CN114747412A - 一种提高星油藤组培苗驯化成活率的方法 - Google Patents
一种提高星油藤组培苗驯化成活率的方法 Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
本发明公开了一种提高星油藤组培苗驯化成活率的方法，星油藤又称为南美油藤、印加果和印加花生等，是大戟科的一种多年生木质藤本油料植物，从其种子榨取的星油藤油可用于食品、药品和化妆品等领域。通过组织培养可在不受季节限制、短期内获得大量星油藤优质种苗，可满足星油藤规模化种植的种苗需求，但也存在组培苗驯化成活率较低的问题。因此，本发明通过合成氨基酸稀土配合物，并应用于星油藤组培苗驯化栽培中，可显著提高星油藤组培苗驯化成活率，驯化成活率达到95％以上，解决了星油藤组培苗驯化成活率低的问题，促进了星油藤规模化种植。
Description
技术领域
本发明涉及植物组织培养方法，具体地涉及一种提高星油藤组培苗驯化成活率的方法。
背景技术
星油藤(Plukenetia volubilis L.)又称为南美油藤、印加果和印加花生等，是大戟科的一种多年生木质藤本油料植物，原产于南美洲安第斯山脉地区热带雨林。从星油藤种子榨取的星油藤油，由于富含多元不饱和脂肪酸且ω-3、ω-6、ω-9三种不饱和脂肪酸高达92％以上，被认为是一种高端植物油，可用于食品、药品和化妆品等领域，对调整血脂、预防心血管疾病、保养肌肤等有显著效果。星油藤于2006年被引进中国科学院西双版纳热带植物园培育种植并获得成功，并云南、贵州等省市进行大规模种植。星油藤种苗繁育主要有播种、嫁接、扦插和组织培养等方式，其中组织培养具有繁殖系数高，虽可在短期内获得大量优质的星油藤种苗，但也存在组培苗驯化成活率较低的问题。
发明内容
本发明的目的在于提供一种提高星油藤组培苗驯化成活率的方法，本发明通过用甘氨酸稀土配合物处理星油藤组培苗，可显著提高星油藤组培苗驯化成活率，从而实现本发明的目的。
本发明的一种提高星油藤组培苗驯化成活率的方法，包括以下的步骤：
(1)将通过组培获得的高约4～7cm、根系粗壮、生长健壮、叶色浓绿的星油藤生根组培苗置于恒温大棚中，打开组培瓶的瓶盖并于25℃在自然光照下炼苗2周；
(2)炼苗结束后，将星油藤组培苗从组培瓶中取出，洗干净根部的培养基后，用10～30μmol/L氨基酸稀土配合物浸泡组培苗10～20s；
(3)氨基酸稀土配合物溶液处理后，将星油藤组培苗移栽培到经消毒处理后的栽培基质中栽培，定期浇水，保持透光度为60～80％，相对湿度85％以上。
栽培基质的配比为炭土:农家肥:河沙＝8:1:1。
步骤(2)所述的氨基酸稀土配合物为甘氨酸稀土配合物RE(Gly)2Cl·6H2O。具体地，所述的氨基酸稀土配合物为甘氨酸镧配合物、甘氨酸铈配合物、甘氨酸钕配合物中的一种。
所述的氨基酸稀土配合物由氯化稀土(RECl3·6H2O)和甘氨酸钠盐(C2H6NNaO3)合成，其合成方法为：首先将RE2O3溶于微过量的1:1盐酸中，加入适量的H2O2加速溶解，溶解完全后微热使残余H2O2分解完全，减压蒸发浓缩至大量晶体析出，抽滤并重结晶3次，所得结晶于P2O5干燥剂中干燥，直至恒重，用EDTA滴定法测定RE3+，用银量法测定Cl-，热重法测结晶水，证实结晶组成为RECl3·6H2O；然后按化学式计量RECl3·6H2O:C2H6NNaO3＝1:2称取，将C2H6NNaO3和RECl3·6H2O分别溶解于水和乙醇中，并在剧烈搅拌下将氯化稀土溶液滴加于甘氨酸钠盐水溶液中，回流搅拌2h，减压浓缩除去乙醇，将溶液转移至锥形瓶中，恒温加热浓缩，直至表面出现膨松晶体，真空干燥得固体配合物；经元素分析、质谱、紫外光谱、红外光谱、差热分析等表征证实氨基酸稀土配合物的组成为RE(Gly)2Cl·6H2O，RE＝La3+、Ce3+、Nd3 +；Gly为甘氨酸。
与现有技术相比，本发明具有的有益效果为：
1.甘氨酸是MS培养基中有机物的组成成分之一，是内源性抗氧化剂还原型谷胱甘肽的组成氨基酸，在农药生产上常用于合成拟除虫菊酯杀虫剂的中间体，也可合成杀虫剂异菌脲和除草剂草甘膦。稀土元素具有特殊的物理化学和生物化学效应，是一种生物微量元素，能与生物体内生物大分子如蛋白质、核酸等形成配合物，参与营养物质的吸收转运、抗逆抗病、激素活性及细胞通透性的调控，可减缓植物因逆境胁迫所产生的不良反应。本发明通过合成甘氨酸稀土配合物，并用其浸泡星油藤组培苗，利用甘氨酸和稀土元素两者配合产生的生物效应提高星油藤组培苗驯化成活率，可显著提高组培苗驯化的成活率。本发明具有成本低、效果好、操作性强等特点，解决了星油藤组培苗驯化成活率低的问题，促进了星油藤规模化种植。
具体实施方式
以下实施例是对本发明的进一步说明，不是对本发明的限制。
实施例一甘氨酸稀土配合物的合成
(1)氯化稀土的制备：将RE2O3(RE＝La3+、Ce3+、Nd3+)溶于微过量的1:1盐酸中，加入适量的H2O2加速溶解，溶解完全后微热使残余H2O2分解完全，减压蒸发浓缩至大量晶体析出，抽滤并重结晶3次，所得结晶于P2O5干燥剂中干燥，直至恒重，用EDTA滴定法测定RE3+，用银量法测定Cl-，热重法测结晶水，证实结晶组成为RECl3·6H2O。
(2)甘氨酸稀土配合物的制备：按化学式计量RECl3·6H2O:C2H6NNaO3＝1：2称取，将C2H6NNaO3和RECl3·6H2O分别溶解于水和乙醇中，并在剧烈搅拌下将氯化稀土溶液滴加于甘氨酸钠盐水溶液中，回流搅拌2h，减压浓缩除去乙醇，将溶液转移至锥形瓶中，恒温加热浓缩，直至表面出现膨松晶体，真空干燥得固体配合物。
(3)甘氨酸稀土配合物的表征：经元素分析、质谱、紫外光谱、红外光谱、差热分析等表征证实氨基酸稀土配合物的组成为RE(Gly)2Cl·6H2O。
实施例二甘氨酸铈配合物[Ce(Gly)2Cl·6H2O]对星油藤组培苗驯化成活率的影响
(1)将通过组培获得的高约4～7cm、根系粗壮、生长健壮、叶色浓绿的星油藤生根组培苗置于恒温大棚中，打开组培瓶的瓶盖并于25℃在自然光照下炼苗2周。
(2)炼苗结束后，将星油藤组培苗从组培瓶中取出，洗干净根部的基后，用10～30μmol/L甘氨酸铈配合物[Ce(Gly)2Cl·6H2O]浸泡组培苗10～20s。
(3)甘氨酸铈配合物溶液处理后，将星油藤组培苗移栽培到经消毒处理后的栽培基质中栽培，定期浇水，保持透光度为60～80％，相对湿度85％。移栽30d后统计驯化苗成活率，成活率达到95％以上。
Ce(Gly)2Cl·6H2O的浓度可以是10μmol/L、15μmol/L、20μmol/L、25μmol/L、30μmol/L。栽培基质的配比为炭土:农家肥:河沙＝8:1:1。
实施例三甘氨酸镧配合物[La(Gly)2Cl·6H2O]对星油藤组培苗驯化成活率的影响
(1)将通过组培获得的高约4～7cm、根系粗壮、生长健壮、叶色浓绿的星油藤生根组培苗置于恒温大棚中，打开组培瓶的瓶盖并于25℃在自然光照下炼苗2周。
(2)炼苗结束后，将星油藤组培苗从组培瓶中取出，洗干净根部的基后，用10～30μmol/L甘氨酸镧配合物[La(Gly)2Cl·6H2O]浸泡组培苗10～20s。
(3)甘氨酸镧配合物溶液处理后，将星油藤组培苗移栽培到经消毒处理后的栽培基质中栽培，定期浇水，保持透光度为60～80％，相对湿度85％。移栽30d后统计驯化苗成活率，成活率达到97％以上。
La(Gly)2Cl·6H2O的浓度可以是10μmol/L、15μmol/L、20μmol/L、25μmol/L、30μmol/L。栽培基质的配比为炭土:农家肥:河沙＝8:1:1。
实施例四甘氨酸钕配合物[Nd(Gly)2Cl·6H2O]对星油藤组培苗驯化成活率的影响
(1)将通过组培获得的高约4～7cm、根系粗壮、生长健壮、叶色浓绿的星油藤生根组培苗置于恒温大棚中，打开组培瓶的瓶盖并于25℃在自然光照下炼苗2周。
(2)炼苗结束后，将星油藤组培苗从组培瓶中取出，洗干净根部的基后，用10～30μmol/L甘氨酸钕配合物[Nd(Gly)2Cl·6H2O]浸泡组培苗10～20s。
(3)甘氨酸钕配合物溶液处理后，将星油藤组培苗移栽培到经消毒处理后的栽培基质(栽培基质的配比为炭土:农家肥:河沙＝8:1:1)中栽培，定期浇水，保持透光度为60～80％，相对湿度85％。移栽30d后统计驯化苗成活率，成活率达到100％。
Nd(Gly)2Cl·6H2O的浓度可以是10μmol/L、15μmol/L、20μmol/L、25μmol/L、30μmol/L。栽培基质的配比为炭土:农家肥:河沙＝8:1:1。
Claims (6)
1.一种提高星油藤组培苗驯化成活率的方法，其特征在于，包括以下步骤：
(1)将通过组培获得的高4～7cm、根系粗壮、生长健壮、叶色浓绿的星油藤生根组培苗置于恒温大棚中，打开组培瓶的瓶盖并于25℃在自然光照下炼苗2周；
(2)炼苗结束后，将星油藤组培苗从组培瓶中取出，洗干净根部的培养基后，用10～30μmol/L氨基酸稀土配合物浸泡组培苗10～20s；
(3)氨基酸稀土配合物溶液处理后，将星油藤组培苗移栽培到经消毒处理后的栽培基质中栽培，定期浇水，保持透光度为60～80％，相对湿度85％以上。
2.根据权利要求1所述的提高星油藤组培苗驯化成活率的方法，其特征在于，所述栽培基质的配比为炭土:农家肥:河沙＝8:1:1。
3.根据权利要求1或2所述的提高星油藤组培苗驯化成活率的方法，其特征在于，所述的氨基酸稀土配合物为甘氨酸稀土配合物。
4.根据权利要求3所述的提高星油藤组培苗驯化成活率的方法，其特征在于，所述的甘氨酸稀土配合物为甘氨酸镧配合物、甘氨酸铈配合物和甘氨酸钕配合物中的一种。
5.根据权利要求3所述的提高星油藤组培苗驯化成活率的方法，其特征在于，所述的甘氨酸稀土配合物由氯化稀土和甘氨酸钠盐合成。
6.根据权利要求5所述的提高星油藤组培苗驯化成活率的方法，其特征在于，所述甘氨酸稀土配合物的合成方法为：将RE2O3溶于微过量的1:1盐酸中，加入H2O2加速溶解，溶解完全后加热使残余H2O2分解完全，减压蒸发浓缩至大量晶体析出，抽滤并重结晶3次，所得结晶于P2O5干燥剂中干燥，直至恒重得RECl3·6H2O；然后按化学式计量RECl3·6H2O:C2H6NNaO3＝1:2称取，将C2H6NNaO3和RECl3·6H2O分别溶解于水和乙醇中，并在剧烈搅拌下将氯化稀土溶液滴加于甘氨酸钠盐水溶液中，回流搅拌2h，减压浓缩除去乙醇，将溶液转移至锥形瓶中，恒温加热浓缩，直至表面出现膨松晶体，真空干燥得固体配合物RE(Gly)2Cl·6H2O。
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| CN106342689A (zh) * | 2016-09-06 | 2017-01-25 | 海南大学 | 一种南美星油藤快速繁殖的方法 |
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