Cucket No. PNA-,3C54-PcT SPECIFICATION ADVENTITIOUS ROOT FORMATION ACCELERATOR, MEDIUM FOR ROOTING CONTAINING THE ADVENTITIOUS ROOT FORMATION ACCELERATOR, AND 5 METHOD OF PRODUCING CLONE SEEDLING USING THE ADVENTITIOUS ROOT FORMATION ACCFERATOR TECHNICAL FIELD [o0: The present invention relates to an adventitious :0 root formazion accelerator, a medium for rooting comprising the adventitious root formation accelerator, and a method of producing a clone seedling using the adventitious root oration accelerator. 15 BACKGROUND '0002 A cutting method, in which a root is Cenerated by planting a scion into a seedbed or a medium and a tissue culture setnod in which a plant tissue is cultured, a shoot is colected and a root :s generated from the shoot, are 20 utilized as a mass production procedure of a Dlant body for producing the homogeneous plant body (seedling) having a trait that meets a purpose in a large amount in fields of agricultural production, forestation, oreeding and others. A rooting ability of the plant tissue has a great effect o0: 25 productivity of clone seedings in these methods. Thus, is important to enhance the rooting ability. 0103: Japanese Patent Aolication Laid-open No. 2001 231335-A (Patent iterature ) describes that the --- ting ability is oromoted by treating a mother tree for taking 33 scions selected from giants belonging to genera Eucaiyptus or Acacia witn paciobutrazol before being employed as a scion. [0304: Meanwhile, Japanese Patent Application Laid-open Docke: No. PNPA-'335--P37 2 No. 2003-1>48-A (Patent Literature 2) describes that a compound having a particular structure has an action to inhibit biosynthesis of abscisic acid in the plant and is useful for regulation of growth and development of the plant. Japanese Patent Aoolication Laid-open No. 2006 117608-A (Patent Literature 3) describes that the compound described in the Patent Literature '2 can nrevest a disease damage of monocotyledons. 10 RELATED ART DOCUMENTS PATENT LITERATURE [0005] Patent Literature 1: Japanese Patent Aolication Laid-open No. 2001-231355-A Patent Literature 2: Jaoanese Patent Apoication laid-open 15 No. 23-i13148-A Patent literature 3: Japanese Patent Application Laid-open No. 2006-117608-A SUMARRY OF TUE INVENTION 20 PROBLEM TO BE SOLVED BY THE INVENTION [OO6 However, when paclobutrazol described in atent Literature i, is used, the scion can be obtained only after several months have passed after treating the mother tree for talking scions with it. Thus, a rapid-acting roCting 25 ac elerator has been required. 0027: The rooting ability in plants is insufficient in the method described in Patent Literature 1. O2 these, when the method is anplied c to plants originally having the low rooting ability such as plants belong 4 ng to genus 30 Eucalyptus, improvement of the rooting ability could not Ce anicipated -. 0008: eanwhi.e, technology described in Patent Literature 2 is technology in which abscisic aid- ha Docket No. PNPA-' 305T rclates :ith growth of plants is controlled, and is different from technology to promote the rooting of the SCion such as the cutting ethod, i.e., promote differentiation alone of a root tissue. A condition to 5 promote the rooting from the scion is generally different from a condition to promote the growth of plants, and a condition to stop the growth often rather promotes 7he rooting. Technology described in Patent Literature 3 is technology concerning the disease damage of monocotyledons, 1C particularly prevention of rice blast, and is different :rom technology concerning the scion and technology concerning the rooting from the scion. [DCC9 -t is an object of the present ilnventiof to provide a compound useful as an active ingredient of an 15 adventitious loot for-ation accelerator that promotes rooting from a olan and enhances its rooting rae is another object of the present invention to provide a rooting method and a meciun for the rooting utilizing the adventitious root formation accelerator comprising the 20 active ingredient in order to enhance a oroductivity of clone seedlings, particularly clone seedlings belonging to plant species having a .. ow rooting ability by methods such as a cutting method and a tissue culture method. 25 MEANS FOR SOLVING PROBLEM 001C] The present invention provides following [: to [4]: 1] An adventitius roo: formation accelerator for a pliant, comprising: abamine. 30 [2] A medium for rooting from a shoot of a plant, comprising: the adventitious root formaton acceerator according to [I]. [3] A method of producing a clone seedling, coorls ing: Douket No. PNPA-' 3 C cu..tIvating a shoot of a nlant in the presence of the advenritious root formation accelerator according to :1]; and generating a root tro:n the shoot. [4] A method of producing a clone seedling, comprising: 5 cultivation a shoot o a plant in the medium for rooting according to [2].; and generating a root from the shoot. EFFECT OF THE INVENTION 001'.] According to the present invention, formation of 10 an adventitious root from a olant can be promoted and a rooting rate can be enhanced. Thus, productivity of clone seedlings can be enhanced. Further, such an effec: is particularly prominent in plant species having low rooting ability. Therefore, the present invention can contribute 15 to mass and rapid Produczion of the cLcne seedings from broac an various plant species, and particularly enables to abundantly and rapnidy produce the clone seedlings even from :he plant species having the lcw rootino abiliv. Thus, it is anticipated to lead a way to its industrial 20 utilization. E-MODIMETS FOR CARRYING OUT THE INVENTION 100121 The adventitious root formation accelerator for the plant of the present invention compcrises abamine as an 25 active ingredient. C313: Abamine is [ '3- (3,4-dimethoxyphenyl) -alyl]- (4 fluorobenzyl) --amino] -acetate methyl ester, and has a structure represented by a following formula (1 Chemical 1P 3cc< No. PNPA-'3054 PCI F -- *(1) N 0 O OCHa 03142 An origin of abamine is no: particuIarly limied, and for example, abamine derived from a natural product and abamine syntnesized by chemical reactions may be used. 5 Examples cf the method of synthesizing abamine _V the cnemical reactions may include the method described in references such as Japanese Patent Application Laid-open No. 20 0 3e- 1314 8r-A. '3C:5: &hQavntiticUS root formation accelerator of -0 the present inventicn only has to comprise abamine, and may combine other ingredients (eg., other adventitious root orationn accelerator, activity accelerator of plant hornone concerning the adventitious root formation, such as auxin'r, if necessary, as long as tneir action is not 15 contrary to the object of the presen: invention. Examples of the other ingredients may include the adventitious root formation accelerator and the auxin activity accelerator described in the SPecification of Japanese Patent Application No. 201C-31641 and the adventitious root 20 formation acceler:-t and the plant hormone activity acceleratCr described in the Specification of Japanese Patent Asplication No. 2010-101642. C016, The adventitious root formatio: accelerator of -he present invention: exerts an effect of promoting the Jcc~e No. PN/-05.PCT 6 adve4titious root formation adding I in cul:ivati on of the plant. [0017] Types of the plants are not particularly limited. The plants oan be classified into woody plants and 5 herbaceous plants. The adventitious root formation accelerator of the present invention is applicable to any of them, preferably applied to the woody plant, and more preferably applied to the woody plant having the rcotino ability inferior to the herbaceous plant. Examples of the 0 woody plants may include: plants belonging no genus Eucalyptus; pants belonging to genus Pinus; plants belonging to genus Cryptomeria such as Japanese cedar (Cryptoneria japonica, Sugi) ; plants belonging to genus Prunus such as flowering cherry (Prunus spp), Japanese 15 apricot (Pr unus muwe) and downy cherry (Prunus toxentosa) plants neiongisg to genus Mangifera such as mango (Mangit4era india); plants belonging to senus Acacia; plants belonging to genus Myrica; plants belonging to genus Quercus such as sawtooth oak (Quercus acu:issima ; plants 20 belonging to genus T/itis; plants belonging no genus Mazus; plans belonging to genus Rosa; plants belonging to genus Camelia such as tea plant (Camellia sinensis) ; plants belonging to genus Jacaranda such as jacaranda (Jacaranda miosifoliia) ; plants belonging to genus Persea such as 25 avocado (Persea america-a) ; plants belonging to genus I'yrus suh as pear (Pyis serotina Rehder, Pyrus pyrifolia) ; and plants belonging to genus Santalon such as sandao wood (Santalum album) . Among them, when -he present inventor is applied to eucalyptus, pine, Japanese cedar, flowerin 3C cherry, mango, avocado, acacia, wax myrtle, sawtooth oak, grape, apple, rose, camellia, tea olant, Japanese apricot, downy cherry, jacaranda, and the like, the effec: of the present ivention is further exerted. n particular, the DoJe! No. NPA P'T plants belonging to genera Eucalyptus, Pinus, Cryptomerza, Camellia, Mangifera, and Persea are preferable. More preferable are for Eucalyptus, pine, Japanese cedar, tea plant, mango and avocado known as their poor rooting 5 ability, and still more preferable are for the e"calypnts and the tea plant. !Col] As Eucalyptus plants, Eucalyptus known as the poor rooting plant is preferable, and Eucalyptus globulus and Silver Dollar Gum are more preferable, and Eucalyptus 10 globulus is still more preferable. [0019] Examples of herbaceous plants to which the adventitious root formation accelerator of the present invention is applicable may include, but are not limited to, plants belonging to families Cruciferae, Solanaceae, 15 Poaceae, Fabaceae, and the like. As described above, the present invention can be applied to food productive pLants that include grains such as rice and wheat, bears and maize together with vegetables such as Solanaceae plants, and thus is greatly anticipated in terms of food productive 20 technology in future. Among the plants belonging to families Cruciferae, Solanaceae, Poaceae, Fabaceae, and the like, Arabidopsis thaliana, Nicotiana tabacum, Oryza sativa, and Lotus corriculatus var. japonicus are broadly and co:conly used as model plants, and thus are greatly 25 anticipated to contribute to progress of research and deveicpment. 0021 The plant may be a whoLe plant body or a part thereof, but is typically the part of the pLant body and preferablvy a shoot in terms of expecting to form the 30 adventitious root. 0021: The shoot refers to a entire :issue having the ability to generate root. Examples of the tissue may include branches, stems, apical buds, axillary buds, Docke: No. :NPA- 3354-PCIT adventitious buds, leaves, cotyledons, hypocotyls, adventitious ge-ms, and shoot primordia, and the rie An oricn of the shoot is not particularly limited, may be a tissue obtained from a plant individual grown in a green 5 house or outdoor, may be a cultured tissue obtained by a tissue culture -ethod, or may be a part of tissue from a native plant body. The shoot can be acquired efficiently from a mother plant for the scion or a multiple shoot. Among them, the scion (scion obtained from the mother .0 plant; , the multiple shoot obtained by sterilaly culturing an organ collected from the mother plant, or stems and leaves obtained by sterilely culturing the above orcan are preferable. [0C22: The sulziple shoot can be induced by cutting off 15 a tissue such as an apical had or an axillarv bud from a planr from which a clone seedling is to be produced by applying the present invention, and cuizuring this. The multiple shoot may be formed by sterilely culturing the organ collected from the mother plant according to the 2C method and the condition described in Japanese ?atent Application Laid-open No. Hei-8-228621-A. The method and condition are approximately as follows. First, a tissue of a bud such as an apical bud or an axillary bud is collected from a plant that is a material, and the surface of the 25 collected tissue Is sterilized by itmersing the tissue in a sodium hyochilorite aqueous solution in which an effective amount of chlorine is about 0.5% to about 4% or a hydrogen peroxide aqueous solution in which an effective amount of chlorine is about 5% to about 13% for about 13 minutes to 3C about 20 minutes. Subsequently, this tissue is washed with sterilized water, and put into a solid medium to open a boll of the bud. Then the multiple shoot is formed by sub culturing growing stems and leaves ir medium having the ?oskc: No. PNPA-13054-PCT same coWosition. When the tissue suc as tne axillarv bud from genus &caLypus or Acacia is used, it is preferable :o use Yurashige and Skoog (hereinafter abbreviated as MS) medium containing 1 to 5% by weigh: of sucrose, about 0.C2 5 mg/L or more and about 1 mg/L or less of benzyladenine (hereinafter abbreviated as BA) as a plant hormone, about 0.2% by weight or more and about 0.3% by weight of gellan gu2 or 0.5% by weight or more and about 1% by wveight or less of agar, or modified MS medium in which contents of 10 anmonium nitrate and potassium nitrate are reduced by half as the solid medium. The shoot actively extends iro. the multiple shoot thus formed. The multiple shoot itself can be maintained and grown by appropriately dividing and oulturing in the same mediun as that used for formation of the multiple shoot. 0023: Meanwhile, the scion may also be used as the shoot. Typic a u y, the adventitious root formation accelerator exerts its effect when given to the scion. The scion may be at least a cart of a plant. Examples of the 20 scion may include: branches such as green branches (branches extended in a current year) and hard branches (branches extended before the previous year); buds such as apical buds and axillary buds; leaves and coyiedons; and hypocotyls. In the case of the woody plants, the green 25 bra5c5 and the hard branch are typically used for the scion and in the case of herbaceous plants, the leaf and the ad are typically used for the scion, but the scion is not iuited thereto. 0024 A method of cultivating the plant in the presence 30 of the adveztitious root formation accelerator of the present invention is not particularly limited, and can be appropriately selected depending on the type, a site, and a condition of the plant. Specifically example of the method Docket No;. PNPA-'3054-POT may include the fcliWng methods: a method or Culturing toe plant (preferably ne shoot) in the medium for rooting continuing the adventitious root formation aCc(ezeator; and a method of contacting a solution containing the adventitious roct formation accelerator with the plant (preferably the shot) - When the cultured tissue obtained by the tissue culture method is used as the shoot, the former is preferable, and when the scion is used as the shoot, both of the former and the latter are preferable. 10 I is of course possible to combine both the methods shown as examples above, i.e., to employ the method of contaclin the solution containing the adventitious root formation accelerator with the plant while the plant is cultured in the medium for rooting containing the advent tious root formation accelerator. When the plant is cultured in the medium for rooting containing toe adventitious root cormation accelerator, a concenttatio Of t-e adventitious root formarion accelerator in the medium for rooting is preferably about 20 0.01 uM or more and about 100 MaY or less, more preferabov about 0.1 l'M or more and about 10 MY or less, and partcru.arly preferably about 0.5 M or more and about 5 uM or less. 0025: When the plant (preferably the shoot) is 25 contactecd with the solution containing the advensitious root formatton accelerator (adventitious root formation accelerator solution),; a .*ethodi for the contact is not particularly rimitedC, and may be appropriately selected openrding on the cype, the site, the condition, and a 30 cuslivation method of the plant. Exaples such a method may include a method of spraying the adventtious root oration accelerator solution directly to the shoot, and a Dooke No. PNPA-'.3054-PT znet::od of wetting the support with the adventous roo: formation accelerator solution. [00262 The adventitious root formation accelerazor solUti-on may be prepared by dissolving the acventitious 5 toot formation accelerator in an appropriate solvent such as water. Exanoles of the water may include deionized water, distilled water, reverse osmotic water, rap water, and the like, and any of them can be used. The concen:ranion of the adventitious root formation 10 accelerator in the adventitious root formation accelerator solution is preferablv about 0.01 pM or more and about 130 M or less, more preferably about 0.1 gM or more and about 10 L or less, and still more preferably about 0.5 uM or more and about 5 MtN or less. 15 :0027 When the adventitious root torai accelerator solution is sraved directly to the plant (preferan y the shoot:, the adventitious root formation accelerator solution may be misted onto a oar: of or all of the plant using a soray. The amount of the advenritious root 20 formation accelerator solution to be sprayed depends on the concentration of the adventitious root formation accelerator in the adventitious root formation accelerator so.t:ion, and car not be defined necessarily, but is preferably about 0.5 ml or more and about 5.0 -l or less 25 and more preferably about 1.0 cL or more and about 3.0 mL or less per one shoot in general. A spray frequency may be once or twice or m:re, but it is preferable to at -east spray upon stating the cultivation. Further dependina on a cultivation condition, the solution may be sprayed 20 additionally and. appropriately (e.g, every several days (2 to 3 days) during the cultivation period. C028: Example of a method of wetting the support with Dncke& N,. 9kMA-I3054"CT 12 the adventitious toot formation accelerator solution may include a method of sprinkling the adventitious root fortarion accelerator solution from above the supoort anc a method of placing the support on a bottom of a vessel 5 filled with the advenritious root formation accelerator solution, which is then poured from a bottom surface. W7hen the adventitious root formation accelerator solution is sorinkled from above the support, the amount of -he adventitious root formation accelerator solution to be 1 sorinkled from the above is preferably about >.0 ml or more and about 19 ml or less and more preferably about 3.0 ml or more and about 5.0 ml or less per one plant (preferably one shoot). When the adventitious root formation accelerator solution is poured from the bottom surface, the 5 adventitious root formation accelerator solution may substantially evenly wet the support. When the support is wetted with the adventitious root formation accelerator solution, the medium for rooting is separately prepared in addition to the adventitious root formation accelerator 20 solution, and the support may be wetted with the both of them. This is as described above. [0029] In the present invention, the medium for rooting means a mediuw used for generating a root from the plant (preferably the shoot). The medium for rooting preferably 25 contains silver ion and/or an antioxidant, and more preferably contains both the silver ion and the antioxidant. The silver ion may be added as a silver compound (silver ion source) such as silver thiosulfate (SS, AcS 4 Os) and silver nitrate into the medium. Among them, SITS is 30 preferable as the silver ion source used in the present invention because when added to the medium to cult-ure the shoot, SITS facilitates heahv roOting and/or extension of the root. It is thought that this is because the silver locke: No. PNPA- 305%-PCI on derived from STS takes a form of thiosulfare silver ion and is negatively charged in the rnedizm. A concentration of the silver ion to be added in the medium for roting varies cepending on the tvne of the silver ion and the 5 other culture condition, but the concentration as the silver ion source is preferably about 0.5 tM or more and about & uM or less and more preferablv about 2 MiY or less and about 6 I or less. [003C] Meanwhile, publicly known antioxidants such as 10 ascorbic acid and sulfite salts may be used as the antioxidant. Among them, the ascorbic acid is preferable as the antioxidant used in zne present invennicn because its residual effect in the medium is low. The concenration of the antioxidant to be added to the medium -or rooting is preferably about 5 mg/L or more And about 200 :g/L or less and more preferably about 20 mg/I or more an -about I00 mg/L or less. 10031: The media for rooting used in the present invention may include inorganic ingredients, carbon sources, 20 vitamins, amino acids, plant hormones, and the like in addition to the above ingredients. 0032: Ex amples of the inorganic ingredient may include elements sucs as nitrogen, phosphorus, potassium, sulur, calcium, magnesium, Iron, manganese, zinc, boron, 2 molybdenum, chlorine, iodine, and cobalt, and inorganic salts containing one or more selected from these elements. Examples of -he inorganic salts may include potassium nitrate, armonium nite,e, anium chre, sodium nitrate, potassium monchydrogen phosphate, sodium 30 dihydrogen phosphate, pozassiumn chloride, magnesIum sul:ate, serrous sulfate, ferric sulfate, manganese sulfate, zinc sulate, copper sulfate, sodium sulfate, calcium chloride, Docket No. PNPA- 3054-CT magnesi L cnloride, boric acid, molybdenum tricxide, lsuod molybdate, potassium iodide, and cobalt chloride, and hydrates thereof. From the above examples, one or any combnnation of two or more may be selected to use as the 5 inorganic ingredient. [0033] :z is preferable to contain nitrogen, phosphorus, and norassium as essential elements in the medam for rooting used in the present invention. Thus, nitrogen, phosphorus, potassium, the inorganic salts con-aining 10 nitrogen, the inorganic salts containing phosphorus, and the inorcanic salts containing potassium are preferable, and nitrogen, phosphorus, potassin, and the inorganic salts containing nitrogen are more preferable among the specific examples of the above inorganic ingredients. When 15 one inorgan'c ingredient is added, its concentration in he ei r rooting is preferably about 0. uY or more and about 103 mM or less and more preferably about I Y cr more anda aout 133 mM or iess. When two or more incredienzs in combination are added, the concentration of each ingredient 20 in. the medium for rooting is preferably about 0.1 X or more and about 100 mY or less and more preferably about 1 WM or more and about 100 mM or less. :00341 Carbohydrates such as sucrose and derivatives -hereof; organic acids such as fatty acids; primary 25 alcohols such as ethanol can be used as the carbon source. From the above specific examples, one or any combination of two or more may be selected to use as the carbon source. The amount of the carbon source to be added into the meiTum for rooting is preferably about - g/I or more and about 100 3S a/ or less and more preferably about 10 g/L or more and about 10 c/ or less. However, when the plant is cultured with supplying carbon dioxide gas, the medium need not 1ockt No. PN 0 4 354-POC containthe Carbon source and it is preferable to contain no carton source. The organic comoud such as sucrose capable of being the carbon source is also the carbon source for microorganisms. Thus, when the medium 5 containing the organic compound is used, it is necessary to culture under a sterile environment. '-owever, by using the medium containing no carbon source, it becomes possible to culture under a non-sterile environen:. [0035] Vitarins such as biotin, thiamine (vitamin B?), 10 pyridoxine (vitamin. 34), pyridoxal, nyridoxamine, calcium pantothenate, inositol, nicotinic acid, nicotinate aide, and/or riboflavin (vitamin B2) may be used. Fro: the above specific examples, one or any combination of two or more .ay be selected to use as the vitamin. When one vitamin is added, its concentration in the medium= for rooting is Dreferablv about 0.01 mg/ or more and about 20C mg/L or :ess and more preferably about '.02 :g/t or more and about 100 nc/P or less. When two or more vitamins in comabination are added, the concentration of each vitamin in the medium 20 for rooting is preferably about 0.01 mg/L or more and abour 150 mg/I or less and more preferably about 0.2 mg/I or sore and about 100 mg/L or less. 0036' Amino acids such as glycine, alanine, glutamic acid, cysteine, phenylalanine, and/or lysine may be used. 25 From the above specific examples, one or any combination two or more may be selected to use as the amino acid. he. one Amino acid is added, its concentrazion in the medium for rooting is preferably Dbous 0.1 mg/or re and about 1000 mg/I or less. When two or snore amino acds in 20 combination are added, the oo::centration or eacs amino acid in the medium for rooting is preferably about 0.2 mg/L or more and about 1000 mg/Ior less. [0037) The plans hormones such as auxins and/or Cocked ro. MNPA-13054I 'I cymokinins may be used. Examples of auxins may include naphthaleneacetic acid (NAA), indoleacetic acid (AA), p chlcrorphenoxyacetic acid, 2,4-diohlorophenoxyacezic acid (2, 4D), indolebutyric acid (TBA), and derivatives thereof. Of 5 these, one or any combination of two or more may be selected from them to use. Examples of cytokinins may include benzyladenine (BA), kine:tin, zeatin, and derivatives thereof. Of these, one or more or any cobi.onation of two or more may be selected to use- Auxins -0 alone, cytokinins alone, or the combination of auxin ard cytokinin may be used as the plant hormone. When one plant hormone is used, its concentration in the medium for rooting I-s preferably about 0.01 mg/I or more and about IC mg/L or less and more preferably abouz 0.02 mg/L or more .5 ana about 10 mg/L or Less. Whe>n two or more plant hormones are used, the concentration of each plant hormone i' the medium for rooting is preferablv about 0.01 mg/L or more ano about 10 mg/l or less and more preferably about 0.02 mg/I or more and about 10 mg/L or less. 20 :0038] :n the present invention, the publicly known medium as 7he medium for culturing the plant tissue may be used as the medium for rooting. The adventitious root formation accelerator, the silver ion and/or the antioxidant, the carbon source, or the olar horwone may be 25 added to the medium approriatey if necessary. Examples of the known medium as the mecium for culturing the plant _issue can include MS eiua, Iinsmaier and skoog medium, White medium, Gambcrg 3-B medium, ard Nisch and Nitsch medium. MS medium and Gam.borg B-5 mediur. are preferable 30 among Them. These media may be apropriately diluted if necessary for the use. [0039] The medium for rooting may be a liquid medium or a socid medium, but the iquid medium is Dreferable in Dock: No. PNAS1304PCT tenrs of work efficiency and less scratch of roots pocn transplant. The liquid medium can be prepared by mixing a medium composition and directly used. The solid medium can be used by solidifying with a solidifying agent such as 5 agar and gellan gum simultaneously with or after preparing by mixing The medium composition as with the liquid medium. The amount of the solidifying agent to be added to the medium varies depending on the type of the solidifying agent and the composition of the medium. When the solidifying agent is agar, the amount to be added is preferably 0.5% by weight or more and 1% by weight or less. When the solidifying agent is gellan gum, the amount to be added is preferably 0.2% by weight or more and 0.3% by weight or less. 15 13401 A method of cutting a outtage of the plant (preferably the shoot) in the medium for rooting can be appropriaoely selected depending on a clture condition sunch as the type of the medium. When the medium for rooting is the solid medium, a base of the shoot may be 20 directly put in the medium for rooting and cultured. meanwhile when the medium for rooting is the liquid medium, the base of the shoot may be put in the support wetted with the medium for rooting as described later to culture the shoot, it is also preferable for enhancing a rooting race 25 to give a physical stimulation such as a scratch the base of the shoot when the shoot is put in the medium for rooting. The base of the shoot means one end of the shoot, which is a region where the root is formed ,chat is, side opposite to an end where the leaf is formed. When the 30 multiple shoot is used as the shoot, the base of the shoot is a region having a cout surface produced when the ultiple shoot is divided. A size (largeness, shape, and the like) of a scratch on the base of the shoot is not particularly Jocket No. PNW~ 30%PCT L8 limi ed. For example, when the multiple shoot is used as the shoot, it is preferable to make a scratch like a cross snape wien the base off the shoot (aforementioned cut surface) is seen from a front direction. A tool such as 5 scissors or a knife can be used for making the scratch. [0041] The support in the present invention is the support to support the plant (preferably the shoo:). When rte medium for rooting (particularly solid median) is used, no support is needed in some cases, but in the case other 10 than it, the support is typically utilized. 0042' The suoorr in which the shoot may be kept to be put during the period of the cultivation is preferable. When the liquid mediim for rooting is used for the cultivation, tse liquid medium is typically used by wet ing 15 the support wih it. Tus, uhe sucort capable oi bein wetted with the liquid is preferable, and among them, the support capable of being substantially evenly wetted with the adventitious toot formation accelerator solution or the qluid median containing the adventitious root formation 20 accelerator is preferabLe. When the liquid medium is used as the medium for rooting, the liquid medium containing no adventitious root formation accelerator solution and the adventiticus root formation accelerator solution may be added separately to the support, or the previcusly prepared 25 liquid medium cotnaining the adventitious root formation accelerator may be added to the support. The support conventionally and commonly ussed can be used as the support, which is not particularly limited. Examples I he suport may include natural soils such as sands and Akadama; 30 artificial soils such as carbonized chaff, coconut fiber, vermicuwite, pearlite, pea7 moss and glass beads; and porous molded articles such as foamed phencl resin and rock wool. Such a support mav be placec in a culture vessel and DOcket No. PNPA-1354-PCT 19 wetted with the adveniiovus root formation accelerator solution or the licuid medium containing the adventitious root fOration accelerator to prepare a rooting bed. When the medium for rooting is the solid medium, the rooting bed 5 may be prepared by directly placing the solid medium in the culture vessel. (0o43] in the present invention, the culture vessel for housing the medium for rooting or the support may be used. The culture vessel conventionally and commonly used can be 10 used as the culture vessel, which is not particularly limited. Examples of the culture vessel may include pots for raising seedlings and plug trays. The culture vessel may be a closed type or an opened type, but the closed Type culture vessel is preferable. The sprayed adventitious 15 root formation accelerator solti on can be retained by using the closed nype of the culture vessel. it also becomes easy zo keep the humiiny in the environment surrounding the shoot and a clone seedling formed therefrom. [0044: When a branch is used as a shoot, it is 20 preferable to use the closed type of the culture vessel as the culture vessel. This makes i easy to place the shoot under the high humidity. Thus, transpiration from leaves on the branch is reduced, and a partial removal of leaves, which has been conventionally performed, may be oitted. 25 :0045- More preferably the carbon dioxide gas can be supplied into the culture vessel. Example of the vessel may include a vessel having an opening covered with a carbon dioxis-permeable membrane. The numidiny in the culture environment can also be controlled easily by the 30 use of The vessel having the opening covered with the carbon dicxide-permeabie membrane. A shape of the opening is not particularly limited. A material for the carbon dioxide-permeable membrane is not particularly limited, and Docket No. PNA'3i4OC7 22 examples of the cerial may include polytetrafluoroethylene and the like. A pore size of the membrane is not particularly limited and examples of :ne membrane may include a re:bzano having the pore size of 5 about 0.1 UM or more and about I LM or les. [C46] A cultivation condition when the plant is cultivated is not partLcularly limited as long as the root can be generated from the plan. The cultivation condition varies depending on the type, the site, the condition of 10 nhe plant, the type of the medium for rooting and the like, and is difficult to be defined complete ly, but, for example, the tertperacure is more preferably about 230C or higher and about 28*C or lower. A light in-ensity is represented as a photosynthetic effective photon tlux density, ard iS preferably about 10 Mnol/m2/s or more and about I mol/m 2 /s or less and more preferably about 50 4mol/:2/s or more and about $00 la/2/s or less. In any case, no rooting from the shoot can be observed in about two weeks or more and within about 5 weeks, 20 :-047] The cultivation is carried out under an irradiation with the light preferably containing a wavelength component of about 650 n or wore and about 673 rm or less and a wavelength component of about 450 nm or more and about 470 lm or less at a ratio of 9:1 to 7:3 and 25 more preferably containing nese wave-ength components a: a atio of 9:1 to 8:2. The rooting frot. the plant 'creferablv the shoot) can further be facilitated by the cultivation under the irradiation with the light contarninn such wavelength components. 33 [0048] Lurther, it is preferable to supply the carbon dioxide gas at typically 330 ppm or more and 2000 popm or less and preferably 800 ppm or more and 1500 ppm or less Docket No PNPA-13054-PCT 21 i e tivation enviro:1en:.Te amount: of th.e carbon dioxide gas to be supplied may be controlled by utilizing an equipment such as an artificial cLimate chamber, a culture vessel having the carbon dioxide-permeable membrane 5 at the opening, and the like. -0049] The humidity can be controlled depending on tre cultivation condition such as the type of the plant. The humidity is typically 50% or more and preferably 60% or more. In the case of the plants belonging to genus 10 ucalyptus, the humidity is preferably 80% or more and more preferably 85% or rore. The humidity within the above range can promote the rooting from the plant-. An upper limit of the humidity is not particularly limited. [0050] Meanwhile, when a scion is used as a shoot, it Is 15 preferaible to shield the light. A light shielding rate is preferably 30% or more and 70% or loss and more preferably 40% or more and 60% or less. L0051> As described above, the adventitious root can be generated from the plant using the adventitious root 20 formation accelerator of the present invention. A plant which has generated the adventitious root is typicallV referred to as a clone seedling. The cultivation is continued for a certain period (dependi no on the condition such as the type, the site, and the state of the plant, and 25 for exarnle, about 10 days or more and 90 days or less in the case of Eucalyptus; using the adventitious root formation accelerator of the -resent invention, the root is grown, and this is transplanted in the vessel for raising the seedling or a nursery and bred to make a seedling 30 capable Qf being used for the prede-ermined purpose such as forestation. The soil, the temperature, and the light intensity upon breeding tne seedling may be set appropriately to be suitable for the plant to be rooted.
Dockot %', PN 0 A-13054-PCT 22 Whe t t is generated from :ne shoo Aeri ved from the cultured tissue such as adventitious bud or shoot primordia, it is typially necessary to pass a process of acclimazion before transplanting to the vessel for raising the seedling 5 or the like. [3052] :Mechanism] In the present invention, the root can be geneted from a shoot of a plant by ouloivating the shoct in the presence of the adventitious root formation accelera-or. A 10 reason for it is unclear. However, since abamine SG has no rooting action in spize of its structural similarity to abamine, it is cresumed that the rooting acceleration effect by the adventitious root formation accelerator would be attributed to no- t'e inhibition of abscisic acid 1: synthesis tha: is the common action of abasise and abamine SG but another action. EXAMPLES :c053] The present invention will be described in more 2C detail below by Examples, [C54: Example 1 and Comparative Example 1: Eucalyptus clone A (G-024) that is a poor rooting strain of Eucailyntus globuLus (hereinafter abbreviated as E. glcbulus simply) was used as a material for a scion. 25 is, a budwood extending to a length o 5 to 22 cm ana having 1 to 3 knots and about 2 to 6 Leaves was cur out from a mother tree for taking scions to prepare a scion. C055: A base of the obtained scion was put into a porous support mace from a foamed phenol resin (trade ra7ie: 30 Oasis suoplied from Smithers-Oasis Co. wetted with 4 tines diluted MS mediu (composition: 412.5 mg/L of ammonium nitrate, 475 ma/L of potassiumt nitrate, 42.52 mg/L of potassium= dihydrogen phosphate, 0.21 mg/L of cotassiurs .Iocke: No. J\A 3:D4-POT 23 iodide, no carbon scarce was added in his medium) to which' 1 wL of abamine and abamine SG (synthesized according to the methods described in Examples 1 to 3 in Japanese Patent Application Laid-open No. 2003-1:3148-A), S M2 STS (AgS 4 0 6 5 as a silver ion source, 50 mo/T of ascorbic acid as an antioxidant, and 2 mg/L of :BA as a plant hormone had been added, and cultured at a carbon dioxide concentration of 1000 ppm at temperature of 2500 under irradiation with red light containing a wavelength comnonent of 650 nm to 670 nm 10 and a wavelength corponent of 450 nm to 470 nm at a ratio of B.2:1.8 and having a photosynthetic effective photon flux density of 51.3 Fjol/m 2 /s for 2 months. A cubic vessel with slightly convexed middle having a maximal dimension of about 0 to 11.5 cm vertical x 12 to 11.5 cm 15 horizontal x 10.0 cm height was used as a Oulture vesseI. A top surface of this culture vessel was provided with one circular opening covered with a membrane havig a pore size of 0.45 pLm and made from colytetrafluoroethylene (trade name: MilliSeal supplied from Millipore). The 20 concentration cf the carbon dcoxide gas in the culture vessel was the concentration of the carbon dioxide gas (about 10C0 ppm) permeated from the membrane of the opening in the culture vessel because the carbon dioxide gas out of the culture vessel permeated the carbon dioxide gas 25 permeable membrase of the opening in the culture vessel. The culture vessel was irradiared with toe red light using COWL Lioht Source Unit (trade name) from Ninpon Medical & Chemical instruments Co., Ltd. manufacturerr name) as a light irradiatior apparatus. The humidity in the culture 30 vessel was controled to be 85% or more by sealing the culture vessel with Parafilm. :00561 Abamine SG is [3-(3,4-dimethoxyphenyl-allyl]-(4- Do:ke: N: \lPA 3C54POT 24 tlcorobenzy )-ainO] -butyrare methyl ester, and a structure represented by the following formula (2) . [0^57] [Chemical 2] F N -.- (2) 0 CH 3Y OCH3 Sixteen scions wer nut in one culture vessel.A rocting rate was calculated from the number of scion sarpes and the number of soots from which The root was generated afLter the culture for 2 months (nwter of 10 rooting) Resxlts are shown in Table 1. [3059] Comparative Example 21 he sample was cultured in the same manner as Example I, except that the medi containing no adventitious root formation accelerator was used. nte ~5 results are shown in Table 1. C20601 [Table 1] Table I . Results of eucalyptus clone A Rooting rate Example 1 Abamnine 66.7% Comparative ExampIe i Abarine SG 28.6% Comnarative Example 2 Additive-free 25.0% 0061 As is evident from Table 1, the rooting rate when 20 abamine was used was prominently increased compared with the case of additive-free. On the other hand, when abamine Dccket No. PNPA-13C54K : 25 SO was used, only the rooting rate that was slightly higher than nhe case of additive-free was obtained. 20062- .Example 2 and Comparative Example 3, The sample was cultured in the same manner as in S Example 1 and Comparative Example 1, except that eucaLyptus clone B (WA-MC2] that was a ultra-poor rooting strain of E. globulus was used as a material for a scion. The results are shown in Table 2. [0063] :Comparative Example 4] 10 The sample was cultured in the same manner as in Example 2, except that the medium containing no adventitious root formation acceLerator was used. The result is shown in Table 2. [0064] :Table 2] 1$ Table 2. Results of eucalyptus clone B Rooting rate Example 2 AbaTine 46.9% Comparative Example 3 Abamine SG 0% ComparativeExanple 4 Additive-free 1.6% :0065] As is evident from Table 2, the rooting rate of the Eucalyptus when abamine was used was prominently increased compared with ithe case of additive-free. On the 20 other hand, when abamine SG was used, no rooting was observed. (0066] :Example 3 One cultivar of tea plant (Camelia sinensis) was used as a material. A budwood extending to a length of 5 to 23 25 cm and having to 2 knots and about 2 to 6 leaves was cut out frot a mother tree for taking scions of the tea p.an: to prepare a scion. [0067] A base of the obtained scion was out ino a support wetted with MS medium diluted to 4 times :n which 30 M abamine and 10 mg/L of 1BA as the plant horsone had been Doce No. PN'A-30tP01T 26 added. here, the sunor: made of a nursery tray (vertical 6 lines and horizontal 11 lines) composed of lined square pyramids of vertical 4 cm x horizontal 4 cm x heigh: 15 cm filled with mixed soil of vermicUite and peat moss was 5 used, one scion was put in one pot, and total 66 scions were put in the cots to subject to the experiment. The scions after being put were cultured at a carbon dioxide concentration of 1000 cm at temperature of 250C and humidity of 6C% under irradiation with the red light !C containing a wavelength component of 650 nn to 670 nm and a wavelength comnonent of 450 n=n to 470 -7 at a ratio of 8.2:1.3 and raving a photosynthetic effective photon flux density of 51.3 LLmol/-2/s for 2 sonchs. The sale was irradiated with the rec light using CCFL Light Source Unit 15 (trade name) from Nippon Medical & Chemical Instruments Co., Ltd. (manufacturer name) as te light irradiation ataratus. 0068: A rooting rate was calculated iro cr number of scion sales and the number of shoots from which the root was generated after the culture for 2 months (n'rber of 20 rooting) The result is shown in Table 3. [0069] -comparative Exarple 5: The samp> was cultured in the same manner as in ExampD le 3, except that the medium containing no adventitious root formation accelerator was used. The 25 result is shown in Table 3. [OC70] :Table 3] Table 3. Results of tea olan< Rooting -ate Example 3 Abamine 97.8% Comparative Example 3 Additive-free i 65.3% C0371: As is evident from Table 3, the rootino rate of 30 the tea plant when abamine was used was prominently Jo:ket lKN. :NPA-13 v54-P0T 27 increased conTared wi:n the case of additive-free. :0072] As described above, it has been demonstrated tha7 the adveonitious root formation accelerator of the present invention exerts the excellent rooting effect regardless of plant species, particularly even in the ultra-poor rooting strain. ic