CN111134125B - 一种生物农药与植物生长调节复合剂及制备方法 - Google Patents
一种生物农药与植物生长调节复合剂及制备方法 Download PDFInfo
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Classifications
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- 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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- 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
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/36—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
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- 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
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
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- 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
- A01N43/06—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 five-membered rings
- A01N43/08—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 five-membered rings with oxygen as the ring hetero atom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
- A01N59/06—Aluminium; Calcium; Magnesium; Compounds thereof
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
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- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
本发明提供了一种生物农药与植物生长调节复合剂,该复合剂来源于鼠李糖脂水解生产鼠李糖的油相副产物,其主要成分含量为:鼠李糖脂0.01‑0.5g/L、混合羟基脂肪酸2.5‑45g/L、鼠李糖0.005‑0.125g/L、氨基酸衍生物0.05‑0.5g/L、5‑甲基糠醛0.05‑1.25g/L、钙离子0.005‑0.025g/L。该复合剂能抑制真菌和病毒引起的植物病害,同时促进蔬菜瓜果的生长与成熟,增加叶绿素的合成。其效果与功能多样性远超已知的活性组分鼠李糖脂和羟基脂肪酸,说明各组分之间存在协同增效作用。此外,该复合剂的原料作为鼠李糖生产的副产物,其几乎无需生产成本,且能节约大量的环保处理费用。该复合剂的有效组分都是天然的,对环境无污染,对人体无毒性,是一种绿色环保的新型产品。
Description
技术领域
本发明涉及一种利用生产废料制备的生物农药与植物生长调节复合剂,属于植物病害防治与生长调节的范畴。
背景技术
在现代农业生产中,广泛使用化学合成物作为农药与植物生长调节剂。化学农药与植物激素的残留会影响人体健康、引起抗药性和损害自然环境,并导致农田植物口感和营养价值差等问题。随着近年来人们对有机食品的需求越来越多,有机农业与无公害农业也随之快速发展,因此,开发新型的生物农药与植物生长调节复合制剂,可大大提高植物性食品的健康性和安全性,顺应时代发展的要求。目前,已有一些专利[US20180327330 A1,CN200910181030等]报道了生物农药与有机肥的复合剂,其成分主要来源于植物提取物、表面活性剂和微生物发酵产物。这种生物农药与有机肥的复合剂,不但同时具有生物农药与有机肥的优点,还可同时施设,降低劳动强度。
目前,已知鼠李糖脂具有抗植物真菌的作用[US 20170265468A1],并增加其他生物农药的效果[CN201510911524]。此外,羟基脂肪酸也具有一定的抗真菌能力(Hou CT&Forman RJ.J Ind Microbiol Biotechnol 2000,24,275-276)。国内也有专利[CN200910086309]报道混合脂肪酸制剂可抑制烟草花叶病毒。
在国内外的各类文献中,生物农药与植物生长调节复合剂的有效组分以商品化的成分为主,其成本较高,未见附加值低的副产物为唯一主料的复合剂。
发明内容
本发明提供了一种生物农药与植物生长调节复合剂。
本发明通过以下技术方案实现:
一种生物农药与植物生长调节复合剂,该复合剂含有:鼠李糖脂0.01-0.5g/L、混合羟基脂肪酸2.5-45g/L、鼠李糖0.005-0.125g/L、氨基酸衍生物0.05-0.5g/L,5-甲基糠醛0.05-1.25g/L,钙离子0.001-0.025g/L。
进一步地,混合羟基脂肪酸包括碳链数目为8-12,碳碳双键数目为0-2的3-羟基脂肪酸。赖氨酸衍生物,甘氨酸衍生物。
一种上述的生物农药与植物生长调节复合剂的制备方法,包括如下步骤:
(1)将鼠李糖脂发酵液调pH至2-4后,静置1-24小时,弃去上清液,得到鼠李糖脂浓度为100-500g/L的浓缩液。
(2)在上述鼠李糖脂浓缩液中加入质量比为1-5%的浓硫酸,在121℃下搅拌水解1-3小时,离心收集其油相。
(3)在步骤(2)收集的油相中加入液碱,调pH至4-8,优选的,调pH至5-7.5。然后加入水,体积比为1:40-1:200,混合均匀获得生物农药与植物生长调节复合剂。
一种生物农药与植物生长调节复合剂的制备方法,鼠李糖脂水解生产鼠李糖的油相副产物中加入液碱,调pH至4-8,优选的,调pH至5-7.5。然后加入水,体积比为1:40-1:200,混合均匀获得生物农药与植物生长调节复合剂。所述油相副产物中含鼠李糖脂2-20g/L、混合羟基脂肪酸500-900g/L、鼠李糖1-5g/L、氨基酸衍生物5-20g/L,5-甲基糠醛5-50g/L,钙离子0.1-1g/L。
所述生物农药与植物生长调节复合剂,其使用方法为:在植物叶面均匀喷洒,用量为20-100ml/m2,每周喷洒1-2次。
所述的生物农药与植物生长调节复合剂,其用途在于:
(1)真菌引起的植物病害防治,例如番茄晚疫病、水稻纹枯病、黄瓜白粉病等;
(2)病毒引起的植物病害防治,例如烟草花叶病毒;
(3)蔬菜瓜果的生长与成熟促进剂;
(4)叶绿素合成的增效剂。
所述的生物农药与植物生长调节复合剂,其优势在于:本发明生物农药与植物生长调节复合剂制备可与鼠李糖生产同时实施,以鼠李糖生产油相副产物为原料,成本极低;并且本发明的生物农药与植物生长调节复合剂来源于天然发酵产物,具有生物可降解、无环境污染、绿色环保、对人体安全的优点。
具体实施方式
下面以具体实施例对本发明进行详细说明。
实施例1:制备鼠李糖脂发酵液
发酵菌种为铜绿假单胞菌ZJU-211(Pseudomonas aeruginosa ZJU-211,菌种保藏号:CCTCC No:M209237),将菌种接种于60ml的种子培养基,温度为38℃,摇床转速200rpm,培养48h得到摇瓶菌种。种子培养基组成如下:一级大豆油,40g/L:NaNO3,4g/L;NaCl,1.5g/L;KCl,1.5g/L;CaCl2·2H2O,0.2g/L;KH2PO4,4g/L;Na2HPO4·12H2O,4g/L;MgSO4,0.3g/L;微量元素,2.5mL/L。所述微量元素组成如下:FeCl3·6H2O 0.08g/L,ZnSO4·7H2O 0.75g/L,CuSO4·5H2O 0.075g/L,MnSO4·H2O 0.75g/L,H3BO3,0.15g/L。
将摇瓶菌种接种到3吨发酵培养基中,接种量2%(v/v),发酵过程温度控制在38℃,通气量为1.0vvm,搅拌转速300rpm,发酵周期4天。发酵培养基组成如下:一级大豆油,70g/L;NaNO3,8g/L;NaCl,1.5g/L;KCl,1.5g/L;CaCl2·2H2O,0.2g/L;KH2PO4,4g/L;Na2HPO4·12H2O,4g/L;MgSO4,0.3g/L;微量元素,2.5mL/L。所述微量元素组成如下:FeCl3·6H2O 0.08g/L,ZnSO4·7H2O 0.75g/L,CuSO4·5H2O 0.075g/L,MnSO4·H2O 0.75g/L,H3BO3,0.15g/L。
发酵结束后,得到含鼠李糖脂50g/L的发酵液约1.6吨。
实施例2:鼠李糖脂发酵液的水解
在实施例1中获得的鼠李糖脂发酵液中加入浓盐酸,调pH至4,搅拌均匀,并静置24小时后,鼠李糖脂沉淀在底部,弃去上清液,得到370L的浓缩鼠李糖脂,其鼠李糖脂浓度为216g/L。在该浓缩鼠李糖脂中加入质量比2.3%的浓硫酸,并在搪瓷罐内加压搅拌水解2小时,水解温度为121℃。
水解结束后,其分层为两相:上层为黑色的油状液体,体积为111L;下层为棕色的水相,体积为259L,其中含有112g/L的鼠李糖,用于后续的鼠李糖生产。
现有的鼠李糖生产技术是将发酵法生产的鼠李糖脂水解后,其分解成含鼠李糖的水相,用于提纯鼠李糖产品,同时,产生了大量的油相副产物,占总体积的1/3。该油相废料被直接丢弃,不但没有产生价值,反而需要高昂的环保处理费用。将本发明与鼠李糖生产同时实施,可有效解决上述问题。
实施例3:油相副产物的成分鉴定
为确定油相中的主要成分,我们对多批次的鼠李糖脂浓缩液进行了水解。通过不同批次鼠李糖生产废料的组分分析,得到各主要成分的含量如下表:
实施例4:生物农药与有机肥复合剂的配置
采用实施例3中的2批次油相废料,分别配置如下:
(1)取20191008批次的油相100L,加入工业液碱6kg,混合均匀后pH被调至4,按1:40的比例,加入自来水4吨,搅拌混合均匀,呈水相的灰黑色液体。其主要成分含量为:鼠李糖脂0.388g/L、羟基脂肪酸17g/L、鼠李糖0.07g/L、氨基酸衍生物0.5g/L、5-甲基糠醛0.455g/L、钙离子0.05g/L。使用前,装入农用便携式喷洒器,在田间直接进行喷洒作业。
(2)取20190930批次的油相100L,加入工业液碱7kg,混合均匀后pH被调至7.1,按1:200的比例,加入自来水20吨,搅拌混合均匀,呈水相的灰黑色液体。其主要成分含量为:鼠李糖脂0.1g/L、羟基脂肪酸2.5g/L、鼠李糖0.0085g/L、氨基酸衍生物0.05g/L、5-甲基糠醛0.079g/L、钙离子0.002g/L。使用前,装入农用便携式喷洒器,在田间直接进行喷洒作业。
实施例5:复合剂抗植物真菌田间实验
采用实施例4中20191008批次油相废料配置的复合剂,并以0.388g/L鼠李糖脂水溶液、17g/L的3-羟基癸酸、0.388g/L鼠李糖脂+17g/L的3-羟基癸酸以及清水作为对照组。对感染番茄晚疫病的番茄植株进行喷洒,剂量均为100ml/m2,7天内共喷洒2次。复合剂组的治疗有效率达到65%,鼠李糖脂组的有效率为35%,羟基癸酸组有效率为5%,鼠李糖脂+羟基癸酸组有效率为52.5%,而清水组的有效率为0%。
采用实施例4中20190930批次油相废料配置的复合剂,并以0.1g/L鼠李糖脂水溶液、2.5g/L的3-羟基癸酸、0.1g/L鼠李糖脂+2.5g/L的3-羟基癸酸以及清水作为对照组。对感染番茄晚疫病的番茄植株进行喷洒,剂量均为100ml/m2,7天内共喷洒2次。复合剂组的治疗有效率达到45%,鼠李糖脂组的有效率为15%,羟基癸酸组有效率为3%,鼠李糖脂+羟基癸酸组有效率为29%,而清水组的有效率为0%。
上述实施例说明复合剂的抑菌效果远高于同浓度的鼠李糖脂或羟基脂肪酸,且高于鼠李糖脂与羟基癸酸合用组,说明复合剂中其他组分也起到了抑菌作用。此外,鼠李糖脂与羟基癸酸合用组的效果优于鼠李糖脂与羟基癸酸的单用,说明鼠李糖脂与羟基脂肪酸之间也存在协同增效作用。
实施例6:复合剂抗植物病毒的田间实验
采用实施例4中20191008批次油相废料配置的复合剂,并以0.388g/L鼠李糖脂水溶液、17g/L的3-羟基癸酸、0.388g/L鼠李糖脂+17g/L的3-羟基癸酸以及清水作为对照组。对感染烟草花叶病毒的烟草植株进行喷洒,剂量均为100ml/m2,7天内共喷洒2次。复合剂组的治疗有效率达到65%,鼠李糖脂组的有效率为5%,羟基癸酸组有效率为32%,鼠李糖脂+羟基癸酸组有效率为59%,清水组的有效率为0%。
采用实施例4中20190930批次油相废料配置的复合剂,并以0.1g/L鼠李糖脂水溶液、2.5g/L的3-羟基癸酸、0.1g/L鼠李糖脂+2.5g/L的3-羟基癸酸以及清水作为对照组。对感染烟草花叶病毒的烟草植株进行喷洒,剂量均为100ml/m2,7天内共喷洒2次。复合剂组的治疗有效率达到55%,鼠李糖脂组的有效率为11%,羟基癸酸组有效率为15%,鼠李糖脂+羟基癸酸组有效率为51%,而清水组的有效率为0%。
上述实施例说明复合剂的抗病毒效果远高于同浓度的鼠李糖脂或羟基脂肪酸单用,且略高于鼠李糖脂与羟基癸酸合用组,因此鼠李糖脂与羟基脂肪酸在抗病毒上具有显著的协同增效作用,且复合剂中的其他组分也具有较弱的抗病毒作用。
实施例7:复合剂促进植株生长的田间实验
采用实施例4中20191008批次油相废料配置的复合剂,并以0.388g/L鼠李糖脂水溶液、17g/L的3-羟基癸酸、0.388g/L鼠李糖脂+17g/L的3-羟基癸酸以及清水作为对照组。对番茄苗进行喷洒,剂量均为20ml/m2,7天内共喷洒2次。喷洒结束后14天,随机测量100株苗的高度增长量,以清水组番茄苗的高度增长量为100%,计算得到复合剂组的番茄苗高度增长了158%,而鼠李糖脂组的高度增长量为105%,羟基癸酸组的高度增长量为102%,与清水组效果差别不大。鼠李糖脂+羟基癸酸组的高度增长量为135%。
采用实施例4中20190930批次油相废料配置的复合剂,并以0.1g/L鼠李糖脂水溶液、2.5g/L的3-羟基癸酸、0.1g/L鼠李糖脂+2.5g/L的3-羟基癸酸以及清水作为对照组。对番茄苗进行喷洒,剂量均为100ml/m2,7天内共喷洒2次。喷洒结束后14天,随机测量100株苗的高度增长量,以清水组番茄苗的高度增长量为100%,计算得到复合剂组的番茄苗高度增长了135%,而鼠李糖脂组的高度增长量为101%,羟基癸酸组的高度增长量为102%,与清水组效果无显著差别。鼠李糖脂+羟基癸酸组的高度增长量为125%.
该实施例说明,复合剂起到了植物生长促进的作用,该作用主要由鼠李糖脂与羟基脂肪酸的合用而产生,其原理可能在于鼠李糖脂促进了植物对羟基脂肪酸的吸收利用。而鼠李糖脂与羟基脂肪酸单用几乎没有该作用。
实施例8:复合剂促进叶绿素合成的田间实验
采用实施例4中20191008批次油相废料配置的复合剂,并以0.388g/L鼠李糖脂水溶液、17g/L的3-羟基癸酸0.388g/L鼠李糖脂+17g/L的3-羟基癸酸以及清水作为对照组。对油菜苗进行喷洒,剂量均为20ml/m2,7天内共喷洒2次。喷洒结束后14天,随机采集100株苗的顶端新鲜叶片合计500g,混合后提取叶绿素,以清水组的叶绿素含量为100%,则复合剂组的叶绿素为145%,鼠李糖脂组为98%,羟基癸酸组为99%,与清水组效果无差异。鼠李糖脂+羟基癸酸组为112%。
采用实施例4中20190930批次油相废料配置的复合剂,并以0.1g/L鼠李糖脂水溶液,2.5g/L的3-羟基癸酸,0.1g/L鼠李糖脂+2.5g/L的3-羟基癸酸,以及清水作为对照组。对油菜苗进行喷洒,剂量均为100ml/m2,7天内共喷洒2次。喷洒结束后14天,随机采集100株苗的顶端叶片,混合后提取叶绿素,以清水组的叶绿素含量为100%,复合剂组的叶绿素为125%,鼠李糖脂组为99%,羟基癸酸组为98%,与清水组效果无差异。鼠李糖脂+羟基癸酸组为109%。
该实施例说明,复合剂起到了植物生长促进的作用,其效果优于鼠李糖脂与羟基脂肪酸合用,且鼠李糖脂与羟基癸酸单用没有该作用。
实施例9:复合剂的生物可降解性
将20191008批次的油相废料进行堆肥实验,采用油相废料与枯秸秆、鸭粪、水按10:10:1:1比例堆放,地衣菌、枯草杆菌用量为25g/m3。经过半个月后,肥堆的温度上升到了48℃,远高于对照组的13℃(冬天的大棚环境温度)。经过1个月的发酵后,肥堆中的油相已经肉眼不可见。经过2个月后,对堆肥取样测定,已经检测不到中长链脂肪酸、鼠李糖脂、氨基酸衍生物和5-甲基糠醛,说明已经完全降解。
该实施例说明复合剂的可降解性,是环境友好,无污染的天然制剂。
应当说明的是,虽然本发明以较佳实施例揭露如上,对本发明实施例中的技术方案进行清楚、完整地描述,然其并非用以限定本发明,在不脱离本发明的精神和范围内,所作出的任何修改、等同替换和改进等,均应包含在本发明的保护范围之内。
Claims (3)
1.一种复合剂在增效叶绿素合成的用途,其特征在于,该复合剂含有:鼠李糖脂0.01-0.5g/L、混合羟基脂肪酸2.5-45g/L、鼠李糖0.005-0.125g/L、氨基酸衍生物0.05-0.5g/L、5-甲基糠醛0.05-1.25g/L、钙离子0.001-0.025g/L;其中,混合羟基脂肪酸包括碳链长度为8-12个碳,碳碳双键数目为0-2的3-羟基脂肪酸。
2.根据权利要求1所述的用途,其特征在于,氨基酸衍生物包括赖氨酸衍生物,甘氨酸衍生物。
3.根据权利要求1所述的用途,其特征在于,所述复合剂通过如下方法制备获得:
往鼠李糖脂水解生产鼠李糖的油相副产物中加入液碱,调pH至4-8;然后加入水,体积比为1:40-1:200,混合均匀获得增效叶绿素合成的复合剂;所述油相副产物中含鼠李糖脂2-20g/L、混合羟基脂肪酸500-900g/L、鼠李糖1-5g/L、氨基酸衍生物5-20g/L,5-甲基糠醛5-50g/L,钙离子0.1-1g/L。
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