JP2019068743A - Method for mass producing matteuccia struthiopteris seedlings using spores at low cost - Google Patents
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Abstract
Description
本発明は、クサソテツ（コゴミ）の胞子を用いた胞子体苗の安価で簡易な大量生産方法に関する。 The present invention relates to an inexpensive and simple mass production method of spore body seedlings using sorghum sorghum.
クサソテツ（Ｍａｔｔｅｕｃｃｉａ ｓｔｒｕｔｈｉｏｐｔｅｒｉｓ）は、日本国内において北海道から九州まで広く分布し、イワデンタ科クサソテツ属のシダ植物であり、一般的には俗称のコゴミとして、ゼンマイやワラビと同様に春の山菜として広く知られている。クサソテツは、山菜特有の食味や風味を有しながら、ゼンマイやワラビに比べアクが少なく一般的な野菜と同様に調理できるため、消費の拡大が期待されている。現在は、東北地方を中心に栽培が行われており、２０１４年は全国で６８．６トンが出荷されている。 Russocks (Matteuccia struthiopteris) are widely distributed from Hokkaido to Kyushu in Japan, and are fern plants of the genus Russetae, and are generally widely known as spring pots and spring pots as well as spring callus and bracken There is. Kisosotetsu is expected to expand consumption because it has the same taste and flavor as wild vegetables, and can be cooked like ordinary vegetables with less moisture compared to spring water and bracken. Currently, cultivation is carried out mainly in the Tohoku region, and in 2014, 68.6 tons were shipped nationwide.
クサソテツの増殖は胞子及びランナー（走出枝）によって行われる。しかし、胞子による増殖は、自然環境下では子株（塊茎の直径がおおむね２cm以下）となるまで約７〜８年、さらに子株から収穫可能な成株（塊茎の直径がおおむね４cm以上）となるまで７年と長い年月を要する。またランナーによる増殖は（１）成株から伸びたランナーの伏せ込み、あるいは（２）ランナーの先に形成される子株の株分けが知られているが、特に西日本では成株を自生地から採取することは困難であり、また成株は安価に入手できないため、（１）及び（２）のいずれの方法も有利な増殖方法とはなり得ない。 The growth of the raspberry takes place by spores and runners. However, growth by spore is about 7 to 8 years until it becomes a offspring (tuber diameter is about 2 cm or less) under natural environment, and until it becomes an mature strain (tuber diameter about 4 cm or more) that can be harvested from offspring. It takes 7 years and long years. In addition, although it is known that (1) runners grow up from the adult strain, or (2) stock split of the offspring formed before the runner, in West Japan, in particular, the adult strain is collected from its own soil Since it is difficult to obtain an adult strain at low cost, neither of the methods (1) and (2) can be an advantageous growth method.
これまでに、幼苗を大量に生育させる方法として、組織培養法及び胞子を用いた増殖法が報告されている。徳島県では、１９９７年に、井内らが、根茎から摘出した茎頂及び若いランナーの先端から植物体を再生する組織培養による大量苗作成方法を確立している（非特許文献１、非特許文献２）。また富山県では、１９９８年に梅林が胞子を用いて２０℃、３，０００ｌｕｘの条件で幼苗を得ることに成功している（非特許文献３）。しかし、いずれの方法も特別な機器（恒温器等）や精密な温度・湿度等の管理が必要なため、苗の生産経費が高価となり、試験場以外の一般的な生産者では取り組めないという問題点があった。山梨県では、２００２年に雨宮ら（非特許文献４）が胞子を用いて特別な機器を使用せずに胞子体を得ることに成功しているが、圃場に定植可能な苗となるまでに１年１０ケ月と長い期間を要し、作出された株数も胞子０．５ｇから２０株程度と苗数が少ないという課題があった。 So far, tissue culture methods and spore propagation methods have been reported as methods for growing seedlings in large quantities. In Tokushima Prefecture, in 1997, Iuchi et al. Established a method for mass seedling production by tissue culture that regenerates a plant from the tip of a stem and a young runner extracted from a rhizome (Non-patent Document 1, Non-patent Document 1) 2). In addition, in Toyama Prefecture, in 1998, Meirin succeeded in obtaining seedlings at a temperature of 20 ° C. and 3,000 lux using spores (Non-Patent Document 3). However, since all methods require special equipment (such as a thermostat) and precise control of temperature and humidity, the cost of producing seedlings is expensive and can not be tackled by general producers other than the test site. was there. In Yamanashi Prefecture, Amamiya et al. (Non-Patent Document 4) succeeded in obtaining spores using spores without using special equipment in 2002, but by the time they become seedlings that can be planted in the field A long period of one year and ten months is required, and the number of plants produced is as low as about 0.5 g to 20 strains of spores, and the number of seedlings is small.
そこで本発明者らは、特別な機器を用いることなく、クサソテツの胞子から簡単かつ大量にクサソテツの胞子苗を生産する方法を開発した。 Therefore, the present inventors have developed a method for producing sorghum seedlings of sassos easily and in a large amount from spore of sassos without using special equipment.
本発明の目的は、胞子を用いたクサソテツ胞子体苗の安価で簡易な大量生産方法を提供することである。 An object of the present invention is to provide an inexpensive and simple method for mass production of X. lavensis spore bodies using spores.
本発明は、以下の特徴を包含する。
［１］胞子を用いたクサソテツ胞子体苗の生産方法であって、（１）クサソテツの胞子から前葉体を得るステップ、（２）前記前葉体から胞子体を得るステップ、及び（３）前記胞子体から定植可能な胞子体苗を得るステップを含む、但しステップ（２）において、ステップ（１）で得られた前葉体を移植することを特徴とする、前記方法。
［２］前記ステップ（１）において、前記胞子を播種床１ｃｍ２あたり約４０μｇ〜約１００μｇの量で播種することを含む、［１］に記載の方法。
［３］前記ステップ（１）で得られる前葉体が、２ｍｍ以上の葉幅を有する、［１］又は［２］に記載の方法。
［４］前記ステップ（１）〜（３）において、前記胞子、前記前葉体及び前記胞子体を光照射の２４時間日長下に置く、［１］〜［３］のいずれかに記載の方法。
［５］前記光照射が、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスの照度である、［４］に記載の方法。
［６］前記ステップ（２）において、前記前葉体を１セル（約５ｃｍ２〜約１０ｃｍ２）あたり数個体ずつ、培養土を含むマルチセルトレイに移植する、［１］〜［５］のいずれかに記載の方法。
［７］前記すべてのステップを保湿下で行う、[１]〜[６]のいずれかに記載の方法。
［８］前記ステップ（３）において、１週間に１回程度の頻度で肥料を施用する、［１］〜［７］のいずれかに記載の方法。
［９］前記ステップ（１）及び（２）のそれぞれにおいて、前記光照射の２４時間日長下に置く期間が、約３０日〜約４５日である、［４］〜［８］のいずれかに記載の方法。
［１０］前記ステップ（３）において、前記光照射の２４時間日長下に置く期間が、約１．８ケ月〜約２．２ケ月である、［４］〜［９］のいずれかに記載の方法。
［１１］前記ステップ（１）〜（３）が、
（１）播種床１ｃｍ２あたり４０μｇ〜１００μｇの量でクサソテツの胞子を、培養土などの培地を充填した光源（例、蛍光灯）付育苗箱に播種し、保湿し、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスの照度で２４時間日長下に約３０日〜約４５日間置き、葉幅２ｍｍ以上であるクサソテツ前葉体を得るステップ、
（２）前記ステップ（１）で得られた前記前葉体を、培養土などの培地を充填したマルチセルトレイに１セル（約５ｃｍ２〜約１０ｃｍ２）あたり数個体（例、２〜５個体）ずつ移植し、保湿し、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスの照度で２４時間日長下に約３０日〜約４５日間置き、クサソテツ胞子体を得るステップ、及び
（３）前記ステップ（２）で得られた前記胞子体に、（例えば、水分を噴霧して）保湿状態を保ちながら（例えば、１週間に１回程度）肥料（例、液肥）を施用し、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスの照度で２４時間日長下に約１．８ケ月〜約２．２ケ月間程度置き、定植可能なクサソテツ胞子体苗を得るステップ、
を含む、［１］〜［１０］のいずれかに記載の方法。
The present invention includes the following features.
[1] A method for producing a spore plant seedling using spores, which comprises the steps of (1) obtaining an anterior body from spores of Saxotetu, (2) obtaining a spore body from the anterior body, and (3) the spore Obtaining the settable spore seedlings from the body, except that in step (2), the anterior lobe obtained in step (1) is transplanted.
[2] The method according to [1], wherein in the step (1), the spores are seeded in an amount of about 40 μg to about 100 μg per 1 cm 2 of the seed bed.
[3] The method according to [1] or [2], wherein the anterior leaf obtained in the step (1) has a leaf width of 2 mm or more.
[4] The method according to any one of [1] to [3], wherein in the steps (1) to (3), the spore, the anterior body and the spore body are placed for 24 hours under light irradiation. .
[5] The method according to [4], wherein the light irradiation has an illuminance of about 2,000 lux to about 3,000 lux immediately below about 20 cm to about 30 cm.
[6] Any one of [1] to [5], wherein in the step (2), the anterior lobe is transplanted to a multi-cell tray containing culture soil for several individuals per cell (about 5 cm 2 to about 10 cm 2 ) The method described in.
[7] The method according to any one of [1] to [6], wherein all the steps are performed under moisturizing.
[8] The method according to any one of [1] to [7], wherein the fertilizer is applied at a frequency of about once a week in the step (3).
[9] In each of the steps (1) and (2), any one of the items [4] to [8], wherein the period of time under which the light irradiation is performed for 24 hours is about 30 days to about 45 days. The method described in.
[10] The method according to any one of [4] to [9], wherein in the step (3), the period of time under which the light irradiation is performed for 24 hours is about 1.8 months to about 2.2 months. the method of.
[11] The steps (1) to (3) are
(1) Seeds of sorghum sorghum in the amount of 40 μg to 100 μg per 1 cm 2 of the seeding bed in a light source (eg, fluorescent light) -bearing nursery box filled with a medium such as culture soil, moisturize, about 20 cm to about 30 cm For about 30 days to about 45 days under an illumination of about 2,000 lux to about 3,000 lux for about 24 to about 45 days, and obtaining an ax forelet having a leaf width of 2 mm or more,
(2) A few individuals (for example, 2 to 5 individuals) per cell (about 5 cm 2 to about 10 cm 2 ) in the multi-cell tray filled with a culture medium such as culture soil, and the anterior lobe obtained in the step (1) Implanting moisture at a time, moisturizing, placing about 20 cm to about 30 cm under about 24 to about 45 days under an illumination of about 2,000 lux to about 3,000 lux at a light intensity of about 2,000 lux to about 3,000 lux for about 30 days to about 45 days; And (3) applying fertilizer (eg, liquid fertilizer) to the spore body obtained in the step (2) while maintaining a moist state (eg, by spraying water) (eg, once a week or so) And about 1.8 months to about 2.2 months under a light intensity of about 2,000 lux to about 3,000 lux under about 20 cm to about 30 cm and under a light intensity of about 1.8 months to about 2.2 months. Step of obtaining seedlings,
The method according to any one of [1] to [10], including
本発明は、胞子を用いたクサソテツ胞子体苗の生産方法において、前葉体を移植することを特徴とし、これによって活着・生着率の向上及び作業の効率化が得られるという利点を提供する。 The present invention is characterized by transplanting the anterior lobe in a method of producing a spore-derived spore body seedling, which offers the advantage that the survival rate and survival rate can be improved and the work efficiency can be obtained.
以下において、本発明をさらに詳細に説明する。 The invention will be described in more detail below.
本発明は、胞子を用いたクサソテツ胞子体苗の生産方法を提供する。 The present invention provides a method of producing a spore seedling using spores.
この方法は、（１）クサソテツの胞子から前葉体を得るステップ、（２）前記前葉体から胞子体を得るステップ、及び（３）前記胞子体から定植可能な胞子体苗を得るステップを含み、ステップ（２）において、ステップ（１）で得られた前葉体を移植することを特徴とする。 The method comprises the steps of: (1) obtaining an anterior body from spores of the plant, (2) obtaining a spore body from the anterior body, and (3) obtaining a spore plant seedling that can be transplanted from the spore body, In step (2), the anterior lobe obtained in step (1) is transplanted.
上記の背景技術に記載されるように、クサソテツの胞子による苗の生産法又は増殖法は、非特許文献３及び４において提案されているが、いずれも胞子体を移植している。これに対して、本発明方法では、前葉体を移植することを特徴としていることから、これらの従来法と異なる。 As described in the above-mentioned background art, although methods for producing or growing seedlings by using spores of Saxoretsu have been proposed in Non-Patent Documents 3 and 4, both are spore body transplanted. On the other hand, the method of the present invention is different from these conventional methods because it is characterized by implanting the anterior lobe.
従来法では、胞子体の２葉展開期に移植が行われるが、湿度が保たれている場合は移植された胞子体の９割程度が活着し生育する一方、室内で湿度が保たれていない場合は２割以下しか活着せず、多くが枯死することが知られている。その原因は、移植の際、根が切れるため、移植後に胞子体が十分な水を吸い上げることができないためと考えられる。 In the conventional method, transplantation is carried out in the 2-leaf development stage of spore bodies, but when humidity is maintained, about 90% of the transplanted spore bodies survive and grow, while humidity is not maintained indoors It is known that only 20% or less survives and many die out. The cause is considered to be that the roots are broken at the time of transplantation, and the spores can not suck up enough water after transplantation.
一方、本発明の方法では、従来法の上記課題を解決する方法として、前葉体の時点（好ましくは、葉幅が約２ｍｍ以上である時点）で移植することによって、前葉体は、短い仮根から必要な水分を確保している状況であり、播種床の表面ごと移植すれば前葉体が枯れることはなく、移植作業による根傷みの発生や活着不良が発生しないため、移植後の生育が順調に進むし、また、従来法のようにセルトレイの培養土深く入っている根を丁寧に掘る必要がないため作業時間の短縮（作業の効率化）にもつながるという利点がある。 On the other hand, in the method of the present invention, as a method of solving the above-mentioned problems of the conventional method, the anterior lobe is short temporary root by transplanting at the time of anterior lobe (preferably, when the leaf width is about 2 mm or more). Since the anterior leaf is not withered if transplanted along the surface of the seed bed, and there is no occurrence of root damage or poor rooting by transplanting work, the growth after transplanting is smooth. In addition, there is an advantage that the working time can be shortened (work efficiency improvement) because it is not necessary to carefully dig the roots in the culture soil of the cell tray as in the conventional method.
以下、胞子体苗生産装置及び胞子体苗生産方法について説明する。 Hereinafter, a spore body seedling production apparatus and a spore body seedling production method will be described.
＜胞子体苗生産装置＞
本発明の方法で使用する胞子体苗生産装置は、育種箱と、この箱の全面を照明する光源とを含む簡易な装置からなる。
<Spore plant seedling production device>
The spore body seedling production apparatus used in the method of the present invention comprises a simple apparatus including a breeding box and a light source for illuminating the entire surface of the box.
「育苗箱」は、クサソテツ胞子の播種床として使用するために培養土、水苔、バーミキュライト、鹿沼土、それらの混合物等の植物育成用培地を充填して用いられるプラスチック製、金属製（例、アルミ、ステンレス等）、木製などの材質の浅い箱であって、箱の底面に排水穴が多数開口しているものを指す。育苗箱には、例えば方形の区画を有する格子又は容器を配置してもよい。育苗箱は、新鮮な培養土を充填した育苗箱と容易に置換できるように台座等に配置されうるし、また、育苗箱は、その中に、マルチセルトレイをセットすることも可能なように成形されてもよい。育苗箱として、例えば、市販の水稲中苗育苗箱を利用することができる。この育苗箱は、水稲用の中苗を作るために汎用されているため、安価にかつ容易に入手可能である。 “Growing box” is made of plastic or metal used by filling plant culture medium such as culture soil, starch syrup, vermiculite, Kanuma soil, mixture of them, etc. for use as a seedbed for sorghum spores. It is a shallow box made of aluminum, stainless steel, etc., wood, etc., and has many drainage holes in the bottom of the box. In the nursery box, for example, a grid or a container having square sections may be arranged. The nursery box can be placed on a pedestal or the like so that it can be easily replaced with a nursery box filled with fresh culture soil, and the nursery box can be shaped so that multi-cell trays can be set therein. May be As a nursery box, for example, a commercially available paddy rice nursery box can be used. This nursery box is widely used to make a medium seedling for rice, so it is inexpensive and easily available.
「マルチセルトレイ」は、多数のセル（例、１００以上又は２００以上）を含む容器、あるいは任意の形状（例、小さいくさび型もしくはＵ字型）のポットが連結して並んでいる育苗パネルであり、本発明では、前葉体を、この中に移植するために使用しうる。トレイの材質はプラスチック製、金属製（例、アルミ、ステンレス等）、木製などであるが、製造のし易さと価格の面でプラスチック製が好ましい。 A “multi-cell tray” is a nursery panel in which a container containing a large number of cells (eg, 100 or more or 200 or more) or pots of arbitrary shape (eg, small wedge or U shape) are connected and aligned. In the present invention, the anterior lobe can be used to implant into this. The tray is made of plastic, metal (eg, aluminum, stainless steel, etc.), wood, etc., but plastic is preferable in terms of ease of production and cost.
光源は、育苗箱に播種された胞子、前葉体、胞子体などに光照射するために必要である。光源として、ＬＥＤや蛍光灯が好ましい。本発明の方法では、上記胞子、前葉体、胞子体などの光照射する照度が、好ましくは、例えば約２，０００ルクス（ｌｕｘ）〜約３，０００ルクス程度必要であるため、光源と照明対象である育種箱との距離を約２０〜３０ｃｍ程度になるように、光源の位置を決定する。 A light source is necessary to light the spores, anterior leaves, spores etc. sown in the nursery box. As a light source, an LED or a fluorescent lamp is preferable. In the method of the present invention, the illumination intensity for light irradiation of the spore, anterior leaf body, spore body, etc. is preferably about, for example, about 2,000 lux (about lux) to about 3,000 lux. The position of the light source is determined so that the distance to the breeding box is approximately 20 to 30 cm.
そのような装置の具体例を図１に示した。
２つのコンテナが所定の間隔を開けて配置され、上記２つのコンテナの間に複数の育種箱（例、水稲中苗育苗箱又はマルチセルトレイ）が配置され、さらに上記２つのコンテナのそれぞれの上面に直管蛍光灯（例、４０Ｗ）の両端が設置され、ここで上記育苗箱の上面が上記蛍光灯の下、所定の距離（例、約２０ｃｍ〜約３０ｃｍ）に位置するように上記蛍光灯が設置される。好ましくは、この装置に設置されている蛍光灯の照度は、約２，０００ルクス〜約３，０００ルクスに調整されうる。コンテナは、収穫物の運搬用途等に使用されうる。
An example of such a device is shown in FIG.
Two containers are arranged at a predetermined interval, and a plurality of breeding boxes (for example, a rice seedling nursery box or a multi-cell tray) are arranged between the two containers, and further on top of each of the two containers. The fluorescent lamp is placed so that both ends of a straight tube fluorescent lamp (e.g., 40 W) are installed, and the upper surface of the seedling box is located below the fluorescent lamp at a predetermined distance (e.g., about 20 cm to about 30 cm) Will be installed. Preferably, the illuminance of the fluorescent light installed in this device can be adjusted to about 2,000 lux to about 3,000 lux. A container can be used for the conveyance application of a crop, etc.
＜胞子体苗生産方法＞
１．クサソテツの胞子から前葉体を得る第１ステップ
このステップでは、クサソテツの胞子を、培養土などの培地を充填した光源付育苗箱に播種し、保湿のために透明材料で被覆し、約２，０００ルクス〜約３，０００ルクスの２４時間日長下に約３０日〜約４５日間置き、葉幅が２ｍｍ以上であるクサソテツ前葉体を得ることができる。
<Spore body seedling production method>
1. First step of obtaining anterior leaves from sorghum spores In this step, spore sorghum are seeded in a light-sourced nursery box filled with a medium such as culture soil, covered with a transparent material for moisturizing, about 2,000 It can be placed for about 30 days to about 45 days under a 24-hour day-length from lux to about 3,000 lux, and it is possible to obtain a raspberry anterior leaf having a leaf width of 2 mm or more.
本明細書において「クサソテツの胞子」は、市販されているクサソテツの苗を生育させて採取した胞子葉や、自然界に自生する胞子葉から得られた胞子であり得る。上記胞子は、温度４℃〜５℃の冷蔵庫中で保存開始後２年以内の胞子葉から得られた胞子であってもよい。 In the present specification, “spores of a sorghum” can be spores obtained by growing commercially available sorghum seedlings and spores obtained from spores that are naturally produced in nature. The spore may be a spore obtained from spore leaves within two years after the start of storage in a refrigerator at a temperature of 4 ° C to 5 ° C.
本明細書において「培地」は、例えば、市販のピートモス、バーミキュライト、土（例、鹿沼土、腐葉土）等の混合土、水苔、バーミキュライトなどの植物育成・栽培用の培地を意味する。培地には、必要に応じて有機肥料（家畜糞、魚粉、米糠、油粕、骨粉、ボカシ肥、草木灰等）及び／又は無機肥料（チッソ、リン、カリ等）を必要量配合してもよいが、本発明の方法の第１ステップ及び次の第２ステップでは、好ましくは肥料を配合しない。なお、本明細書では、断らない限り「培地」を「培養土」と互換的にかつ便宜的に使用することがある。 In the present specification, the “medium” refers to, for example, a culture medium for growing and cultivating plants such as commercially available peat moss, vermiculite, mixed soil such as soil (eg, Kanuma soil, humus), water stalk, vermiculite and the like. In the culture medium, necessary amounts of organic fertilizer (animal dung, fish meal, rice bran, rice bran, oil meal, bone meal, bokashi manure, grass ash, etc.) and / or inorganic fertilizer (eg, chisso, phosphorus, potassium etc.) may be blended. In the first and second steps of the method of the invention, preferably no fertilizer is incorporated. In the present specification, "medium" may be used interchangeably and conveniently with "cultured soil" unless otherwise specified.
本発明では、胞子の播種量が前葉体発生量に影響するため重要である。好ましくは、胞子を播種床１ｃｍ２あたり約４０μｇ〜約１００μｇ、さらに好ましくは約５０μｇ〜約８０μｇの量で播種するのがよい（後述の表２参照）が、条件の違いなどによっては、この範囲以外の播種量を使用することもできる。 In the present invention, the amount of spore inoculation is important because it affects the amount of anterior leaf development. Preferably, the spores should be seeded in an amount of about 40 μg to about 100 μg, more preferably about 50 μg to about 80 μg, per 1 cm 2 of the seeding bed (see Table 2 below). Other seeding rates can also be used.
胞子を光照射するときには、２４時間日長が前葉体発生数との関係で最も好ましい（後述の表１参照）。本明細書において「２４時間日長」とは、人工光源を用いて、連続２４時間光を照射することを意味する。 When the spores are irradiated with light, a 24-hour day length is most preferred in relation to the number of frontal plate development (see Table 1 below). As used herein, “24-hour day-length” means irradiating light continuously for 24 hours using an artificial light source.
またこのとき、光照射の上記照度が、好ましくは、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスであるが、照度は、前葉体が発生する限り、この範囲外の明るさでもよい。 At this time, preferably, the above-mentioned illuminance of light irradiation is about 2,000 lux to about 3,000 lux under about 20 cm to about 30 cm, but the illuminance is out of this range as long as the anterior lobe is generated. It may be brightness.
本発明の方法では、すべてのステップを保湿下で行うことが必要である。そのためには、簡単には、透明フィルム、透明ガラスなどの「透明材料」を用いて育苗箱の上面を覆うことができる。この方法では、クサソテツの胞子及び培養土等の保湿状態を保つことが重要である。透明材料として、育苗箱の上面を覆うように密着させることが可能であり、且つ光をほぼ完全に透過させることができるフィルム状の素材が好ましく、例えば市販の農業用ビニール、食品用ラップフィルム等が挙げられる。 In the method of the present invention, it is necessary to carry out all the steps under moisturizing. For that purpose, the upper surface of the nursery box can be easily covered using a "transparent material" such as a transparent film, transparent glass or the like. In this method, it is important to maintain the moisture condition of the spore and culture soil of the rasp. As the transparent material, a film-like material which can be in close contact so as to cover the upper surface of the nursery box and can transmit light almost completely is preferable. For example, commercially available vinyl for agriculture, wrap film for food, etc. Can be mentioned.
上記光照射の２４時間日長下に置く期間は、好ましくは約３０日〜約４５日、さらに好ましくは約３５日〜約４０日である。 The period of time under which the light irradiation is performed for 24 hours is preferably about 30 days to about 45 days, more preferably about 35 days to about 40 days.
第１ステップで胞子から前葉体が得られるが、前葉体は、好ましくは少なくとも２ｍｍの葉幅を有する。２ｍｍ程度の葉幅をもつ前葉体であれば、短い仮根を有しているため水を吸い上げることが可能である。 In the first step the anterior body is obtained from spores, but the anterior body preferably has a leaf width of at least 2 mm. If it is a front lobe having a leaf width of about 2 mm, it has a short temporary root and can suck up water.
本明細書における「前葉体」は、クサソテツの胞子が発芽して形成される配偶体であって、腹面に造精器と造卵器の両方を有し、さらに下部に短い仮根を有する、葉幅が少なくとも２ｍｍ（もしくは２ｍｍ以上）の植物体である。 In the present specification, “an anterior body” is a gametophyte formed by germinating sorghum sorghum, which has both an animator and an egg generator on its ventral surface and a short temporary root at its lower portion. It is a plant whose leaf width is at least 2 mm (or 2 mm or more).
第１ステップでは、具体的な例示として、クサソテツの胞子を、培養土などの培地を充填した光源（例、蛍光灯）付育苗箱に播種し、保湿のために透明材料で被覆し、約２，０００ルクス〜約３，０００ルクスの２４時間日長下に約３０日〜約４５日間置き、葉幅が２ｍｍ以上であるクサソテツ前葉体を得ることができる。 In the first step, as a specific example, spores of Saxorectus are sown in a light source (eg, fluorescent light) -reared seedling box filled with a medium such as culture soil, and covered with a transparent material for moisturizing, about 2 It can be placed for about 30 days to about 45 days under a 24-hour day length of about 1,000 lux to about 3,000 lux to obtain a raspberry anterior leaf having a leaf width of 2 mm or more.
２．前葉体から胞子体を得る第２ステップ
このステップでは、具体的な例示として、第１ステップで得られた前葉体を、培養土などの培地を充填したマルチセルトレイに１セル（約５ｃｍ２〜約１０ｃｍ２）あたり数個体ずつ移植して透明材料で被覆し、約２，０００ルクス〜約３，０００ルクスの２４時間日長下に約３０日〜約４５日間置き、クサソテツ胞子体を得ることができる。
2. Second Step of Obtaining Spore Bodies from Anterior Lobes In this step, as a specific example, the anterior leaves obtained in the first step are placed in one cell (about 5 cm 2 to about 4 cells) in a multi-cell tray filled with a culture medium such as culture soil. Several individuals per 10 cm 2 ) are transplanted and covered with a transparent material, and placed for about 30 days to about 45 days under a 24-hour day length of about 2,000 lux to about 3,000 lux to obtain a scorpion sporophyte it can.
上記前葉体を、第１ステップの場合と同様に光照射の２４時間日長下に置くことが重要である。 As with the first step, it is important to place the anterior lobe under a 24 hour day of light irradiation.
またこのとき、光照射の上記照度が、好ましくは、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスであるが、照度は、胞子体が発生する限り、この範囲外の明るさでもよい。 At this time, preferably, the above-mentioned illuminance of light irradiation is about 2,000 lux to about 3,000 lux under about 20 cm to about 30 cm, but the irradiance is out of this range as long as spores are generated. It may be brightness.
さらに、上記前葉体を１セル（約５ｃｍ２〜約１０ｃｍ２（例、３個程度のとき５ｃｍ２〜５.５ｃｍ２））あたり数個体、好ましくは２〜５個、さらに好ましくは３〜４個体、最も好ましくは３個ずつ、培養土などの培地を含むマルチセルトレイに移植することが好ましい（後述の表３参照）。ここで、「数個体」は、２〜１０個体を意味する。 Further, the anterior lobe body 1 cell number per population (about 5 cm 2 ~ about 10 cm 2 (eg, three approximately 5cm 2 ~5.5cm 2) when), preferably 2-5, more preferably 3-4 Individuals, most preferably three, are preferably implanted in a multi-cell tray containing a medium such as culture soil (see Table 3 below). Here, "several individuals" means 2 to 10 individuals.
また、第１ステップと同様に、このステップでも保湿下で行うことが必要である。上記のように透明材料でトレイを被覆することによって保湿することができる。 Also, as in the first step, this step also needs to be performed under moisturizing. It can be moisturized by covering the tray with a transparent material as described above.
上記光照射の２４時間日長下に置く期間は、好ましくは約３０日〜約４５日、さらに好ましくは約３５日〜約４０日である。 The period of time under which the light irradiation is performed for 24 hours is preferably about 30 days to about 45 days, more preferably about 35 days to about 40 days.
３．胞子体から定植可能な胞子体苗を得る第３ステップ
このステップでは、具体的な例示として、第２ステップで得られた胞子体に、（例えば、水分を噴霧して）保湿状態を保ちながら適宜肥料（例、液肥）を施用し、約２，０００ルクス〜約３，０００ルクスの２４時間日長下に、好ましくは約１．８ケ月〜約２．２ケ月間置き、定植可能なクサソテツ胞子体苗を得ることができる。
3. Third Step for Obtaining Settable Spore Plant Seedlings from Spore Bodies In this step, as a specific example, the spore bodies obtained in the second step are kept moist (for example, by spraying water), as appropriate Fertilizer (eg, liquid fertilizer) is applied and placed on a 24-hour day length of about 2,000 lux to about 3,000 lux, preferably about 1.8 months to about 2.2 months. You can get body seedlings.
上記胞子体を、上記第１ステップ及び第２ステップと同様に、光照射の２４時間日長下に置くことが重要である。 As with the first and second steps, it is important to place the spores under a 24-hour light irradiation period.
またこのとき、光照射の上記照度が、好ましくは、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスであるが、照度は、胞子体苗が発生する限り、この範囲外の明るさでもよい。 Also, at this time, the above-mentioned illuminance of light irradiation is preferably about 2,000 lux to about 3,000 lux under about 20 cm to about 30 cm, but the irradiance is out of this range as long as spore body seedlings are generated. The brightness of the
このステップでは肥料（例、液肥）を使用するが、肥料を施用する頻度は、好ましくは、１週間に１回程度である。肥料は、上記の有機肥料又は無機肥料であり、例えば、「ハイポネックス」（例、Ｎ−Ｐ−Ｋ＝６−１０−８）等の市販の肥料であってよい。 Fertilizer (eg, liquid fertilizer) is used in this step, but the frequency of applying the fertilizer is preferably about once a week. The fertilizer is the above-mentioned organic fertilizer or inorganic fertilizer, and may be, for example, a commercially available fertilizer such as "Hyponex" (e.g., N-P-K = 6-10-8).
また、光照射の２４時間日長下に置く期間は、好ましくは、上記のとおり、約１．８ケ月〜約２．２ケ月であり、さらに好ましくは約２ケ月間であるが、胞子体苗が発生する限り、この期間以外でもよい。 Also, as described above, the period for which the 24 hour photoperiod of light irradiation is performed is preferably about 1.8 months to about 2.2 months, and more preferably about 2 months, but the spore body seedlings The period may be outside this period as long as
さらに、このステップもまた、保湿下で行う必要がある。植物体のサイズが大きくなるため、「（例えば、水分を噴霧して）保湿状態を保つ」ことが好ましく、具体的には、播種床である育苗箱に充填した培養土などの培地が乾かないように、霧吹き等を利用して培地に水を吹きかけ、その後育苗箱の上面を農業用ビニールや食品用ラップのような透明材料で覆うことがよい。 Furthermore, this step also needs to be done under moisturizing. Because the size of the plant increases, it is preferable to keep moist (for example, by spraying water). Specifically, the culture medium such as culture soil filled in the seedling box, which is the seeding bed, does not dry As described above, it is preferable to spray water on the culture medium using atomization or the like and then cover the upper surface of the nursery box with a transparent material such as agricultural vinyl or food wrap.
本明細書における「定植可能なクサソテツ胞子体苗」は、クサソテツ胞子体の発生から約２ヶ月等の上記期間で得られる、草丈が高く（例、５ｃｍ以上）、葉幅が広く（例、２ｃｍ以上）、葉色が濃い、良好に生育した胞子体苗が好ましい。 In the present specification, the "plantable xenopus spore plant seedling" has a high plant height (eg, 5 cm or more) and a wide leaf width (eg, 2 cm) obtained in the above period such as about 2 months from the development of the Or above), foliar seedlings having a deep leaf color and grown well are preferred.
本明細書における「クサソテツ胞子体」は、配偶体である前葉体から得られた雌雄の配偶子が融合して複相（二倍体）の接合子となり、これが細胞***を繰り返して多細胞体となったものである。 In the present specification, the "Xusomatus spore body" is a multi-phase body in which the male and female gametes obtained from the anterior body, which is the gamete, are fused to form a multiphase (diploid) zygote, which repeats cell division. It is
４．好適な方法の具体例
本発明の方法は、上記１〜３のステップを含み、各ステップの具体例を組み合わせた好適な実施形態は、以下の方法を含む。
4. Specific Example of the Preferred Method The method of the present invention includes the steps 1 to 3 above, and the preferred embodiment combining the specific examples of the steps includes the following method.
すなわち、好適な方法の例は、前記ステップ（１）〜（３）が、
（１）播種床１ｃｍ２あたり４０μｇ〜１００μｇの量でクサソテツの胞子を、培養土などの培地を充填した光源（例、蛍光灯）付育苗箱に播種し、保湿し、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスの照度で２４時間日長下に約３０日〜約４５日間置き、葉幅２ｍｍ以上であるクサソテツ前葉体を得るステップ、
（２）前記ステップ（１）で得られた前記前葉体を、培養土などの培地を充填したマルチセルトレイに１セル（約５ｃｍ２〜約１０ｃｍ２）あたり数個体（例、２〜５個体）ずつ移植して保湿し、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスの照度で２４時間日長下に約３０日〜約４５日間置き、クサソテツ胞子体を得るステップ、及び
（３）前記ステップ（２）で得られた前記胞子体に、（例えば、水分を噴霧して）保湿状態を保ちながら（例えば、１週間に１回程度）肥料（例、液肥）を施用し、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスの照度で２４時間日長下に約１．８ヶ月〜約２．２ケ月間置き、定植可能なクサソテツ胞子体苗を得るステップ、
を含む、上記方法である。
That is, in the example of the preferred method, the steps (1) to (3) are
(1) Seeds of sorghum sorghum in the amount of 40 μg to 100 μg per 1 cm 2 of the seeding bed in a light source (eg, fluorescent light) -bearing nursery box filled with a medium such as culture soil, moisturize, about 20 cm to about 30 cm For about 30 days to about 45 days under an illumination of about 2,000 lux to about 3,000 lux for about 24 to about 45 days, and obtaining an ax forelet having a leaf width of 2 mm or more,
(2) A few individuals (for example, 2 to 5 individuals) per cell (about 5 cm 2 to about 10 cm 2 ) in the multi-cell tray filled with a culture medium such as culture soil, and the anterior lobe obtained in the step (1) Step by step to moisturize, place under about 20 cm to about 30 cm under an illumination of about 2,000 lux to about 3,000 lux, for about 24 hours to about 30 days to about 45 days, to obtain a scorpion scorpion, And (3) applying fertilizer (eg, liquid fertilizer) to the spore body obtained in the step (2) while maintaining a moist state (eg, by spraying water) (eg, once a week or so) The plant can be planted for about 22 months to about 2.2 months for 24 hours under a light intensity of about 2,000 lux to about 3,000 lux under about 20 cm to about 30 cm. To get
Is the above method.
実施例を参照しながら本発明をさらに具体的に説明するが、本発明の範囲は、当該実施例に限定されないものとする。 The present invention will be more specifically described with reference to examples, but the scope of the present invention is not limited to the examples.
以下の各実施例において、胞子については、タキイ種苗株式会社から購入したクサソテツ苗を２０１２年４月に研究所内の圃場に定植し、生育させた株から発生した胞子葉を２０１４年１０月に採取し、胞子嚢群に切り分けてジッパー付の食品保存用袋に入れ、温度５℃の冷蔵庫で４ケ月保存したものを使用した（胞子は採取後２年以内であれば発芽する）。播種床については、縦３０ｃｍ×横６０ｃｍの水稲中苗育苗箱に市販の培養土（ピートモス、バーミキュライト、鹿沼土等の混合土で肥料は含まない）を充填したものを用いた。胞子の播種の直後に、この水稲中苗育苗箱を、蛍光灯とコンテナを組み合わせた簡易な装置の下に置いた（図１）。 In the following examples, with regard to spores, sssp. Saplings purchased from Takii Seed Co., Ltd. were planted in a field in the laboratory in April 2012, and spore leaves generated from the grown strains were collected in October 2014 Then, they were divided into spore sac groups, placed in a zippered food storage bag, and stored for 4 months in a refrigerator at a temperature of 5 ° C. (the spores germinate within 2 years after collection). As the seeding bed, used was one obtained by filling a commercially available culture soil (a mixed soil such as peat moss, vermiculite, Kanuma soil and the like, which does not contain a fertilizer) in a 30 cm × 60 cm paddy rice seedling rearing box. Immediately after sowing of spores, the rice seedling nursery box was placed under a simple device combining a fluorescent lamp and a container (FIG. 1).
[実施例１]日長時間が前葉体の発生数に及ぼす影響
２０１５年３月２０日に、１００ｍｇの胞子を十分に散水した水稲中苗育苗箱（播種床）へ均一に播き、保湿のためこの水稲中苗育苗箱を食品用ラップで包んだ。その直後、自然光が当たらない段ボール内で、上記装置の４０Ｗ直管蛍光灯（ＦＬＲ４０Ｓ・Ｗ／Ｍ）の下２０ｃｍ（２５００ｌｕｘ）に育苗箱を置き、日長時間が異なる区（０時間／日、６時間／日、１２時間／日、２４時間／日）を設け、２０１５年５月７日に、葉幅（以下「幅」という）２ｍｍ以上の前葉体の発生数について、水稲中苗育苗箱内で２か所（縦１０ｃｍ×横１０ｃｍ）を調査して平均値を求めた。
[Example 1] The effect of long time duration on the number of anterior leaf development On March 20, 2015, 100 mg of spores were sown evenly in a seedling nursery box (seeded bed) sufficiently watered with water, for moisturizing The rice seedling nursery box was wrapped in a food wrap. Immediately after that, place the nursery box in 20 cm (2500 lux) under the 40 W straight tube fluorescent lamp (FLR 40 S · W / M) of the above device in a cardboard box that is not exposed to natural light, 6 hours / day, 12 hours / day, 24 hours / day), and on May 7, 2015, the number of frontal leaves with a width of 2 mm or more (hereinafter referred to as “width”) Two places (10 cm in length x 10 cm in width) were investigated inside, and the average value was calculated.
胞子播種から約２週間程度で前葉体が発生し始め、４０日程度経過すると、移植に適した幅２ｍｍ程度の大きさとなった。 The anterior lobe began to develop about 2 weeks after spore inoculation, and after about 40 days, it became a size about 2 mm wide suitable for transplantation.
表１に示したとおり、幅２ｍｍ以上の前葉体の発生数は、日長時間が２４時間、１２時間、６時間、０時間の順に多かった。６時間では幅２ｍｍ以下の前葉体の発生は確認できたが、０時間は前葉体の発生自体が確認できなかった。 As shown in Table 1, the number of frontal lobes having a width of 2 mm or more was larger in the order of 24 hours, 12 hours, 6 hours, and 0 hours for a long time. At 6 hours, the occurrence of anterior lobes of 2 mm or less in width could be confirmed, but at 0 hours, the occurrence of anterior lobe itself could not be confirmed.
日長時間１２時間より日長時間２４時間の方が、幅２ｍｍ以上の前葉体の数が多かった。このことから、前葉体の発生及びその生育には、光及び日長時間が影響している。効率よく前葉体を得るためには、日長時間は２４時間が最適である。 The number of anterior lobes with a width of 2 mm or more was larger for 24 hours a day than for 12 hours a day. From this fact, the development of the anterior body and its growth are affected by light and long hours. In order to obtain an anterior body efficiently, 24 hours is suitable for long hours.
[実施例２]胞子の播種量が前葉体の発生数に及ぼす影響
２０１５年３月５日に、１０ｍｇ、５０ｍｇ、１００ｍｇ、５００ｍｇ、１，０００ｍｇに計量した胞子を、水稲中苗育苗箱（播種床）にそれぞれ均一に播き、保湿のためこの水稲中苗育苗箱を食品用ラップで包んだ。その直後、育苗箱に直射日光があたらない室内で、２４時間日長の４０Ｗ直管蛍光灯の下２０ｃｍ（２，５００ｌｕｘ）に置き、２０１５年４月１５日に、幅２ｍｍ以上の前葉体の発生数について、水稲中苗育苗箱内で３か所（縦１０ｃｍ×横１０ｃｍ）を調査し、平均値を求めた。
[Example 2] The effect of the spore seeding rate on the incidence of anterior lobes On March 5, 2015, the spore, which was weighed to 10 mg, 50 mg, 100 mg, 500 mg, and 1,000 mg, was used to The seeds were sown evenly on the floor, and the rice seedling nursery box was wrapped with a food wrap for moisturizing. Immediately after that, the nursery box was placed under 20 cm (2,500 lux) of 40 W straight fluorescent light with a 24 hour day length in a room where direct sunlight is not exposed, and on April 15, 2015, the frontal body of 2 mm or more in width With regard to the number of outbreaks, three places (10 cm long × 10 cm wide) were examined in the rice seedling nursery box, and an average value was determined.
前葉体（２ｍｍ以上）の発生数は、表２に示したとおり、１００ｍｇ、５０ｍｇ、１０ｍｇ、５００ｍｇ、１，０００ｍｇの順に多かった。１，０００ｍｇ播種した場合、調査日の２０１５年４月１５日においては前葉体の発生は確認できたが、幅は２ｍｍ以下であった。 The number of frontal lobes (2 mm or more) occurred in the order of 100 mg, 50 mg, 10 mg, 500 mg and 1,000 mg as shown in Table 2. When sowed at 1,000 mg, the occurrence of anterior lobe was confirmed on April 15, 2015 on the survey date, but the width was 2 mm or less.
水稲中苗育苗箱（縦３０ｃｍ×横６０ｃｍ）に対し、胞子播種量が１０ｍｇ、５０ｍｇの場合は、均一に播種しても播種床に前葉体が発生していないスペースが見られた。胞子播種量が５００ｍｇ、１，０００ｍｇの場合は、播種床いっぱいに前葉体が重なり合うように発生し、生育は不良であった。 When the spore seeding rate was 10 mg and 50 mg for a rice seedling nursery box (30 cm long × 60 cm wide), a space where no anterior leaf was generated in the seeding bed was seen even if the seeding was carried out uniformly. When the spore seeding rate was 500 mg and 1,000 mg, the anterior leaf occurred so as to overlap the entire seed bed, and the growth was poor.
胞子播種量が１００ｍｇの場合は、前葉体が発生していないスペースは少なく、前葉体の重なりも少なく、生育は良好であった。このことから、水稲中苗育苗箱（縦３０ｃｍ×横６０ｃｍ）に対しては、胞子量１００ｍｇ及びその前後が最適量である。 When the spore seeding rate was 100 mg, there was little space where no anterior lobe was generated, little overlap of anterior lobes, and the growth was good. From this fact, the spore amount of 100 mg and the amount before and after that are the optimum amount for a rice seedling-growing box (length 30 cm × width 60 cm).
[実施例３]前葉体の移植数及び移植したセルトレイの大きさが、胞子体発生セル率及び胞子体の生育に及ぼす影響
２０１５年４月１５日に、試験２の前葉体（胞子量１００ｍｇで播種し、２４時間日長下で生育させた前葉体）を、２箱の２８８穴セルトレイの全穴及び２箱の１２８穴セルトレイの全穴にそれぞれ１個体ずつ、さらに、２箱の２８８穴セルトレイの全穴及び２箱の１２８穴セルトレイの全穴にそれぞれ３個体ずつ移植し、５月下旬に、胞子体の発生後、灌水を１日２回行い、５００倍希釈の市販の液肥（ハイポネックス６−１０−８）を１週間に１回施用し、６月３０日に、胞子体苗が発生したセルの割合（胞子体が１個以上発生しているセルの割合）について調査し、平均値を求めた。なお、セルトレイは、１セルあたり５.３ｃｍ２の面積を有した。
[Example 3] Effect of transplanted number of anterior lobe and size of transplanted cell tray on spore generation cell rate and growth of sporophyte On April 15, 2015, anterior lobe of test 2 (spore amount: 100 mg) The anterior leaf (seed body) sown and grown under a 24-hour photoperiod), one each for each hole of two 288-well cell trays and one each of two 128-well cell trays, and two more 288-well cell trays Three wells are implanted in each of the whole hole and the two holes of the 128-well cell tray, and after sporangia are generated in late May, irrigation is performed twice a day, and a 500-fold dilution of commercially available liquid fertilizer (Hyponex 6) -10-8) was applied once a week, and on June 30, the percentage of cells in which spore seedlings were generated (percentage of cells in which one or more spores were generated) was investigated, and the average value I asked for. The cell tray had an area of 5.3 cm 2 per cell.
表３に示したとおり、胞子体の発生セル率は、前葉体を３個体ずつ移植した方が、１個体ずつ移植するより高くなった。また、１２８穴セルトレイ及び２８８穴セルトレイに前葉体を３個体ずつ移植した場合、表４に示したとおり、胞子体発生セル率及び生育に大きな差はなかった。 As shown in Table 3, the developmental cell rate of spore bodies was higher when three anterior lobes were transplanted than when they were implanted one by one. In addition, when three anterior lobes were implanted in each of the 128-well cell tray and the 288-well cell tray, as shown in Table 4, there was no significant difference in spore-producing cell rate and growth.
上記のとおり、前葉体の段階で移植を行うことにより、２ｍｍ程度の前葉体は短い仮根を有するために必要な水分を確保することが可能であり、播種床の表面ごと移植すれば前葉体は枯死せず、移植作業による根傷みの発生や活着不良が発生しないため、移植後順調に生育し得た。また、セルトレイの奥深くに入っている根を丁寧に掘る必要がないため、作業時間を短縮することもできた。 As described above, by performing transplantation at the stage of anterior lobe, the anterior lobe of about 2 mm can secure water necessary for having a short temporary root, and the anterior lobe if transplanted along the surface of the seed bed. Since the plant did not wither and did not develop root damage or poor survival due to the transplanting work, it could grow smoothly after transplantation. In addition, it was also possible to reduce the working time because it is not necessary to carefully dig the roots deep in the cell tray.
１つのセル穴に対する移植数が１個体の場合には胞子体発生セル率は４２．４％と低かったが、１つのセル穴に対する移植数を３個体に増やすことによって、胞子体発生セル率を高めることができる。定植までの期間が胞子体発生から２ケ月と短いこと、ある程度密植であってもクサソテツの光飽和点が低いことから、２８８穴セルトレイで育苗することが可能である。 Although the spore generation cell rate was as low as 42.4% when the number of transplants to one cell hole was one individual, the spore body generation cell rate was increased by increasing the number of transplants to one cell hole to 3 individuals. It can be enhanced. It is possible to raise seedlings in a 288-well cell tray because the period until planting is as short as two months after sporulation and the light saturation point of the plant is low even if densely planted to some extent.
[実施例４]液肥施用が胞子体の生育に及ぼす影響
２０１５年３月５日に、水稲中苗育苗箱に１００ｍｇ／箱の胞子を播種し、その後２４時間日長処理して発生した前葉体を、４月１５日に２箱の２８８穴セルトレイに３個体ずつ移植し、胞子体の発生後、灌水を１日２回行い、５月２３日から１週間に１回の割合で液肥（ハイポネックス６−１０−８）を５００倍希釈、１，０００倍希釈、２，０００倍希釈に分けて施用する区（１回に１．５ｍＬ／セルを施用）と、無施用区（水のみで管理）とを設け、６月３０日に、２８８穴セルトレイの各処理区１０セルの２カ所で胞子体の葉数４枚目の草丈、葉幅（最も広い長さ）について調査し、平均値を求めた。なお、１セルから胞子体が１株以上発生した場合は１株に間引いた。
[Example 4] The effect of liquid fertilizer application on the growth of spore bodies On March 5, 2015, 100 mg / box of spores were sown in a rice seedling nursery box, and then the treatment was performed for 24 hours for an anterior body generated. , 3 individuals in 2 boxes of 288-well cell trays on April 15 and after spore formation, perform irrigation twice a day, and liquid manure (Hyponexus) once a week from May 23 6-10-8) divided into 500-fold dilution, 1,000-fold dilution and 2,000-fold dilution (1.5 mL / cell applied at a time) and no application (only water control) And, on June 30, examine the plant height and leaf width (the widest length) of the fourth leaf of the spore body at two places of 10 cells in each treatment area of 288-well cell tray on June 30, and average value I asked. When one or more strains of spores were generated from one cell, they were thinned to one strain.
表５に示したとおり、胞子体の生育は、液肥を施用した方が、無施用の場合と比較して、草丈が高く葉幅が広くなった。また液肥の希釈倍数による生育の差はほとんどなかった。 As shown in Table 5, the growth of spore bodies was higher in plant height and wider in leaf width when liquid fertilizer was applied as compared with no application. In addition, there was almost no difference in growth due to dilution of liquid fertilizer.
液肥を施用しない場合は葉色が薄くなりやや黄色くなっていたことから、この場合には肥料切れとなって胞子体の生育が抑制されたと推定される。液肥の施用による生理障害は生じなかったため、肥料が不足することがないように、液肥の希釈倍数は５００倍が適当である。 In the case where liquid fertilizer was not applied, the leaf color became pale and became yellowish in this case, and in this case, it is presumed that the growth of the spore body was suppressed due to lack of fertilizer. Since no physiological disorder was caused by the application of liquid fertilizer, the dilution factor of liquid fertilizer is suitably 500 times so as not to run out of fertilizer.
本発明の、クサソテツの胞子を用いた胞子体苗の安価で簡易な大量生産方法は、栄養豊富でアクが少なく調理が容易な食用山菜としての、また薬用原料としてのクサソテツの人工栽培に有用である。 The inexpensive and simple mass production method of spore body seedlings using sorghum of scorpionfish of the present invention is useful for artificial cultivation of scorpionfish as an edible vegetable which is rich in nutrients and easy to prepare with little or no mashing, and as a medicinal material. is there.
Claims (11)
（１）播種床１ｃｍ２あたり４０μｇ〜１００μｇの量でクサソテツの胞子を、培養土などの培地を充填した光源付育苗箱に播種し、保湿し、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスの照度で２４時間日長下に約３０日〜約４５日間置き、葉幅２ｍｍ以上であるクサソテツ前葉体を得るステップ、
（２）前記ステップ（１）で得られた前記前葉体を、培養土などの培地を充填したマルチセルトレイに１セル（約５ｃｍ２〜約１０ｃｍ２）あたり数個体ずつ移植して保湿し、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスの照度で２４時間日長下に約３０日〜約４５日間置き、クサソテツ胞子体を得るステップ、及び
（３）前記ステップ（２）で得られた前記胞子体に、保湿状態を保ちながら肥料を施用し、約２０ｃｍ〜約３０ｃｍ直下で約２，０００ルクス〜約３，０００ルクスの照度で２４時間日長下に約１．８ケ月〜約２．２ケ月間置き、定植可能なクサソテツ胞子体苗を得るステップ、
を含む、請求項１〜１０のいずれか１項に記載の方法。 The steps (1) to (3) are
(1) The sorghum sorghum sorghum in the amount of 40 μg to 100 μg per 1 cm 2 of the seeding bed is sown in a lighted nursery box filled with a medium such as culture soil, moisturized, about 2,000 lux under about 20 cm to about 30 cm Step for obtaining an axillary leaf anterior leaf that is placed for about 30 days to about 45 days under a photoperiod of 24 hours under an illuminance of about 3,000 lux, and a leaf width of 2 mm or more,
(2) The anterior lobe obtained in the step (1) is moistened by transferring several individuals per cell (about 5 cm 2 to about 10 cm 2 ) to a multi-cell tray filled with a culture medium such as culture soil, 20 to about 30 cm right under a light intensity of about 2,000 lux to about 3,000 lux under a 24-hour day length for about 30 days to about 45 days to obtain a scorpion scorpion, and (3) the step (2) Fertilizers are applied to the spore bodies obtained in 1.) under moisturizing condition, and under about 20 cm to about 30 cm, about 2,000 lux to about 3,000 lux, for about 24 hours under a photoperiod of about 1 hour. 8 months to about 2.2 months, a step of obtaining plantable xenopus spore plant seedlings,
The method according to any one of claims 1 to 10, comprising
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