WO2018122860A1 - Système et procédé de détection précoce pour la prévention d'un accident de réfrigération post-récolte - Google Patents

Système et procédé de détection précoce pour la prévention d'un accident de réfrigération post-récolte Download PDF

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WO2018122860A1
WO2018122860A1 PCT/IL2018/050007 IL2018050007W WO2018122860A1 WO 2018122860 A1 WO2018122860 A1 WO 2018122860A1 IL 2018050007 W IL2018050007 W IL 2018050007W WO 2018122860 A1 WO2018122860 A1 WO 2018122860A1
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temperature
volatile compound
mango
fruit
storage
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PCT/IL2018/050007
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Alkin NOAM
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Ministry Of Agriculture
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q3/00—Condition responsive control processes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/497—Physical analysis of biological material of gaseous biological material, e.g. breath
    • G01N33/4977—Metabolic gas from microbes, cell cultures or plant tissues

Definitions

  • the present invention relates to the field of postharvest treatment. More particularly, the invention relates to a system and methods for early detection for prevention of postharvest deterioration of quality of agricultural product in storage.
  • Postharvest cold storage is widely used to prolong the storage time of fresh produce.
  • Cold storage is considered the most effective method of prolonging storage of fresh produce, because it maintains fruit quality and defers fruit deterioration. Cold storage slows down cellular respiration rate and ripening-related metabolic processes in fruit. Thus, as the fruit is stored at a lower temperature the fruit storage period is extended and fruit deterioration is inhibited. Optimal cold storage conditions can reduce weight loss and fruit rot, however storage at suboptimal temperatures disrupts a number of metabolic processes.
  • mango is consumed worldwide for its delicious taste and aroma and nutritional qualities.
  • Mango fruit is stored at 10-12 °C; however storage at lower temperature causes chilling injury (CI), reduced fruit quality and even fruit loss.
  • CI symptoms include lenticel discoloration and pitting on the peel, internal breakdown, uneven ripening, poor color development, and reduced aroma and flavor.
  • the fruit with CIs ripens with reduced development of normal flavor and aroma.
  • the following table illustrates some known storage temperatures acceptable for number of agricultural product s, although there are variations among various cultivars and different ripening stages.
  • Metabolisms in agricultural product may be affected by post-harvest conditions, such as temperature, irradiation, the surrounding gaseous composition, and the chemicals being used. Consideration has been given to a possible use of various additives in attempt to enhance resistance to decay. For example, some fruits showed longer shelf-life times with acceptable quality in response to addition of hexanal, probably as consequence of activation of natural defense response in the harvested fruit. Hexanal may act as signaling intra-plant and/or inter- plant. Thus, for example, when leaves are being damaged by herbivores or pathogens, they begin to produce hexanals, some of which may induce defense responses. However treating agricultural products by additives is not related, thus it has no effect on responding to realtime metabolic processes occurring in agricultural product during cold storage. BRIEF DESCRIPTION OF DRAWINGS
  • Figure 1 schematically illustrates upregulation of ⁇ -linolenic acid-oxidation pathway genes in response to suboptimal cold storage
  • FIG. 1 Heat map of relative expression of genes in the ⁇ -linolenic acid-oxidation pathway at two different storage temperatures (5 °C and 12 °C) at different sampling times (2, 7 and 14 days) compared to harvest time.
  • Figure 3 shows table of transcripts relative expression values of ⁇ -linolenic acid-oxidation pathway and their abbreviations
  • Figure 4 (A-E). shows a table of volatile compounds concentrations in ⁇ g/g FW, found in mango fruit peels at harvest, and during cold storage at 12°C and suboptimal (5°C) temperatures
  • FIG. schematically illustrates changes in C 6 and C9 volatile concentrations during cold storage at 12°C and suboptimal (5°C) temperatures
  • the present invention is intended to overcome the deficiencies of the background art by a real-time monitoring of agricultural product quality during storage relying on early detection of metabolisms occurring in the stored products in response to suboptimal temperatures which may lead to deterioration of quality of crops, e.g., mango, pomegranate, avocado, citrus, pepper, lattice, apple, pear, cherry, peach, grape, banana, and tomato.
  • an early detection of onset of metabolism, associated with crop response to chilling and quality deterioration is based on changes in production of volatile compounds by a product being exposed to stress conditions during storage. The presence of volatile compounds related with early phases of such metabolisms, could be detectable before the symptoms of such deterioration become notable.
  • Deterioration in agricultural product quality may include appearance of Chilling Injury (CI).
  • the evaluation and determination of agricultural product quality may include the extent of the appearance of CI symptoms which are frequently caused by sub-optimal storage temperature.
  • a system for prevention of post-harvest CI may include at least one sensor capable of detecting a presence or measuring concentrations of at least one volatile compound which is indicative of CI and/or product quality deterioration.
  • volatile compounds may include at least one alkene, or at least one oxylipin e.g, 1- hexanal, (Zj-3-hexenal, (Zj-3-hexenol, (EJ-2-hexenal, nonanal, heptanal, octanal..
  • one computerized module capable of controlling an operation of a conditioning system.
  • the terms “computerized module”, “computerized control module”, “control module”, “module” and “controller” may be used interchangeably hereinafter.
  • An operation of a conditioning system in a depot typically includes an adjustment of storage temperature.
  • storage temperature temperature in a depot
  • the control module processes input concerning presence and/or concentrations of at least one volatile compound which indicate agricultural product quality which may be associated with CI.
  • the adjustment of the temperature in a depot may include elevation of this temperature whenever the above mentioned volatile compound is detected.
  • the adjustment of the temperature may be related to changes in concentrations of the above mentioned one or more volatile compounds.
  • the conditioning system when the concentration of such volatile compound increases, the conditioning system would receive a command from the control module to elevate the storage temperature. Lowering storage temperature can prolong storage time to as long as no CI occurs.
  • the conditioning system may reduce depot temperature when there is no detection of the above mentioned volatile compound.
  • altering of temperature may include reducing depot temperature whenever the presence of at least one volatile compound drops below a threshold level.
  • reduction of storage temperature may be implemented when measurements indicate that the onset of rise of concentration of such a volatile compound decreases.
  • Some aspects of the present invention include methods for prevention of post-harvest chilling injury (CI).
  • Such methods include detecting a presence or measuring concentrations of at least one volatile compound which is indicative for CI and/or agricultural product quality such as mango, pomegranate, avocado, citrus, pepper, apple, pear, cherry, peach, grape, banana, and tomato.
  • the above methods may be particularly useful during cold storage in order to keep the storage temperature as low as possible while avoiding CI and/or quality deterioration.
  • such methods may include also processing inputs concerning detection or measuring concentrations of the aforementioned volatile compound. Such processing of input may be utilized for altering conditions in a depot where agricultural product is being stored in order to respond and adjust conditions to avoid deterioration in agricultural product quality.
  • Altering conditions in a depot may include according to some embodiments of the above aspects, elevation of storage temperature whenever detection of presence of the above mentioned volatile compound is occurred. Similarly the adjustment of the temperature may be according to changes in concentrations of the above mentioned volatile compound. For example, when a concentration of such volatile compound increases, the conditioning system would receive a command from the control module to elevate the storage temperature. According to some embodiments of the present invention the conditioning system may reduce depot temperature when there is no detection of the above mentioned volatile compound. Similarly reduction in storage temperature may be done when measurements are indicating that the concentration of such a volatile compound decreases. According to some embodiments of the above aspects a target value for storage temperature may be set according to certain requirements such as the type of crop in storage. In some examples embodying the above aspects, alteration of the storage temperature ranges between one Celsius degree below the target value to three degrees Celsius above the target value, (e.g., 10°C, 7°C, 5°C, 3°C, and 2°C).
  • the present invention is intended to overcome the deficiencies of the background art by realtime monitoring of crop CI and/or quality during storage relying on early detection of metabolites that may lead to or indicate a CI and/or deterioration of quality of stored vegetable products, e.g., mango, pomegranate, avocado, citrus, pepper, lattice, apple, pear, cherry, peach, grape, banana, and tomato.
  • mango pomegranate, avocado, citrus, pepper, lattice, apple, pear, cherry, peach, grape, banana, and tomato.
  • aspects of the present invention include rely on response of vegetable or fruit to stress conditions such as suboptimal temperature. Such response may lead to activation of some metabolic pathways that can induce synthesis of volatile compounds which may serve according to embodiments of the present invention as signals which indicate beginning deterioration of fruit quality. The releasing of such volatiles compounds may be detected long before CI symptoms become visible.
  • a system with at least one sensor capable of detecting a presence or measuring concentrations of at least one volatile compound which indicates chilling stress in fruit or vegetables. Output generated by such a sensor may be received by at least one control module which performs analysis and generates output signals to effect the operation of depot conditioning system to alter storage temperature.
  • average storage temperatures may be reduced in comparison to existing practice (by minimizing of unnecessary safety margins), relying on implementing a real-time monitoring of agricultural product quality based on detection of changes in volatile compounds produced by the agricultural product, and on a real-time response of a computerized control module governing an operation of storage conditioning system.
  • systems as described above may obviate a need to use additives to prolong shelf life.
  • the adjustment of the temperature may include elevation of this temperature whenever detection of presence of the above mentioned volatile compound is occurred.
  • the adjustment of the temperature may be according to changes in concentrations of the above mentioned volatile compound. For example, when a concentration of such volatile compound increases, the conditioning system would receive a command from the control module to elevate the storage temperature.
  • the conditioning system may reduce depot temperature when there is no detection of the above mentioned volatile compound.
  • reduction in storage temperature may be done when measurements are indicating that concentration of such a volatile compound decreases. Lowering storage temperature can prolong shelf life as long as no CI occurs.
  • Some aspects of the present invention include methods for prevention of post-harvest chilling injury (CI).
  • methods may include also processing inputs concerning detection or measuring concentrations of the aforementioned volatile compound.
  • processing of input may be utilized for altering conditions in a depot where agricultural product is being stored in order to respond and adjust conditions to avoid deterioration in agricultural product quality.
  • Altering conditions in a depot may include according to some embodiments of the above aspects, elevation of storage temperature whenever detection of presence of the above mentioned volatile compound is occurred.
  • the adjustment of the temperature may be according to changes in concentrations of the above mentioned volatile compound. For example, when a concentration of such volatile compound increases, the conditioning system would receive a command from the control module to elevate the storage temperature.
  • the conditioning system may reduce depot temperature when there is no detection of the above mentioned volatile compound.
  • reduction in storage temperature may be done when measurements are indicating that the concentration of such a volatile compound decreases.
  • a target value for storage temperature may be set according to certain requirements such as the type of agricultural product in storage.
  • alteration of the storage temperature ranges between one Celsius degree below the target value to three degrees Celsius above the target value, (e.g., 10°C, 7°C, 5°C, 3°C, and 2°C).
  • Mango flavor and aroma are determined by the composition of the fruit volatiles and differ among cultivars.
  • Subtropical fruit as mango is considered to be susceptive to non- optimal conditions during cold storage.
  • Mango fruit are typically stored at 10-12 °C. It was found that concentrations of volatile constituents in mango fruit peel and pulp in various cultivars may change in response to various postharvest conditions, such as temperature, vapor treatment, methyl jasmonate (MeJA) presence, fruit fly, fruit ripening and diseases. A reduction in production of aroma volatile compounds for example in 'Kensington Pride' mango pulp was observed in response to CI.
  • MeJA methyl jasmonate
  • Lipid peroxidation is a key metabolic process that is activated in response to chilling. Lipoxygenase is considered to play an important role in the peroxidation of unsaturated lipids, a possible cause of changes in membrane lipid composition, resulting in decreased membrane fluidity, membrane dysfunction and altered cellular homeostasis. Transcriptomic analysis of 'Keitt' mango fruit in response to suboptimal temperature storage revealed the activation of several pathways, including oxidation of a-linolenic acid and glycerophospholipid metabolism, resulting in lipid peroxidation. Lipoxygenase catalyzes the first step of a- linolenic acid oxidation, leading to synthesis of C 6 and C9 aldehydes.
  • Oxylipins are a diverse class of lipid metabolites derived from the oxidation of unsaturated fatty acids that act as signaling molecules. Hydroperoxide lyase (HPL) catalyzes the cleavage of hydroperoxides to generate C 6 volatiles, also referred to as green leaf volatiles that may have a role in during plant defense signaling.
  • HPL Hydroperoxide lyase
  • Mango fruit (Mangifera indica L., cv. Keitt) were obtained 1-2 h postharvest from a commercial orchard (Mor Hasharon storage house, Israel), and transported (1 h) to the Agricultural Research Organization (Israel). Uniform, unblemished fruit weighing 424 ⁇ 16 g were selected, washed with tap water and air-dried. On the same day (day of harvest), six biological replicates, each with 10 fruit, were stored at 5 °C, 8 °C, 12 °C for 19 days in the cold-storage rooms, and a further 7 days at 20 °C (shelf-life storage).
  • the temperature in the cold-storage room was monitored by a DAQ tool (double-strand wire logger/data-acquisition control system; T.M.I. Barak Ltd., Ramat-Gan, Israel).
  • Fruit core temperature was monitored using a MicroLite data logger LITE5032P-EXT-A (Fourier Technologies, Ramat-Gan, Israel), by inserting the probe to 5 cm depth near the fruit calyx.
  • the experiments were repeated in three consecutive seasons— 2013, 2014 and 2015— and gave similar results. The presented experiment is of cv. Keitt in 2014.
  • RNA-Sequencing (RNA-Seq)
  • RNA extraction from mango fruit peel tissue, cDNA library preparation, and the RNA- Seq protocol using the Illumina Hiseq2000 system were as described previously.
  • the raw reads of 14 libraries were subjected to quality trimming and filtering using Trimmomatic software, sequences were mapped to a reference mango transcriptome using the Bowtie2 software alignment protocol.
  • Abundance estimates were calculated for each mango transcript by the RSEM software package.
  • the Bioconductor EdgeR package of the Bioconductor R packages was used to identify differentially expressed transcripts for each pair of samples, based on the count estimations for each transcript. Transcripts that were more than fourfold differentially expressed with a false discovery rate (FDR)-corrected statistical significance smaller than le-5 were considered differentially expressed.
  • FDR false discovery rate
  • Samples were prepared by randomly slicing Mango fruit peel tissues (2 mm deep) from six fruits. Each sample was kept for 19 days in cold storage at 5 °C or 12 °C. Specimens were taken of each sample after the second day, and after days 7, 14 and 19 of the cold storage. After 19 days of cold storage the samples were stored at 20 °C. Specimens were taken after the first day and the seventh day of storage at 20 °C. This experiment was repeated for three times..
  • Peel tissue (2 g) of each sample and its replicates were immediately collected in a 20- mL amber vial (LaPhaPack, Langerwehe, Germany) prepared in advance with 5 mL of 20% (w/v) NaCl (Sigma-Aldrich, St Louis, MO), 0.6 g NaCl, and 50 of 10 ppm 5-2-octanol (Sigma-Aldrich) added as an internal standard. Samples were stored at -20 °C until analysis. On the day of analysis, samples were prewarmed for 1 h at 30 °C on an orbital shaker at 250 rpm.
  • a solid-phase microextraction (SPME) holder (Agilent, Palo Alto, CA) assembled with fused silica fiber (Supelco, Bellefonte, PA) coated with polydimethylsiloxane (50/30 ⁇ thickness) were used to absorb the volatile compounds. Absorption and desorption of the aromatic compounds were performed on a Agilent gas chromatograph series 7890A fitted with an Agilent HP-5MS fused silica capillary column (30 mm long x 0.25 mm ID x 0.25 ⁇ film thickness), coupled to a 5975C MS detector (Agilent).
  • SPME solid-phase microextraction
  • RNA deep sequencing was conducted using Illumina HiSeq 2000 on samples extracted from the peel part of 'Keitt' mango fruit stored at 12 °C or 5 °C for 2, 7 and 14 days, as described previously.
  • the upregulated clusters were BLASTed against the KEGG database (http://www.genome.jp/kegg/), identified and collated to the upregulated pathways.
  • the ⁇ -linolenic acid-oxidation pathway was activated in mango in response to chilling stress ( Figure 1).
  • Key genes of the a-linolenic acid metabolic pathway such as those encoding 13S- lipoxygenase (LOX), allene oxide synthase (AOS), allene oxide cyclase (AOC) and 12- oxophytodienoate reductase (OPR), were significantly upregulated at 5 °C compared to fruit on day of harvest and those stored at 12 °C.
  • the mango transcriptome revealed 10 LOX genes, of which 4 13S-LOX genes were upregulated.
  • LOXii and LOXiii were upregulated 2.3- and 2.5-fold, respectively, after 7 days of storage at 5 °C vs. 12 °C.
  • AOS isoform-encoding genes (compl 1945, comp26483, comp29600) were upregulated 4.2-, 3.1- and 4.6-fold, respectively, in response to 2 days of cold storage at 5 °C vs. 12 °C storage.
  • AOC (compl3545) was upregulated at 5 °C vs. 12 °C at all-time points, with a maximum 9.2-fold increase after 7 days in cold storage.
  • OPR (compl 8454) was upregulated at 5 °C vs. 12 °C at all-time points, with a maximum 2.4-fold increase after 7 days of cold storage.
  • a correlation analysis of all 43 compounds showed strong coactivation among various monoterpenes and sesquiterpenes, among alcohols (i-butanol, 3-methyl-i-butanol and 2- methyl-i-butanol), and among the aldehydes and their alcohol derivatives (i-hexanal, (Z)-3- hexenal, (Zj-3-hexenol, (E)-2 -hexenal, nonanal and octanal).
  • the volatiles (5-3-carene, (Z)- ?-ocimene and terpinolene were found at higher concentrations in fruit at harvest and stored at 12 °C than in fruit stored at 5 °C.
  • the concentration of C 6 and C9 aldehydes (i- hexanal, fZJ-3-hexenal, (Zj-3-hexenol, (EJ-2-hexenal and nonanal) during 19 days of cold storage (5 °C or 12 °C) and an additional 7 days of shelf life (20 °C) showed that all of these compounds increased significantly after 2 days of cold storage at 5 °C and remained elevated until fruit transfer to 20 °C.
  • the transfer of mango fruit to the higher temperature of 20 °C was accompanied by a sharp decrease in the concentration of those C 6 and C9 aldehydes in fruit stored previously at 5 °C and a slight increase for fruit stored previously at 12 °C.
  • Lipid peroxidation is a characteristic metabolism in fruits suffering from CIs. An elevation in lipid peroxidation was observed by luminescence and MDA biochemical analysis in 'Keitt' mango fruit after 14 days of storage at 5 °C, just before the visual CI symptoms appeared. However, the transcripts in the metabolic pathway of fatty-acid oxidation of a-linolenic acid metabolism were activated after 2 days of storage at 5 °C. In this regard, the oxidation of a- linolenic acid is connected not only to fatty acid oxidation, but also to much more basic defense responses to abiotic stress— JA synthesis and oxylipin signaling— which further activated the fruit defense response.
  • JA is a major compound regulating the global plant response to abiotic stress. JA is also known to activate chilling resistance in various fruits.
  • LOX is a key gene in the response to chilling as it initiates the first step of a-linolenic acid oxidation and the synthesis of MeJA. It was found that four LOX transcripts were upregulated in response to chilling ( Figure 2). Downstream of LOX in the JA biosynthesis pathway is AOS, which is an important enzyme in the defense response to wounding that acts as a key enzyme in oxylipin metabolism. It was found that mango AOS transcripts were activated in response to chilling (compl l945, comp26483, comp29600).
  • oxidation and degradation of ⁇ -linolenic acid also lead to the release of oxylipins, such as the volatile alkenes C 6 and C 9 .
  • This reaction is mediated by HPL, which was consistently expressed during storage at 12 °C and 5 °C.
  • the resultant C 6 and C 9 oxylipins which are known as green leaf volatiles or oxylipins, are released mainly in response to wounding and biotic stress.
  • Transcriptomic evaluation of mango fruit peel for example suggested that storage at suboptimal temperature upregulates genes of the ⁇ -linolenic acid-oxidation pathway which leads to synthesis of C 6 and C 9 aldehydes before CI symptoms become visible as demonstrated in gas chromatography-mass spectrometry (GC-MS) analysis of the volatile profile of mango fruit peel.
  • GC-MS gas chromatography-mass spectrometry
  • a presence of oxylipin volatiles of C6 and C9 aldehydes may serve according to aspects of the present invention as signals which indicate a beginning of metabolisms which are associated with CI and/or deterioration of fruit quality. Such an early indication may be utilized for monitoring CI and/or fruit quality and for real-time adjustment of storage conditions.

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Abstract

Des aspects de la présente invention comprennent la détection de composés volatils liés à la réponse d'un légume ou d'un fruit (par exemple, une mangue) suite à une contrainte comme une température sous-optimale. Une telle réponse peut conduire à l'activation de certaines voies métaboliques qui peuvent induire la synthèse de composés volatils qui, selon des modes de réalisation de la présente invention, peuvent servir de signaux qui indiquent l'apparition d'un accident de réfrigération et/ou la détérioration de la qualité des fruits. La libération de ces composés volatils peut être détectée longtemps avant que les symptômes d'accident de réfrigération puissent être constatés. Selon certains modes de réalisation de l'invention, la sortie de capteurs détectant la présence de composés volatils indiquant que les fruits commencent à souffrir d'une contrainte de réfrigération est reçue par au moins un module de commande qui fait fonctionner le système de conditionnement de façon à modifier des conditions de stockage pour éviter un accident de réfrigération et/ou une détérioration de la qualité de fruits ou de légumes.
PCT/IL2018/050007 2017-01-02 2018-01-02 Système et procédé de détection précoce pour la prévention d'un accident de réfrigération post-récolte WO2018122860A1 (fr)

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Publication number Priority date Publication date Assignee Title
WO2021153513A1 (fr) * 2020-01-27 2021-08-05 日本電気株式会社 Procédé de détection d'hormone végétale et procédé de détection précoce d'une infection par une maladie dans une plante utilisant ce dernier
JP7359227B2 (ja) 2020-01-27 2023-10-11 日本電気株式会社 植物ホルモンのセンシング方法、及びそれを用いた植物の病気感染の早期検出方法

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