CN102511509A - 一种从幌伞枫叶中提取对抗薇甘菊的化感物质的方法 - Google Patents
一种从幌伞枫叶中提取对抗薇甘菊的化感物质的方法 Download PDFInfo
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Abstract
本发明公开了一种从幌伞枫叶中提取对抗薇甘菊的化感物质的方法。该方法具体步骤为:取幌伞枫叶,加5~20倍量的55~100体积%乙醇,回流提取1~5次,每次提取30分钟~90分钟,合并提取液,过滤,减压浓缩,加10~20倍量的水制成悬浮液,用10~30倍于悬浮液的乙酸乙酯进行萃取即得。本发明的幌伞枫提取物对薇甘菊的防治效果优于现有技术制备的提取物,具有广泛的应用前景。
Description
技术领域
本发明属于入侵植物生物防控技术领域,具体涉及一种从幌伞枫叶中提取对抗薇甘菊的化感物质的方法。
背景技术
薇甘菊(Mikaina micrantha H.B.K.)属菊科(Compositae)假泽兰属,是广布于我国华南地区的主要入侵杂草。薇甘菊在种间竞争中排挤当地植物,极易形成单优种群,表现出很强的竞争优势,给粤东地区的农林业生产及自然生态系统造成严重威胁,是国家环境保护公布的首批入侵我国的外来物种之一。对于薇甘菊的防治,现一般采取人工清除,喷洒农药等方法进行清除。植物化感作用(allelopathy)是指植物(含微生物)通过向环境中释放化学物质而对其他植物或微生物产生有益或有害的作用,包括自毒作用(Rice E L. Allelopathy(2 edition),New York:Academic Press,1984)。植物化感作用是通过向环境释放化学物质来实现的,这些起作用的化学物质称为花干物质(allelochemical)。从目前的研究结果来看,酚类和萜类两类次生物质是高等植物的主要化感物质。用其他植物的化感物质来防治入侵种的范例是有的,且大多都处于开发阶段。近年来,国内外对薇甘菊的生物学特性、生物和化学防除等方面开展了大量研究,如徐高峰等(5种植物对薇甘菊化感作用研究,西南农业学报,2009年22卷5期:1439-1443)用水作为溶剂对云南省薇甘菊发生地伴生的常见本地物种异型莎草(Cyperus difformis)、水蓼(Polygonum hydropiper)和外来物种空心莲子草(Alternanthera philoxcroides)、三叶鬼针草(Bidens pilosaL.)以及农粮作物红薯(Ipomoea batatas)地上部分进行提取,后将各提取液配制成一定质量浓度的营养液培养薇甘菊幼苗,12 d后测其对薇甘菊幼苗的生长抑制情况。结果表明,5种植物化感作用的综合抑制效应总体表现为:空心莲子草>异型莎草>三叶鬼针草,水蓼和红薯对薇甘菊幼苗生长的抑制程度相似,但强度明显弱于前3种植物。
中国专利公开号为1826864的发明公开了一种完全遏制恶性杂草薇甘菊新个体生长的方法,是建造人工植物群落的方法,通过采用种植面积不小于56.25m2幌伞枫纯林以抑制薇甘菊的生长,达到了只用二个生长季的抚育就能使新的薇甘菊幼苗或个体不再出现的效果。但是种植林木在适用范围上有局限性,且抑制效果慢。
幌伞枫(Heteropanax fragrans (Roxb.) Seem),又名大蛇药、五加通等,为五加科幌伞枫属乔木植物。赵厚本等(华南地区几种常见植物对薇甘菊的化感作用研究,生态环境,2007,16(1):130-134)用幌伞枫的水提液或乙醇提取液配制成一定质量浓度的营养液,培养已生根的薇甘菊(Mikania micrantha)枝条,1个月后测其生长情况。具体水提方法是采集地上新鲜部分风干后粉碎称取125g加一定量蒸馏水浸泡24h,过滤,连续浸泡3次,合并滤液即得水提液;醇提方法是将植物地上新鲜部分风干粉碎后称取125g,加入一定量体积分数为95%的乙醇浸泡24h,过滤,连续浸泡3次,合并滤液,将乙醇减压蒸干,得深褐色浸膏,将浸膏放入烧杯加蒸馏水剧烈搅拌,得醇提液。用上述水提液和醇提液配上营养液培养薇甘菊,结果发现幌伞枫水提液培养的的薇甘菊插条无一成活;幌伞枫醇提液培养的薇甘菊插条成活率只有15.56%,表明幌伞枫对薇甘菊生长具有化感抑制作用。
宋任华等采用化学反应及广谱数据分析了幌伞枫的化学成分(大蛇药化学成分的研究,云南植物研究,1988,,1(4):457-465),发现其根茎的正丁醇提取物中含有化合物Ⅲ(其结构为mclaleucic acid-28-[α-L-rhamnopyranosyl-(1-4)- β-D-glucopyranosyl-(1-6)- β-D- glucopyranosyl),其粗苷的水解部分含有齐墩果酸、白千层酸、胡萝卜苷和化合物Ⅱ(其结构为3β,23-二羟基-20(29)-羽扇烯-27,28-二羧酸),另外还有化合物Ⅰ,为无色针状晶体,mp>300℃,对Liebermann试剂呈紫红色,示为三萜,质谱测定分子离子峰为486,分子式为C30H46O5。
发明内容
本发明的目的在于通过改进现有的幌伞枫提取物的提取方法,提供一种新的从幌伞枫叶中提取对抗薇甘菊的化感物质的方法,该方法提取到的物质对薇甘菊杀灭或抑制效果更强烈。
本发明通过以下技术方案实现上述目的:
一种从幌伞枫叶中提取对抗薇甘菊的化感物质的方法,其制备方法步骤如下:
取幌伞枫叶,加5~20倍量的55~100体积95%乙醇,回流提取1~5次,每次提取30分钟~90分钟,合并提取液,过滤,滤液减压浓缩,加10~20倍量的水制成悬浮液,用10~30倍于悬浮液的乙酸乙酯进行萃取。
幌伞枫叶可再生、易获得,且风干粉碎过程简单,用时短,是幌伞枫整株植物中优选的原料,在制备过程中,优选用经过干燥(如风干),粉碎处理的幌伞枫叶。在提取步骤中,我们发现乙酸乙酯层所提取的物质对于薇甘菊的萌发起到的抑制作用是最大的,这一步大大提升了产品的品质。
上述乙醇体积百分浓度优选为95%,在这一提取浓度下,提取的物质更为纯净。乙醇加入量优选为幌伞枫叶粉的15倍,可以在最大限度提取幌伞枫有效物质的前提下节省乙醇用量。
上述步骤中减压浓缩后优选加入15倍量的水制成悬浮液,以使得浓缩物能够充分溶解,利于下一步的操作。
乙酸乙酯用量优选为悬浮液的20倍,利于进行充分提取。
上述任意一种从幌伞枫叶中提取对抗薇甘菊的化感物质的方法提取得到的物质。
与现有技术相比,本发明具有以下有益效果:
经过实验证明,对经过乙醇提取后浓缩的物质再进行乙酸乙酯萃取,可以去除大部分杂质而使有效物质聚集,验证实验证明,经过乙酸乙酯萃取后的化感物质对薇甘菊的化感作用更为强烈,得到的提取物对入侵植物薇甘菊的化感作用明显优于现有技术中的提取方法。通过幌伞枫化感物质的提取,可以探索开发天然除草剂的使用,并且这项技术将大大降低了人力财力的消耗,以及对生态环境的破坏。
具体实施方式
下面结合实施例进一步解释本发明,实施例中除作特殊说明外,均为本领域常规实验技术手段,实施例中乙醇浓度均为体积百分浓度。
实施例1
取新鲜幌伞枫叶,用搅碎机对其进行搅碎处理,将搅碎处理后的汁液用纱布、滤纸类的物质进行过滤,取滤液,加入15倍量的95%乙醇回流提取3次;合并提取液后过滤,减压浓缩,得乙醇浸膏,加入15倍的蒸馏水制成悬浮液;用20倍的乙酸乙酯进行萃取,将萃取液减压浓缩,从而使得乙酸乙酯蒸发,得乙酸乙酯浸膏。
实施例2
取幌伞枫叶,风干,粉碎,取10kg,加入15倍量(150L)的95%乙醇回流提取3次;合并提取液后过滤,减压浓缩,得乙醇浸膏900g,加15倍的蒸馏水(13.5L)制成悬浮液;用20倍(270L)的乙酸乙酯进行萃取,将萃取液减压浓缩,从而使得乙酸乙酯蒸发,得乙酸乙酯浸膏20g。
实施例3
其他步骤如实施例2,制备成悬浮液后用不同的溶剂(乙醇、石油醚、乙酸乙酯、水)进行萃取,将乙醇浸膏、石油醚浸膏、乙酸乙酯浸膏、水相浸膏溶解于对应的溶剂中,配成0.005g/ml,0.001g/ml,0.0002g/ml的浓度。精确吸收5ml该液体,加入到放有两层滤纸的9cm培养皿中,自然挥干溶剂,再加入5ml蒸馏水,对照(ck)为对应有机溶剂(水相浸膏对照为水),每个培养皿放置30粒薇甘菊种子,每个梯度设3个重复。放入人工气候箱中培养。培养条件:光照时间12h,光照强度为4500lx,温度为30℃;黑暗时间12h,温度为28℃;湿度为80%,每天补充损失的水分。以胚根或胚轴突破种皮1mm为萌发标准,每天统计萌发率。结果显示如下:
表1 四种浸膏对薇甘菊种子萌发率的影响
0.005g/ml | 0.001g/ml | 0.0002g/ml | ck | |
乙醇浸膏 | 57.22±10.84a | 63.33±7.60ab | 69.44±4.43b | 59.79±11.09ab |
石油醚浸膏 | 31.11±6.94a | 53.33±5.77b | 64.44±12.62b | 62.22±1.92b |
乙酸乙酯浸膏 | 13.33±3.33a | 56.67±5.77b | 50±6.67b | 72.22±7.70c |
水相浸膏 | 60±6.67a | 66.67±3.33a | 58.74±10.68a | 70.52±3.71a |
*数值为平均值±标准差,同一行中含至少一个相同字母的组间在0.05水平无显著差异。
从表1中可以看出,不同浓度的乙醇浸膏溶液对薇甘菊种子萌发率的影响并不显著。0.005g/ml浓度的石油醚浸膏溶液对薇甘菊种子萌发率的抑制与对照相比达到了显著水平,而乙酸乙酯浸膏溶液的各个浓度对薇甘菊种子萌发率的抑制与对照相比都达到了显著水平。各个浓度的水相部分浸膏溶液对薇甘菊种子萌发率的影响与对照相比都没有显著差异。从上述结果可知,幌伞枫乙酸乙酯浸膏对薇甘菊种子的萌发率抑制最为显著。
选择幌伞枫叶提取步骤中的其他制备参数时,在相同实验条件下,不同的有机溶剂萃取得到的终产物在抑制薇甘菊种子萌发时,乙酸乙酯的萃取物抑制效果也明显优于其他溶剂的萃取结果,因实验步骤极其相似,此处不再复述。
Claims (7)
1.一种从幌伞枫叶中提取对抗薇甘菊的化感物质的方法,其特征在于步骤如下:
取幌伞枫叶,加5~20倍量的55~100体积%乙醇,回流提取1~5次,每次提取30分钟~90分钟,合并提取液,过滤,滤液减压浓缩,加10~20倍量的水制成悬浮液,用10~30倍于悬浮液的乙酸乙酯进行萃取。
2.根据权利要求1所述从幌伞枫叶中提取对抗薇甘菊的化感物质的方法,其特征在于所述幌伞枫叶经过干燥、粉碎处理。
3.根据权利要求1所述从幌伞枫叶中提取对抗薇甘菊的化感物质的方法,其特征在于所述乙醇体积百分浓度为95%。
4.根据权利要求1所述从幌伞枫叶中提取对抗薇甘菊的化感物质的方法,其特征在于所述乙醇加入量为幌伞枫叶粉的15倍。
5.根据权利要求1所述从幌伞枫叶中提取对抗薇甘菊的化感物质的方法,其特征在于减压浓缩后加入15倍量的水制成悬浮液。
6.根据权利要求1所述从幌伞枫叶中提取对抗薇甘菊的化感物质的方法,其特征在于乙酸乙酯用量为悬浮液的20倍。
7.权利要求1~6任意一项从幌伞枫叶中提取对抗薇甘菊的化感物质的方法提取得到的物质。
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CN104255363A (zh) * | 2014-08-29 | 2015-01-07 | 云南省林业科学院 | 一种利用杉木控制薇甘菊的方法 |
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CN106235326A (zh) * | 2016-07-21 | 2016-12-21 | 天峨县全盛蜂业科技有限公司 | 一种枫叶汁的提取方法及应用 |
CN117678462A (zh) * | 2024-02-01 | 2024-03-12 | 云南省农业科学院农业环境资源研究所 | 一种基于红薯化感作用的除草剂减施增效方法 |
CN117678462B (zh) * | 2024-02-01 | 2024-04-19 | 云南省农业科学院农业环境资源研究所 | 一种基于红薯化感作用的除草剂减施增效方法 |
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