TWI768374B - 含有胺基酸或其鹽與寡醣之植物活力劑及其使用 - Google Patents
含有胺基酸或其鹽與寡醣之植物活力劑及其使用 Download PDFInfo
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- TWI768374B TWI768374B TW109120003A TW109120003A TWI768374B TW I768374 B TWI768374 B TW I768374B TW 109120003 A TW109120003 A TW 109120003A TW 109120003 A TW109120003 A TW 109120003A TW I768374 B TWI768374 B TW I768374B
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- plant
- oligosaccharide
- aforementioned
- amino acid
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Abstract
本發明課題在於控制、強化植物的生理過程,並改善農作物的活力、產量、品質及收成後的保存性。
解決方式為將以含有胺基酸或其鹽與寡醣為特徵的植物活力劑施用於植物。
Description
本發明係有關於一種含有胺基酸或其鹽與寡醣之植物活力劑及使用該植物活力劑之植物的栽培・生產方法。
植物常會因日照時間、氣溫及降雨量等非生物壓力(stress),以及病蟲害等生物壓力而導致收成量減少。尤其是為了增加農業作物的收成量,迄今使用各種的肥料及農藥。肥料為植物生長所需的營養源,但不具有緩和壓力的機能。農藥雖可直接驅除寄生於植物的病蟲害而排除生物壓力,但使用農藥時,雖可充分確認其安全性,但有過量攝取而對人體或環境造成影響之虞;尤其是藉由化學合成法所製成的農藥等藥劑,若一旦進行噴灑,則有長期間殘留於土壤中等的疑慮,可行的話期望藉由其他方法對生物壓力賦予抗性。由此,近年來,除此等之外,作為對人體或環境均安全的物質,生物刺激素(biostimulants)的利用備受矚目。
「生物刺激素」又稱「生物刺激劑」或「植物活力劑」等,係指含有任意的物質群・微生物,其係施用於植物體或其根體系時,藉由刺激在自然狀態之作物體內亦發生的一系列過程,而能夠提升養分吸收、提高施肥效率或賦予抗壓性而提升品質者,且對病蟲害未顯示直接的作用,因此未分類為任何殺蟲/殺菌劑者。亦即,係指存在於自然界之成分(包含微生物),非為植物激素或營養分,但即使極少量即可刺激植物的活力而促進生長的物質。一般而言,藉由將生物刺激素施用於植物,可提高植物的養分吸收與養分利用率,促進生長,使農作物的產量與品質變佳。農業用生物刺激素中,為了控制、強化作物的生理過程,而含有施用於植物或土壤之化合物、物質及其他製品的各種製劑。生物刺激素,為了改善作物的活力、產量、品質及收成後的保存性,係透過與營養素不同的路徑而對植物的生理產生作用。
如此,藉由生物刺激素,不會發生習知農藥或肥料所造成的問題,可刺激植物原本所具有的能力,而促進其成長。
作為與此種生物刺激素有關者,迄今有人報導:組合幾丁寡醣與具有抗菌活性之幾丁聚醣等而成的植物活力劑(專利文獻1);在食用醋中摻混寡醣類及植物萃取成分而成的植物活力劑(專利文獻2);含有纖維素的植物成長促進劑(專利文獻3);含有六呋喃糖(hexofuranose)衍生物的植物生長調節劑(專利文獻4);使用經低分子化之幾丁質或幾丁聚醣來提高植物的耐病性之方法(專利文獻5);含有幾丁質及/或幾丁聚醣等的肥料(專利文獻6);以及含有麩胺酸與脯胺酸作為構成胺基酸的植物成長促進劑(專利文獻7)。
[先前技術文獻]
[專利文獻]
[專利文獻1] 日本特開平9-143013號公報
[專利文獻2] 日本特開2001-64112號公報
[專利文獻3] 日本特開2002-114610號公報
[專利文獻4] 日本特開2013-151438號公報
[專利文獻5] 日本特開2015-48436號公報
[專利文獻6] 日本特開2017-95352號公報
[專利文獻7] 日本特開2019-006774號公報
[專利文獻8] 國際公開第2017/104687號
[發明所欲解決之課題]
本發明所欲解決之課題在於控制、強化植物的生理過程,並改善農作物的活力、產量、品質及收成後的保存性。
[解決課題之手段]
本案發明人等為解決上述課題而致力進行研究並重複多次實驗的結果發現意外的創見,即胺基酸或其鹽與寡醣之組合可促進植物的生長且可增大植物的誘引劑活性,終至完成本發明。
本發明係如下所述。
[1] 一種植物活力劑,其特徵為含有胺基酸或其鹽與寡醣。
[2] 如1之植物活力劑,其中前述寡醣為選自由幾丁寡醣、纖維寡醣及木寡醣所成群組的至少1種。
[3] 如1或2之植物活力劑,其中作為前述寡醣,係含有幾丁寡醣、纖維寡醣及木寡醣之3種全部。
[4] 如1~3中任一項之植物活力劑,其中前述胺基酸為選自由甘胺酸、麩胺酸及組胺酸所成群組的至少1種。
[5] 如1~4中任一項之植物活力劑,其中前述胺基酸為甘胺酸。
[6] 如1~5中任一項之植物活力劑,其中前述植物活力劑中之前述寡醣的總含量為0.05~10質量%。
[7] 如1~6中任一項之植物活力劑,其中前述植物活力劑中之前述胺基酸或其鹽的總含量為2.5~30質量%。
[8] 如1~7中任一項之植物活力劑,其進一步含有展開劑。
[9] 一種植物栽培方法,其係包含將如1~8中任一項之植物活力劑施用於植物。
[10] 如9之方法,其係包含將前述植物活力劑,以前述寡醣的總含量成為0.1~500質量ppm的濃度施用於植物。
[11] 如9或10之方法,其中前述植物活力劑對植物的施用係藉由葉面噴灑來進行。
[12] 一種生產方法,其係與未施用如1~8中任一項之植物活力劑的情形相比,具有增大之誘引劑活性的植物或其一部分的生產方法,其係包含藉由如9~11中任一項之方法來栽培植物。
[13] 如12之方法,其中前述誘引劑活性係藉由測定植物中的聚葡萄糖酶(glucanase)產生量來決定。
[14] 一種肥料組成物,其係含有如1~8中任一項之植物活力劑。
[發明之效果]
根據本發明,不會發生習知農藥或肥料對人體或環境造成影響等問題,可控制、強化植物的生理過程,並改善農作物的活力、產量、品質及收成後的保存性。
[實施發明之形態]
於本發明第一樣態中,提供一種植物活力劑,其特徵為含有胺基酸或其鹽與寡醣。
本發明之「植物活力劑」係不僅包含具有與植物的生長有關之溫度、光、水及鹽等非生物壓力的緩和作用,亦包含具有病蟲害等生物壓力的緩和作用者。
本發明中使用之寡醣宜為選自由幾丁寡醣、纖維寡醣及木寡醣所成群組的至少1種。本發明之植物活力劑,更佳的是,作為寡醣,含有選自由幾丁寡醣、纖維寡醣及木寡醣所成群組的2種以上,再更佳為含有幾丁寡醣、纖維寡醣及木寡醣之3種全部。
於本發明中,幾丁寡醣係作為源自植物以外的生物之物質,例如真菌、昆蟲、源自甲殼類之成分的誘引劑(即「外生誘引劑」)使用;而且,纖維寡醣及木寡醣則是作為源自植物之物質的誘引劑(即「內生誘引劑」)使用。
誘引劑係可對高等植物的組織或培養細胞誘導生物防禦反應之物質的總稱,在植物的免疫機構中可誘導抗病性。植物可以存在於葉面等的受體感測誘引劑,而觸發抗病原反應。藉此,對各種病原菌引起分泌各種化合物的生物防禦作用(免疫)。誘引劑若作用於植物,則可誘導植物防禦素或感染特異性蛋白質的合成/累積、活性氧生成、活性氮生成、過敏感反應性細胞死亡、基因表現變化等的防禦反應;認為藉由此等反應,可保護植物免於病原菌的侵害而提高耐病性。
植物防禦素係藉由誘引劑的作用而在植物體內合成、累積之抗菌性化合物,生產之抗菌性化合物係隨植物物種而異。代表性的植物防禦素可舉出類黃酮、萜類、脂肪酸衍生物等。活性氧具有殺死病原微生物的作用,而且,活性氧及活性氮可發揮作為單獨或共同觸發各種防禦反應的訊號之機能。此種誘引劑效果所產生的抗病性,從對於廣泛的病害可增強其抗性等來看,可望利用於農業上。
幾丁寡醣係包含部分脫乙醯基化之幾丁聚醣寡醣,且數個N-乙醯葡萄糖胺相連而成的寡醣類,一般而言,可藉由將源自甲殼類等之幾丁質水解等而得;其亦稱寡聚-N-乙醯葡萄糖胺。
亦即,幾丁寡醣可藉由將幾丁質以化學或酵素方式部分水解而得,其中該幾丁質係由蟹、蝦等甲殼類的殼等,藉由常用方法所調製而成。幾丁寡醣較佳使用選自N-乙醯殼二糖、N-乙醯殼三糖、N-乙醯殼四糖、N-乙醯殼五糖、N-乙醯殼六糖、N-乙醯殼七糖、N-乙醯殼八糖等的一種或多種的混合物。此等當中,尤以N-乙醯殼五糖、N-乙醯殼六糖、N-乙醯殼七糖係誘引劑效果較高。
本發明中所使用之幾丁寡醣特佳為具有下述化學結構者:。
此外,亦包含式中的乙醯基(COCH3
)部分脫落而成為NH2
者。此種脫乙醯基化的比例較佳為幾丁寡醣全體的30%以下,更佳為20%以下,再更佳為15%以下。
纖維寡醣係多個葡萄糖藉由β-糖苷鍵聚合而成的低聚糖類,近年來發現保濕性、抑制黏膩、賦予清淡味、減少澱粉老化、抑制蛋白質變性等機能性,而可望利用於醫藥、化妝品、食品、飼料領域。尤其是葡萄糖的聚合度為3以上的纖維寡醣,在上述增大機能性、賦予新穎機能性方面有著更大的期待。目前工業上利用的纖維寡醣係藉由酵素反應所製造而成,主成分為葡萄糖與二聚物之纖維二糖,幾乎不含有三聚物之纖維丙糖以上的寡聚物。然而,近年來藉由本案申請人等報導一種纖維寡醣之製造方法,其係在使用碳觸媒之植物性生質的水解反應中,藉由控制升溫速度、冷卻速度、反應溫度、反應時間使其水熱反應,而含有葡萄糖的聚合度為3~6的寡聚物(專利文獻8)。
藉由纖維素的水解反應獲得纖維寡醣時,作為纖維素原料,較佳使用Avicel(Merck公司製)等的結晶性微粉纖維素、或棉絨紙漿。
木寡醣係數個木糖經由β-糖苷鍵聚合而成的低聚糖類,一般而言,係藉由屬半纖維素之主成分的聚木糖的水解而得到,主要係作為食品用途販售。
植物活力劑僅含有幾丁寡醣及木寡醣此2種作為寡醣時,幾丁寡醣與木寡醣的含量之質量比較佳為幾丁寡醣:木寡醣=1:1~1:5,更佳為幾丁寡醣:木寡醣= 1:1.5~1:4。
植物活力劑僅含有幾丁寡醣及纖維寡醣此2種作為寡醣時,幾丁寡醣與纖維寡醣的含量之質量比較佳為幾丁寡醣:纖維寡醣=1:1~1:5,更佳為幾丁寡醣:纖維寡醣= 1:1.5~1:4。
植物活力劑含有幾丁寡醣、纖維寡醣及木寡醣之3種作為寡醣時,各寡醣相對於幾丁寡醣、纖維寡醣與木寡醣的總含量的比例,較佳為幾丁寡醣10~50質量%且纖維寡醣10~50質量%且木寡醣10~60質量%。各寡醣的比例更佳為幾丁寡醣20~40質量%且纖維寡醣20~40質量%且木寡醣20~55質量%。
本發明中所使用之胺基酸較佳為選自由甘胺酸、麩胺酸及組胺酸所成群組的至少1種,更佳為甘胺酸。
本發明中所使用之胺基酸之鹽不特別限定,可舉出例如胺基酸之鈉鹽、鉀鹽等。胺基酸之鹽的具體例可舉出麩胺酸鈉等。
本發明之植物活力劑能以粉狀、顆粒狀、液狀等任一種形態製品化,一般而言較佳調成容易噴灑的液狀。本發明之植物活力劑能以使胺基酸或其鹽與寡醣以高濃度溶解於水等溶劑的原液之形式供給。於一實施樣態中,植物活力劑原液中之前述寡醣的總含量較佳為0.05~10質量%,更佳為0.1~8質量%,再更佳為0.5~6質量%。於另一實施樣態中,植物活力劑原液中之前述寡醣的總含量較佳為1~15質量%,更佳為3~12質量%,再更佳為5~10質量%。
於一實施樣態中,本發明之植物活力劑原液中之前述胺基酸或其鹽的總含量較佳為2.5~30質量%,更佳為5~20質量%,再更佳為9~16質量%。於另一實施樣態中,前述胺基酸或其鹽的總含量較佳為10~70質量%,更佳為15~60質量%,再更佳為20~50質量%。
本發明之植物活力劑亦可進一步含有活性成分之胺基酸或其鹽與寡醣以外的其他成分,例如防腐劑、展開劑、懸浮劑、增黏劑、賦形劑。防腐劑可舉出山梨酸鉀、對羥基苯甲酸酯、安息香、脫氫乙酸鈉、日本扁柏油、苯氧乙醇、聚胺基丙基雙胍、聚離胺酸等。展開劑係以界面活性劑為主成分的黏稠液體,只要可作為植物活力劑之展開劑使用則不特別限制,可舉出例如聚氧乙烯壬基苯基醚、山梨醇酐脂肪酸酯、聚氧乙烯己糖醇酐脂肪酸酯等。懸浮劑可舉出聚磷酸或聚磷酸之鹽類、或聚羧酸型高分子界面活性劑等。增黏劑可舉出羧甲基纖維素(CMC)、聚丙烯醯胺、澱粉等水溶性高分子、或廢糖蜜、酒精發酵濃縮廢液、胺基酸發酵濃縮廢液等。賦形劑可舉出乳糖或澱粉等。
於本發明第二樣態中,提供一種植物栽培方法,其係包含將本發明之植物活力劑施用於植物。
本發明之植物活力劑所施用的對象植物不特別限制,典型上為農作物,可舉出菊科、茄科、十字花科、禾本科、豆科、薔薇科、葫蘆科、旋花科、藜科、百合科、繖形科、錦葵科、薑科、蓮科等的植物。
具體而言,可舉出白菜、高麗菜、青花菜、花椰菜類、小松菜、日本蕪菁、白蘿蔔、蕪菁等十字花科植物、馬鈴薯、番茄、茄子、青椒、辣椒、獅頭辣椒、煙草等茄科植物、春菊、萵苣、葉萵苣、牛蒡、款冬等菊科植物、西瓜、哈密瓜、南瓜、黃瓜、苦瓜、絲瓜、瓠瓜等葫蘆科植物、菠菜、莙薘菜、菾菜、無翅豬毛菜、甜菜等藜科植物、胡蘿蔔、芹菜、香芹、鴨兒芹等繖形科植物、大豆(毛豆)、紅豆、菜豆、蠶豆、豌豆、四稜豆、花生等豆科植物、番薯、空心菜等旋花科植物、韭菜、蔥類、洋蔥、大蒜、蘆筍等百合科植物、草莓、蘋果、梨子、枇杷等薔薇科植物、秋葵、棉等錦葵科植物、生薑等薑科植物、蓮等蓮科植物、玉米、米、大麥、小麥、甘蔗等禾本科植物等。
其中,較佳為高麗菜、小松菜等十字花科植物、番茄、茄子等茄科植物、萵苣、葉萵苣等菊科植物、草莓、蘋果等薔薇科植物;其中更佳為小松菜、番茄。
本發明之植物活力劑,一般係對其原液添加水等稀釋成期望的濃度(例如稀釋成1000倍)而使用,係以植物活力劑中之前述寡醣的總含量較佳成為0.1~500質量ppm的濃度施用於植物。以植物活力劑中之前述寡醣的總含量更佳為0.5~200質量ppm,再更佳為1~100質量ppm的濃度施用於植物。
植物活力劑對植物的施用可藉由本業界熟習的方法來進行,噴灑方法亦不特別限定,可為例如直接噴灑於植物的葉、莖等之方法、噴灑於供栽培植物的培養基或土壤中之方法、摻混於肥料等之後再噴灑於培養基或土壤中之方法等任一種。此外,摻混於肥料中時,作為肥料有含有氮、磷酸、鉀之化學肥料、油炸渣、魚渣、骨粉、海藻粉末、醣類、維生素類等有機質肥料等,其種類不特別限定。就噴灑方法而言,尤以藉由葉面噴灑來進行,在可有效顯現誘引劑活性上係較佳者。葉面噴灑可藉由本業界熟習的手法,例如動力噴霧器、掛肩噴霧器、撒播機、噴灑器、有人或無人直升機、煙霧器、手動噴霧器等來進行。
將植物活力劑摻混於肥料中噴灑時,相對於肥料組成物中的固含量100質量%,前述寡醣的含量較佳為10~50質量%,更佳為15~40質量%。相對於肥料組成物中的固含量100質量%,前述胺基酸或其鹽的含量較佳為10~50質量%,更佳為15~40質量%。肥料組成物,除前述寡醣及前述胺基酸或其鹽以外,較佳含有選自氮、磷酸、鉀的至少1種營養素,更佳含有氮、磷酸及鉀之3種營養素全部。若為液狀肥料時,肥料組成物較佳含有70~99質量%,更佳含有75~99質量%的水,較佳在噴灑前將此原液稀釋100倍至1000倍來使用。
藉此方法來栽培植物,與未施用植物活力劑時相比,成為可生產具有誘引劑活性的植物或其一部分(例如根、莖、葉、花、果實、種子、組織、細胞等),而且可改善農作物的活力、產量、品質及收成後的保存性。
諸如上述,誘引劑效果係屬抗病性的指標之一而為重要,而本案發明人等此次發現,根據屬誘引劑效果的訊號之一的聚葡萄糖酶產生量,藉由測定其酵素活性,可評定誘引劑活性。採取栽培中之植物的部分葉片,並分析其聚葡萄糖酶活性,可隨時間經過對同一個體進行評定。
以下示出誘引劑活性之評定方法中的程序的概要:
(i)進行植物的取樣及前處理;(ii)以BSA作為蛋白質標準作成檢量線(利用根據色素結合法之波長(600nm)的吸光度);(iii)測定(i)中所調製之檢體的蛋白質濃度;(iV)測定(i)中所調製之檢體的聚葡萄糖酶活性。具體而言,係以藉由聚葡萄糖酶使可溶性低分子分解物游離便會發色的B-HS試劑,以波長590nm的吸光度值評定其活性;(v)算出每單位蛋白質的聚葡萄糖酶活性。
關於此種誘引劑活性之評定方法的具體程序,係於下述實施例中詳細加以說明。
藉由以下實施例,對本發明更具體地加以說明,惟本發明非限定於此。
[實施例]
[1.寡醣的準備]
(1)幾丁寡醣
將幾丁質(FUJIFILM Wako Pure Chemical股份有限公司製,純化幾丁質)10g分散於含有磷酸1.2g的水30mL中後,將經減壓乾燥的粉末與直徑5mm的氧化鋁球100g共同裝入容量250mL的氧化鋁罐中,裝設於行星式球磨機(FRITSCH公司製,PULVERISETTE6)上以500rpm連續進行6小時處理而取得反應物。此外,溫度係於室溫下開始,剪切放熱所產生的溫度上升係順其自然發展。
接著將反應物懸浮於水中並以氫氧化鈣中和而得到漿液,使用5B濾紙將此漿液藉由吸濾器進行過濾,並將回收之濾液冷凍乾燥而取得幾丁寡醣粉末。
(2)纖維寡醣
以下實施例1、2及比較例1、4中使用藉由「製造方法1」所製造的纖維寡醣;實施例6、8、10中使用藉由「製造方法2」所製造的纖維寡醣。
(製造方法1:自結晶性微粉纖維素的取得)
將Avicel(Merck公司製結晶性微粉纖維素)10g和活性碳BA50(Ajinomoto Fine-Techno(股)製)1.5g,與直徑1.5cm的氧化鋁球2000g共同裝入容量3600mL的陶瓷罐磨機中,裝設於桌上型罐磨機旋轉台(日陶科學(股)製,桌上型罐磨機型式ANZ-51S)上,以60rpm進行48小時處理而取得反應原料。此外,溫度係於室溫下開始,剪切放熱所產生的溫度上升係順其自然發展。
接著,將反應原料0.374g與水40mL置入高壓反應器(內容積100mL,Omlabo(股)製高壓釜,HASTELLOY C22製)後,一邊以600rpm攪拌一邊以10~30℃/分鐘(平均升溫速度11.3℃/分鐘)加熱至230℃而至反應溫度後,隨即停止加熱,將反應器以10~30℃/分鐘(平均降溫速度16.7℃/分鐘)以風冷冷卻而製成反應液。
接著藉由離心分離裝置回收反應液而得到上清液,並將此上清液冷凍乾燥而取得纖維寡醣粉末。
(製造方法2:自棉絨紙漿的取得)
將棉絨紙漿(TOKOKOSEN股份有限公司,纖維素含有率97%)271g(含水率1.8%,乾燥質量266g),使用食品摻合機(型號:HBF500S,Hamilton Beach公司製)與85質量%磷酸(FUJIFILM Wako Pure Chemical股份有限公司製特級試劑)38g混合,而取得反應原料309g(含水率3.4%,磷酸含有率10.4%)。
接著,將反應原料309g與ϕ3/4吋碳鋼珠13kg一併投入振動磨機(裝置名:MB-1型,中央化工機股份有限公司製,罐尺寸5L)中,以總振幅8mm、振動數16.2Hz、夾套流通水溫度75℃的條件,藉由乾式粉碎進行水解反應24小時後,回收反應粉體。
將此反應粉體10g與離子交換水90g置入200L燒杯中,使用磁力攪拌器以25℃進行攪拌1小時,而取得纖維素水解物的萃取液。
接著,由對萃取物添加40質量%氫氧化鈣水溶液1.3g,使用磁力攪拌器以25℃進行攪拌1小時而調製的中和液中,藉由離心分離裝置回收上清液後,進行冷凍乾燥而取得纖維寡醣粉末。
(3)木寡醣
以下實施例1、2及比較例1、4中使用下述「市售品」之木寡醣;實施例6~9、11中則是使用藉由「製造方法」所製造的木寡醣。
(製造方法:自玉米芯粉末的取得)
將Acremonium Cellulolyticus TN株(FERM P-18508)在裝有液體培養基(Avicel50g/L、KH2
O4
24g/L、硫酸銨5g/L、酒石酸鉀1/2H2
O 4.7g/L、尿素4g/L、Tween80 1g/L、MgSO4
・7H2
O 1.2g/L、ZnSO4
・7H2
O 10mg/L、MnSO4
・5H2
O 10mg/L、CuSO4
・5H2
O 10mg/L)100mL的500mL燒瓶中以30℃、搖晃培養6天而得到培養液的離心分離上清液50mL,使玉米芯粉末5g懸浮,以50℃進行攪拌反應72Hr而得到反應液的離心分離上清液,予以冷凍乾燥而取得木寡醣原料粉末。
(市售品)
使用B Food Science股份有限公司製木寡醣95P。
[2.胺基酸的準備]
(1)甘胺酸:昭和電工股份有限公司製
(2)離胺酸:FUJIFILM Wako Pure Chemical股份有限公司製
(3)麩胺酸鈉(MSG):FUJIFILM Wako Pure Chemical股份有限公司製
(4)組胺酸:FUJIFILM Wako Pure Chemical股份有限公司製
(5)丙胺酸:FUJIFILM Wako Pure Chemical股份有限公司製
(6)精胺酸:FUJIFILM Wako Pure Chemical股份有限公司製
[3.結球萵苣之根乾燥重量的測定]
(1)植物活力劑的調製
將[1.寡醣的準備]中所準備之各寡醣及[2.胺基酸的準備]中所準備之各胺基酸,以成為下述表中之比較例1-2及實施例1、6-7中的植物活力劑濃度(質量ppm)的1000倍的方式,以各組成比率進行攪拌器攪拌而溶解於水中後,以0.45μm濾器進行除菌後調成植物活力劑原液。
(2)栽培試驗
於試驗區,係對供實施水耕栽培用而準備的養液150L添加各條件之植物活力劑原液150g並使其分散(稀釋1000倍),在溫度20~22℃下開始各120株的栽培。栽培合計實施60天,開始追加植物活力劑原液150g後每1週合計進行5次,並測定結球萵苣的根乾燥重量。
根乾燥重量的測定係切除根的部分,並藉由恆溫乾燥機使切除的根以50℃乾燥12小時後測定重量來進行。
此外,表1中的根乾燥重量係表示將未添加植物活力劑之無添加區的根乾燥重量設為100%時的比例(%)。
[4.小松菜之誘引劑活性的評定]
(1)植物活力劑的調製
將[1.寡醣的準備]中所準備之各寡醣及[2.胺基酸的準備]中所準備之各胺基酸,以成為下述表中之比較例4-10及實施例2、8-9中的植物活力劑濃度(質量ppm)的1000倍的方式,以各組成比率進行攪拌器攪拌而溶解於水中後,以0.45μm濾器進行除菌後調成植物活力劑原液。將此原液以水稀釋成1000倍,使用於以下栽培試驗。
(2)MS培養基的調製
小松菜的培育係使用村重-斯科格(MS)洋菜培養基。將調製之植物活力劑添加於MS培養基,使其成為各比較例及實施例所示之最終濃度,其後,以121℃、20分鐘進行高壓釜殺菌。
(3)播種及生長方法
於無塵實驗台上,將經高壓蒸氣滅菌的MS培養基移至苗木箱(plant box)中,充分冷卻後,各播種10粒小松菜(Wakami,SAKATA SEED)的種子。其後,於亮處以22℃、24小時,在長日照條件下使其生長6天。
(4)蛋白質萃取
調製以下組成的蛋白質萃取用緩衝液。
在BioMasher(Nippi股份有限公司)隨附之1.5ml微離心管(tube)中添加上述製作的蛋白質萃取用緩衝液300μl,於其中置入以剪刀剪成4×4mm左右而取樣的葉(植物體)。將此操作針對各試樣進行5次。接著,用手使攪拌棒旋轉,將植物體弄碎至眼中可見之固體成分大致消失的程度。在15,000×g、10分鐘、4℃的條件下進行離心分離,在新的1.5ml微離心管中回收水層,而調製萃取液。
(5)蛋白質濃度的調整
將純度已知之牛血清白蛋白(BSA)2mg/ml進行循序稀釋(稀釋成1/2、1/4、1/8、1/16、1/32、1/64)而製成標準品。使用製作之標準品,取600nm下的吸光度(Abs600)的平均值,作成檢量線(n=3)。將考馬斯亮藍(CBB)液300μl分注於96孔盤中,添加6μl之上述調製之萃取液。測定Abs600。空白試樣係使用MilliQ。將萃取液的吸光度代入藉由循序稀釋之BSA2mg/ml所作成的檢量線,求出蛋白質濃度。
此外,當萃取液的Abs600測定值偏離檢量線時,以超純水(MilliQ)適當稀釋並進行再測定而求出蛋白質濃度。
利用求得的值,進行稀釋使萃取液的濃度成為一定,使用於以下的聚葡萄糖酶活性測定。
(6)聚葡萄糖酶活性的測定
在1.5ml微離心管中混合將1錠B-HS試劑(Megazyme公司)懸浮於MilliQ 10ml而成的B-HS基質溶液100μl、0.2M磷酸緩衝液(pH6.0)50μl及上述調製之稀釋液或超純水(空白試樣)50μl,而調製各比較例及實施例之試樣。每15分鐘充分搖晃試樣,同時在30℃的水浴中,以1小時30分鐘進行酵素反應。添加100μl(合計300μl)的反應停止液0.2N NaOH而停止反應。在15,000rpm、5分鐘的條件下進行離心分離,將上清液200μl分注於96孔盤中,為了評定聚葡萄糖酶活性而測定590nm下的吸光度(Abs590),並與未施用植物活力劑的情形(比較例3)比較。
[5.小松菜之生物體重量的測定]
(1)植物活力劑的調製
將[1.寡醣的準備]中所準備之各寡醣及[2.胺基酸的準備]中所準備之各胺基酸,以成為下述表中之比較例12-14及實施例3-5、10-11中的植物活力劑濃度(質量ppm)的1000倍的方式,以各組成比率進行攪拌器攪拌而溶解於水中後,以0.45μm濾器進行除菌後調成植物活力劑原液。將此原液以水稀釋成1000倍,使用於以下栽培試驗。
(2)生物體重量的測定
準備作為作物之小松菜(Wakami,SAKATA SEED)、作為培養土之有機蔬菜用土(廣田商店)、作為肥料之新多木有機液肥3號(多木化學股份有限公司)。
在濾紙上進行作物的發芽後,對裝有培養土與肥料的25孔連結罐(全體280×280mm,一孔50×50×50mm)進行定植。定植後以一週的間隔一邊對小松菜進行各植物活力劑的葉面噴灑一邊於培養室中培育34天後,測定生物體重量,與未施用植物活力劑的情形(比較例11)比較。
Claims (14)
- 一種植物活力劑,其特徵為含有胺基酸或其鹽與寡醣,前述寡醣為選自由幾丁寡醣、纖維寡醣及木寡醣所成群組的至少1種。
- 如請求項1或2之植物活力劑,其中作為前述寡醣,係含有幾丁寡醣、纖維寡醣及木寡醣之3種全部。
- 如請求項1或2之植物活力劑,其中前述胺基酸為選自由甘胺酸、麩胺酸及組胺酸所成群組的至少1種。
- 如請求項1或2之植物活力劑,其中前述胺基酸為甘胺酸。
- 如請求項1或2之植物活力劑,其中前述植物活力劑中之前述寡醣的總含量為0.05~10質量%。
- 如請求項1或2之植物活力劑,其中前述植物活力劑中之前述胺基酸或其鹽的總含量為2.5~30質 量%。
- 如請求項1或2之植物活力劑,其進一步含有展開劑。
- 一種植物栽培方法,其係包含將如請求項1~8中任一項之植物活力劑施用於植物。
- 如請求項9之方法,其係包含將前述植物活力劑,以前述寡醣的總含量成為0.1~500質量ppm的濃度施用於植物。
- 如請求項9或10之方法,其中前述植物活力劑對植物的施用係藉由葉面噴灑(foliar spraying)來進行。
- 一種生產方法,其係與未施用如請求項1~8中任一項之植物活力劑的情形相比,具有增大之誘引劑活性的植物或其一部分的生產方法,其係包含藉由如請求項9或10之方法來栽培植物。
- 如請求項12之方法,其中前述誘引劑活性係藉由測定植物中的聚葡萄糖酶產生量來決定。
- 一種肥料組成物,其係含有如請求項1~8中任一項之植物活力劑。
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- 2020-06-15 WO PCT/JP2020/023472 patent/WO2020255934A1/ja unknown
- 2020-06-15 KR KR1020217036854A patent/KR20210151190A/ko not_active Application Discontinuation
- 2020-06-15 JP JP2021528238A patent/JPWO2020255934A1/ja active Pending
- 2020-06-15 EP EP20827345.8A patent/EP3984361A4/en active Pending
- 2020-06-15 US US17/619,821 patent/US20220369569A1/en active Pending
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2024
- 2024-04-05 JP JP2024061800A patent/JP2024088736A/ja active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1792164A (zh) * | 2006-01-06 | 2006-06-28 | 胡小安 | 一种植物生长调节剂 |
JP2008044854A (ja) * | 2006-08-10 | 2008-02-28 | Kyoei Seika:Kk | 植物成長活性剤およびその植物成長活性剤を用いた植物生育方法 |
TW201524357A (zh) * | 2013-12-16 | 2015-07-01 | Richin Life Code Biotechnology Co Ltd | 一種提高植物抗旱的植物生長調節組合物 |
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EP3984361A4 (en) | 2023-08-09 |
JP2024088736A (ja) | 2024-07-02 |
KR20210151190A (ko) | 2021-12-13 |
KR20240063161A (ko) | 2024-05-09 |
US20220369569A1 (en) | 2022-11-24 |
EP3984361A1 (en) | 2022-04-20 |
WO2020255934A1 (ja) | 2020-12-24 |
CN113993379A (zh) | 2022-01-28 |
JPWO2020255934A1 (zh) | 2020-12-24 |
TW202107998A (zh) | 2021-03-01 |
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