CN114946312B - 一种发芽玉米中类胡萝卜素高效富集的方法 - Google Patents
一种发芽玉米中类胡萝卜素高效富集的方法 Download PDFInfo
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- A—HUMAN NECESSITIES
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- A01C1/08—Immunising seed
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- Life Sciences & Earth Sciences (AREA)
- Soil Sciences (AREA)
- Environmental Sciences (AREA)
- Pretreatment Of Seeds And Plants (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
本发明公开了一种发芽玉米中类胡萝卜素高效富集的方法,属于农产品加工技术领域。本发明以黄玉米籽粒为原料,置于0.05~0.1mmol/L水杨酸溶液中浸种处理7h后停止浸泡1h,并更换新的水杨酸溶液继续浸泡2次,随后在24~26℃黑暗条件下正常发芽72~96h后,制得富集类胡萝卜素的发芽玉米。本发明技术操作简单易行、成本较低,且生长出来的发芽玉米中总类胡萝卜素含量高达22.8~28.6μg/g DW,其中叶黄素和玉米黄质含量分别为11.0~12.4μg/g DW和7.4~8.5μg/g DW,发芽玉米的DPPH和ABTS自由基清除能力分别提高1.2~1.5和1.4~1.8倍,类胡萝卜素合成关键基因PSY表达量提高1.9~2.4倍,CYP97C表达量提高2.9~3.5倍,BCH1表达量提高2.1~3.2倍。
Description
技术领域
本发明涉及的是一种发芽玉米中类胡萝卜素高效富集的方法,属于农产品加工技术领域。
背景技术
玉米(Zea mays L.)是富含维生素A源的主要粮食作物之一,也是我国普遍种植的一种农作物,主要用于淀粉制造、酿酒、饲料和食品工业。黄玉米籽粒包含六种主要的类胡萝卜素:α-胡萝卜素、β-胡萝卜素、α-隐黄质、β-隐黄质、叶黄素和玉米黄质。大力开发玉米中的营养成分-类胡萝卜素,能够为玉米种植、淀粉生产等行业带来高额的产品附加值。类胡萝卜素是一种天然的食品着色剂,也可作为生产保健产品的添加剂,其中叶黄素和玉米黄质对预防和治疗心血管疾病、白内障以及老年性黄斑变性等疾病具有明显功效。
芽苗菜是用可食用种子培育出来的嫩芽、芽苗、芽球等的总称。研究表明,发芽可以进一步增强种子的营养价值和药用价值。在发芽的过程中,大分子的碳水化合物、蛋白质发生分解和降解,同时小分子的糖、游离氨基酸含量增加。此外,一些次生代谢产物,如类胡萝卜素、多酚含量增加。研究发现,在植物籽粒发芽过程中,施加外源诱导物可促进一些代谢产物的合成,激活关键酶的活力及其基因表达量。水杨酸(Salicylic acid,SA)是广泛存在于植物体内的一种小分子酚类物质,也是一种植物内源激素,具有独特的生理作用。八氢番茄红素合成酶(PSY)是类胡萝卜素合成的第一步限速酶,胡萝卜素β环羟化酶(BCH)和胡萝卜素ε-环羟化酶(CYP97C)是催化α-胡萝卜素和β-胡萝卜素合成叶黄素和玉米黄质的关键酶。因此,在玉米籽粒浸种过程中进行SA浸种处理,可激活类胡萝卜素合成关键基因的表达,促使发芽玉米合成更多的类胡萝卜素,同时生长出来的发芽玉米具有较高的自由基清除能力。
发明内容:
本发明所要解决的技术问题:本发明提供一种发芽玉米中类胡萝卜素高效富集的方法,技术工艺简便、成本较低,同时生长出的发芽玉米的类胡萝卜素含量及DPPH和ABTS自由基清除能力均显著提高。其技术方案如下:
一种发芽玉米中类胡萝卜素高效富集的方法,包括以下步骤:
(1)筛选:用清水淘洗玉米籽粒,剔除干瘪,破损的玉米籽粒;
(2)消毒:用0.5%NaClO溶液浸泡20min消毒;
(3)水杨酸浸种:将步骤(2)中消毒后的玉米籽粒用去离子水淘洗,清洗表面残余的NaClO溶液;用水杨酸溶液于25℃下浸泡24h,期间每浸泡处理7小时,停止浸泡1小时,并更换新的水杨酸溶液;
(4)发芽:将浸泡后的玉米籽粒置于发芽机中,每隔4h喷洒去离子水,在24~26℃黑暗条件下发芽72~96h,即得富含类胡萝卜素的发芽玉米。
优选地,所述水杨酸浓度为0.05~0.1mmol/L。
优选地,所述每次浸种水杨酸用量为2~3倍体积。
本发明的有益效果:
(1)本发明采用水杨酸浸种处理,显著提高了发芽玉米中类胡萝卜素含量,其含量达22.8~28.6μg/g DW,是对照组的1.26~1.58倍,其中叶黄素的含量为11.0~12.4μg/gDW,玉米黄质含量为7.4~8.5μg/g DW。经水杨酸浸种处理后,发芽玉米中DPPH和ABTS自由基清除能力分别提高1.2~1.5和1.4~1.8倍。水杨酸浸种处理激活了类胡萝卜素合成关键基因PSY、CYP97C和BCH1的表达量,分别提高1.9~2.4倍,2.9~3.5倍,和2.1~3.2倍。
(2)本发明具有投资少,方便易行,效果显著的特点,比化学合成和基因工程方法更具有安全性。
具体实施方式
实施例1:
(1)筛选:用清水淘洗玉米籽粒,剔除干瘪,破损的玉米籽粒;
(2)消毒:用2倍体积的0.5%NaClO溶液浸泡20min消毒;
(3)水杨酸浸种:将步骤(2)中消毒后的玉米籽粒用去离子水淘洗,清洗表面残余的NaClO溶液;用2倍体积的0.05mmol/L水杨酸溶液于25℃下浸泡24h,期间每浸泡处理7小时,停止浸泡1小时,并更换新的水杨酸溶液;
(4)发芽:将浸泡后的玉米籽粒置于发芽机中,在26℃黑暗条件下发芽96h,每隔4h喷洒去离子水。
(5)HPLC测定发芽玉米籽粒中总类胡萝卜素含量为22.8μg/g DW,是对照组的1.26倍,其中叶黄素和玉米黄质含量分别为11.0和7.4μg/g DW,分别是对照组的1.19和1.21倍,发芽玉米DPPH和ABTS自由基清除能力分别提高1.2和1.4倍,荧光定量PCR分析后类胡萝卜素合成关键基因PSY、CYP97C和BCH1表达量分别提高1.9、2.9和2.1倍。
实施例2:
(1)筛选:用清水淘洗玉米籽粒,剔除干瘪,破损的玉米籽粒;
(2)消毒:用2倍体积的0.5%NaClO溶液浸泡20min消毒;
(3)水杨酸浸种:将步骤(2)中消毒后的玉米籽粒用去离子水淘洗,清洗表面残余的NaClO溶液;用2倍体积的0.05mmol/L水杨酸溶液于25℃下浸泡24h,期间每浸泡处理7小时,停止浸泡1小时,并更换新的水杨酸溶液;
(4)发芽:将浸泡后的玉米籽粒置于发芽机中,在25℃黑暗条件下发芽72h,每隔4h喷洒去离子水。
(5)HPLC测定发芽玉米籽粒中总类胡萝卜素含量为28.6μg/g DW,是对照组的1.58倍,其中叶黄素和玉米黄质含量分别为12.4和8.5μg/g DW,分别是对照组的1.34和1.39倍,发芽玉米DPPH和ABTS自由基清除能力分别提高1.5和1.8倍,荧光定量PCR分析后类胡萝卜素合成关键基因PSY、CYP97C和BCH1表达量分别提高2.4、3.5和3.2倍。
实施例3:
(1)筛选:用清水淘洗玉米籽粒,剔除干瘪,破损的玉米籽粒;
(2)消毒:用2倍体积的0.5%NaClO溶液浸泡20min消毒;
(3)水杨酸浸种:将步骤(2)中消毒后的玉米籽粒用去离子水淘洗,清洗表面残余的NaClO溶液;用3倍体积的0.1mmol/L水杨酸溶液于25℃下浸泡24h,期间每浸泡处理7小时,停止浸泡1小时,并更换新的水杨酸溶液;
(4)发芽:将浸泡后的玉米籽粒置于发芽机中,在24℃黑暗条件下发芽84h,每隔4h喷洒去离子水。
(5)HPLC测定萌发后玉米籽粒中总类胡萝卜素含量为25.4μg/g DW,是对照组的1.40倍,其中叶黄素和玉米黄质含量分别为11.5和7.9μg/g DW,分别是对照组的1.24和1.30倍,发芽玉米DPPH和ABTS自由基清除能力分别提高1.3和1.6倍,荧光定量PCR分析后类胡萝卜素合成关键基因PSY、CYP97C和BCH1表达量分别提高2.1、3.1和2.5倍。
以上所属实施例仅仅是对本发明的优选实施方式进行描述,并非对本发明的构思和范围进行限定,在不脱离本发明涉及方案前提下,本领域技术人员对本发明的技术方案作出的各种显而易见的变型和改进,均应落入本发明的保护范围,本发明请求保护的技术内容,已全部记载在权力要求书中。
Claims (6)
1.一种发芽玉米中类胡萝卜素高效富集的方法,其特征在于,以黄玉米籽粒为原料,经NaClO溶液浸泡消毒,置于0.05~0.1mmol/L水杨酸溶液中浸种处理24h后,正常发芽72~96h,并在发芽过程中喷淋去离子水,制得富集类胡萝卜素的发芽玉米。
2.根据权利要求1所述的一种发芽玉米中类胡萝卜素高效富集的方法,包括以下具体步骤:
(1)筛选:用清水淘洗玉米籽粒,剔除干瘪,破损的玉米籽粒;
(2)消毒:用0.5%NaClO溶液浸泡20min消毒;
(3)水杨酸浸种:将步骤(2)中消毒后的玉米籽粒用去离子水淘洗,清洗表面残余的NaClO溶液;用水杨酸溶液于25℃下浸泡24h,期间每浸泡处理7小时,停止浸泡1小时,并更换新的水杨酸溶液;
(4)发芽:将浸泡后的玉米籽粒置于发芽机中,每隔4h喷淋去离子水,在24~26℃黑暗条件下发芽72~96h,即得富含类胡萝卜素的发芽玉米。
3.根据权利要求1或2所述的一种发芽玉米中类胡萝卜素高效富集的方法,其特征在于,制得的发芽玉米籽粒中总类胡萝卜素含量为22.8~28.6μg/g DW,是对照组的1.26~1.58倍,其中叶黄素的含量为11.0~12.4μg/g DW,玉米黄质含量为7.4~8.5μg/g DW,与对照组相比,分别提高1.19~1.34倍和1.21~1.39倍。
4.根据权利要求1所述的一种发芽玉米中类胡萝卜素高效富集的方法,其特征在于,所述水杨酸溶液每次浸种用量为2~3倍体积。
5.根据权利要求1所述的一种发芽玉米中类胡萝卜素高效富集的方法,其特征在于,水杨酸溶液浸泡处理后的发芽玉米中DPPH和ABTS自由基清除能力分别提高1.2~1.5和1.4~1.8倍。
6.根据权利要求1所述的一种发芽玉米中类胡萝卜素高效富集的方法,其特征在于,水杨酸溶液浸泡处理后发芽玉米中类胡萝卜素合成关键基因PSY表达量提高1.9~2.4倍,CYP97C表达量提高2.9~3.5倍,BCH1表达量提高2.1~3.2倍。
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