JP5959103B2 - Spontaneous dormant arousal judgment method for low temperature demanding deciduous trees - Google Patents

Description

  The present invention relates to a method for determining a dormant awakening period of a low-temperature-requiring deciduous tree, and more particularly to a method for determining whether a spontaneous dormant awakening period has been reached using the content of commutating sugars in duct fluid as an index.

At present, in the cultivation site of deciduous fruit trees, it is desired to speed up shipment by covering trees as early as possible from the viewpoint of unit sales price of fruits.
Specifically, in order to accelerate fruit ripening and early shipment, simple covered cultivation is carried out to cover and heat a tree that has entered the spontaneous dormancy awakening period after the spontaneous dormancy period.

In middle to high latitude areas, a deciduous tree in a spontaneous dormancy period needs to encounter a certain amount of low temperature in order to wake up from the spontaneous dormancy (see, for example, Non-Patent Document 1, Patent Document 1, 2, and the like).
However, due to the recent warming of the climate and accompanying weather irregularities, the temperature in winter often changes at a relatively high temperature, making it difficult to determine the dormancy awakening time of trees.
For this reason, if the covering time is too early, the buds are not sufficiently awakened, and the subsequent growth becomes asymmetric, and in severe cases, there is a problem that flowering and fruiting cannot be secured. On the other hand, when the covering time is delayed, there are increasing cases in which a sufficient growth promoting effect cannot be obtained.
Ensuring the yield and quality of fruit trees has become a serious problem for farmers, and therefore development of a method for reliably determining the spontaneous dormancy arousal period has been desired (see Non-Patent Document 1, for example).

Conventionally, the most common method for determining the spontaneous dormancy awakening period is to collect branches, insert them into water, heat them, and observe the presence or absence of germination.
However, the biggest problem with this determination method is that it takes at least one week (several weeks if it is long) to actually confirm the sprouting. Therefore, in many cases, the spontaneous dormancy awakening period has already been reached when the determination is completed.

In addition, a temperature-growth model (DVI model) was developed that uses temperature information such as AMeDAS data to diagnose whether a tree has encountered a low temperature above a certain level necessary for dormancy awakening and to estimate the stage of bud development. (For example, see Non-Patent Document 1).
However, only a few useful fruit trees are being developed for DVI models, and at this stage, they are not applicable to any deciduous tree.
The DVI model generally uses wide-area temperature data such as AMeDAS data, and the sleep / wakefulness stage cannot be determined by garden or tree. In order to use the DVI model in more detail, such as at a park, temperature data at that point is required.
In addition, even if the DVI model is applied in units of gardens, the dormancy stage of flower buds varies from tree to tree due to the influence of the growing place (for example, sunlight, slope slope) in the same park. Therefore, the DVI model has a problem that it is impossible to accurately grasp the dormancy status of flower buds in each tree unit.

JP 2005-176728 A (Spontaneous Dormancy Awakening Method and Spontaneous Dormancy Awakening Agent for Deciduous Trees) JP 2010-047514 A (Spontaneous Dormancy Awakening Agent and Spontaneous Dormancy Awakening Method for Deciduous Fruit Trees)

Sugiura, Motosu, 1997, Agricultural Meteorology (J.Agric.Meteol.) 53 (4): p285-290, Elucidation and modeling of spontaneous dormancy arousal and temperature in Japanese pear Gyllenstrand, N., Clapham, D., Kallman, T. and Lagercrantz, U. 2007. A Norway spruce FLOWERING LOCUS T homolog is implicated in control of growth rhythm in conifers.Plant Physiology 144: 248-258. Hasebe, M., Kofuji, R., Ito, M., Kato, M., Iwatsuki, K. and Ueda, K. 1992. Phylogeny of gymnosperms inferred from rbcL gene sequences. Bot. Mag. Tokyo 105: 673-679 .

  The present invention solves the above-mentioned problems and develops a method for determining a swift and surely per-tree reliable spontaneous dormancy awakening period of a low-temperature demanding deciduous tree without being affected by climate warming or abnormal weather For the purpose.

As a result of intensive research, the present inventor has found that the content of commutating sugar, which has remained low during spontaneous dormancy, in the deciduous tree conduit fluid, rises rapidly in line with the time when the spontaneous dormancy awakening period is reached. I found it.
That is, by using the content of commutating sugar in the conduit fluid as an index, it was found that it was possible to determine that the period when the content was low was still in the spontaneous dormancy period, and that the high period had reached the spontaneous dormancy awakening period. . In addition, the said determination was a technique which can be determined on the same day for the tree unit to be determined.

Furthermore, the present inventor has found that a method for indirectly determining the content of commutated sugar can be employed in order to more quickly determine whether the spontaneous dormancy awakening period has been reached.
That is, it has been found that (i) a method of judging by the Brix instruction value and (ii) a method of judging by the value of the total organic carbon content can be adopted.
In addition, when the invert sugar is a reducing sugar, it has been found that (iii) a method of mixing with a potassium permanganate solution at a certain ratio and judging by the difference in color tone after the color reaction can be adopted. It was.

The present invention has been made based on these findings.
The present invention according to [Claim 1] is characterized by determining whether or not the spontaneous dormancy awakening period has been reached, using the content of commutating sugar in the conduit fluid as an index, and spontaneous dormancy of a low-temperature demanding deciduous tree The present invention relates to an awakening period determination method.
The present invention according to [Claim 2] relates to the determination method according to claim 1, wherein the content of the invert sugar is the total content of all sugars constituting the invert sugar.
The present invention according to [Claim 3] relates to the determination method according to claim 1, wherein the content of the invert sugar is the content of the main sugar of the invert sugar.
The present invention according to [Claim 4] relates to the determination method according to any one of Claims 1 to 3, wherein the low-temperature required deciduous tree is a low-temperature required deciduous tree belonging to a seed plant.
-The present invention according to [Claim 5] relates to the determination method according to any one of claims 1 to 4, wherein the low temperature requirement deciduous tree is a low temperature requirement deciduous tree belonging to angiosperms or Ginkgo biloba. is there.
-The present invention according to [Claim 6] relates to the determination method according to any one of Claims 1 to 5, wherein the index is a rapid increase in the content of the invert sugar. It is.
-The present invention according to [Claim 7] is characterized in that, in the Japanese pear as the low-temperature-requiring deciduous tree, the total content of all sugars constituting the commutated sugar or the content of the main commutated sugar is 7. The determination method according to claim 1, wherein it is determined that the spontaneous dormancy arousal period has been reached when the dose is 21 mg / mL or more.
The present invention according to [Claim 8] has reached the spontaneous dormancy awakening period when the determination satisfies the following conditions (A) and (B) for Japanese pears as the low temperature requirement deciduous tree: It is related with the determination method in any one of Claims 1-6 which is determined.
(A) A condition in which the total content of all sugars constituting the invert sugar is 17 mg / mL or more.
(B) A condition in which the total content of all the sugars constituting the invert sugar is at least twice the content of the invert sugar at the time described in (C) below.
(C) The time when the buds in which the bud scales are loose and appear green when the temperature is maintained at 25 ° C. for 12 hours for 21 days are 0%.
-The present invention according to [Claim 9] has reached the spontaneous dormancy arousal period when the determination satisfies the following conditions (D) and (E) for Japanese pears as the low temperature requirement deciduous tree: It is related with the determination method in any one of Claims 1-6 which is determined.
(D) A condition in which the total content of all sugars constituting the invert sugar is 6 mg / mL or more.
(E) A condition in which the total content of all the sugars constituting the invert sugar is at least three times the content of the invert sugar at the time described in (F) below.
(F) The time when 0% of the buds are in a state in which the scales of the buds are loose and green when subjected to isothermal cultivation for 21 days at 25 ° C. for 12 hours.
· Present according to [Claim 10] invention, the low temperature requirement deciduous is a low temperature requirement deciduous belonging to Rosaceae; the commutation sugar major sugar, sorbitol; of claims 1 to 6 The method relates to any one of the determination methods.
The present invention according to [Claim 11] is one in which the conduit fluid is recovered by centrifuging a branch collected from the low temperature requirement deciduous tree; or a branch collected from the low temperature requirement deciduous tree Recovered from the other end by pressurizing one end of the end; or recovered from the other end by aspirating from one end of a branch taken from the low temperature demanding deciduous tree It is related with the determination method in any one of Claims 1-10.
-The present invention according to [Claim 12] is the determination according to any one of claims 1 to 11, wherein the determination is performed by estimating the content of commutated sugar from the Brix indication value of the conduit fluid. It relates to a determination method.
-The present invention according to [Claim 13] is that the determination is performed by estimating the content of commutated sugar from the value of the total organic carbon content of the conduit fluid. The method relates to any one of the determination methods.
In the present invention according to [Claim 14], the determination is performed by mixing the conduit liquid and the potassium permanganate solution, and estimating the content of reducing sugar from the color tone after the color reaction of the mixed liquid. The present invention relates to a determination method according to any one of claims 1 to 11 .

The present invention makes it possible to provide a method for determining the swift, dormant and awakening period of a low-temperature demanding deciduous tree 'quickly' and 'in units of trees' without being affected by climate warming or abnormal weather. .
Thus, the present invention makes it possible to provide information for determining the start time of appropriate covering and warming cultivation in the cultivation of deciduous fruit trees, and to carry out covering and warming cultivation that can guarantee the yield and quality of fruits. Make it possible.
In addition, it is possible to provide information for determining the appropriate start time for covering and heating cultivation even in the early cultivation of ornamental flower buds of deciduous woody nature, and covering and heating that can guarantee the production and quality of cut flowers Enables cultivation.

In addition, the present invention can provide a method for determining whether a tree has reached the spontaneous dormancy awakening period, easily by anyone at the cultivation site without requiring an analyzer such as liquid chromatography. To do.

In Example 1, it is the figure which showed the relationship of a time-dependent change of DVI value and sorbitol content in a conduit | pipe liquid. The symbol “○” in the figure indicates the measurement result in FY2008, “■” indicates the measurement result in 2009, and “◆” indicates the measurement result in FY2010. In Example 2, it is the figure which showed the influence which the difference in the site | part in one branch has on the change of sorbitol content. The symbol “■” in the figure indicates the measurement result of the branch tip, “●” indicates the middle of the branch, and “□” indicates the trunk side of the branch. In Example 4, the results showing the color tone after the color reaction of each xylem and KMnO ₄ solution collected in 2010. In Example 5, it is a graph which shows the correlation of the sorbitol content of each conduit | pipe liquid extract | collected in 2010, and a Brix indicator value. It is the figure which showed the evolutionary bifurcation relationship of the main deciduous trees including the various low temperature requirement deciduous trees made into the measurement object in Example 7,8. The branching relationship was described according to the 3rd edition of the APG classification system. It is the photograph image figure which classified the germination stage of Japanese pear (variety name: Kosui) into 7 stages. Numbers 1 to 7 described below each bud in the figure are numbers representing stages of the budding stage. It is the photograph image figure which classified the germination stage of the apple (variety name: Fuji) into 7 stages. Numbers 1 to 7 described below each bud in the figure are numbers representing stages of the budding stage. It is the photograph image figure which classified the germination stage of the peach (variety name: Akatsuki) into 7 steps. Numbers 1 to 7 described below each bud in the figure are numbers representing stages of the budding stage. It is the photograph image figure which classified the germination stage of a chestnut (variety name: Porotan) into 7 steps. Numbers 1 to 7 described below each bud in the figure are numbers representing stages of the budding stage. It is a photographic image diagram showing the sprouting stages of oysters (variety name: Ehime) classified into 7 stages. Numbers 1 to 7 described below each bud in the figure are numbers representing stages of the budding stage. It is the photograph image figure which classified the germination stage of the ginkgo (variety name: Kusu) into 7 stages. Numbers 1 to 7 described below each bud in the figure are numbers representing stages of the budding stage. In Example 7, it is the figure which showed the relationship of the time-dependent change of the germination rate of a Japanese pear (variety name: Kosui), and the content of commutation sugar. In Example 7, it is the figure which showed the relationship of the time-dependent change of the germination rate of apple (variety name: Fuji), and the content of commutation sugar. In Example 7, it is the figure which showed the relationship of the time-dependent change of the germination rate of peach (variety name: Akatsuki), and the content of commutation sugar. In Example 7, it is the figure which showed the relationship of the time-dependent change of the germination rate of persimmon (variety name: Ehime), and the content of commutation sugar. In Example 7, it is the figure which showed the relationship of the time-dependent change of the germination rate of ginkgo (variety name: Kusu) and the content of commutation sugar. In Example 8, it is the figure which showed the relationship of the time-dependent change of the germination rate of peach (variety name: Akatsuki), and the content of commutation sugar. In Example 8, it is the figure which showed the relationship of the time-dependent change of the germination rate of chestnut (variety name: Polotan), and the content of commutation sugar. In Example 8, it is the figure which showed the relationship of the time-dependent change of the germination rate of oyster (variety name: Ehime), and the content of commutation sugar. In Example 8, it is the figure which showed the relationship of the time-dependent change of the germination rate of ginkgo (variety name: Kusu) and the content of commutation sugar.

Hereinafter, embodiments of the present invention will be described in detail.
The present invention relates to a method for determining a spontaneous dormant awakening period of a low-temperature demanding deciduous tree.

[Low-temperature deciduous tree]
The determination method of the present invention determines, for all deciduous trees belonging to angiosperms and gymnosperms, whether or not the tree has reached the spontaneous dormant awakening period by using the content of commutating sugar in the conduit fluid as an index. It becomes possible.
The 'low temperature requirement deciduous tree' to be judged according to the present invention is a plant belonging to a so-called seed plant, which is a deciduous tree that has fallen awakened by encountering a low temperature for a certain period of time after falling under short days or low temperature conditions. Point to.
Here, the seed plants include extinct fern seed plants, but in the present ones, they indicate a taxon that combines angiosperms and gymnosperms.

Here, 'diapause arousal due to low temperature requirement deciduous trees' is widely used in the taxonomic groups of almost all angiosperms listed below (almost all eyes and families) and deciduous trees of ginkgo and pine in gymnosperms. It is a phenomenon that can be seen.
In addition, many types of angiosperms 'Evergreens', such as German spruce (Picea abies), which is a gymnosperm 'Evergreen', do not fall off, but sprout due to low temperature requirements. The growth initiation phenomenon itself has been confirmed.
In addition, regarding the germination phenomenon from dormancy, homology of the Flowering Locus T gene is involved in its regulation in both angiosperms and gymnosperms, and universality at the gene level has been confirmed (for example, Non-Patent Document 2 (see N. Gyllenstrand et al., 2007).
From these facts, it is recognized that the mechanism of diapause by the low temperature requirement of deciduous trees belonging to angiosperms and gymnosperms is caused by a common mechanism (in evolution, it is a co-derived trait).

-Gymnosperm The gymnosperm (Gymnosperms) which is the object of the present invention refers to a taxonomic group of seed plants having a female reproductive organ with an exposed ovule. The plant group is a taxonomic group that is most closely related to angiosperms described later in the modern plant. It does not have petals like angiosperms, and pollen is sprayed by an air medium.
Specific examples of gymnosperms include ginkgo biloba, cycad, pine and gnetsum. According to recent molecular phylogenetic analysis, these four taxa are considered to be taxonomic groups separated from a single ancestor plant (for example, see Non-Patent Document 3 (M. Hasebe et al., 1992)). ).
Among the gymnosperms, deciduous trees that require low temperature are known as Ginkgo biloba ginkgo (Ginkgoceae genus) and pine larch (Pinus pine). Therefore, in this invention, a ginkgo and a larch can be made into a suitable determination object in a gymnosperm.
In particular, ginkgo is a deciduous tree that is useful as an application target of the determination method of the present invention because it can produce ginkgo.

Angiosperm Angiosperm (Angiosperm), which is the subject of the determination of the present invention, is a group of seed plants having a female reproductive organ with an ovule covered with percutaneous skin, branched from a single ancestor plant Refers to a taxon.
Many angiosperms have the characteristic of attaching so-called 'flowers', which are reproductive organs adapted to pollen insects and avian media. In addition, some angiosperms have the characteristic of forming fruit adapted for bird and animal spraying of seeds.
The common ancestor plant of angiosperms is thought to have appeared about 140 million years ago (Jurassic).

Specific examples of angiosperms are Euchycots, which are the true dicotyledonous species, Euphoridae, Azaleas (Oysteraceae, Trichophyceae), Gallia, Glycidae, Perilla (Lepidoptera, Ganoderma), The plants of the genus Coleaceae, Cleoptera, Cleidae), eggplant, celery, echaceae, chrysanthemum, and scorpiona (Lordaceae) can be mentioned.
In addition, roses of the true roses that are also true dicotyledons (Rosaceae, Mulberryaceae, Currantaceae, Chrysanthemumaceae, Hydrangeaceae), Cucurbitaceae, Beechidae (Beechaceae, Walnutidae, etc.) Family), legume, quintranoid (Coleoptera), ostrichid, Brassicaceae, blue-eyed, euphorbiaceae (Apiaceae, Citrudidae), oleoptera (Apiaceae), auropoda, crosomaidae, aphididae , Plants.
It is also a true dicotyledonous but is considered to be a different strain from the true roses and true chrysanthemum, Lepidoptera, Grapeaceae (Vineaceae), Biwamoduki, Berberidopsis, Sandalid, Gundera, Gunnera, Mention may be made of the plants of the order of the order of the order of the order of the order of the order of the order of the order of the order of the order of the order of the order of the order of the order of the order of the order of the order is the order of the order of the order of the order of the order of the genus Porphyra, Lepidoptera, Porphyridae (Gumidae), Aburabidae, Pinaceae, Ranunculaceae (Aceraceae).

In addition, monocots, which are taxonomic groups different from the above-mentioned true dicotyledons, are the order of Coleoptera, Omodaceae, Cleoptera, Dioscorea, Lily, Octopus, Dasipogonidae, Coconut, Rice, Cirrus, ginger, and so on.
Furthermore, in the evolutionary line of angiosperms, magnolias (Bolemicaceae, Magnoliaceae), Camellia, Pepper, Canela, Ganoderma, Austrobailea (Papaveraceae), water lily, Menaceae of the order of Amborella.

-Plants that produce commercially useful fruit trees and flower buds In the present invention, any deciduous tree belonging to these angiosperms or gymnosperms and requiring low temperature can be used as a determination target.
In addition, examples of plants that are particularly important in the present invention include deciduous trees that are capable of producing commercially important fruits and florets (particularly cut flowers).

  Specific examples of fruit trees belonging to the order of roses include: Rosaceae, Japanese pear (Pear genus), Pear (Pear genus), Chugokunashi (Pear genus), Apple (Apple genus), Sweet cherry (Cherry genus) , Ume (Prunus), Plum (Prunus), Spinosa Prunus (Prunus), Apricot (Prunus), American Cherry [Black Cherry] (Prunus), Summit Cherry (Prunus), Almond (Prunus), Yusraume (Prunus genus), Peach (Peach genus), Karin (Boke genus), Quince (Quince genus), Prunus quince (Prunus genus), June berry (Pleurotus genus), Raspberry genus (Raspberry genus), Blackberry , Raspberries.

  In addition, other fruit trees belonging to the order of rosaceae include mulberry (mulberry), fig (ficus genus), currant [currant] (currant genus), currant [gooseberry] (currant genus), red currant [Red currant] (currant genus);

Examples of the fruit tree belonging to the order of the beech include the beech family Chestnut (Chestnut genus);
Further, examples of fruit trees belonging to the order of the order Pomegranate are Pomegranates (Pomegranates) of the family Misohagi.
Moreover, examples of fruit trees belonging to the order of Mucurodia include citrus family Salamander (Sangusa genus);
In addition, examples of fruit trees belonging to the order of Quintrano can include Euphorbiaceae, A. japonicum (Apiaceae).
In addition, examples of fruit trees belonging to the order of grapes include grapes of the Grapeaceae family (Grapes).

In addition, fruit trees belonging to the order of the azaleas include blueberries of the azalea family, bilberries (genus genus), cranberries (genus genus), bilberries (genus genus); , Kiwi fruit (genus Matabia) and Sarnashi (genus Matatabi).
In addition, examples of fruit trees belonging to the order of the scorpionae include a lotus cup of the honeysuckle family (the genus Honeysuckle).
Examples of fruit trees belonging to the order of the genus Porphyra include Gummy (Gummy genus).
In addition, examples of fruit trees belonging to the order of Ranunculaceae include Akebidae (Akebi).
Further, examples of fruit trees belonging to the order of the magnolia include Banglei (Banleci) and Popo (Popoe).
Further, examples of fruit trees belonging to the order of the Austrobailea include the pine family of the family Euphorbia.
Examples of fruit trees belonging to gymnosperms include Ginkgo biloba ginkgo (Ginkgoaceae ginkgo genus).

On the other hand, examples of deciduous trees used for production of ornamental florets include the following roses.
Specific examples include: Rosaceae (Chrysanthemum), Ume (Chrysanthemum), Peach (Peach), Prunus (Prunus), Yukiyanagi (Prunus spp.), Boke (Genus Peculiar); (Hydrangea), can be mentioned.

-Rosaceae Deciduous Trees As described above, particularly important judgment targets in the present invention include rose family plants including many types suitable for production of commercially important fruit trees and flower buds.
Specific examples of the Rosaceae plant include the subfamily Rose (genus Rose, Genus Strawberry, Yamabuki, etc.), the subfamily Shimosuke (genus Shimotake, Genus Sakura, etc.), and the subfamily Pear (genus Papida, apple, Biwa) Etc.), a taxon that consists of plants belonging to the subfamily Sakura (genus Sakura).
Among these, in this invention, it can apply especially suitably with respect to the cultivar of the fruit (or seed) of these deciduous trees.

In particular, examples of trees to which the determination target of the present invention can be suitably applied include fruit trees belonging to the genus Pyrus.
Examples of the pear genus include P.pyrifolia var. Culta (Japanese pear), P.pyrifolia var. Pyrifolia (Yamanashi), P.communis (Pear), P.bretschneideri (Chugokunashi), P.ussuriensis var. Aromatica ( Examples include Iwate Yamanashi), P.ussuriensis var. Hondoensis (Aonashi), P.calleryana (Penix), P.betulifolia (Penanthus pneumoniae), P.amygdaliformis (Almond Pear).
Among these, as a determination target of the present invention, it is particularly assumed that the present invention is suitable for determination of fruit trees belonging to P.pyrifolia (Japanese pear, Yamanashi). In addition, any varieties belonging to the species (for example, Kosui, Hosui, 20th century, Gold 20th century, Shintaka, Emerging, Minamisui, Chojuro, Ehime, Kikusui, Bansankichi etc.) ) And can be determined by the present invention.

[Spontaneous Dormancy Awakening Period]
The deciduous tree to be determined according to the present invention is a woody plant (tree) requiring low temperature. Low temperature requirement refers to the property that a tree that has entered a spontaneous dormancy period in the middle to high latitudes needs to accumulate low temperature encounters for a certain period or more in order to reach the dormancy awakening period.
It should be noted that the temperature and duration required for accumulating low temperature encounters are slightly different in temperature required by plant species.
For example, in the case of Japanese pear, the required low temperature is recognized as 9 ° C or less.

Here, the “spontaneous dormant awakening period” refers to a developmental stage in which the bud has acquired the ability to fully bloom or leave.
Specifically, 50% or more when the tree (or branch) is cultivated for 21 days at 25 ° C under the condition of 12 hours of day length (including constant temperature treatment with the cuts inserted in water) When the bud scales of the buds are in a loose green state (with a germination rate of 50% or more), the tree (or branch) is surely in a spontaneous dormant awakening period (the buds are fully flowering or leaving leaves) It is recognized that it has reached the development stage that acquired the ability).
In the case of Japanese pear, it is defined as a developmental stage that has acquired the ability to bloom 70% or more of flower buds (not including leaf buds) of trees (or branches) when cultivated at 25 ° C (for example, non-patented) Reference 1).

Here, when artificially warming or covering cultivation of a tree (or branch) that has reached the 'spontaneous dormancy awakening period' due to low temperature encounter above a certain level, the buds sprout quickly and most of them bloom or emerge. It becomes possible to let leaves.
On the other hand, trees that are still in the “spontaneous dormancy period” are in a developmental stage where the buds have not sufficiently acquired the ability to blossom or leave. Therefore, even if the tree (or branch) in the spontaneous dormancy period is artificially heated or covered, it takes a long time to sprouting, and even if many buds do not bloom and leave leaves It cannot be grown.

[Conversion sugar content in conduit liquid]
The kind and composition of sugar molecules constituting commutated sugar in the conduit fluid vary depending on the kind of plant.

Here, the “commutated sugar” is a sugar molecule synthesized based on a photosynthesis product (glucose or the like) photo-synthesized in a leaf or a saccharide (starch or the like) accumulated in a storage tissue. It refers to a sugar molecule that is used to transport the whole plant or other organs through a conduit.
Specific examples of the invert sugar include sucrose, glucose, sorbitol, mannitol, raffinose, stathiose, fructose, galactose, and myo-inositol.

In the present invention, as the 'inverted sugar content', (i) the content of all sugar molecules composing the invert sugar (that is, the total content of all sugars composing the invert sugar) is used as an index. Thus, it is possible to determine whether or not the spontaneous dormancy awakening period has been reached with the highest sensitivity. That is, the method using the total sugar content in the conduit fluid as an index is the most preferred embodiment in the present invention.
In addition, (ii) the case where the content of sugar molecules (the content of main sugars) serving as the main component of commutating sugar is used as an index is preferable because accurate determination is possible.
In addition, it is also possible to use (iii) the content of sugar molecules other than the main component constituting the invert sugar (the content of sugars other than the main sugar) as an index. Among them, sugar molecules having the second highest content are preferable. However, when a saccharide with a small composition ratio is used as an index, there may be a case where the kinetic fluctuation of the saccharide content is small and the determination is difficult.

-Inverted sugar in angiosperms In most types of angiosperms, the main sugar in invert sugar is sucrose. Even if it is not the main sugar, it accounts for a large proportion of the sugars that make up the invert sugar.
Therefore, in almost all types of deciduous trees belonging to angiosperms, it is possible to determine whether or not the spontaneous dormancy awakening period has been reached by using the sucrose content in the conduit fluid as an index.

・ Rose family inversion sugar In the case of Rosaceae, the main sugar of the inversion sugar is sorbitol. Therefore, in the case of the deciduous tree of the said plant, it becomes possible to determine whether it has reached the spontaneous dormancy awakening period by using sorbitol content as an index.
In addition, the invert sugar of the plant includes sucrose and glucose. Although the accuracy is lowered, it is also possible to determine the spontaneous dormancy awakening period using sucrose or glucose as an index.

・ Terucinaceae of the family Oleaceae and the pine family, Oleaceae, and Verbanaceae produce mannitol and stathiose as the main sugars of the inverted sugar. Therefore, in the case of deciduous trees of these plants, it is possible to determine whether or not the spontaneous dormancy arousal period has been reached by using the mannitol or stathiose content as an index.
In addition, the invert sugars of these plants include raffinose and sucrose. Although accuracy is reduced, it is also possible to determine the spontaneous dormancy arousal period using raffinose or sucrose as an index.

・ Crystals of the asteraceae In the asteraceae (Celastraceae), galactose is produced as the main invertible sugar. Therefore, in the case of the deciduous tree of the plant, it is possible to determine whether or not the spontaneous dormancy awakening period has been reached by using the galactose content as an index.
In addition, the invert sugar of the plant includes stathiose and sucrose. Although accuracy is reduced, it is also possible to determine the spontaneous dormancy wakefulness period using stathiose or sucrose as an index.

-Tumbled sugar of the scorpionaceae family Paplownia tomentosa, which belongs to the Scrophulariaceae plant, produces sucrose and statiose as the main sugars of the tumble sugar. Therefore, in the case of the said plant, it becomes possible to determine whether it has reached the spontaneous dormancy wakefulness period by using sucrose or stathiose content as an index.
In addition, the invert sugar of the plant also contains raffinose. Although the accuracy is lowered, it is also possible to determine the spontaneous dormancy awakening period using raffinose as an index.

・ Two-headed quailaceae, terrestrial terrestrial tumble sugar In genus quailaceae (Bignoniaceae), teraceae (Buddleiaceae), stathiose is produced as the main sugar of the inverting sugar. Therefore, in the case of the plant, it is possible to determine whether or not the spontaneous dormancy awakening period has been reached by using the statioose content as an index.
Moreover, sucrose and raffinose are also contained in the invert sugar of the plant. Although accuracy is reduced, it is also possible to determine the spontaneous dormancy-wakefulness period using these sugars as an index.

・ Inverted sugar in the family Tabata Family Myo-inositol is produced as an invert sugar in addition to sucrose in Kiwifruit (Actinidia deliciosa), Sarcinia (Actinidia arguta), etc. Therefore, in the case of the said plant, it becomes possible to determine whether it has reached the spontaneous dormancy awakening period by using myo-inositol or sucrose content as an index.

-Ginkgo biloba plant inversion sugar In ginkgo biloba ginkgo biloba ginkgo, the main sugar of inversion sugar is glucose. Therefore, in the case of the said plant, it becomes possible to determine whether it has reached the spontaneous dormancy awakening period by using glucose content as a parameter | index.
Moreover, fructose and sucrose are included in the invert sugar of the plant. Although accuracy is reduced, it is also possible to determine the spontaneous dormancy awakening period using fructose or sucrose as an index.

[Judgment index]
In the present invention, the content of commutating sugar in the conduit fluid is used as an index, and the value of the content of commutating sugar that has been changing at a low value within a certain range is used as an index. is there.

  (1) Regarding the determination of the present invention, specifically, the total content of the invert sugar or the content of the main sugar of the invert sugar is 21 mg / mL or more, preferably 22 mg / mL or more, further 23 mg / mL. If it is above, or more than 24 mg / mL, or more than 25 mg / mL, it can be determined that the spontaneous dormancy-wakefulness period has been reached with the value of the content only (single measurement) .

(2) In addition, even if the total content of invert sugar is less than the value described in (1) above, if both the following conditions (A) and (B) are satisfied, spontaneous dormancy arousal It can be determined that the period has been reached.
Specifically, the condition (A) in which the total content of invert sugar total sugar is 17 mg / mL or more, preferably 18 mg / mL or more, further 19 mg / mL or more, more preferably 20 mg / mL or more; and Satisfying both of the conditions (B), wherein the invert sugar content is at least 2 times, preferably at least 2.5 times, and even at least 3 times the commutation sugar content at the time of germination rate 0% In this case, it can be determined that the spontaneous dormancy awakening period has been reached.

(3) In addition, even when the total content of invert sugar is less than the values described in (1) and (2) above, both of the following conditions (D) and (E) are satisfied: It can be determined that the spontaneous dormancy awakening period has been reached.
Specifically, the condition (D), wherein the total content of invert sugar is 6 mg / mL or more, preferably 6.5 mg / mL or more, further 7 mg / mL or more, and further 8 mg / mL or more; And the condition (E) where the content of commutating sugar is 3 times or more, preferably 4 times or more, and even 5 times or more than the content of commutating sugar at the time of germination rate 0%. When it satisfies, it can be determined that the spontaneous dormancy awakening period has been reached.

Here, a specific type of the determination is shown.
In the determination of the present invention, for example, (i) when the total sugar content of the commutated sugar in the conduit fluid is extremely low (for example, 1 mg / mL), the bud development stage is still in the spontaneous dormancy period. Can be determined.
(Ii) Even if the total sugar content of the commutated sugar in the conduit fluid is somewhat high (for example, 10 mg / mL), it does not increase compared to the content at the time of germination rate 0%. It can be determined that the bud development stage is still in the spontaneous dormancy period.

On the other hand, (iii) the case where the total sugar content of the commutated sugar in the conduit fluid is somewhat high (for example, 10 mg / mL) and more than a predetermined amount compared to the content at the time when the germination rate is 0% (for example, If it has risen to 3 times or more), it can be determined that the bud development stage has reached the spontaneous dormancy awakening period.
In addition, (iv) when the total sugar content of the commutated sugar in the conduit fluid is extremely high (for example, 25 mg / mL), it may be determined that the bud development stage has reached the spontaneous dormancy-wakefulness period. it can.

It should be noted that the principle of this judgment is that the content of commutating sugars in the conduit fluid in low-temperature demanding deciduous trees changes at a low value when (i) is in spontaneous dormancy, and (ii) is the time when the spontaneous dormancy awakening period is reached. It is based on the phenomenon of a sudden rise in agreement.
Here, when the content of commutating sugar rapidly rises, in plants where the DVI model (model for estimating growth status from local temperature data: see Non-Patent Document 1) is established, the value reaches 1.0. It coincides with the time (= the time when many of the target trees in the area are estimated to have reached the spontaneous dormancy awakening period in the natural state).
In addition, after awakening germination, sugar is consumed by metabolism for germination growth of buds, so that the content of commutated sugar starts to decrease.

[Collecting conduit fluid]
In the present invention, it is necessary to collect the conduit fluid as a method for measuring the content of commutating sugar in the conduit fluid.

-Collection site | part Here, any site | part can be employ | adopted as a site | part which collects conduit fluid. You may extract from any part, such as a branch, a trunk, and a root. In particular, it is preferable to collect from “branches”, and felled or pruned branches can be suitably used.
Strictly speaking, the measured commutation sugar content indicates the measured dormancy stage of the branch, but there is no significant difference in the commutation sugar content in the whole tree.
Therefore, in the present invention, any branch of a tree can be used to determine the dormancy stage of the entire tree.

  In addition, in order to reliably determine whether the commutation sugar content has entered the rising period using the branches, the conduit fluid is removed from about half of the branches ahead (particularly the tip of the branch). It is desirable to collect. This is because the branch tip side portion is delayed in the increase of the commutated sugar content as compared with the trunk side portion, and thus the dormancy stage can be determined more reliably.

-Collection method Any method can be adopted as a method for collecting the conduit fluid. For example, it is possible to employ a method of directly punching an injection needle or the like into a branch or the like, and preferably, using the fact that the conduit is a tube having a cylindrical shape, the centrifugation method described in detail below, It is preferable to employ a pressure method, a suction method, or the like.

In particular, in the present invention, as a method for recovering the conduit liquid simply and quickly, a method of recovering the liquid from the branch conduit by centrifugation can be employed.
Specifically, the branches taken from the target tree are shredded and placed in a container (for example, a centrifuge tube, a conical tube, etc.) (so that the angle between the branches and the centrifugal force is acute or perpendicular). A method of centrifuging with a centrifugal force of 50 × g or more can be employed. Further, the upper limit value of the centrifugal force may be in a range where the tissue is not broken, and examples thereof include 10,000 × g or less. In view of actual operation, a range of 500 to 5,000 × g is a substantial range and is preferable.
Any device or apparatus can be used for the centrifugation as long as it can be centrifuged, and it is possible to use not only a normal centrifuge but also a dehydration mode of a household washing machine, for example. .
It is also possible to obtain the centrifugal force by attaching a string or the like to the centrifuge tube and swinging it manually with a string. Specifically, it is possible to obtain the centrifugal force (g) required to recover the above-mentioned conduit liquid by attaching a string so that the centrifugal radius is 50 cm or more, preferably 80 cm or more. It is.
For example, it is possible to use a shape in which two or more holes are made in a portion near the opening of the centrifuge tube and the string is passed through the string, and the centrifuge tube and the string form a ring.
Moreover, the thing of the shape which attached the string to the adapter which can attach or detach a centrifuge tube can also be used.

In the centrifugation, cells and the like are not crushed, so that only the liquid in the conduit (cylindrical tube) can be collected at the bottom of the container.
Here, as the shredding of the branches, it may be usually shredded (for example, with a length of 3 cm or more). However, when the shredding process is performed so that the cells are crushed, Characters are mixed and not suitable for the following measurement. Also, it is not preferable to perform pulverization or grinding because the cells are crushed.
The upper limit of the length is not particularly limited, but may be any length as long as it can enter the centrifuge tube. For example, it can be 20 cm or less, preferably 15 cm or less, and further 13 cm or less.

・ Pressurization method In addition to the above-mentioned centrifugal method, as a method of easily collecting the conduit fluid, a method of collecting the conduit fluid from the other end by pressurizing one end of the collected branch can be adopted. .
Specifically, a method of connecting a tube or the like to one end of a branch obtained by chopping both ends of a branch, sending compressed air through the tube or the like, and extruding and recovering a conduit liquid from the other (opposite) end Can be mentioned.
As a means for pressurizing, any method may be used. For example, compressed air can be sent by using a pressure chamber, an air inlet, a syringe or the like.

-Suction method Moreover, the method of collect | recovering from the said one end can be employ | adopted by attracting | sucking (reducing pressure) one end of the extract | collected branch contrary to a pressurization method.
Specifically, mention is made of a method of connecting a tube or the like to one end of a branch obtained by chopping both ends of the branch, sucking through the tube or the like, and sucking and collecting the conduit liquid from the one end. Can do.
Here, any means may be used for suction (decompression), but suction can be performed using, for example, a syringe, an aspirator, a vacuum cleaner, or the like.

It is possible to measure the content of commutated sugar by subjecting the collected conduit fluid to analysis as it is.
For example, fractionation using paper chromatography, thin layer chromatography, high performance liquid chromatography, gas chromatography, capillary electrophoresis, etc., and mass spectrometer, electrochemical detector, evaporator light scattering detector, etc. are connected to these. By quantifying, the total sugar content and the contents of various sugars can be measured.
And the growth stage of a bud can be determined as mentioned above by using the value of the measured content as an index.

[Simple measurement method]
Furthermore, in the present invention, it is possible to determine the spontaneous dormancy wakefulness more easily and quickly using another value correlated with the commutation sugar content as an index. For example, the following method can be employed.

-Method using the Brix instruction value as an index In the present invention, it is possible to determine the spontaneous dormancy awakening period using the Brix instruction value of the conduit fluid as an index.
The Brix instruction value is a value (degree,%) that represents the total amount of solids contained in the solution by utilizing the property that the refractive index of the solution changes depending on the solid content concentration.
In the deciduous tree conduit liquid component, the ratio of commutated sugar is very high, so the Brix indication value and the content of commutated sugar show a very high correlation (a linear function with a linear slope).

In the present invention, the principle can be used to estimate the commutation sugar content from the Brix instruction value, and to determine the spontaneous dormancy-wakefulness period.
In the present invention, the index of the Brix instruction value that can be determined to have reached the spontaneous dormancy awakening period can be obtained with reference to the measurement results shown in FIG.
For example, in the case of Japanese pear whose main sugar of the invert sugar is sorbitol, the Brix indicator value corresponding to a sorbitol content of 10 mg / mL is about 1.3, and the Brix indicator value corresponding to a sorbitol content of 20 mg / mL is about 2.3. is there.

  In addition, although the Brix instruction | indication value used as the parameter | index in the said determination can be measured using a refractometer, it can also be measured using the simple refractometer and sugar content meter for measuring the sugar content of a drink.

-Method using the total organic carbon content as an index In the present invention, it is possible to determine the spontaneous dormancy awakening period using the value of the total organic carbon (TOC) content in the conduit fluid as an index.
The TOC content is a value indicating the carbon content of the total organic matter, but in the deciduous tree conduit liquid component, since the proportion of commutated sugar is very high, the TOC content and the commutated sugar content are A very high correlation (relation of a linear function whose slope is a straight line) is shown.

In the present invention, the principle can be used to estimate the commutation sugar content from the TOC content, and to determine the spontaneous dormancy wakefulness period.
Note that the value of the TOC content as an index in the determination can be measured using any measurement kit or apparatus that can detect and quantify organic carbon of 2 mg / mL or more. That is, determination using a simple measurement kit is possible.

-Method using the color tone of the KMnO ₄ solution as an index In the present invention, for the deciduous tree in which the main component sugar of the invert sugar is a reducing sugar, or for a deciduous tree that contains a large amount of reducing sugar as the constituent sugar of the invert sugar It is possible to employ a method of mixing with potassium permanganate (KMnO ₄ ) solution at a certain ratio, estimating the reducing sugar content based on the color difference after the color reaction, and determining the spontaneous dormancy wakefulness period.
The KMnO ₄ solution is one of redox reaction titrants and is an effective reagent for quantifying oxidizable substances. Therefore, the amount of reducing sugar that is an oxidizable substance can be easily measured.
The principle utilizes the property that the color tone after color reaction of KMnO ₄ and reducing sugar exhibits brown when there is little reducing sugar, and brown, red, red-orange, and orange as reducing sugar increases. Is.
In addition, examples of the reducing sugar that is used in the invert sugar include sorbitol, glucose, mannitol, galactose, and fructose. In particular, sorbitol and glucose can be suitably targeted from the relationship with the content.

In the present invention, in order to facilitate the determination of spontaneous dormant arousal period by colorimetry, conduit fluid having a reducing sugar content of 5 to 30 mg / mL, preferably 5 to 25 mg / mL, more preferably 10 to 20 mg / mL. When it is used for the determination, it is desirable to exhibit red to reddish orange.
For example, a method of mixing the KMnO ₄ solution so that the final concentration of KMnO ₄ after mixing with the conduit liquid is 0.75 to 6%, preferably 1 to 5%, more preferably 2.25 to 4.5%. Can do.
In order to determine more accurately, the background of the mixed solution in a transparent container is white, and a colorimetric table ("color tone after color reaction of KMnO ₄ and reducing sugar" and "reducing sugar content" It is desirable to make a determination in comparison with a card or paper showing a relative relationship.

· In the simple measurement kit present invention, by using a color principle of reducing sugars and KMnO ₄ solution, it is possible to provide anyone easily feasible spontaneous dormancy period determination kit in cultivation site.
For example, (a) a container in which KMnO ₄ solution is enclosed as a stock (reagent bottle), (b) a container that performs a transparent or translucent color reaction (for example, containers of glass, polyethylene, polyethylene terephthalate, polystyrene, polypropylene, etc.) , (C) `` Spontaneous dormant awakening period '', which is a package of `` color tone after color reaction of KMnO ₄ and reducing sugar '' and a colorimetric table showing the relative relationship between `` reducing sugar content in conduit fluid '' A determination kit "can be provided.
In addition, (d) instruments necessary for collecting conduit fluid (for example, syringes, tubes, adapters for centrifuge tubes with strings, etc.) can be packed together in a kit.
By using this kit, a predetermined amount of KMnO ₄ solution is put into a transparent container, and a predetermined amount of the collected conduit fluid is added and mixed to compare with the colorimetric table. Can be determined.

Further, as an embodiment of the kit, an embodiment in which the above (a) and (b) are combined can be adopted. Specifically, a mode in which the KMnO ₄ solution is sealed in advance in a transparent or translucent container (preferably a pack test container) having a stopper or a lid is possible.
In this kit, it is possible to determine the spontaneous dormancy arousal period only by directly adding and mixing the conduit fluid to the KMnO ₄ solution and comparing it with the colorimetric table.

  EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated, the scope of the present invention is not limited by these.

[Example 1] "Relationship between DVI value and sorbitol content in conduit fluid"
Measure the time course of commutation sugar content (sorbitol) during the period from spontaneous dormancy to dormancy awakening in Japanese pear grown in open-air cultivation, and investigate the correlation with DVI value, which is a temperature index in climate It was.

(1) Measurement of sorbitol content in conduit fluid Take a branch from a Japanese pear (variety: Kosui) planted in the field of the Fruit Tree Research Institute, cut it into 5-10cm, put it into a centrifuge tube, 3000rpm (1,000x c) for 10 minutes. The liquid obtained from the branch to the centrifuge tube was recovered as a conduit liquid.
About the obtained conduit | pipe liquid, 'sorbitol content' was measured by the liquid chromatography.
The measurement was carried out over a period of three years from 2008 to 2010, from October, from the spontaneous dormancy period to the dormancy awakening period, from February to the following February.
Then, using the AMeDAS data of Tsukuba City, calculate the 'DVI value' (an index representing the stage of flower bud growth predicted from the temperature data) in the area including the sampling point, and calculate the sorbitol content in the conduit fluid. The correlation was investigated. The results are shown in Figure 1.

(2) Results Here, “when the DVI value reaches 1.0” is a time defined as “a time when the flower buds have reached the spontaneous dormancy awakening period” (see Non-Patent Document 1). In Tsukuba, 2008, 2009, and 2010, DVI reached 1.0 on December 26, December 25, and December 27, respectively.
As a result of the above measurement, the sorbitol content in the conduit fluid was kept low until the DVI value was less than 1.0 (until mid December), but from the time when the DVI value exceeded 1.0 (late December) It became clear that it rose rapidly and remained at a high level (see Figure 1). In addition, this phenomenon was confirmed in any of the surveyed years (2008-2010).
From these, it was suggested that 'the time when the sorbitol content in the duct fluid suddenly rises' and 'the time when the spontaneous dormancy-wakefulness period is reached' agree very well.

[Example 2] “Difference in sorbitol content due to differences in branch sites”
It was investigated whether the change of sorbitol content would be affected by one branch site.

(1) Measurement of sorbitol content in conduit fluid
Branches (approx. 100cm) collected from Japanese pear (variety: Kosui) in 2010 were divided into three equal parts at the tip, middle, and trunk, and the sorbitol content in the conduit fluid was the same as in Example 1. 'Measured.
The measurement was performed over time from November, which is the period from spontaneous dormancy to dormancy awakening, to February of the following year. The result is shown in figure 2.

(2) Results As a result, the sorbitol content in the conduit fluid suddenly increases and stays at a high level at the time when the DVI value reaches 1.0, regardless of which part of the branch is used. (See Figure 2). That is, it was shown that any part of the branch tip part, the intermediate part, and the trunk side part can be used for the measurement of the sorbitol content of the present invention.
However, the sorbitol content in the trunk side of the branch increased more rapidly than the branch tip and the middle. On the other hand, the sorbitol content increased more slowly at the branch tip than at the middle and trunk sides.
From this, in order to reliably determine the dormancy arousal time, it was considered desirable to use the middle part of the branch where the sorbitol content gradually increases (particularly the branch point part).

[Example 3] "Relationship between spontaneous dormancy awakening period and sorbitol content in conduit fluid"
It was actually verified whether the sorbitol content in the duct fluid changes depending on whether or not the spontaneous dormancy arousal period is reached.

(1) Measurement of sorbitol content in conduit fluid and calculation of germination arrival days
In fiscal 2010, four annual seedlings of Japanese pear (variety: Kosui) were cultivated in the usual field on the field of the Fruit Tree Research Institute.
In the cultivation period, from October to February of the following period from the spontaneous dormancy period to the dormancy awakening period, the period (i) DVI value is 0.8 (period when the spontaneous dormancy awakening period is not reached in the natural state), and (ii) DVI At the time when the value is 1.2 (the time when the spontaneous dormancy awakening period is reached in the natural state), branches are taken from two pot seedlings, and the sorbitol content in the conduit fluid is measured in the same manner as in Example 1. did.
In addition, after collecting the branches, the pot seedlings (the same tree as the tree from which the branches were collected) were collected and examined for “days of germination”. The number of days to germinate is calculated as the number of days until 90% of the spiked buds are sprouting when the collected pot seedlings are cultivated in an artificial weather chamber at 15 ° C. did. We also observed the flowering status of the sprouted flower buds. These results are shown in Table 1.

(2) Results As a result, in the trees collected when the DVI value was 0.8, it took more than 40 days for the buds to reach, many buds did not bloom, and the spontaneous dormancy arousal period had not been reached (no spontaneous dormancy was not observed). It is sufficient). The sorbitol content in the conduit fluid at the time of pot seed collection was a low value of 7.4 mg / mL.
On the other hand, in the trees collected when the DVI value is 1.2, the number of days of germination reached only 23 days, most of the flower buds bloomed and reached the spontaneous dormancy awakening stage (sufficient spontaneous dormancy awakening is sufficient) It was shown that. The sorbitol content in the conduit fluid at the time of pot seed collection was extremely high at 40.0 mg / mL.
From these results, it was demonstrated that the pear fluid content in the duct fluid was significantly increased in pears that reached the spontaneous dormancy awakening period, compared to before the period.

[Summary of Examples 1 to 3]
The sorbitol content in the pear duct fluid was kept low when the germination was in the spontaneous dormancy period, but it was found that the sorbitol content rapidly increased in accordance with the time when the spontaneous dormancy awakening period was reached.
From this finding, it was shown that it is possible to determine the spontaneous dormancy-wakefulness period in each tree unit using the value of sorbitol content in the conduit fluid and the degree of increase as an index.

[Example 4] "Study 1 on simple measurement method of sorbitol content in conduit fluid"
Potassium permanganate (KMnO ₄ ) solution, which is one of redox reaction titrants, is an effective reagent for the quantification of oxidizable substances. Therefore, it was examined whether KMnO ₄ solution can be used to easily measure the amount of sorbitol, a reducing sugar.

(1) Coloration with KMnO ₄ solution For each conduit fluid collected in Example 1 in FY 2010, mix so that conduit fluid: KMnO ₄ solution = 3: 1 (final concentration of KMnO ₄ : 2.25%), The color tone after the color reaction of the mixture was observed. The results are shown in FIG. Table 2 shows the correlation between the color tone and the sorbitol content and the determination of whether there is spontaneous dormancy arousal.
The date when the DVI value in fiscal 2010 reached 1.0 was December 27.

(2) Results As a result, the color tone of the mixed solution was brown (color tone when the content of reducing sugar was low) in the duct fluid before and after December 24, before reaching the spontaneous dormancy awakening period. On the other hand, in the duct fluids on and after January 4th when the spontaneous dormancy awakening period was reached, reddish orange to orange (color tone when the content of reducing sugar was high) was exhibited.
The color change in the color reaction from brown to red-orange was consistent with the increase in sorbitol content.
From this, it was shown that by examining the color tone of the mixed solution of the conduit fluid and the KMnO ₄ solution, it can be determined whether the bud flower bud is in the spontaneous dormant awakening period.

[Example 5] “Study 2 on simple measurement method of sorbitol content in conduit fluid”
It was investigated whether the sorbitol content of the conduit fluid could be easily measured using the Brix indicator value, which can easily measure the total solute content in the solution.

(1) Measurement by Brix indicated value For each conduit fluid collected in Example 1, the relationship between the sorbitol content of conduit fluid and the Brix indicated value of conduit fluid was examined using a Brix saccharimeter. The results are shown in FIG.

(2) Results As a result, it was shown that the Brix indication value and the sorbitol content of the conduit fluid can be expressed by a function represented by a linear function and correlate very high. From this, it was shown that it is possible to determine the sorbitol content and the spontaneous dormancy time by measuring the Brix indication value.

[Example 6] “Examination of simple recovery method for conduit fluid”
We examined a method that can easily collect the conduit liquid on site without using a centrifuge.

As a centrifuge tube, drill two holes on the side close to the lid of the conical tube (50 mL), tie it through a string of about 160 cm (a twine used when pruning and pruning fruit trees), and swing it around the looped part. A tube with a string having a centrifuge radius of about 80 cm was prepared.
Branches were collected from adult Japanese pear (variety: Kosui) planted in the field of Fruit Tree Research Institute, and 5 branches cut to about 7-10 cm were placed in a conical tube.
It was confirmed that about 0.5 mL (sufficient amount to be used for the above measurement) of conduit liquid could be collected at the bottom of the tube when it was shaken for 1 minute at a rotation speed of 150 rpm with a string tip.
From the results, it was shown that the conduit liquid can be easily and immediately recovered without using any special equipment at the cultivation site.

[Example 7] “Application to various low-temperature demanding deciduous trees”
In the above examples, sorbitol in the conduit fluid that serves as a determination index for the spontaneous dormancy-wakefulness period is the main sugar of the invert sugar in the Rosaceae plant.
Therefore, in other low-temperature demanding deciduous trees, we verified whether it is possible to determine the spontaneous dormancy-wakefulness period using the value of the commutation sugar content in the duct fluid and the degree of increase as an index.

(1) Measurement of sugar content in conduit fluid In addition to Japanese pears of the genus Rosaceae (variety: Kosui) studied in the above example, apples are low-temperature-required deciduous trees belonging to the same Rosaceae family as pears Apples of the genus (variety: Fuji), peaches of the peach genus (variety: Akatsuki), oysters of the genus Oysteraceae (variety: Ehime), which are different in the type of invert sugar from the rose family, and other than angiosperms As a plant, we investigated ginkgo biloba ginkgo (growth: Kusu), which is a gymnosperm plant (see Fig. 5).

From these five types of low-temperature-required deciduous trees (all of which are planted in the field of the Fruit Tree Research Institute), branches are collected at intervals of about 3 weeks from the spontaneous dormancy period to the dormancy awakening period of each plant. did.
The collected branch was cut to about 5 to 10 cm, put into a centrifuge tube, and centrifuged at 3000 rpm (1,000 × g) for 10 minutes to collect the conduit liquid accumulated at the bottom of the centrifuge tube.
About the obtained conduit | pipe liquid, after refine | purifying with a cartridge column, the total sugar content was measured by HPLC. In addition, the content of main sugars of the invert sugar of each plant was also measured.
Here, the main sugars of the invert sugar are sorbitol in the Japanese pears, apples, and peaches, sucrose in the oysters in the oyster family, and glucose in the ginkgo biloba. The measurement results are shown in FIG.

(2) Calculation of sprouting rate Part of the above-collected branches are cut to about 10-30 cm so as to include the tip part, and the cut part is inserted into water, and 25 ° C., 12 hours a day in an artificial weather device. Under the long condition, the heating treatment (constant temperature treatment) for 21 days was performed.
After the heating treatment, the ratio of the buds that reached a state where the scales were loose and green (buds having 4 or more germination stages shown in FIG. 6) among all the buds of the branches was calculated and used as the germination rate.
Here, when the germination rate is 0.5 (50%) or more, it can be determined that the branches subjected to the germination treatment have surely reached the spontaneous dormancy awakening period.
As buds here, flower buds were used for rose family plants (Japanese pears, apples, peaches), and leaf buds were used for other deciduous trees (oysters, ginkgo).
The calculated germination rate values are shown in FIG.

(3) Results ・ Content of the main sugars of tumble sugar As a result, the bud germination rate was maintained as low as 0 (0%) in all of the five types of low temperature requirement deciduous trees investigated (spontaneous dormancy period) At a certain time), the content of the main sugar of the commutated sugar was also kept low.
However, it became clear that the content of the main sugars of the commutated sugar also increased rapidly, coincident with the time when the germination rate of the buds increased rapidly.
And when the germination rate reached 0.5 (50%) or more (a time when the spontaneous dormancy awakening period was reached), the content of the main sugars of the invert sugar was also high.

In addition to the main taxonomic groups of angiosperms (Authentic Dicotyledons), these target plants include the rose family of real roses (Japanese pears, apples, peaches), and the true asteraceae family of oysters (oysters). It includes ginkgo of the ginkgo biloba which is a gymnosperm which is an outer group of the whole angiosperms.
Therefore, the knowledge about spontaneous dormancy awakening is recognized as a property common to all seed plants (a taxonomic group consisting of angiosperms and gymnosperms) (common derived traits) (see FIG. 5).
From this, it was recognized that it was possible to determine the spontaneous dormancy wakefulness period of all low-temperature demanding deciduous trees using as an index the content value and the degree of increase of the commutated sugar main sugar in the conduit fluid.

-Content of total sugar in tumble sugar The content of total sugar in tumble sugar in the conduit fluid was also shown to rise sharply in line with the increase in germination rate. In addition, the rate of increase was shown to increase more sensitively than the main sugar content.
From this, it was recognized that the spontaneous dormant arousal period of all low-temperature-requiring deciduous trees can be determined by using the content value and the degree of increase of the total sugar in the conduit fluid as an index.

[Example 8] “Judgment using the non-major sugars of invert sugar as an index”
We verified whether it is possible to determine the spontaneous dormancy / wakefulness period even when the non-major sugar of the commutating sugar (the commutating sugar having the second highest composition ratio) is used as an index among the constituent sugars of the commutating sugar.

(1) Measurement of sugar content in conduit fluid In addition to peach (variety: Akatsuki), oyster (variety: Ehime), ginkgo (variety: Kusu) investigated in Example 7, A survey was conducted on (variety: Polotan).
The collection of the conduit fluid and the measurement of the content of commutation sugar were performed in the same manner as described in Example 7 (1). The commutated sugars measured here were those having the second highest composition ratio.
The second most common invert sugar was sucrose for peaches, glucose for oysters, fructose for chestnuts, and fructose for ginkgo biloba. The measurement results are shown in FIG.

(2) Calculation of germination rate The germination rate was calculated in the same manner as in the method described in Example 7 (2), and the measurement results are shown in FIG.

(3) Results ・ Contents of the main sugars of tumble sugar As a result, the bud germination rate was kept low at 0 (0%) in all four types of low temperature requirement deciduous trees investigated (spontaneous dormancy period) At the same time, the content of commutating sugar with the second highest composition ratio was kept low.
However, after that, it became clear that the content of the sugar also increased in line with the time when the germination rate of buds rose rapidly.
And when the germination rate reached 0.5 (50%) or higher (a time when the spontaneous dormancy awakening period was reached), the sugar content was also high.
Based on this, it is possible to determine the spontaneous dormancy-wakefulness period of low-temperature demanding deciduous trees using as an index the content value and the degree of increase in the content of non-major sugars in commutation sugars (the commutation sugars with the second highest composition ratio) in the conduit fluid. Was suggested.

By this invention, it becomes possible to grasp | ascertain correctly the covering warming time of a deciduous tree for every tree. Therefore, it is possible to carry out early cultivation at a higher unit price while ensuring high quality and yield at the cultivation site for farmer's fruit and flower production.
It is also expected to contribute as a technology that significantly reduces the risk of misrecognizing the spontaneous dormancy awakening period due to the effects of climate change and global warming.

Claims (14)

  A method for determining a spontaneous diapause wakefulness period of a low-temperature demanding deciduous tree, characterized by determining whether a spontaneous diapause wakefulness period has been reached using the content of commutating sugars in the conduit fluid as an index.   2. The determination method according to claim 1, wherein the content of the invert sugar is a total content of all sugars constituting the invert sugar.   2. The determination method according to claim 1, wherein the content of the invert sugar is the content of a main sugar of the invert sugar.   The determination method according to any one of claims 1 to 3, wherein the low-temperature required deciduous tree is a low-temperature required deciduous tree belonging to a seed plant.   The determination method according to any one of claims 1 to 4, wherein the low-temperature required deciduous tree is a low-temperature required deciduous tree belonging to angiosperms or Ginkgo biloba.   6. The determination method according to any one of claims 1 to 5, wherein the index uses a rapid increase in the content of the invert sugar as an index. For the Japanese pear as the low-temperature-requiring deciduous tree , the determination is that the total content of all sugars constituting the invert sugar, or the content of the main invert sugar is 21 mg / mL or more, spontaneous dormancy awakening The determination method according to claim 1, wherein it is determined that the period has been reached. The determination is that for the Japanese pear as the low-temperature demanding deciduous tree, it is determined that the spontaneous dormancy awakening period has been reached when the following conditions (A) and (B) are satisfied. The determination method according to any one of the above.
(A) A condition in which the total content of all sugars constituting the invert sugar is 17 mg / mL or more.
(B) A condition in which the total content of all the sugars constituting the invert sugar is at least twice the content of the invert sugar at the time described in (C) below.
(C) The time when the buds in which the bud scales are loose and appear green when the temperature is maintained at 25 ° C. for 12 hours for 21 days are 0%. For the Japanese pear as the low-temperature demanding deciduous tree , the determination is that it is determined that the spontaneous dormancy arousal period has been reached when the following conditions (D) and (E) are satisfied: The determination method according to any one of the above.
(D) A condition in which the total content of all sugars constituting the invert sugar is 6 mg / mL or more.
(E) A condition in which the total content of all the sugars constituting the invert sugar is at least three times the content of the invert sugar at the time described in (F) below.
(F) The time when 0% of the buds are in a state in which the scales of the buds are loose and green when subjected to isothermal cultivation for 21 days at 25 ° C. for 12 hours. The determination method according to any one of claims 1 to 6 , wherein the low-temperature-required deciduous tree is a low-temperature-required deciduous tree belonging to the family Rosaceae; and the invert sugar main sugar is sorbitol.   The conduit fluid is recovered by centrifuging a branch taken from the low temperature demanding deciduous tree; or by pressurizing one end of a branch taken from the low temperature demanding deciduous tree, the other 1 1. Collected from one end; or collected from one end of the branch taken from the cold-requiring deciduous tree by suction from one end; The determination method of crab.   12. The determination method according to claim 1, wherein the determination is performed by estimating the content of commutating sugar from the Brix indication value of the conduit fluid.   12. The determination method according to claim 1, wherein the determination is performed by estimating the content of commutated sugar from the value of the total organic carbon content of the conduit fluid.   12. The determination according to any one of claims 1 to 11, wherein the determination is performed by mixing the conduit liquid and the potassium permanganate solution and estimating the content of reducing sugar from the color tone after the color reaction of the mixed liquid. The determination method of crab.