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    • 专利摘要:
    Procedure to promote the abscission of the olive and facilitate its collection by applying N-nitro-l-arginine methyl ester (l-name). The present invention relates to a method to promote the abscission of the olive, especially to facilitate its collection by applying l-name in the form of foliar spray before the olive harvest. The treatment of olive trees with l-name reduces the retention force of the olive to the tree and, in this way, facilitates that these can be collected economically with little manual or mechanical assistance within the appropriate time of collection and this, without causing defoliation of the olive trees (Machine-translation by Google Translate, not legally binding)
    公开号:ES2613583A1
    申请号:ES201631636
    申请日:2016-12-21
    公开日:2017-05-24
    发明作者:María Carmen GÓMEZ JIMÉNEZ
    申请人:Universidad de Extremadura；
    IPC主号:
    专利说明:

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    PROCEDURE TO PROMOTE THE ABSCISION OF THE OLIVE AND FACILITATE ITS COLLECTION THROUGH THE APPLICATION OF METAL N-NITRO-L-ARGININE
    ESTER (L-NAME)
    DESCRIPTION
    TECHNICAL SECTOR
    This invention relates to a new procedure, in the area of agriculture, to promote abscission of the fruit by treatment with N-Nitro-L-arginine methyl ester (L-NAME). Specifically, the invention relates to a method to reduce the retention force of the olive to the tree and facilitate the collection of the olive manually or mechanically.
    STATE OF THE TECHNIQUE
    Abscission is a complex process, genetically programmed and regulated during development, through which multicellular organs such as leaves, flowers, or fruits become separated from the body of the plant (AB Bleecker and SE Patterson. The Plant Cell 9 ( 1997) 1169; JE Taylor and CA Whitelaw. New Phytologist 151 (2001) 323; JA Roberts, KA Elliot, and ZH Gonzalez-Carranza. Annual Review of Plant Biology. 53 (2002) 131). Abscission occurs at predetermined sites called the abscission zones (ZA), which consist of some layers of small cells that respond in different ways to neighboring cells such as hormonal or environmental signals (JA Roberts, KA Elliot, and ZH Gonzalez- Carranza Annual Review of Plant Biology 53 (2002) 131; ZH Gonzalez-Carranza, AA Shahid, L. Zhang, Y. Liu, U. Ninsuwan and JA Roberts. Plant Physiology 160 (2012) 1342).
    In the abscission of the fruit, knowledge of the mechanisms involved is essential to develop control strategies and improve harvesting. Advances in genetic and molecular analyzes on abscission of fleshy fruits have been limited by the lack of appropriate experimental genetic material (genotypes or mutants of abscission) and by the small amount of ZA cells available in the model species for these analyzes . Likewise, abscission of the fleshy fruit is a highly variable process according to the species and varieties. In recent years, the combination of classic physiological, genetic and genomic studies have demonstrated the implication of multiple routes
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    and coordinated processes in abscission (Gonzalez-Carranza, AA Shahid, L. Zhang, Y. Liu, U. Ninsuwan and JA Roberts. Plant Physiology 160 (2012) 1342; LH Estornell, J. Agusti, P. Merelo, M. Talon and FR Tadeo, Plant Science199 (2013) 48; C. Niederhuth, SK Cho, K Seitz and JC Walker. Journal of Integrative Plant Biology 55 (2013) 1251). Among these routes and processes involved is the hormonal signaling of ethylene, auxin, abscessic acid, and jasmonic acid (M. Ogawa, P. Kay, S. Wilson, SM Swain. Plant Cell 21 (2009) 216), as well as the pathway of HAE HSL2, the transmembrane traffic and subsequent responses regulated by these signals (C. Niederhuth, SK Cho, K Seitz and JC Walker. Journal of Integrative Plant Biology 55 (2013) 1251). However, to clarify the network (s) that integrates these different routes and processes, genomics studies have been required, such as microarrays and RNAseq studies used in Arabidopsis (S. Cai and CC Lashbrook. Plant Physiology 146 (2008 ) 1305; CC Lashbrook and S. Cai Plant Signal. Behav. 3 (2008) 733; C. Niederhuth, SK Cho, K Seitz and JC Walker. Journal of Integrative Plant Biology 55 (2013) 1251). In other plant species other than Arabidopsis, functional genomic approaches are being applied to study abscission. Microarrays have been used to study changes in the genetic expression of ethylene-induced abscission in citrus fruits (J. Agusti, P. Merelo, M. Cercos, FR Tadeo and M. Talon. BMC Plant Biol. 23 (2009) 127) , apple (A. Botton, G. Eccher, C. Forcato, A. Ferrarini, M. Begheldo, M. Zermiani, S. Moscatello, A. Battistelli, R. Velasco, B. Ruperti, A. Ramina. Plant Physiol. 155 (2011) 185; H. Zhu, CD Dardick, EP Beers, AM Callanhan, R. Xia and R. Yuan. BMC Plant Biol. 11: (2011) 138) and tomato (S. Meir, S. Philosoph-Hadas , S. Philosoph-Hadas, S. Sundaresan, KSV Selvaraj, S. Burd, R. Ophir, B. Kochanek, MS Reid, C. Jiang and A. Lers. Plant Physiol. 154 (2010) 1929; T. Nakano, M. Fujisawa, Y. Shima and Y. Ito. BMC Plant Biol. 9 (2013) 13). These studies have revealed changes in the enzymes that modify the cell wall, as well as hormonal interaction and factors such as nutritional stress in abscission. Approaches based on transcriptome sequencing have also been used to analyze fruit abscission (C. Zhou, AN Lakso, TL Robinson and S. Gan. Mol. Genet. Genom. 280 (2008) 83; J. Corbacho, F. Romojaro, JC Pech, A. Latche and MC Gomez-Jimenez. PLoS One. 8 (2013) e58363).
    In olive trees (Olea europaea L.), a crop of great economic interest in Spain, being in the first place worldwide in surface area and production of olive oil and table olives, the regulation of the abscission of the fruit constitutes an agronomic problem. Currently, there are olive varieties, such as "Manzanilla Sevillana" (variety of table olives) and "Arbequina" (variety of oil), whose fruits have a high retention force at the time of collection and therefore, the effectiveness of mechanical collection is very
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    low, causing an increase in the costs of harvesting the fruit, which represents, today, a funny cost to the farmer. The use of chemicals that facilitate the abscission of olives has been under investigation for over forty years (HT Hartmann, A. Tombesi and J. Whisler. J. Amer. Soc. Hort. Sci. 95 (1970) 635) . The best products to facilitate the abscission of olives have been ethylene releasers (HT Hartmann, A. Tombesi and J. Whisler. J. Amer. Soc. Hort. Sci. 95 (1970) 635; GC Martin. Acta Hortic. 356 (1994) 284; S. Bartolini, C. Cantini, and C. Vitagliano Acta Hortic. 329 (1993) 249; J. Burns, L. Ferguson, K. Glozer, WH Krueger, RC Rosecrance. Hortscience 43 (2008) 1449; MC Parra-Lobato and MC Gomez-Jimenez.J Exp Bot. 62 (2011) 4447), such as: 2- chloroethylphosphonic acid (ethephon, ethrel), 2-chloroethyl-tris- (2-methoxy-ethoxy) - silane (Alsol) and 2-chloroethylsulfonic acid, which release ethylene, thus causing a decrease in the retention strength of the olive, and consequently facilitating its collection. However, these ethylene-releasing products exert negative side effects for the olive grove, such as leaf fall (G. Lang and G. Martin G. Journal of the American Society for the Horticultural Science, 114 (1989) 134), the acceleration of the ripening of the olive (AI Ozguven, F. Ozguven, O. Gezerel, H. Tath and C. Yilmaz. Acta Hortic. 463 (1998) 359), reduction in the flowering index of the following year (HT Hartmann , A. Tombesi and J. Whisler, J. Amer. Soc. Hort. Sci. 95 (1970) 635) and also, these products have their greatest disadvantage in their own toxicity and that of certain products of their degradation, by releasing the ethylene.
    Carboxylic 1-amino-cyclopropane acid (ethylene precursor) is known to be capable of inducing olive abscission (see, for example, ES Pat. No. 496303). It is also known that the addition of certain natural substances to olive trees can favor the abscission of the fruit (see, for example, ES Pat. No. 328941). However, these chemicals have their greatest disadvantage in the activation of the ripening of the fruit, which is not desirable in the collection of table olive varieties, such as the "Manzanilla Sevillana".
    Currently, to facilitate the abscission of the olive and the collection of table olive varieties, a product called CLOUD (ethephon 48% [SL] P / V) has been marketed by the company SAPEC AGRO. However, this treatment continues to have negative effects on the olive grove already mentioned above, such as activation of leaf fall, activation of fruit ripening and environmental toxicity. In recent years, experiments have been carried out by spraying foliar ethephon with monopotassium phosphate verifying that this product causes abscission of the fruit with less fall of olive leaves (D. Barranco, O. Arquero, C. Navarro, and HF Rapoport. HortScience 39 ( 2004) 1313, however, the results are highly dependent on weather conditions and
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    crop status, and therefore, they have not been reproducible (S. Castro-Garcla, GL Blanco Roldan, F. Jimenez-Jimenez, JA Gil-Ribes, L. Ferguson, K. Glozer, WH Krueger, EJ Fichtner, JK Burns, JA Miles and UA Rosa, Acta Hortic. 965 (2012) 29; I. Zipori, A. Dag and Y. Tugendhaft. Hortscience 49 (2014) 55). Therefore, an active agent for the abscission of the fruit has not yet been found to facilitate mechanized harvesting in table olive varieties. In this sense, the sector is currently demanding the use of compounds that promote the abscission of the fruit to improve the mechanized harvesting of varieties of table olives, and therefore, to reduce the harvesting costs of the harvest, without negative effects on the olive grove.
    The procedure object of this patent application comes to solve this problem by using Ww-Nitro-L-arginine methyl ester hydrochloride, also known as L-NG-Nitroarginine methyl ester or L-NAME. L-NAME has the molecular formula C7H15N5O4 • HCl and the molecular weight 269.69 g / mol (CAS #: 51298-62-5). L-NAME, a competitive inhibitor of the enzyme nitric oxide synthase (NOS), is an arginine analog that inhibits the production of nitric oxide. The chemical structure of L-NAME is shown in Figure 1.
    Nitric oxide, a product of nitrogen metabolism, has emerged as an important intercellular and intracellular messenger controlling numerous physiological processes (A. Besson-Bard, A. Pugin and D. Wendehenne. Annual Review of Plant Biology 59 (2008) 21) . In plants, nitric oxide synthesis is mediated mainly by (i) nitrite reduction by nitrate reductase (NR; WM Kaiser, H. Weiner, A. Kandlbinder, CB Tsai, P. Rockel, M. Sonoda and E Planchet, Journal of Experimental Botany 53 (2002) 875), mitochondrial electron transfer (E. Planchet, K. Jagadis Gupta, M. Sonoda and WM Kaiser WM. The Plant Journal 41 (2005) 732), or a form non-enzymatic (PC Bethke, MR Badger and RL Jones. The Plant Cell 16 (2004) 332) and (ii) the oxidation of arginine by the enzyme NOS (FQ Guo, M. Okamoto and NM Crawford NM Science 302 (2003) 100 ). However, there is still some uncertainty about the nature and timing of nitric oxide synthesis by these various mechanisms. Nitric oxide has been shown to perform multiple biological functions in plants, particularly in development processes such as germination, root organogenesis, stomatal closure, flowering and senescence (L. Lamattina, C. Garcla-Mata , M. Graziano and G. Pagnussat Annual Review of Plant Biology 54 (2003) 109); ace! as in stress responses (M. Arasimowicz-Jelonek, J. Floryszak-Wieczorek and J. Kubis. Plant Science 177 (2009) 682; W. Ma, W. Xu, Y. Chen, Z. He and M. Ma. Floor 232 (2010) 325). However, little is known about nitric oxide
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    endogenous during the abscission process. We have demonstrated (MC Parra-Lobato and MC Gomez-Jimenez. Journal of Experimental Botany. 62 (2011) 4447) a correlation between the abscission of the fruit and a significant decrease in the emission of nitric oxide in olive trees, however, they are unknown Enzymes that participate in nitric oxide metabolism during abscission.
    DESCRIPTION OF THE INVENTION
    The present invention relates to a procedure to promote the abscission of the olive, in particular, to facilitate its collection by the application of L-NAME in the form of foliar pulverization before the olive harvest. The treatment of the olive trees with L-NAME reduces the retention force of the olive to the tree and thus, facilitates that these can be economically collected with little manual or mechanical assistance within the appropriate time of collection and without causing olive tree defoliation .
    The collection is the most expensive operation in the olive grove, exceeding 50% of the total cultivation expenses. In the last decades the collection has been mechanized, although for varieties of table olives and oil varieties of very early collection, the effectiveness of demolition is very low due to the high retention force of the fruit and therefore, the collection is forced manual raising collection costs. In this sense, the production of table olives worldwide is becoming unsustainable due to the stagnation in economic profitability and the increase in manual collection costs. With the need to mechanize the collection of table olives due to the existing problems of manual collection costs, the sector demands the use of chemical compounds that cause the abscission of the fruit, not affecting the olive tree as much as possible.
    The main technical problem to which this invention is directed, takes into account that at the moment there is no definitive solution to facilitate the collection of the olive in table olive varieties and in oil varieties that are collected very early, whether Harvesting is manual as if it is carried out mechanically, and consequently, to reduce the high costs of olive harvesting.
    The present invention is directed to solve the need of the olive sector of an agent that favors the abscission of the olive, especially, in varieties of table olives
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    ("Manzanilla Sevillana") and in varieties of oil that are collected very early ("Arbequina) without causing damage to the olive tree, such as defoliation and acceleration of the ripening of the fruit, to thereby reduce the costs of olive harvest . It has been found, surprisingly, that L-NAME at a given concentration has a favorable action on the abscission of the olive applied days before harvesting without causing damage to the olive tree. The invention provides a solution to the existing need through the use of L-NAME. Therefore, the present invention aims to present a new procedure to favor the abscission of the olive, as well as a new product that activates the abscission of the fruit, especially for varieties of table olives and varieties of very early harvest oil. In addition, a product has been found for the abscission of the olive that does not have the disadvantages specified above.
    The application of this product allows the collection of the fruit is carried out at the appropriate time, so that the fruit is in optimal conditions for consumption as a table olive, being able to make the collection stepwise, as the mills admit it. This product is totally harmless and its application is very easy, it can be done in the form of rain with any of the devices used in the pulverization of insecticidal products.
    The main advantages of the present invention are:
    - Significantly reduced collection costs, especially in table olive varieties and oil varieties that are harvested very early.
    - Reduction of damage to the olive tree (defoliation).
    - Allows harvesting at the optimal time.
    - Vibration time reduction.
    - Increase in the percentages of demolition of olives.
    - Increase of the profitability of the exploitation.
    BRIEF DESCRIPTION OF THE CONTENT OF THE FIGURES
    Figure 1 represents the chemical structure of L-NAME
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    Figure 2 shows the branches of the "Manzanilla Sevillana" variety where:
    a) Water treated branches (control) for 3 days.
    b) Water treated branches (control) for 6 days.
    c) Branches treated with 2500 ppm of L-NAME for 3 days.
    d) Branches treated with 2500 ppm of L-NAME for 6 days.
    Figure 3 shows the branches of the variety "Arbequina" where:
    a) Water treated branches (control) for 3 days.
    b) Water treated branches (control) for 6 days.
    c) Branches treated with 2500 ppm of L-NAME for 3 days.
    d) Branches treated with 2500 ppm of L-NAME for 6 days.
    The method according to the invention is characterized in that olives bearing olives are treated before harvesting with L-NAME. The use of L-NAME according to the invention allows such fruits to be detached with little manual or mechanical assistance due to the decrease in the retention force of the fruit and therefore, can be collected economically without causing broth of the leaf in the olive tree.
    L-NAME can be applied by atomizing trees with an aqueous solution containing L-NAME according to the invention at the concentration 2500 ppm (parts per million). Aqueous solutions of L-NAME with 2500 ppm concentration are prepared. The treatment of olive trees with L-NAME solutions is carried out 3 to 6 days before harvesting and causes a decrease in the retention force of the fruit, facilitating the release of the olive. It is advantageous to spray or spray the leaves and fruits of the olive trees at a rate of about 5-10 L / tree. After 3-6 days after the treatment, the branches of the tree are shaken, manually or mechanically, to cause the broth of the fruits.
    It follows that the invention brings together all the necessary characteristics and properties, so that the fruit falls naturally in the time set in advance. Describe sufficiently the nature and scope of the present invention, as! as the way to be able to put it into practice, it is stated that the terms in which it is written is
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    memory are true and faithful reflection of the object described, should be taken with a broad character and never in a limiting way.
    EXAMPLE OF EMBODIMENT OF THE INVENTION
    Example 1.
    “PROCEDURE TO PROMOTE THE ABSCISION OF THE OLIVE AND FACILITATE ITS COLLECTION THROUGH THE APPLICATION OF N-NITRO-L-ARGININA METIL ESTER (L-NAME)“ in olive trees of the variety “Manzanilla Sevillana”.
    To determine the action of L-NAME on the abscission of the olives, the following test was carried out in the field:
    The field test was carried out on 20-25 year old trees of the "Manzanilla Sevillana" olive variety grown under drip irrigation and irrigation with suitable fertilizers in solution in an Olivenza (Badajoz) farm. The treatment was carried out on a total of 20 olive trees of the "Manzanilla Sevillana" variety, selected by size uniformity and fruit load.
    For the preparation of the treatment of the aqueous solution of 2500 ppm of L-NAME, 5 g of L-NAME (Sigma-Aldrich) was dissolved in water to a volume of 2000 mL.
    The olives are sprayed in the form of an aqueous solution (2000 mL per branch) once at 6 days before the olive harvest. It is advantageous to spray or spray the aqueous solution of the leaves and fruits of the trees. In each olive tree, two uniform branches were selected: a control branch (application of 2000 mL of water) and a treated branch (application of 2000 mL of 2500 ppm of L-NAME) in a total of 20 olive trees of the variety “Manzanilla Sevillana "
    In order to get a measurable idea of the process of abscission of the fruit at 3 and 6 days after treatment, the retention force of the fruit (FRF), in kg force (kgf), was measured by a dynamometer (Correx, Switzerland). For this, the FRF measurements were made at 50 fruits for each branch of the 20 trees selected at 3 days and 6 days after treatment. Therefore, the control fruits were treated with water at 6 days, and collected from the branches of the same trees treated with L-NAME at 3 and 6 days after treatment. Figure 2 shows the branches of the “Manzanilla Sevillana” variety, where a) are the branches treated with water (control) for 3 days; b) branches treated with water (control) for 6 days of treatment; c) the branches treated with 2500 ppm of L-NAME for 3 days and d) the branches treated with 2500 ppm of L-NAME for 6 days of treatment.
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    The following table indicates the average value of the FRF measured in the fruits of the control branches (water treatment) and the treated branches (treatment with 2500 ppm L-NAME) of the variety "Manzanilla Sevillana" at 3 and 6 days of the treatment (100 fruits / tree) The application of L-NAME reduced the FRF of the olives of the "Manzanilla Sevillana" variety and did not cause defoliation in the olive tree.
    Table 1. Force of retention of the fruit and% defoliation in olive trees of the variety "Manzanilla Sevillana" at 3 and 6 days after the treatment with 2500 ppm of L-NAME and with water (control). The results are mean ± SD ( n = 1000).
     Treatment time (dlas) Fruit retention force (kgf) Defoliation (%)
     Control  3 0.70 ± 0.04 0
     6  0.68 ± 0.06 0
     2500 ppm L- NAME  3 0.32 ± 0.08 0
     6  0.30 ± 0.08 0
    After 3-6 days after the treatment, the tree branches are shaken, manually or mechanically, to cause the fruit broth and the olive is harvested.
    Example 2
    "PROCEDURE TO PROMOTE THE ABSCISION OF THE OLIVE AND FACILITATE ITS COLLECTION THROUGH THE APPLICATION OF N-NITRO-L-ARGININE METAL ESTER (L-NAME)" in olive trees of the variety "Arbequina".
    The field test was carried out on 20-25 year old trees of the "Arbequina" olive variety grown under drip irrigation and irrigation with suitable fertilizers in solution in an Olivenza (Badajoz) farm. The treatment was carried out on a total of 20 olive trees of the "Arbequina" variety, selected by size uniformity and fruit load.
    For the preparation of the treatment of the aqueous solution of 2500 ppm of L-NAME, 5 g of L-NAME (Sigma-Aldrich) was dissolved in water to a volume of 2000 mL.
    The olives are sprayed in the form of an aqueous solution (2000 mL per branch) once at 6 days before the olive harvest. It is advantageous to spray or spray the aqueous solution of the leaves and fruits of the trees. In each olive tree, two uniform branches were selected: a control branch (application of 2000 mL of water) and a treated branch
    (application of 2000 mL of 2500 ppm of L-NAME) in a total of 20 olive trees of the “Arbequina” variety.
    In order to get a measurable idea of the process of abscission of the fruit at 3 and 6 days after treatment, the retention force of the fruit (FRF), in kg force (kgf), was measured by means of a dynamometer (Correx, Switzerland) . For this, the FRF measurements were made at 50 fruits for each branch of the 20 trees selected at 3 days and 6 days after treatment. Therefore, the control fruits were treated with water at 6 days, and collected from the branches of the same trees treated with L-NAME at 3 and 6 days after treatment. Figure 3 shows the branches of the “Arbequina” variety, where a) are the branches treated with water 10 (control) for 3 days; b) branches treated with water (control) for 6 days of treatment; c) the branches treated with 2500 ppm of L-NAME for 3 days and d) the branches treated with 2500 ppm of L-NAME for 6 days of treatment.
    The following table shows the average value of the FRF measured in the fruits of the 15 control branches (water treatment) and the treated branches (treatment with 2500 ppm L-NAME) of the “Arbequina” variety at 3 and 6 days of treatment (100 fruits / tree). The application of L-NAME reduced the FRF of the olives of the “Arbequina” variety and did not cause defoliation in the olive tree.
    20 Table 2. Force of retention of the fruit and% defoliation in olive trees of the variety "Arbequina" at 3 and 6 days after the treatment with 2500 ppm of L-NAME and with water (control). The results are mean ± SD (n = 1000).
     Treatment time (dlas) Fruit retention force (kgf) Defoliation (%)
     Control  3 0.64 ± 0.10 0
     6  0.62 ± 0.08 0
     2500 ppm L- NAME  3 0.40 ± 0.06 0
     6  0.38 ± 0.04 0
    25 After 3-6 days after the treatment, the tree branches are shaken, manually or mechanically, to cause fruit broth and the olive is harvested.
    权利要求:
    Claims (3)
    [1]

    5 1. PROCEDURE TO PROMOTE THE ABSCISION OF THE OLIVE AND FACILITATE
    ITS COLLECTION THROUGH THE APPLICATION OF N-NITRO-L-ARGININE METHYL ESTER (L-NAME), characterized in that the olive trees are treated, 3 to 6 days before the olives are harvested, by spraying or spraying foliar with an aqueous solution of L-NAME, which causes a decrease in the retention force of the olive to the tree, 10 subsequently proceeding to manual or mechanical agitation of the tree.
    [2]
    2. A method, according to claim 1, characterized in that the concentration of L-NAME in the solution is 2500 parts per million.
    [3]
    3. A procedure, according to claim 1, wherein the olive trees are the olive trees of the "Manzanilla Sevillana" variety.
    15 4. A procedure, according to claim 1, wherein the olive trees are the olive trees of the variety
    "Arbequina."
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