CN107439550B - 一种提高酸雨胁迫下大麦种子萌发及幼苗抗酸能力的海藻糖保护剂及其制备方法与应用 - Google Patents
一种提高酸雨胁迫下大麦种子萌发及幼苗抗酸能力的海藻糖保护剂及其制备方法与应用 Download PDFInfo
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Abstract
本发明涉及一种提高酸雨胁迫下大麦种子萌发及幼苗抗酸能力的海藻糖保护剂，及其制备方法与应用，该保护剂为种子浸泡剂，以水为溶剂，每升水中含有海藻糖1.0～6.0g。本发明的海藻糖保护剂可以提高酸雨胁迫下大麦种子的发芽率、增强酶活性，保持细胞膜的稳定性，大大增强了幼苗抗酸能力，同时具有抗干旱、盐胁迫的能力。
Description
技术领域
本发明涉及一种提高酸雨胁迫下大麦种子萌发及幼苗抗酸能力的海藻糖保护剂及其制备方法与应用，属于农作物抗逆制剂技术领域。
背景技术
大麦是一种适应性广、抗逆性强、产量高的禾谷类作物，在保健食品、酿造和饲料行业中广泛使用，在世界各大洲均有种植。
酸雨是指由于空气中的SO2，NOx等超标引起的pH值小于5.6的酸性降水。近年来，随着我国国民经济和工业的迅猛发展，对化石燃料的需求日益增加，其燃烧后产生的酸性物质的排放严重影响了大气质量，导致我国酸雨问题日趋严重，酸雨面积超过国土面积的29％，而且面积仍在扩大，降水酸性继续升高。
随着酸雨污染的日益加剧，对酸雨危害的研究也越来越广泛，目前已有很多关于酸雨对植物影响的报道。迄今为止，研究者们逐步系统地开展了酸雨对植物宏观和显微伤害、膜脂过氧化、膜保护酶活性与自由基平衡、物质代谢、光合与呼吸作用等方面的影响研究。酸雨伤害所诱发的群落演替以及种群波动，都被认为与植物宏观表象下隐藏的种子微观萌发过程相关，而萌发是植物生活史的起点，也是植物生物量形成的关键。因此，国内外研究者开展了大量关于酸雨胁迫对种子萌发过程的影响研究。
以花生种子为试材，研究了不同值的酸雨对其萌发的影响，结果发现花生种子发芽率、胚根长度、胚根直径和活力指数等均随着酸雨强度的增加而减少；以水稻、小麦和油菜种子为研究对象，探讨了不同强度的模拟酸雨对其种子萌发的影响，结果表明pH2.0时种作物种子均不发芽，pH2.5时水稻和小发异状发芽，pH≥3.0时，水稻、小麦和油菜种子的发芽率、发芽势、发芽(活力)指数均随着酸雨强度的降低而降低，并呈现显著差异；大量研究表明，酸雨抑制作物种子萌发的原因包括很多方面，具体表现为种子透性发生变化，离子动态平衡失调，呼吸代谢和物质代谢素乱、酶活性发生改变等。
海藻糖是一种可靠、安全的天然糖类，含海藻糖的生物在高温脱水，高渗透压条件下，避免细胞脱水所造成的营养损耗和细胞损伤，使机体具有较强的抗旱性。研究者发现盐胁迫下通过使用海藻糖可以降低小麦、黄瓜、番茄叶片细胞电解质渗透率和丙二醛含量，防止叶绿素的降解，并且番茄根系活力，干物质积累和植物生长速率则均有提高。海藻糖预处理也可提高黄瓜幼苗的抗冷性和抗旱性。大量研究表明，植物体内海藻糖在植物抵御干旱和高盐、高温等非生物胁迫的过程中发挥着重要作用，但关于海藻糖在植物抵御酸雨胁迫方面的研究还未见任何文献报道。
发明内容
本发明提供一种提高酸雨胁迫下大麦种子萌发及幼苗抗酸能力的海藻糖保护剂及其制备方法与应用,该海藻糖保护剂可以提高酸雨胁迫下大麦种子的发芽率、增强酶活性，保持细胞膜的稳定性，大大增强了幼苗抗酸能力，同时具有抗干旱、盐胁迫的能力。
本发明的技术方案如下：
一种提高酸雨胁迫下大麦种子萌发及幼苗抗酸能力的海藻糖保护剂，该保护剂为种子浸泡剂，以蒸馏水为溶剂，每升水中含有海藻糖1.0～6.0g。
根据本发明优选的，所述的海藻糖保护剂每升水中含有海藻糖1.5～5.7g。
进一步优选的，所述的海藻糖保护剂每升水中含有海藻糖3.0～5.1g。
最为优选的，所述的海藻糖保护剂每升水中含有海藻糖3.4g。
提高酸雨胁迫下大麦种子萌发的海藻糖保护剂的制备方法，将海藻糖按比例加入蒸馏水中，溶解后搅拌均匀即得。
根据本发明，海藻糖保护剂的应用，作为大麦种子保护剂使用，用于提高酸雨胁迫下大麦种子的萌发率。
根据本发明优选的，具体应用方法如下:
将大麦种子消毒、冲洗，沥干水分，浸入海藻糖保护剂中置于18-23℃恒温下浸泡6～24h，沥干水分，置于酸雨胁迫下进行发芽。
根据本发明进一步优选的，浸泡时间为10～16h。
根据本发明优选的，所述的消毒为用质量分数3％H2O2溶液消毒10min，所述的冲洗为用蒸馏水冲洗3-6次。
根据本发明优选的，浸泡时每升海藻糖保护剂中加入0.2～0.8kg大麦种子。
根据本发明优选的，酸雨pH为3.0～5.6。
本发明的海藻糖保护剂中海藻糖的含量至关重要，对酸雨胁迫下种子的萌发以及幼苗的抗酸性起着决定性的影响，海藻糖的含量过低起不到保护作用，海藻糖的含量过高对种子的萌发有抑制作用，因此，严格控制海藻糖的含量。
本发明的优点及效果：
1、本发明的海藻糖保护剂可以提高酸雨胁迫下大麦种子的发芽率、增强酶活性，保持细胞膜的稳定性，大大增强了幼苗抗酸能力，同时具有抗干旱、盐胁迫的能力。
2、本发明的海藻糖保护剂及浓度，可以使酸雨胁迫下的种子及幼苗得到有力保护，在种子发芽7天后，发芽率比蒸馏水处理增加50.5％，株高比蒸馏水提高25.9％，鲜重比蒸馏水处理提高28.8％，MDA比蒸馏水处理降低41.1％，CAT活性比蒸馏水处理提高15.6％，POD活性比蒸馏水处理提高138.6％，幼苗叶绿素含量比蒸馏水提高69.5％，根细胞受到的损伤明显降低。
附图说明：
图1为酸雨胁迫下不同处理剂处理对大麦种子萌发率影响柱状图；
图2为酸雨胁迫下不同处理剂处理下Evans blue对大麦根染色对比图，颜色越深代表大麦根细胞的损伤越大。
具体实施方式
下面结合实施例对本发明的技术方案做进一步说明，但本发明的保护范围不限于此。
实施例1：
一种提高酸雨胁迫下大麦种子萌发及幼苗抗酸能力的海藻糖保护剂，该保护剂为种子浸泡剂，以蒸馏水为溶剂，每升水中含有海藻糖1.7g。
制备方法：将海藻糖按比例加入蒸馏水中，溶解后搅拌均匀即得。
模拟酸雨的配置：用分析纯H2SO4、HNO3和HCl的摩尔比为3.56:1：1.42配制成pH值为1.0的酸雨母液(1L母液中分别加入H2SO4 2.78ml,KNO3 2.78g,CaCl21.0g,(NH4)2SO47.48g)，使用时将酸雨稀释成不同pH的模拟酸雨溶液。
应用：将大麦种子用质量分数3％H2O2溶液消毒10min，再用蒸馏水冲洗5次，沥干水分。将大麦种子浸入海藻糖保护剂中置于20℃恒温水浴锅浸泡12h，每升海藻糖保护剂中加入0.2kg大麦种子。然后沥干水分，置于模拟酸雨胁迫的发芽机中，由发芽机每隔1h喷洒pH为3.5的模拟酸雨1次，喷洒时间持续1min，发芽机的种子托盘为网状漏水式。
实施例2：
一种提高酸雨胁迫下大麦种子萌发及幼苗抗酸能力的海藻糖保护剂，该保护剂为种子浸泡剂，以蒸馏水为溶剂，每升水中含有海藻糖3.4g。制备方法同实施例1。
应用方法同实施例1，不同之处在于：每升海藻糖保护剂中加入0.4kg大麦种子。
实施例3：
一种提高酸雨胁迫下大麦种子萌发及幼苗抗酸能力的海藻糖保护剂，该保护剂为种子浸泡剂，以蒸馏水为溶剂，每升水中含有海藻糖5.1g。制备方法同实施例1。
应用方法同实施例1，不同之处在于：每升海藻糖保护剂中加入0.6kg大麦种子。
对照例：
将大麦种子用质量分数3％H2O2溶液消毒10min，再用蒸馏水冲洗5次，沥干水分。将大麦种子浸入蒸馏水中置于20℃恒温水浴锅浸泡12h，每升蒸馏水中加入0.6kg大麦种子，然后沥干水分，置于模拟酸雨胁迫的发芽机中，由发芽机每隔1小时喷洒pH为3.5的模拟酸雨1次，发芽机的种子托盘为网状漏水式。
应用例1：
将实施例1中海藻糖保护剂处理的大麦种子，于20℃置于模拟酸雨胁迫的发芽机中，由发芽机每隔1小时喷洒模拟酸雨1次，进行发芽率、株高、鲜重、MDA、幼苗叶绿素含量测试；与对照例进行对照，每个处理包含5个重复组，每组有100粒种子；与对照例相比，在种子发芽7天后，发芽率比对照增加21.5％，株高比对照提高20.5％，鲜重比对照提高23.2％，MDA比对照降低15％，CAT活性比对照提高13.3％，POD活性比对照提高129.7％，幼苗叶绿素含量比对照提高70.9％，Evans blue对根染色的结果见图2所示，从图中可以看出根细胞受到的损伤明显降低。
应用例2：
将实施例2中海藻糖保护剂处理的大麦种子，于20℃置于模拟酸雨胁迫的发芽机中，由发芽机每隔1小时喷洒模拟酸雨1次，进行发芽率、株高、鲜重、MDA、幼苗叶绿素含量测试；与对照例进行对照，每个处理包含5个重复组，每组有100粒种子；与对照例相比，在种子发芽7天后，发芽率比对照增加50.5％，株高比对照提高25.9％，鲜重比对照提高28.8％，MDA比对照降低41.1％，CAT活性比对照提高15.6％，POD活性比对照提高138.6％，幼苗叶绿素含量比对照提高69.5％，Evans blue对根染色的结果见图2所示，从图中可以看出根细胞受到的损伤明显降低。
应用例3：
将实施例3中海藻糖保护剂处理的大麦种子，于20℃置于模拟酸雨胁迫的发芽机中，由发芽机每隔1小时喷洒模拟酸雨1次，进行发芽率、株高、鲜重、MDA、幼苗叶绿素含量测试；与对照例进行对照，每个处理包含5个重复组，每组有100粒种子；与对照例相比，在种子发芽7天后，发芽率比对照增加53.8％，株高比对照提高21.6％，鲜重比对照提高24.2％，MDA比对照降低34.5％，CAT活性比对照提高26.1％，POD活性比对照提高220.4％，幼苗叶绿素含量比对照提高63.3％，Evans blue对根染色的结果见图2所示，从图中可以看出根细胞受到的损伤明显降低。
Claims (2)
1.一种提高酸雨胁迫下大麦种子萌发及幼苗抗酸能力的海藻糖保护剂的应用，该保护剂为种子浸泡剂，以蒸馏水为溶剂，每升水中含有海藻糖3.4g；作为大麦种子保护剂使用，用于提高酸雨胁迫下大麦种子的萌发率；具体应用方法如下:
将大麦种子消毒、冲洗，沥干水分，浸入海藻糖保护剂中置于18-23℃恒温下浸泡10-16h，沥干水分，置于酸雨胁迫下进行发芽，浸泡时每升海藻糖保护剂中加入0.2~0.8 kg大麦种子；所述的消毒为用质量分数3%H2O2溶液消毒10 min，所述的冲洗为用蒸馏水冲洗3-6次，酸雨pH为3.0~5.6。
2.根据权利要求1所述的海藻糖保护剂的应用，其特征在于，提高酸雨胁迫下大麦种子萌发及幼苗抗酸能力的海藻糖保护剂的制备方法，将海藻糖按比例加入蒸馏水中，溶解后搅拌均匀即得。
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