clusters. More acidic alpha-tubulin isoforms were detected in buds, while more basic alpha-isoforms were prevalently found in tendrils and flowers. Similarly, more acidic beta-tubulin isoforms were used in the bud stage while a basic beta-tubulin isoform was essentially used in leaves and two central beta-tubulin isoforms were characteristically used in tendrils and flowers. Acetylated alpha-tubulin was not detected in any sample while tyrosinated alpha-tubulin was essentially found in large latent buds and in bursting buds in association with a distinct subset of tubulin isoforms. The implication of these data on the use of different tubulin isoforms during grapevine development is discussed.

To further understand post-translational modifications (PTMs) of plant alpha-tubulin, post-translationally modified alpha-tubulin isoforms from selected tissues of Zea mays L. were examined using two-dimensional electrophoresis and immunoblotting. Except for polyglycylated tubulin, tyrosinated, detyrosinated, acetylated and polyglutamylated alpha-tubulin isoforms were all present in maize tissues. Tyrosinated alpha-tubulin was the predominant variant in all cases, with isoforms alpha1-alpha4 (alpha5) being the most common components. Leaves exhibited a striking difference in PTM patterns of alpha-tubulin isoforms compared to other tissues examined. In leaves, several major specific isoforms were highly modified by detyrosination, acetylation and polyglutamylation. In pollen and anthers, only the most abundant isoform alpha3 was acetylated to an appreciable extent, and no acetylated isoform was found in roots. Similarly, in pollen, anthers and roots, only alpha3 was appreciably polyglutamylated. Additionally, a detyrosinated isoform alpha6 was present in anthers and in leaves, while the tyrosinated isoform alpha6 seemed to be pollen specific. These results indicate that certain types of PTM of plant alpha-tubulin preferentially occur in a tissue-specific way.