Expression analysis of phytochromes A, B and floral integrator genes during the entry and exit of grapevine-buds from endodormancy

A common molecular regulatory pathway that involves PHYA, PHYB and floral integrator genes CONSTANS (CO), FLOWERING LOCUS T (FT) and SUPRESSOR OF OVEREXPRESSION OF CO1 (SOC1) has been suggested to participate in the regulation of photoperiod dependent processes such as flowering and dormancy. In grapevines (Vitis vinifera L.), decreasing photoperiod and low temperatures trigger the transition of buds into endodormancy (ED), a process that is accompanied by drastic changes in gene expression of VvPHYA and B in leaves. To analyse the relationship of VvPHYA, VvPHYB, and grape homologues of Arabidopsis floral integrator genes VvCO, VvFT, VvMADS8, with ED, a comparative expression analysis of these genes was performed in grapevine-leaves and buds before, during and after the transition of buds into ED. The expression of all the above genes in the bud-tissue, and the fact that photoperiod regulates differently the expression of VvPHYA and B in buds than in leaves, suggests that the bud might be an autonomous or semi-autonomous organ in perceiving and transducing the photoperiod signal. On the other hand, the coordinated down-regulation of VvFT in leaves and buds during the transition of buds into ED, and its subsequent up-regulation following the application of dormancy-breaking compounds, hydrogen cyanamide (HC) and sodium azide, suggests that VvFT could play a key role in stimulating bud-growth by repressing their entry into ED.     

Auxin and cytokinin related gene expression during active shoot growth and latent bud paradormancy in Vitis riparia grapevine

Cultural practices for canopy management in grapevines rely on intensive manipulation of shoot architecture to maintain canopy light levels. In contrast to common model plant systems used to study regulation of branch outgrowth, the grapevine has a more complex architecture. The node contains first, second and third order axillary meristems. The prompt bud (N+1) develops into a summer lateral and a latent compound bud develops in the basal node of the summer lateral (N+2, N+3(1,2)). The outgrowth potential of latent buds was determined using common canopy management treatments (shoot tip decapitation and removal of summer laterals and leaves) and monitoring the rate of latent bud outgrowth. Two shoot node regions (apical and basal) with differential outgrowth potential were characterized and it was noted that the shoot tip, summer laterals and leaves in addition to node position contributed to the inhibition of latent bud outgrowth. To advance the understanding of the molecular regulation of bud outgrowth and paradormancy in the complex shoot architecture of grapevines, the expression of auxin and cytokinin genes involved in branching (amidase (VrAMI1), PINFORMED-3 (VrPIN3) and isopentenyl transferase (VrIPT)) were monitored in shoot tips and differentially aged buds of Vitis riparia grapevine shoots. In addition, Histone 3 (VrH3) and a hexose transporter (VrHT1) expression were monitored as a measure of tissue activity. The expression of VrAMI1 and VrPIN3 remained constant in actively growing shoot tips and decreased significantly with increasing bud maturation in paradormant buds. VrHT1 expression was greater in buds than in any other plant tissue tested. VrHT1 may have the potential to be used as an indicator of paradormancy status in grapevines. These characterizations in the complex architecture of the grapevine provide an excellent model system for molecular analysis of bud outgrowth and shoot architecture development.     

Possible role of catalase in post-dormancy bud break in grapevines

Changes in the activity of catalase (Cat) and in the levels of H2O2 were followed throughout dormancy in buds of grapevines (Vitis vinifera L.). In grapevines grown in the Elqui valley in Chile, a region with warm-winters, the activity of Cat increased during the recess period of buds, reaching a maximum and thereafter decreased to less than one third of its maximal activity. Three isoforms of Cat were detected in extracts of buds by native PAGE analysis, and the extracted activity was inhibited competitively by hydrogen cyanamide (HC), a potent bud-break agent. Furthermore, HC applications to field-grown grapevines in addition to the expected effect on advancing bud break, reduced the Cat activity during bud dormancy. Similar reductions were observed during dormancy in buds of grapevines grown in the Central valley in Chile, a region with temperate winters, suggesting that HC and winter chilling inhibits the activity of the main H2O2 degrading enzyme in grape buds. A transient rise in H2O2 levels preceded the release of buds from endodormancy, moreover, the peak of H2O2 and the onset of bud break occurred earlier in HC treated than in control grapevines, suggesting the participation of H2O2 as a signal molecule in the release of endodormancy in grape buds. The relationship between Cat inhibition, rise in H2O2 levels and initiation of bud break are discussed.     

Alterations in the pattern of peroxidase isoenzymes and transient increases in its activity and in H2O2 levels take place during the dormancy cycle of grapevine buds: the effect of hydrogen cyanamide

Polymorphism of peroxidase (Px) and changes in its activity and in H₂O₂ content were studied in buds of grapevine during dormancy. Three isoforms of Px were detected in bud-extracts, two basic and one acidic, however, the pattern of Px isoenzyme changed with the progress of dormancy. Thus, basic Px isoenzymes disappeared from extracts previous to the onset of bud-break, while acidic isoenzymes remained relatively unaltered throughout the whole dormancy period. Furthermore, transient increases in the activity of Px and in the content of H₂O₂ occurred previous to endodormancy release, when buds were fully dormant. Hydrogen cyanamide (H₂CN₂), a potent bud breaking agent in grapevines advanced as expected bud-break, but also advanced the occurrence of Px and H₂O₂ peaks and the changes in Px isoenzymes pattern. The results suggests that H₂O₂ could function as a signalling molecule inducing endodormancy release, and changes in Px polymorphism could be a useful marker to study endo/ecodormancy phase transition in buds of grapevines.     

Peroxidase Isoenzyme Patterns in Cell Cultures derived from Cotyledon, Stem, Leaf and Fruit from Grapevine (Vitis vinifera cv. Monastrell)

The expression of peroxidase isoenzymes capable of oxidizing4-hydroxystilbenes was studied during the establishment of cellcultures derived from different tissues (cotyledon, stem, leafand fruit) of Vitis vinifera cv. Monastrell vines. This wascarried out in order to elucidate whether different tissuesof the same plant maintain persistent tissue-specific patternsof gene expression during in vitro culture or whether in vitrocultures are characterized by identical patterns of gene expressionirrespective of the tissue's origin. The results illustratedthat both the isozyme patterns and the substrate specificityof the peroxidase activity secreted to the medium are analogousfor the profile of acidic (Prx A) and basic (Prx B) peroxidaseisoenzymes, only quantitative differences being shown in theneutral peroxidase isoenzyme (Prx N) pattern. These resultssuggest that in vitro cultures of grapevines are characterizedby similar patterns of gene expression, no matter what theirtissue of origin.Copyright 1995, 1999 Academic Press Grapevine, vitis vinifera, cell cultures, 4-hydroxystilbene oxidizing peroxidase isoenzyme, substrate specificity     

Expression of cucumber stress-related anionic peroxidases during flower development or a cryptic infective process

The striking diversity in the expression pattern of the stress-related anionic peroxidase was observed during development of female cucumber flower. While the isoenzyme Prx3 was accumulated constitutively in the course of flower development, the expression patterns of other two isoenzymes (Prx1 and Prx2) were restricted to the period after flower opening. The virus infection was simulated by careful opening of the intact female flower buds 3 d before anthesis followed by exposition to the glasshouse environment for 3 d. The results obtained in this experiment revealed a marked accumulation of the isoenzyme Prx1 and Prx2 at anthesis. Under normal flower development, the pistils did not accumulate these isoenzymes at this stage. In contrast, the pattern of expression of Prx3 as well as of the pistil-specific peroxidase isoenzyme remained unchanged, confirming a constitutive type of expression. Beside the pistil, a 3-d exposition of the stripped flowers resulted in a marked accumulation of Prx1 and Prx2 isoenzymes also in both adjacent flower organs - the ovary and the pedicel. At the same time of the normal development of female flower these organs did not accumulate these isoenzymes.     

Expression of defence-related peroxidases Prx7 and Prx8 during abiotic stresses in barley roots

The expression of defence-related peroxidases Prx7 and Prx8 in barley roots grown under selected abiotic stress conditions (toxic metals: Cd, Al, Co, Cu, Hg; drought, salinity, extreme temperatures: heat, cold) and compounds activating (2,4-D) or inhibiting (SHAM) POD activity as well as H₂O₂ and H₂O₂ scavenger (DTT) was characterized. Strong Cd concentration dependent expression of Prx8 peroxidase gene was observed, which correlated with root growth inhibition induced by Cd- and some other stress factors (heavy metals, heat and salinity). Application of H₂O₂ did not cause changes in expression of Prx8, but H₂O₂ scavenger (DTT) as well as the inhibitor (SHAM) and the activator (2,4-D) of PODs induced increase in Prx8 expression. Our results demonstrate that root growth inhibition during any disturbance of active oxygen species (AOS) in root tissue is correlated with up-regulation of Prx8 gene expression in barley roots.