CN115336586B - 一种提高白藜发芽率和芽苗中叶酸含量的方法 - Google Patents
一种提高白藜发芽率和芽苗中叶酸含量的方法 Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- 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
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
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Abstract
本发明涉及农业技术领域,具体是一种提高白藜发芽率和芽苗中叶酸含量的方法,是采用浓度为50‑200mg/L天冬酰胺溶液处理白藜种子。本发明提供了一种通过氨基酸处理提高白藜发芽率和芽苗中营养品质的方法,具体是通过添加外源天冬酰胺提高白藜发芽率和芽苗中叶酸含量,经过50‑200mg/L天冬酰胺溶液处理后,发芽率,苗长,根长,鲜物重和芽苗中叶酸含量显著优于未经天冬酰胺处理的材料。本发明的发芽方法简单,快速高效,将为利用藜麦芽苗进行产业化利用提供支撑。
Description
技术领域
本发明涉及农业技术领域,具体地说,是一种通过添加外源天冬酰胺处理提高白藜发芽率和芽苗中叶酸含量的方法。
背景技术
藜麦(Chenopodium quinoa Willd.),为苋科藜亚科藜属植物,是唯一一种可满足人体基本营养需求的单体植物,被称为全球10大健康营养食品之一。联合国粮农组织(FAO)认为藜麦是最适宜人类的全营养食品,享有“超级谷物”、“未来食品”和“营养黄金”的美誉。2013年,联合国设为“国际藜麦年”,旨在让世界关注藜麦在提供粮食和营养安全、消除贫困以及在支持千年发展目标等方面所能发挥的作用。藜麦不仅富含蛋白质、维生素、矿物质元素、氨基酸、纤维素等有益化合物和矿物质元素,具有极高的营养价值,还含有黄酮、多酚、皂苷、植物甾醇等多种生物活性物质,具有抗氧化、降血压、降血脂、抗肿瘤、提高人体免疫力、预防心血管疾病、糖尿病等多种生理功效,是广受欢迎的药食同源作物。同时,它生长于高海拔山区,具有抗寒、抗旱、抗贫瘠和高盐碱土的特点。
藜麦为起源于5000年前安第斯山脉地区的一种假谷物,性状为小、平、圆三种类型,市面上常见的颜色为红、黑和白。从口感上来说,白色藜麦口感软糯,易于消化,红黑色藜麦更有嚼劲,从价格上说,白色藜麦生长周期短,种植成本低,市场售价略低于红黑色藜麦。从营养成分来说,有报道指出,白色藜麦的水分含量在7.52%~8.33%之间,灰分含量在2.38%~2.47%之间,脂肪含量在3.71%~3.89%之间,蛋白质含量在20.88%~20.95%之间;红色藜麦的水分含量在8.24%~9.15%之间,灰分含量在2.05%~2.19%之间,脂肪含量在4.19%~4.20%之间,蛋白质含量在16.75%~17.03%之间。
藜麦种子较小,千粒重普遍仅1.4-3.0g,顶土能力差,极易受到天气、土壤、温度和湿度等条件的影响。藜麦种子萌发是它生命活动最旺盛的时期,对植株形态建成及后期的生长发育起着决定性的作用。其中,发芽率、苗高、根长和鲜物重是评价种子萌发能力的常用指标。因此在种植技术上,如何提高藜麦种子的发芽率和出苗率是首先要解决的问题,以保证田间苗齐苗壮,为藜麦丰产打下坚实的基础。
已有报道通过添加外源物质(10-15mg/L的胺鲜酯(DA-6);0.1g/ml-0.15g/ml的PEG溶液;低浓度的盐溶液(NaCl、CaCl2、KCl、MgCl2)等可以促进种子的萌发。前期曾有针对酰胺类新化合物肉桂酰胺-a,肉桂酰胺季铵盐-A,山梨酰胺(简称SLXA)、山梨酰胺季铵盐(简称SLJA)、萘酐酰胺(简称NAXA)、萘酐酰胺季铵盐(简称NAJA)对小麦种子发芽的影响,发现这些化合物在质量浓度为62.5mg/L或125mg/L时,能够明显促进种子发芽,提高种子活力。
植物的天冬酰胺主要通过天冬酰胺合成酶转移谷氨酰胺的氨基到天冬氨酸形成的。有报道指出天冬酰胺介导的氮代谢的调节在调控植物应对生物和非生物胁迫过程中同样发挥着重要的作用(Hwang I S,An S H,Hwang B K.Pepper asparagine synthetase 1(CaAS1)is required for plant nitrogen assimilation and defense responses tomicrobial pathogens[J].The Plant Journal,2011,67(5):749-762.Maaroufi-DguimiH,Debouba M,Gaufichon L,et al.An Arabidopsis mutant disrupted in ASN2encoding asparagine synthetase 2exhibits low salt stress tolerance[J].PlantPhysiology and Biochemistry,2011,49(6):623-628.)。Lam等通过分子生物学手段直接证实了天冬酰胺合成酶在植物体内氮代谢和氮的源库关系调节中的重要作用,通过表达拟南芥天冬酰胺合成酶基因ASN1,显著增加了韧皮部中天冬酰胺的含量,促进了天冬酰胺从原组织(叶和茎)到库组织(花和果实)的运输,进而改善了拟南芥种子中氮营养的状况(LamH M,Wong P,Chan H K,et al.Overexpression of the ASN1 gene enhances nitrogenstatus in seeds of Arabidopsis[J].Plant Physiology,2003,132(2):926-935.)。但是,利用天冬酰胺提高种子发芽率的研究还未见报道,尤其是通过添加天冬酰胺提高了白藜芽苗中叶酸的含量。
藜麦叶片中含有叶酸,它是一类水溶性B族维生素,人体不能自身合成,只能从饮食中摄取。为满足正常生命活动的需要,卫健委推荐成人叶酸摄入量400ug/天,孕妇摄入量为600ug/天。目前,居民叶酸平均摄入量为180.9ug/天,远低于标准水平,摄入充足的人群不足9%。现已证实叶酸摄入不足会引发胎儿神经管发育缺陷,巨幼红细胞贫血,老年痴呆症,心血管疾病和多种癌症等疾病。因此,通过添加天冬酰胺提升白藜种子的萌发率和芽苗中叶酸的营养价值具有重要的现实意义。
发明内容
本发明的目的在于提供一种提高白藜发芽率和营养品质的方法,具体是通过添加外源天冬酰胺提高白藜发芽率和芽苗中叶酸含量。
本发明的第一方面,提供天冬酰胺在提高白藜发芽率和芽苗中叶酸含量中的应用。
进一步的,所述的白藜为籽粒大小均匀、饱满、无虫眼、无病害的白藜种子。
进一步的,所述的天冬酰胺提高白藜发芽率和芽苗中叶酸含量的溶液浓度为50-200mg/L。
更进一步的,所述的天冬酰胺提高白藜发芽率和芽苗中叶酸含量的溶液最适浓度为100mg/L。
本发明的第二方面,提供一种提高白藜发芽率和芽苗中叶酸含量的方法,是采用浓度为50-200mg/L天冬酰胺溶液处理白藜种子。
进一步的,所述的方法包括以下步骤:
(a)选种:选取大小均匀、籽粒饱满、色泽光亮和无虫眼的白藜种子;
(b)消毒:利用10%次氯酸钠消毒种子30min;
(c)清洗:清水洗净2次;
(d)浸泡:浸泡在50-200mg/L的天冬酰胺溶液中,1-2小时;
(e)将种子均匀铺在有一层灭菌纱布的托盘上,纱布也用50-200mg/L天冬酰胺溶液浸泡;
(f)将育苗盘放置于温度22±2℃,湿度60-75%,光照为600-800μmolm-2s-1的环境中发芽。
进一步的,步骤d和步骤e中天冬酰胺溶液浓度为100mg/L。
进一步的,步骤f中的发芽育苗盘上覆盖一层保鲜膜,锁住水分。
进一步的,检测发芽率,苗长,根长和鲜物重等指标;检测芽苗中叶酸等营养指标。
本发明实验发现,在50mg/L和200mg/L天冬酰胺提供下,发芽率相比未经天冬酰胺处理的材料有显著性差异,在100mg/L天冬酰胺提供下,发芽率相比未经天冬酰胺处理的材料有极显著差异。
根长在25mg/L,50mg/L,100mg/L和200mg/L天冬酰胺浓度下相比未经天冬酰胺处理的材料显著较长,苗高在100mg/L天冬酰胺下相比未经天冬酰胺处理的材料显著较高,鲜物重在50mg/L和100mg/L天冬酰胺下相比未经天冬酰胺处理的材料显著较大。天冬酰胺浓度50mg/L,100mg/L和200mg/L下,芽苗中叶酸含量显著优于未经天冬酰胺处理的材料。
在天冬酰胺浓度100mg/L时,发芽率,苗长,根长,鲜物重和芽苗中叶酸含量显著优于未经天冬酰胺处理的材料。
本发明优点在于:
1、本发明提供了一种通过氨基酸处理提高白藜发芽率和营养品质的方法,具体是通过添加外源天冬酰胺提高白藜发芽率和芽苗中叶酸含量,经过50-200mg/L天冬酰胺溶液处理后,发芽率,苗长,根长,鲜物重和芽苗中叶酸含量显著优于未经天冬酰胺处理的材料。
2、本发明的发芽方法简单,快速高效,将为利用藜麦芽苗进行产业化利用提供支撑。
附图说明
图1:不同外源天冬酰胺浓度下白藜芽菜种子发芽率;其中,*代表天冬酰胺浓度和对照之间有P<0.05显著性差异;**代表天冬酰胺浓度和对照之间有P<0.01极显著差异。
具体实施方式
下面结合实施例对本发明提供的具体实施方式作详细说明。
实施例1:
取籽粒大小均匀、饱满、无虫眼、无病害的白色格藜种子用10%次氯酸钠消毒30min,然后用自来水冲洗干净,再用蒸馏水漂洗2~3遍。将漂洗后的种子分别置于5个天冬酰胺梯度的浓度溶液中浸泡1-2小时,即0、25mg/L、50mg/L、100mg/L和200mg/L,其中,0mg/L天冬酰胺浓度为空白对照(CK),每个处理设置3次重复。采用铺有一层灭菌纱布的苗盘为发芽床,且提前将纱布置于了不同天冬酰胺浓度的梯度溶液中进行30min浸泡,然后将浸泡后的供试种子均匀置于苗盘中,每个苗盘种子2000粒(1g,300粒左右,称重6g),将苗盘放于小人工气候室中培养,培养条件为22±2℃,光暗周期为16h/8h,光照强度为600-800μmolm-2s-1。以种子的胚芽长度达到白藜种子长度的一半视为种子发芽,每天在固定时间观察记录种子发芽数。在第7天统计种子发芽率;第10天每个处理随机选择10株正常发育的幼苗,分别测定幼苗根长、苗高,称量鲜重,营养指标、托盘的总白藜苗产量。各指标计算公式如下:
发芽率(GE)=(7d内发芽种子数量/供试种子数)×100%
结果发现,在50mg/L和200mg/L天冬酰胺提供下,发芽率相比0mg/L有显著性差异,在100mg/L天冬酰胺提供下,发芽率相比0mg/L有极显著差异。
根长在25mg/L,50mg/L,100mg/L和200mg/L天冬酰胺浓度下相比0mg/L显著较长,苗高在100mg/L天冬酰胺下相比0mg/L显著较高,鲜物重在50mg/L和100mg/L天冬酰胺下相比0mg/L显著较大。
表1:不同天冬酰胺浓度下白藜芽苗的根长,苗高和鲜物重比较
注:表中数据均为平均值±标准差,同列不同小写字母表示处理间差异达到显著水平(P<0.05)。
实施例2:
白藜芽苗中叶酸含量的测定方法参考GB5009.211-2014,步骤简单总结如下:
标准储备液(20.0ug/ml)的配置:精确称取20.0mg叶酸标准品,用氢氧化钠乙醇溶液溶解并转移至1000ml容量瓶中,定容。
标准中间液(0.200ug/ml):准确吸取1.00ml叶酸标准储备液置于100ml棕色容量瓶中,用氢氧化钠乙醇溶液稀释并定容至刻度,混匀后储存于瓶中2℃-4℃冰箱可保存1年。
标准工作液(0.200ng/ml):准确吸取1.00ml叶酸标准中间液置于1000ml棕色容量瓶中,用水稀释并定容至刻度,混匀,现用现配。
菌种和接种液的制备参考标准。准确测定试样2-5g,转入100ml锥形瓶中,加30ml磷酸缓冲液,振摇5min后,具塞,于121℃(0.10MPa-0.12MPa)高压水解15min。试样取出后冷却至室温,加入1ml蛋白酶-淀粉酶液,混合。加入3滴-5滴甲苯后,置于37℃±1℃恒温培养箱内酶解16h-20h。取出,转入100ml容量瓶,加水定容至刻度,过滤。另取一只锥形瓶,同试样操作,定容至100ml,过滤。作为酶空白液。
稀释:根据试样中叶酸含量用水对试样提取液进行适当稀释,使试样稀释液中叶酸含量在0.2ng/ml-0.6ng/ml范围内。
测定:将培养好的标准系列管、试样和酶空白系列管用旋涡混匀器混匀。用厚度为1cm比色杯,于540nm处,依次测定,公式参考标准如下:
式中:
X——试样中叶酸含量,单位为微克每百克(μg/100g);
——试样稀释液中叶酸浓度平均值,单位为纳克每毫升(ng/mL);
f——试样提取液稀释倍数;
——酶空白液中叶酸浓度平均值,单位为纳克每毫升(ng/mL);
V——试样提取液定容体积,单位为毫升(mL);
m——试样质量,单位为克(g);
——由纳克每克(ng/g)换算为微克每百克(μg/100g)的系数。
表2
注:表中数据均为平均值±标准差,不同小写字母代表不同天冬酰胺浓度下,叶酸含量的差异达到显著水平(P<0.05)。
测定结果显示,在100mg/L天冬酰胺时,白藜芽苗中叶酸含量显著优于其他处理浓度。
以上已对本发明创造的较佳实施例进行了具体说明,但本发明创造并不限于所述实施例,熟悉本领域的技术人员在不违背本发明创造精神的前提下还可做出种种的等同的变型或替换,这些等同的变型或替换均包含在本申请权利要求所限定的范围内。
Claims (8)
1.天冬酰胺在提高白藜发芽率和芽苗中叶酸含量中的应用;所述的天冬酰胺提高白藜发芽率和芽苗中叶酸含量的溶液浓度为50-200mg/L。
2.根据权利要求1所述的天冬酰胺在提高白藜发芽率和芽苗中叶酸含量的应用,其特征在于,所述的白藜为籽粒大小均匀、饱满、无虫眼、无病害的白藜种子。
3.根据权利要求1所述的天冬酰胺在提高白藜发芽率和芽苗中叶酸含量中的应用,其特征在于,所述的天冬酰胺提高白藜发芽率和芽苗中叶酸含量的溶液浓度为100mg/L。
4.一种提高白藜发芽率和芽苗中叶酸含量的方法,其特征在于,是采用浓度为50-200mg/L天冬酰胺溶液处理白藜种子。
5.根据权利要求4所述的提高白藜发芽率和芽苗中叶酸含量的方法,其特征在于,所述的方法包括以下步骤:
(a)选种:选取大小均匀、籽粒饱满、色泽光亮和无虫眼的白藜籽粒;
(b)消毒:利用10%次氯酸钠消毒种子30min;
(c)清洗:清水洗净2次;
(d)浸泡:浸泡在50-200mg/L的天冬酰胺溶液中,1-2小时;
(e)将种子均匀铺在有一层灭菌纱布的托盘上,纱布也用50-200mg/L天冬酰胺溶液浸泡;
(f)将育苗盘放置于温度22±2℃,湿度60-75%,光照为600-800μmolm-2s-1的环境中发芽。
6.根据权利要求5所述的提高白藜发芽率和芽苗中叶酸含量的方法,其特征在于,步骤d和步骤e中天冬酰胺溶液浓度为100mg/L。
7.根据权利要求5所述的提高白藜发芽率和芽苗中叶酸含量的方法,其特征在于,步骤f中的发芽育苗盘上覆盖一层保鲜膜,锁住水分。
8.根据权利要求5所述的提高白藜发芽率和芽苗中叶酸含量的方法,其特征在于,检测发芽率,苗长,根长,鲜物,叶酸含量。
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CN1420729A (zh) * | 1999-06-18 | 2003-05-28 | 阿尔比恩国际公司 | 用氨基酸螯合物强化的谷物颗粒核仁 |
WO2014160354A1 (en) * | 2013-03-14 | 2014-10-02 | Georgia State University Research Foundation, Inc. | Inhibiting or reducing fungal growth |
CN104160802A (zh) * | 2014-08-01 | 2014-11-26 | 兰州大学 | 一种提高四翅滨藜种子发芽率的方法 |
CN108718605A (zh) * | 2018-06-11 | 2018-11-02 | 滨州学院 | 利用曼陀罗叶水提液控制藜种子萌发和幼苗生长的方法 |
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CN1420729A (zh) * | 1999-06-18 | 2003-05-28 | 阿尔比恩国际公司 | 用氨基酸螯合物强化的谷物颗粒核仁 |
WO2014160354A1 (en) * | 2013-03-14 | 2014-10-02 | Georgia State University Research Foundation, Inc. | Inhibiting or reducing fungal growth |
CN104160802A (zh) * | 2014-08-01 | 2014-11-26 | 兰州大学 | 一种提高四翅滨藜种子发芽率的方法 |
CN108718605A (zh) * | 2018-06-11 | 2018-11-02 | 滨州学院 | 利用曼陀罗叶水提液控制藜种子萌发和幼苗生长的方法 |
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