.)Raffinose family oligosaccharides (RFOs) have been implicated in mitigating the effects of environmental stresses on plants. In seeds, proposed roles for RFOs include protecting cellular integrity during desiccation and/or imbibition, extending longevity in the dehydrated state, and providing substrates for energy generation during germination. A gene encoding galactinol synthase (GOLS), the first committed enzyme in the biosynthesis of RFOs, was cloned from tomato (Lycopersicon esculentum Mill. cv Moneymaker) seeds, and its expression was characterized in tomato seeds and seedlings. GOLS (LeGOLS-1) mRNA accumulated in developing tomato seeds concomitant with maximum dry weight deposition and the acquisition of desiccation tolerance. LeGOLS-1 mRNA was present in mature, desiccated seeds but declined within 8 h of imbibition in wild-type seeds. However, LeGOLS-1 mRNA accumulated again in imbibed seeds prevented from completing germination by dormancy or water deficit. Gibberellin-deficient (gib-1) seeds maintained LeGOLS-1 mRNA amounts after imbibition unless supplied with gibberellin, whereas abscisic acid (ABA) did not prevent the loss of LeGOLS-1 mRNA from wild-type seeds. The presence of LeGOLS-1 mRNA in ABA-deficient (sitiens) tomato seeds indicated that wild-type amounts of ABA are not necessary for its accumulation during seed development. In all cases, LeGOLS-1 mRNA was most prevalent in the radicle tip. LeGOLS-1 mRNA accumulation was induced by dehydration but not by cold in germinating seeds, whereas both stresses induced LeGOLS-1 mRNA accumulation in seedling leaves. The physiological implications of LeGOLS-1 expression patterns in seeds and leaves are discussed in light of the hypothesized role of RFOs in plant stress tolerance.The raffinose family oligosaccharides (RFOs) are soluble galactosyl-Suc carbohydrates that constitute a significant component of phloem-transported sugars in some plants (Haritatos et al., 2000). Their accumulation in plants is associated with stressful environmental conditions such as cold, heat, or dehydration (Santarius, 1973; Santarius and Milde, 1977; Hinesley et al., 1992; Ashworth et al., 1993; Wiemken and Ineichen, 1993; Bachmann et al., 1994; Taji et al., 2002). The potential role of RFOs in stress tolerance has been intensively studied in seeds, particularly with respect to desiccation tolerance and longevity in the dehydrated state. RFOs are abundant in most mature desiccation-tolerant ("orthodox") seeds and are often rare or absent in "recalcitrant" seeds that cannot withstand desiccation (Lin and Huang, 1994; Sun et al., 1994). In seeds of many species, RFO accumulation coincides with the development of desiccation tolerance during seed maturation (Koster and Leopold, 1988; Leprince et al., 1993; Bewley and Black, 1994; Horbowicz and Obendorf, 1994; Black et al., 1996; Brenac et al., 1997a Brenac et al., , 1997b, and RFO content has been positively correlated with seed longevity in storage (Bernal-Lugo and Leopold, 1992; Horbowicz and Obendorf, 1994; Lin ...
The proliferating cell nuclear antigen (PCNA) is a protein factor required for processive DNA synthesis that is associated with G(1) cell cycle proteins. It has been demonstrated previously that, in germinating maize (Zea mays) embryonic axes, PCNA forms protein complexes with two Cdk-A proteins (32 and 36 kDa) and with a putative D-type cyclin. These complexes exhibit protein kinase activity on histone H1 and on the maize homologue of the pRB (retinoblastoma) protein. Flow cytometry has been used to study the influence of the phytohormones benzyladenine (BA) and abscisic acid (ABA) on cell cycle advancement during maize germination. It was found that, while BA accelerates the passage of cells from G(1) to G(2), ABA delays cell cycle events so that most cells seem to remain in G(1). The amounts of PCNA and Cdk-A proteins also vary according to the hormone treatment. In embryonic axes, PCNA increases rapidly during early germination in BA, compared with a gradual increase in water, while ABA treatment had only a marginal effect. However, of the two Cdk-A proteins, the 32 kDa protein is strongly reduced after 15 h of imbibition in water while this occurs later when axes are imbibed in BA or ABA. The PCNA-associated protein kinase activity in the BA and ABA treatments falls after 3 h of imbibition compared with activity in the control; however, while kinase activity in the BA treatment continues to decline during imbibition, it remains relatively constant until 24 h of imbibition in the ABA treatment. By contrast, a p13(Suc1)-associated Cdk-A kinase is activated after 15 h of imbibition under all treatments, particularly in ABA. These results suggest that, in maize, ABA delays the germination process by affecting cell cycle advancement, stopping cells mostly in a G(1) state.
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