CN109315403A - 提高经济作物与农作物营养功能成分的促进剂 - Google Patents
提高经济作物与农作物营养功能成分的促进剂 Download PDFInfo
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
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- Life Sciences & Earth Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
本发明公开了一种提高经济作物与农作物营养功能成分的促进剂,其是分子量为0.4‑10 kDa壳寡糖、分子量为0.5‑11 kDa低聚甲壳素、分子量为0.3‑10 kDa海藻寡糖中的一种或三种的任意比混合物;本发明原料来源广泛,加工成本低廉,使用简单,作用效果十分显著,可以有效提高经济作物与农作物内代谢产物的含量,且环境友好无污染,无有害化学残留。
Description
技术领域
本发明涉及植物代谢物促进剂技术领域,涉及一种提高经济作物与农作物营养功能成分促进剂。
背景技术
随着我国经济的快速发展,人们对经济作物与农作物的产量与质量要求越来越高,本功能营养成分促进剂不仅能促进植物的抗逆生长,还能非常有效地提高植物中多酚等营养功能物质的含量,增加经济作物和农作物的经济和营养功能价值。
我国海洋领土面积辽阔,海洋资源丰富,我国越来越重视对海洋资源与环境的开发利用。甲壳素是地球上储量仅次于纤维素的第二大可更新的有机资源,且我国东南部地区大面积临海,利用水产加工废弃物进行甲壳素的工业化生产,具有丰富的原料来源,与甲壳素和壳聚糖的大分子相比,低聚甲壳素水溶性好、无毒、更易被人体吸收,研究表明,低聚甲壳素具有抗肿瘤、增强免疫力、抗菌、保湿及促进植物抗逆生长等多种生理功能。壳寡糖是通过降解壳聚糖得到的低聚糖,由2~10个氨基葡糖通过β-1,4-糖苷键连接而成,由于其聚合度小,且含较多氨基,故水溶性较好,易吸收。壳寡糖具有抗病毒、调节免疫力、抗肿瘤和抗菌等作用,作为一种激发子,在植物病害防治上具有抑制病毒侵染的作用。海藻生活在开放的水体中,易受到病原侵袭和环境改变等因素带来的各种威胁,大规模的病害会给海藻养殖业带来巨大的经济损失。
目前国内外学者对多糖的降解研究较多,其方法主要有物理降解、化学降解、酶降解等。但是绝大多数多糖在水中的溶解性很差,就算在稀酸性水中的溶解性也不好,会造成成团、絮凝等情况,从而导致在后续降解过程中反应不均一,降解产物分子量大小差异很大,进而影响后续产品的质量和品质稳定性,同时由于需要反复降解提升了生产成本。
发明内容
本发明的目的是克服现有技术存在的缺点,提供一种植物营养功能成分促进剂,能够有效地增强经济作物和农作物内营养功能成分的含量,从而提升作物的经济价值;本发明提高经济作物与农作物营养功能成分的促进剂为分子量为0.4-10kDa壳寡糖、分子量为0.5-11kDa低聚甲壳素、分子量为0.3-10 kDa海藻寡糖中的一种或三种的任意比混合物。
本发明促进剂为常规市售产品或按常规方法制得的,例如可以通过以下方法制备得到:在功率为200-500 W的超声条件下,分别将壳聚糖、甲壳素、海藻多糖分散并溶解在(0.5-5%,m/v)苹果酸或枸椽酸等有机酸溶液中,然后加入有机酸溶液质量的1-3%的过氧化氢,在75-100℃、微波条件下反应0.5-2 h,分别制得分子量为0.4-10kDa壳寡糖、分子量为0.5-11kDa的低聚甲壳素、分子量为0.3-10kDa的海藻寡糖。
本发明促进剂使用时,将上述制备的分子量为0.4-10 kDa的壳寡糖,0.5-11 kDa的低聚甲壳素和0.3-10 kDa的海藻寡糖的三种寡糖分别或先后加入水中搅拌均匀,使壳寡糖为浓度10-1000mg/L、低聚甲壳素浓度为10-1000mg/L和海藻寡糖浓度为50-1000mg/L,即可制得经济作物与农作物营养功能成分促进剂。
本发明与现有技术相比,本发明具有如下优点:
1、本发明采用超声-微波辅助降解制备一种植物功能营养成分促进剂,可以有效解决多糖在稀酸溶液中成团和絮凝情况,降解反应均一,降解产物分子量大小差异较小;
2、通过实验发现本功能营养成分促进剂能够提高植物的代谢物生成量,例如总酚、总黄酮和二萜酚含量,大量研究表明植物酚类物质具有抗氧化、延缓衰老和预防退行性疾病等功效,故施用本营养功能成分促进剂后可以提高作物的经济价值;
3、其原料来源广泛,成本低廉,使用简单,作用效果十分显著,可以有效提高经济作物与农作物代谢产物的含量,且环境友好无污染,无有害化学残留。
附图说明
图1是迷迭香经不同浓度三种寡糖复合物处理后HPLC图(对照组);
图2是迷迭香经不同浓度三种寡糖复合物处理后HPLC图(低浓度组);
图3是迷迭香经不同浓度三种寡糖复合物处理后HPLC图(高浓度组)。
具体实施方式
下面结合具体实施实例对本发明作进一步说明,这些实验的事例仅用于说明本发明,不用于限制本发明的应用范围。在阅读了本发明的记载以后,本领域的科技人员对其等效的各种改动、修改和修饰,都属于本发明权利要求所限定的范围。
实施例1:
首先是在超声的辅助下(300W),将壳聚糖均匀分散并溶解于苹果酸溶液(1%,m/v)中,在苹果酸溶液中加入其质量1%的过氧化氢,在80℃、微波条件下反应1h,制得分子量为0.4-10kDa的低分子量壳寡糖,再将获得的低分子量壳寡糖溶于蒸馏水中,使得壳寡糖的浓度为10-1000 mg/L,即制得壳寡糖植物营养功能成分促进剂。
实施例2:
首先是在超声的辅助下(200W),将甲壳素均匀分散并溶解于苹果酸溶液(2%,m/v)中,在苹果酸溶液中加入其质量2%的过氧化氢,在85℃、微波条件下反应0.5h,制得0.5-11 kDa的低分子量甲壳素,再将获得的低分子量甲壳素溶于蒸馏水中,使得低聚甲壳素的浓度为10-1000 mg/L,即制得低聚甲壳素植物营养功能成分促进剂。
实施例3:
首先是在超声的辅助下(200W),将海藻糖均匀分散并溶解于枸椽酸溶液(2%,m/v)中,在枸椽酸溶液中加入其质量1%的过氧化氢,在85℃、微波条件下反应1h,制得0.3-10 kDa的低分子量甲壳素,再将获得的低分子量海藻糖溶于蒸馏水中,使得低聚甲壳素的浓度为50-1000mg/L,即制得海藻寡糖植物营养功能成分促进剂。
实施例4:
将上述制备的分子量为0.4-10 kDa的壳寡糖,0.5-11 kDa的低聚甲壳素和0.3-10 kDa的海藻寡糖的三种寡糖先后加入水中搅拌均匀,使其壳寡糖为浓度10-1000 mg/L、低聚甲壳素浓度为10-1000 mg/L和海藻寡糖浓度为50-1000 mg/L,即可制得经济作物与农作物营养功能成分促进剂。
实施例5:本实施例在种子萌芽时期对大豆种子用实施例1-4中所制壳寡糖、低聚甲壳素和海藻寡糖的一种或三种寡糖的组合复配进行浸种处理,具体实验如下:
将大豆种子消毒后用蒸馏水冲洗干净并晾干,挑选饱满、大小均匀的黄豆种子分别用实施例1-4中制得的壳寡糖200 mg/L和/或低聚甲壳素200 mg/L和/或海藻寡糖500 mg/L一种或三种寡糖的组合复配处理。
实验方法:将大豆种子进行五组不同处理的浸种实验,浸泡20-30 min,每组100粒,于发芽机中发芽培养96 h后测定总酚含量和总黄酮含量,用UHPLC-ESI-HRMS/MS测定其次级代谢产物组成变化。培养条件为:温度20-25℃,相对湿度75%-95%。第一组为壳寡糖200mg/L,第二组为低聚甲壳素200 mg/L,第三组为海藻寡糖500 mg/L,第四组为壳寡糖200mg/L、低聚甲壳素200 mg/L和海藻寡糖500 mg/L复配,第五组为对照组,用蒸馏水处理。
本发明所得提取物中次级代谢物组成鉴定:采用UHPLC-ESI-HRMS/MS对大豆种子中次级代谢产物的组成变化进行分析,具体操作如下:UHPLC分离条件为:分离柱为ZORBAXSB-C18 column (2.1 × 100 mm, 1.8 μm, 美国);流动相A为0.1%甲酸水,B为乙腈;分离梯度为:0-1 min,5% B;1-5 min,5-15% B;5-10 min,15-25% B;10-15 min,25-45% B;15-16 min,45-5% B;16-20 min,5% B;流速为0.2 mL/min;进样量为3 μL。质谱鉴定条件为:高分辨率质谱为Q-ExactiveOrbitrap质谱(Thermo Fisher Scientific,德国),主要技术参数为:全扫范围为50-1200 m/z;鞘气流速为,32 L/min;辅助气体流速,8 L/min;吹扫气体流速4 L/min;负离子模式喷雾电压,3.3 kV;离子传输管温度,320℃;S-lens RF水平,50%。
种子经各组处理0,24,48,72 h,96 h后,总酚含量、总黄酮含量变化如表1所示;经壳寡糖200 mg/L、低聚甲壳素200 mg/L、海藻寡糖500 mg/L及三种复配处理的大豆经72 h处理后总酚、总黄酮含量有显著提升,其中经三种寡糖复配处理72 h后的大豆总酚、总黄酮含量提升最为明显,分别提升了36.32%,39.87%;经UHPLC-ESI-HRMS/S检测发现,其酚类物质组成种类无变化,主要酚类物质如表2 所示。多酚是一类广泛存在于植物体内含有多个羟基的酚类植物成分的总称,是植物体内重要的次生代谢产物。本植物营养功能成分促进剂可促进经济作物与农作物的初级及次级代谢物的生成量,从而提升经济作物与农作物的的营养价值和功能活性,进而提升作物的经济价值。
表1:不同处理组中大豆种子的总酚、总黄酮含量变化
表2大豆主要次级代谢物酚类物质的组成鉴定
。
实施例6:本实施例在种子萌芽时期对大豆种子用实施例1-4中所制壳寡糖、低聚甲壳素和海藻寡糖的一种或三种寡糖的组合复配进行浸种处理,考察不同处理组在萌发过程中其提取物清除自由基的能力变化,具体实验如下:
将大豆种子消毒后用蒸馏水冲洗干净并晾干,挑选饱满、大小均匀的黄豆种子分别用实施例1中制得的壳寡糖200mg/L和/或低聚甲壳素200mg/L和/或海藻寡糖500mg/L一种或三种寡糖的组合复配处理。
实验方法:同实施例5;
待种子经各组处理0,24,48,72h,96 h 后,不同提取物对DPPH和ABTS自由基清除能力如表3所示;所有处理组的72h提取物对DPPH和ABTS的清除能力均显著增强,特别是经三种寡糖复配处理72h的大豆,其提取物对DPPH和ABTS自由基清除能力提升最为明显,分别达77.48 ± 2.45%,83.71 ± 3.73%。而食用经本营养功能成分促进剂处理后的农作物或经济作物的能够更有效地清除人体内过多的自由基,降低人体氧化应激水平,从而可以降低心血管疾病、退行性疾病和老化等疾病的患病风险。
表3不同处理组中大豆种子的DPPH、ABTS自由基清除能力
。
实施例7:用实施例1-4中所制的三种寡糖复配处理在幼苗时期对迷迭香进行喷洒,具体实验如下:
分别用低浓度(壳寡糖200 mg/L、低聚甲壳素200 mg/L和海藻寡糖500 mg/L)和高浓度(壳寡糖400 mg/L、低聚甲壳素400 mg/L和海藻寡糖1000 mg/L)三种寡糖复合物喷洒处于叶片生长关键期的迷迭香,每隔5天进行一次喷洒,连续喷洒60天,同时设立蒸馏水喷洒的对照组。
取10 g迷迭香叶片粉碎后溶于200 mL的80%异丙醇中,于40℃超声提取30 min,取1 mL迷迭香提取物加入4 mL 80%甲醇经0.22 μm滤膜过滤于进样瓶中,用HPLC分析低浓度(壳寡糖200 mg/L、低聚甲壳素200 mg/L和海藻寡糖500 mg/L)、高浓度(壳寡糖400 mg/L、低聚甲壳素400 mg/L和海藻寡糖1000 mg/L)及对照组中迷迭香次级代谢产物的含量变化情况。HPLC方法具体操作如下:HPLC分离条件为:分离柱为Amethyst C18-H (4.6 × 250mm, 5μm, 美国);流动相A为0.1%甲酸水,B为乙腈;分离梯度为:0-10 min,25% B;10-15min,25-75% B;15-25 min,25-75% B;25-26 min,75-25% B;26-35 min,75-25% B;流速为1mL/min;进样量为20 μL。
由图1、2、3,表4可看出,迷迭香经不同浓度的三种寡糖复合物喷洒后,其叶片中主要活性物质迷迭香酸含量、鼠尾草酚含量与对照组相比显著提高(p< 0.05),且二者含量随寡糖浓度增大而升高,而鼠尾草酸含量无明显变化,可能是在提取过程中鼠尾草酸分解为鼠尾草酚所致;以上结果表明壳寡糖三种寡糖复合物能够显著提升迷迭香中主要活性次级代谢产物的含量,从而可以提升迷迭香的营养和经济价值。
表4不同浓度三种寡糖复配处理后中迷迭香中酚类物质含量变化
注:含量单位为:μg/g
综上所述,可以确定本发明中三种寡糖及其复合物可以有效地提高经济作物和农作物中营养功能成分的含量,提高植物原材料提取物的自由基清除能力,从而可以有效提高作物的营养价值和经济价值。
Claims (1)
1.一种提高经济作物与农作物营养功能成分的促进剂,其特征在于:是分子量为0.4-10 kDa壳寡糖、分子量为0.5-11 kDa低聚甲壳素、分子量为0.3-10 kDa海藻寡糖中的一种或三种的任意比混合物。
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