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Garlic extract improves budbreak of the 'Niagara Rosada' grapevines on sub- tropical regions/Extrato de alho melhora a superacao da dormencia de videiras 'Niagara Rosada' em regioes subtropicais.

INTRODUCTION

The 'Niagara Rosada' grapevine (Vitis labrusca) is originated from a natural mutation of 'Niagara', an American grapevine introduced from Alabama (USA) that occurred in 1933, in Sao Paulo State, Brazil. This mutant grapevine supplanted the original 'Niagara' and is one of the most important table grape cultivar in Brazil (TECCHIO et al., 2009).

The grapevine, typical deciduous species of temperate climate, is broadly cultivated in tropical and subtropical areas, like the zones of Sao Paulo State, Southeastern region of Brazil. In such areas, the chilling requirement for budbreak are not fulfilled, leading to a late leafing, reduced and no regular sprouting and, consequently, decrease in yield, causing significant economic loses. Chilling exposing necessary for normal bud growth ranges between 50 and 400h at temperatures of 7.0[degrees]C, varying for each grapevine cultivar and 'Niagara Rosada' grapevines needs about 100 h (DOKOOZLIAN et al., 1998),

Previous literature indicates that the exposition of the buds to chilling inhibits the activity of the catalase, enzyme present in aerobics cells that decomposes the hydrogen peroxide ([H.sub.2][O.sub.2]) in molecular oxygen and [H.sub.2]O (NIR et al., 1986). Its physiologic function eliminates the excess of [H.sub.2][O.sub.2] produced during the cellular metabolism, avoiding its accumulation and consequent cellular damage. However, evidences have shown that [H.sub.2][O.sub.2] acts as a chemical signal in response to biotic and abiotic stresses (BARTOSZ, 1997; FOYER et al., 1997).

After that, the plant leads to [H.sub.2][O.sub.2] deintoxication through a sequence of reactions connected to the pentose phosphate pathway, leading to an increase of reduced nucleotides NAD(P)H, raising the metabolism and the induction of dormancy termination, bud burst and rapid growth (NIR et al., 1986). According to PINTO et al. (2007), these metabolic changes has as consequence the increase on the levels of AMP/ATP intracellular relation that induces proteinkinases SNF, that plays a role in the transduction signal system for endodormancy end of buds.

Nowadays, both calcium cyanamide (CaC[N.sub.2]) and hydrogen cyanamide ([H.sub.2]C[N.sub.2]) are recommended in Brazil for budbreak induction of grapevines, but [H.sub.2]C[N.sub.2] is the mostly used in Brazil for all temperate zone fruits (HAWERROTH et al., 2010). For the Jundiai region, in Brazil, CaC[N.sub.2] 200g [L.sup.-1] or [H.sub.2]C[N.sub.2] 25g [L.sup.-1] promoted budbreak of the grapevine 'Niagara Rosada' in 100% of the buds (PIRES et al., 1985).

In United States and in Italy the use of [H.sub.2]C[N.sub.2] was estimated in 112,490 and 36,287kg, respectively, mainly in grapes. However, hydrogen cyanamide is highly toxic. The Environmental Protection Agency of the United States classifies this compound in the highest toxicity category (category I), and it has been under regulatory review by European Union authorities (SETTIMI et al., 2005). Accordingly to exposed, there is a necessity of new agents for dormancy break that are easily available, effective, low toxic and used in low concentrations.

Searching for new alternatives for budbreak, KUBOTA & MIYAMUKI (1992) verified that garlic paste applied to cane cut surfaces of 'Muscat of Alexandria' grapevines, immediately after pruning, was more efficient than calcium cyanamide (CaC[N.sub.2]), a substance typically used for vines in Japan. Satisfactory results were also obtained with 20% garlic oil in 'Pione' and 'Thompson Seedless' grapevines (KUBOTA et al., 2000). BOTELHO et al. (2007), observed 37% and 75% sprouted buds in cuttings of grapevines cv. Cabernet Sauvignon sprayed with garlic extract 3%, submitted to 0 and 168 chilling hours (= 7, 0[degrees]C), respectively, but this treatment were less effective than hydrogen cyanamide.

This study aimed to investigate the effect of the garlic extract, compared to the calcium cyanamide and hydrogen cyanamide on budbreak of 'Niagara Rosada' grapevines.

MATERIAL AND METHODS

The experiment was carried out in two commercial 'Niagara Rosada' (Vitis labrusca) vineyards in Sao Paulo State, Southeastern region of Brazil: one located in Jundiai, (23[degrees]06'S and 46[degrees]55'W, 715m a.s.l.); and another in Indaiatuba (23[degrees]05'S and 47[degrees]13'W, and 630m a.s.l.). The nine-year-old vines on the rootstock IAC-766 'Campinas' (106-8Mgt x Vitis caribaea) were spaced 2.0 by 1.0m and trained to a unilateral cordon. The canes, six per cordon, were pruned to a single-bud spur in 26 July 2007, in Jundiai vineyard and; in 11 July 2007, in Indaiatuba.

The treatments were composed of the following commercial products: Bioalho[R] (garlic extract 700mL [L.sup.-1], Natural Rural Co.), Dormex[R] ([H.sub.2]C[N.sub.2] 490g [L.sup.-1], Basf Co.) and Hiraguen-S[R] (Nonyl phenoxy poly (ethylene oxy) ethanol 200g [L.sup.-1], Hirarabras Co.). Immediately after pruning, the following treatments were sprayed to 'drip point' at dormant bud stage using a hand driven sprayer: garlic extract (GE) at 0, 14, 28, 42, 56 e 70mL [L.sup.-1]; calcium cyanamide (CaC[N.sub.2]) at 200g [L.sup.-1] and hydrogen cyanamide ([H.sub.2]C[N.sub.2]) at 25g [L.sup.-1]. In all the solutions were added 20mL [L.sup.-1] of a no ionic adjuvant.

In each vineyard, the trial was laid out using a completely randomized design with eight treatments and six one-whole-plant replicates. Each vine plant was examined for the following variables: 1) sprouting time: number of days between pruning and begging of sprouting; 2) sprouting percentage: considered when no more evolution in bud sprouting was verified; 3) cycle length: number of days between pruning and harvest, 4) number of clusters per plant. Budbreak was regarded when a green tinge was seen beneath the bud scales; this evaluation was performed weekly. The harvest was carried out when the total soluble solids content of most grapes reached 15oBrix.

All data were analyzed statistically using the ANOVA in the SANEST statistical package (ZONTA & MACHADO, 1987). Meaningful comparisons were generated using Tukey's test (5% level) and regression analysis.

RESULT AND DISCUSSION

In both vineyards, the time for sprouting beginning after pruning was linearly reduced in function of garlic extract doses, anticipating 30 days in Indaiatuba and 6 days in Jundiai for the highest dose (70mL [L.sup.-1]) (Figures 1A and 1B). The conventional treatments with CaC[N.sub.2] and [H.sub.2]C[N.sub.2] were the most effectives for accelerate bud sprouting, but they did not differ of the GE at 56 and 70mL [L.sup.-1] in Indaiatuba vineyard (Figure 1A) and, GE at 56mL [L.sup.-1] did not differ from [H.sub.2]C[N.sub.2], in Jundiai vineyard (Figure 1B) .

The sprouting percentage was linearly increased with garlic extract doses, in Indaiatuba and Jundiai vineyards (Figure 2A and 2B). The highest values for bud sprouting was verified for the treatments with CaC[N.sub.2] and [H.sub.2]C[N.sub.2], reaching 100%, although they were similar to the garlic extract 70mL [L.sup.-1], that attained 89 and 94% of sprouted buds compared to only 56 and 75% in control plants, in Indaiatuba and Jundiai, respectively.

The cycle between pruning and harvest was linearly decreased in both vineyards in relation to garlic extract doses, but the shortest cycles were observed for the conventional treatments with CaC[N.sub.2] and [H.sub.2]C[N.sub.2], completing 124 and 130 days for both treatments, in Indaiatuba and Jundiai, respectively. In control plants the cycle was 185 days in Indaiatuba and 150 days in Jundiai (Figures 3A and 3B, respectively).

The number of clusters per plant was improved with increasing doses of garlic extract showing a quadratic effect in both vineyards locations (Figures 4A and 4B). The conventional treatments for budbreak with CaC[N.sub.2] and [H.sub.2]C[N.sub.2] showed the highest values for number of clusters, but they did not differ from the treatment with GE at 70mL [L.sup.-1], in Indaiatuba and Jundiai.

In this experiment, the best results for budbreak were verified for CaC[N.sub.2] and [H.sub.2]C[N.sub.2] treatments. Similarly, BOTELHO et al. (2002) verified that applications of 3.0% [H.sub.2]C[N.sub.2] to nonchilled Centennial Seedless grapevines induced about 70% budbreak after two weeks from treatment date, while buds on control vines attained only 30%. In Cabernet Sauvignon grapevines, MIELE (1991) observed maximum percentage of bud burst with concentrations between 1.8% and 1.9% hydrogen cyanamide in Rio Grande do Sul State, Brazil. PEREZ & LIRA (2005) demonstrated that [H.sub.2]C[N.sub.2] inhibited competitively the activity of catalase. In this case, a shortening in bud endodormancy could be related to catalase inhibition through [H.sub.2][O.sub.2] accumulations in bud tissues. An excess of [H.sub.2][O.sub.2] could generate an oxidative stress or act as chemical signal that triggers the expression of genes related to endodormancy release.

Garlic extract treatments also promoted bud sprouting, similar then the standard treatments with cyanamides. Similar results were verified with the use of garlic-based compounds in grapevines, but using much more concentrated solutions (from 20 to 100%) (KUBOTA & MIYAMUKI, 1992; KUBOTA et al., 1999a,b, 2000). According to KUBOTA et al. (2002) the active substances in garlic responsible for breaking bud dormancy are volatile compounds containing sulfur and an allyl group (C[H.sub.2]CHC[H.sub.2]), specially the diallyl dissulfide, which is the most abundant sulfide in garlic. Exposure of grapevine cuttings to volatiles from grated garlic and commercial garlic oil promoted budbreak (KUBOTA et al., 1999a) and exposures to volatiles of diallyl di- and tri-sulfides were also effective (KUBOTA et al., 1999b).

However, the physiological role of these compounds on the breaking bud dormancy in no chilled grapevines was not established (KUBOTA et al., 2000). Probably, these substances would actuate by the same mechanism proposed by PINTO et al. (2007) as a result of oxidative stress through accumulation of [H.sub.2][O.sub.2]. LEMAR et al. (2005) observed that the application of garlic extract caused an oxidative stress in cells of Candida albicans, but in this case this would lead to the inhibition of the growth of this fungus colonies and the destruction of its cellular components.

Bioalho[R] (garlic extract) is a natural product recommended as insect repellent for organic production systems, according to current Brazilian legislation. It is obtained from cold drawing out of garlic extract by press and this formulation is totally soluble in water. The GE at 70mL [L.sup.-1] promoted 'Niagara Rosada' grapevines budbreak and could be recommended for agroecologic viticulture. Although, cyanamide treatments were more effective; these products show high toxicity and is not allowed for organic fruit production.

In spite of the promising results showed by the treatment with GE in field conditions, the results are still preliminary, needing new researches, mainly aiming the increase of economical viability and the reduction of doses, for example with the use of different adjuvants. According to DOKOOZLIAN et al. (1998), the scales of the buds vines, that protect them against dry up and extreme climatic conditions, are a barrier for products used for the dormancy break. In this case, the adjuvants could increase the penetration of the products in the bud, increasing its efficiency.

CONCLUSION

The effect of the treatment with garlic extract 70mL [L.sup.-1] on budbreak of the 'Niagara Rosada' grapevines was similar to the conventional treatments with CaC[N.sub.2] 200g [L.sup.-1] or [H.sub.2]C[N.sub.2] 25g [L.sup.-1] in both vineyard locations. These treatments also reduced the time for beginning of sprouting and the cycle from pruning to harvest and increased the number of cluster per plant. Garlic extract could be used in substitution of CaC[N.sub.2] or [H.sub.2]C[N.sub.2] in agroecologic vineyard.

REFERENCES

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FOYER, C.H. et al. Hydrogen peroxide and glutathione associated mechanisms of acclamatory stress tolerance and signaling. Physiologia Plantarum, Hoboken v.100, n.2, 241-254, 1997. Available from: <http://www3.interscience.wiley.com/journal/119168585/issue>. Accessed: May 13, 2009. doi: 10.1111/j.1399-3054.1997.tb04780.x.

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KUBOTA, N.; MIYAMUKI, M. Breaking bud dormancy in grapevines with garlic paste. Journal of the American Society for Horticultural Science, Alexandria, v.117, n.6, p.898-901, 1992. Available from: http://journal.ashspublications.org/cgi/reprint/117/6/898?>. Accessed: Jan 15, 2009.

KUBOTA, N. et al. Breaking bud dormancy in grapevine cuttings with garlic volatiles. Journal of the Japanese Society for Horticultural Science, Kioto, v.68, n.5, p.927-931, 1999a. Available from: <http://www.journalarchive.jst.go.jp/english/jnlabstract_en.php?cdjournal=jjshs1 925&cdvol=68&noissue=5&sta rtpage=92>. Accessed: Jul 18, 2009. doi:10.2503/jjshs.68.927

KUBOTA, N. et al. Identification of active substances in garlic responsible for breaking bud dormancy in grapevines. Journal of the Japanese Society for Horticultural Science, Kioto, v.68, n.6, p.1111-1117, 1999b. Available from: <http://www.journalarchive.jst.go.jp/jnlpdf.php?cdjournal=jjshs1925&cdvol=68&noi ssue=6&startpage=11 11&lang=en&from=jnltoc>. Accessed: Sep 20, 2009. doi:10.2503/jjshs.68.1111.

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LEMAR, K.M. et al. Allyl alcohol and garlic (Allium sativum) extract produces oxidative in Candida albicans. Microbiology, Reading, n.10, v. 151, p.3257-3265, 2005. Available from: <http://mic.sgmjournals.org/cgi/reprint/151/10/3257>. Accessed: Dec 20, 2009. doi: 10.1099/mic.0.28095-0.

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PIRES, E.J.P. et al. Acao de calciocianamida e alzodef na epoca de brotacao e produtividade da videira 'Niagara Rosada'. Anais da Escola Superior de Agricultura "Luiz de Queiroz", v.42, n.2, p.469-479, 1985.

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TECCHIO, M.A. et al. Efeito do acido naftalenoacetico e do cloreto de calcio na reducao das perdas pos-colheita em uva 'Niagara Rosada'. Revista Brasileira de Fruticultura, Jaboticabal, v.31, n.1, p.53-61, 2009. Available from: <http://www.scielo.br/scielo.php?script=sci_arttext&pid=S010029452009000100009&l ng=pt&nrm=iso>. Accessed: Sep 01, 2010. doi: 10.1590/S010029452009000100009.

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Renato Vasconcelos Botelho (I) Erasmo Jose Paioli Pires (II) Mara Fernandes Moura (II) Maurilo Monteiro Terra (II) Marco Antonio Tecchio (II)

(I) Departamento de Agronomia, Universidade Estadual do Centro-Oeste (Unicentro). Rua Simeao Varella de Sa, 03, 85040- 080, Guarapuava, PR, Brasil. E-mail: rbotelho@unicentro.br. Autor para correspondencia.

(II) Instituto Agronomico de Campinas (IAC). Campinas, SP, Brasil.

Received 04.23.10 Approved 10.07.10 Returned by the author 10.27.10 CR-3479
Figure 1--Sprouting time (days) between pruning and begging of
sprouting of 'Niagara Rosada' grapevines sprayed with different
treatments for budbreak in two vineyard locations: Indaiatuba
(CV=36.7%) (A) and Jundiai (CV=34.0%) (B).

(A)

v = 55.905 - 0.4163x, [r.sup.2] = 0.877**

garlic extract (mL [L.sup.-1])

0       56a
14      53ab
28      37cd
42      43bc
56      53cde
70      26de
CaCN2   21e
H2CN2   23e

(B)

garlic extract (mL [L.sup.-1])

y = 40.333 - 0.1, [r.sup.2] = 0.8833 **

0       40a
14      30a
28      38a
42      37a
56      33ab
70      34a
CaCN2   23c
H2CN2   28bc

Note: Table made from Bar graph.

Figure 2--Percentage of sprouting of 'Niagara Rosada' grapevines
sprayed with different treatments for budbreak in two vineyard
locations: Indaiatuba (CV=26.3%) (A) and Jundiai (CV=9.7%) (B).

garlic extract (mL [L.sup.-1])

(A) y - 56.38 + 0.351x, [r.sup.2] = 0.7020**

0       56b
14      64b
28      69b
42      67b
56      67b
70      89ab
CaCN2   100a
H2CN2   100a

Note: Table made from Bar graph.

Figure 3--Cycle (days) between pruning and harvest of 'Niagara
Rosada' grapevines sprayed with different treatments for budbreak in
two vineyard locations: Indaiatuba (CV=22.2%) (A) and Jundiai
(CV = 1.1%) (B).

(A)  y = 186.29 - 0.9653x, [r.sup.2] = 0.9463**

Cycle (days)

garlic extract (mL [L.sup.-1])

0       58a
14      53a
28      37cd
42      43bc
56      33cde
70      26de
CaCN2   21e
H2CN2   23e

(B)

Cycle (days)

garlic extract (mL [L.sup.-1])

0       150a
14      150a
28      142b
42      137c
56      137c
70      137c
CaCN2   130d
H2CN2   130d

Note: Table made from Bar graph.

Figure 4--Number of clusters per plant of 'Niagara Rosada' grapevines
sprayed with different treatments for budbreak in two vineyard
locations: Indaiatuba (CV=66.6%) (A) and Jundiai (CV=11.8%) (B).

(A)

v = 2.5429 - 0.0007 x + 0.0007 [x.sup.2], [r.sup.2] = 0.7242 *

0       1.8c
14      4bc
28      3.1bc
42      3.2bc
56      4.2bc
70      6.4abc
CaCN2   9.5ad
H2CN2   11a

garlic extract (mL [L.sup.-1])

(B)

0       8.5c
14      8.3c
28      8.5c
42      8.8bc
56      9.5bc
70      10.8ab
CaCN2   12a
H2CN2   12a

garlic extract (mL [L.sup.-1])

Note: Table made from Bar graph.
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Author:Botelho, Renato Vasconcelos; Pires, Erasmo Jose Paioli; Moura, Mara Fernandes; Terra, Maurilo Montei
Publication:Ciencia Rural
Date:Nov 1, 2010
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