CN117024931A - 一种抗菌保水凝胶及其制备方法和应用 - Google Patents

一种抗菌保水凝胶及其制备方法和应用 Download PDF

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CN117024931A
CN117024931A CN202311008319.4A CN202311008319A CN117024931A CN 117024931 A CN117024931 A CN 117024931A CN 202311008319 A CN202311008319 A CN 202311008319A CN 117024931 A CN117024931 A CN 117024931A
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antibacterial
acid
grafting
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CN117024931B (zh
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曲浩
罗梓文
龙亚芹
陈洪云
龙丽雪
陈龙
陈林波
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Tea Research Institute Yunnan Academy of Agricultural Sciences
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Abstract

本发明公开了一种抗菌保水凝胶及其制备方法和应用，属于植物嫁接技术领域。本发明的抗菌保水凝胶，包括以下质量份数的原料：水苏糖20～30份、聚乳酸12～15份、裙带菜提取物8～10份、明胶10～13份、水杨酸1.5～2.5份、吲哚丁酸0.5～0.8份、复硝酚钠1～2份、黄腐酸2～3份和改性甘露聚糖4～5份。本发明的抗菌保水凝胶具备良好的透气性、生物相容性、保水性和抑菌性，是一种优良的植物嫁接用材料，可以促进嫁接接口的快速愈合，同时具有一定的透气性，在为嫁接植物提供水分的同时，还可以避免因不透气造成的嫁接接口腐烂的问题，可以显著提升植物嫁接的成活率。

Description

一种抗菌保水凝胶及其制备方法和应用
技术领域
本发明涉及植物嫁接技术领域，特别是涉及一种抗菌保水凝胶及其制备方法和应用。
背景技术
嫁接是植物的人工繁殖方法之一，即把一种植物的枝或芽，嫁接到另一种植物的茎或根上，使接在一起的两个部分长成一个完整的植株。嫁接的方式分为枝接和芽接，嫁接是利用植物受伤后具有愈伤的机能来进行的，并且嫁接不仅能够保持接穗品种的优良性状，还能利用砧木的有利特性，达到早结果、增强抗寒性、抗旱性、抗病虫害的能力，在经济利用繁殖材料的同时增加苗木数量。常用于果树、林木、花卉的繁殖上；也用于瓜类蔬菜育苗上。
影响嫁接苗成活的因素主要有砧木和接穗之间的亲和力、嫁接时间、砧木和接穗质量、温度和水分等。而接穗一般是枝条或者芽，其与母体分离后和砧木之间建立疏导组织需要至少两三周的时间，此时，仅靠接穗自身的营养和水分，远远不能实现愈合。因此，就需要在嫁接之后采取一些提高愈合速度、提供营养成分和保水的措施来提高嫁接的成活率。目前都是采用涂抹愈合液或营养液的方式提高愈合速度、提供营养成分，但该方式容易滋生病菌；而为了保水常常利用薄膜缠绕接穗与砧木的连接处，但薄膜不透气，包扎太紧密造成湿度过大，包扎不紧密起不到保持水分的作用，植物嫁接的成活率并没有得到显著的提高。
发明内容
本发明的目的是提供一种抗菌保水凝胶及其制备方法和应用，以解决上述现有技术存在的问题。
为实现上述目的，本发明提供了如下方案：
本发明的技术方案之一：一种抗菌保水凝胶，包括以下质量份数的原料：水苏糖20～30份、聚乳酸12～15份、裙带菜提取物8～10份、明胶10～13份、水杨酸1.5～2.5份、吲哚丁酸0.5～0.8份、复硝酚钠1～2份、黄腐酸2～3份和改性甘露聚糖4～5份。
进一步地，所述裙带菜提取物的制备方法，包括以下步骤：
(1)将裙带菜孢子叶粉碎后加水混合均匀，调节pH至5.5～6.5，然后加入复合酶保温酶解后收集上清液；
(2)将上清液浓缩后用乙醇溶液沉淀，得到所述裙带菜提取物。
进一步地，所述裙带菜孢子叶和水的质量/体积比为1g:(8～10)mL；所述复合酶的添加量为裙带菜孢子叶质量的2.0～2.5％；所述乙醇溶液的浓度为95vol.％。
进一步地，所述保温酶解的温度为45～55℃，时间为2.5～3.5h；所述复合酶，包括以下质量份数的组分：纤维素酶2～3份、胰蛋白酶1～2份和木瓜蛋白酶1～2份。
裙带菜提取物具有抗菌活性，可以抑制病原菌的生长。
进一步地，所述改性甘露聚糖的制备方法，包括以下步骤：
将谷氨酸、甘露聚糖和水混合均匀，加热回流反应后浓缩、干燥，得到所述改性甘露聚糖。
甘露聚糖经谷氨酸改性后具有优异的抑菌活性，二者协同作用，可以显著抑制病原菌的生长繁殖。
进一步地，所述加热回流反应的温度为80～90℃，时间为8～10h。
本发明的技术方案之二：一种上述抗菌保水凝胶的制备方法，包括以下步骤：
将水苏糖加入水中加热溶解，然后加入聚乳酸、裙带菜提取物和明胶，进行一次搅拌后加入水杨酸、吲哚丁酸、复硝酚钠、黄腐酸和改性甘露聚糖，二次搅拌后高速剪切，得到所述抗菌保水凝胶。
进一步地，所述水苏糖和水的质量比为1g:(45～55)mL；所述加热溶解的温度为80～85℃；所述一次搅拌的转速为80～100r/min，时间为30～40min；所述二次搅拌的转速为150～180r/min，时间为20～30min；所述高速剪切的转速为3500～3800r/min，时间为5～8min。
水苏糖、聚乳酸、裙带菜提取物和明胶搅拌反应后可以形成具备良好的透气性、生物相容性的凝胶材料，该凝胶材料还具有良好的保水性和抑菌性。
抗菌保水凝胶中的水杨酸、吲哚丁酸、复硝酚钠、黄腐酸和改性甘露聚糖可以促进植物伤口的愈合，抑制病原菌的生长繁殖。
本发明的技术方案之三：一种上述抗菌保水凝胶在茶树嫁接中的应用。
进一步地，所述应用的方法，具体包括：将所述抗菌保水凝胶涂敷在嫁接接口处，静置2～3min，然后包裹纱布。
本发明公开了以下技术效果：
(1)本发明的抗菌保水凝胶具备良好的透气性、生物相容性、保水性和抑菌性，是一种优良的植物嫁接用材料，可以促进嫁接接口的快速愈合，同时具有一定的透气性，在为嫁接植物提供水分的同时，还可以避免因不透气造成的嫁接接口腐烂的问题，可以显著提升植物嫁接的成活率。
(2)本发明克服了现有嫁接技术用薄膜缠绕接穗与砧木的连接处，造成的接口污染、接口愈合慢、砧木与接穗不紧密、嫁接成活率低的问题。
(3)本发明的抗菌保水凝胶可以提升砧木与接穗的连接效果，加快愈合，提升嫁接成活率。
(4)本发明的抗菌保水凝胶可以应用于多个品种茶树嫁接，可以提高砧木和接穗的融合速度，提升茶树嫁接成活率。
具体实施方式
现详细说明本发明的多种示例性实施方式，该详细说明不应认为是对本发明的限制，而应理解为是对本发明的某些方面、特性和实施方案的更详细的描述。
应理解本发明中所述的术语仅仅是为描述特别的实施方式，并非用于限制本发明。另外，对于本发明中的数值范围，应理解为还具体公开了该范围的上限和下限之间的每个中间值。在任何陈述值或陈述范围内的中间值以及任何其他陈述值或在所述范围内的中间值之间的每个较小的范围也包括在本发明内。这些较小范围的上限和下限可独立地包括或排除在范围内。
除非另有说明，否则本文使用的所有技术和科学术语具有本发明所述领域的常规技术人员通常理解的相同含义。虽然本发明仅描述了优选的方法和材料，但是在本发明的实施或测试中也可以使用与本文所述相似或等同的任何方法和材料。本说明书中提到的所有文献通过引用并入，用以公开和描述与所述文献相关的方法和/或材料。在与任何并入的文献冲突时，以本说明书的内容为准。
在不背离本发明的范围或精神的情况下，可对本发明说明书的具体实施方式做多种改进和变化，这对本领域技术人员而言是显而易见的。由本发明的说明书得到的其他实施方式对技术人员而言是显而易见的。本申请说明书和实施例仅是示例性的。
关于本文中所使用的“包含”、“包括”、“具有”、“含有”等等，均为开放性的用语，即意指包含但不限于。
以下实施例所述的“份”均为“质量份”。
实施例1
一种抗菌保水凝胶的制备方法：
抗菌保水凝胶，由以下质量百分数的原料：水苏糖25份、聚乳酸12份、裙带菜提取物10份、明胶12份、水杨酸2.0份、吲哚丁酸0.6份、复硝酚钠1.5份、黄腐酸2份和改性甘露聚糖4.5份。
(1)裙带菜提取物的制备方法
A.将裙带菜孢子叶(新鲜)粉碎后加水(裙带菜孢子叶和水的质量/体积比为1g:10mL)混合均匀，调节pH至6.0，然后加入复合酶(复合酶的添加量为裙带菜孢子叶质量的2.2％)保温酶解(保温酶解的温度为50℃，时间为3h)后收集上清液；
复合酶，由以下质量份数的组分组成：纤维素酶2.5份、胰蛋白酶2份和木瓜蛋白酶1.5份。
B.将上清液浓缩至原体积的1/3后用浓度为95vol.％的乙醇溶液沉淀，沉淀经干燥(50℃)后得到裙带菜提取物。
(2)改性甘露聚糖的制备方法
将谷氨酸2份、甘露聚糖10份和水(甘露聚糖和水的质量/体积比为1g:10mL)混合均匀，加热回流反应(温度为90℃，时间为8h)后浓缩、干燥，得到所述改性甘露聚糖。
(3)抗菌保水凝胶的制备方法
将水苏糖加入水(水苏糖和水的质量/体积比为1g:50mL)中加热(85℃)溶解，保持温度不变，然后加入聚乳酸、裙带菜提取物和明胶，进行一次搅拌(转速为100r/min，时间为30min)后加入水杨酸、吲哚丁酸、复硝酚钠、黄腐酸和改性甘露聚糖，二次搅拌(转速为180r/min，时间为20min)后高速剪切(转速为3600r/min，时间为6min)，得到抗菌保水凝胶。
实施例2
一种抗菌保水凝胶的制备方法：
抗菌保水凝胶，由以下质量百分数的原料：水苏糖30份、聚乳酸15份、裙带菜提取物8份、明胶10份、水杨酸1.5份、吲哚丁酸0.8份、复硝酚钠1份、黄腐酸2.5份和改性甘露聚糖4份。
(1)裙带菜提取物的制备方法
A.将裙带菜孢子叶(新鲜)粉碎后加水(裙带菜孢子叶和水的质量/体积比为1g:8mL)混合均匀，调节pH至5.5，然后加入复合酶(复合酶的添加量为裙带菜孢子叶质量的2.5％)保温酶解(保温酶解的温度为55℃，时间为2.5h)后收集上清液；
复合酶，由以下质量份数的组分组成：纤维素酶3份、胰蛋白酶1份和木瓜蛋白酶1份。
B.将上清液浓缩至原体积的1/3后用浓度为95vol.％的乙醇溶液沉淀，沉淀经干燥(50℃)后得到裙带菜提取物。
(2)改性甘露聚糖的制备方法：
将谷氨酸2份、甘露聚糖10份和水(甘露聚糖和水的质量/体积比为1g:10mL)混合均匀，加热回流反应(温度为80℃，时间为8h)后浓缩、干燥，得到所述改性甘露聚糖。
(3)抗菌保水凝胶的制备方法
将水苏糖加入水(水苏糖和水的质量/体积比为1g:45mL)中加热(82℃)溶解，保持温度不变，然后加入聚乳酸、裙带菜提取物和明胶，进行一次搅拌(转速为80r/min，时间为35min)后加入水杨酸、吲哚丁酸、复硝酚钠、黄腐酸和改性甘露聚糖，二次搅拌(转速为150r/min，时间为30min)后高速剪切(转速为3800r/min，时间为5min)，得到抗菌保水凝胶。
实施例3
一种抗菌保水凝胶的制备方法：
抗菌保水凝胶，由以下质量百分数的原料：水苏糖20份、聚乳酸13份、裙带菜提取物10份、明胶13份、水杨酸2.5份、吲哚丁酸0.5份、复硝酚钠2份、黄腐酸3份和改性甘露聚糖5份。
(1)裙带菜提取物的制备方法
A.将裙带菜孢子叶(新鲜)粉碎后加水(裙带菜孢子叶和水的质量/体积比为1g:9mL)混合均匀，调节pH至6.5，然后加入复合酶(复合酶的添加量为裙带菜孢子叶质量的2.0)保温酶解(保温酶解的温度为45℃，时间为3.5h)后收集上清液；
复合酶，由以下质量份数的组分组成：纤维素酶2份、胰蛋白酶2份和木瓜蛋白酶2份。
B.将上清液浓缩至原体积的1/3后用浓度为95vol.％的乙醇溶液沉淀，沉淀经干燥(50℃)后得到裙带菜提取物。
(2)改性甘露聚糖的制备方法：
将谷氨酸2份、甘露聚糖10份和水(甘露聚糖和水的质量/体积比为1g:10mL)混合均匀，加热回流反应(温度为85℃，时间为10h)后浓缩、干燥，得到所述改性甘露聚糖。
(3)抗菌保水凝胶的制备方法
将水苏糖加入水(水苏糖和水的质量/体积比为1g:50mL)中加热(80℃)溶解，保持温度不变，然后加入聚乳酸、裙带菜提取物和明胶，进行一次搅拌(转速为90r/min，时间为35min)后加入水杨酸、吲哚丁酸、复硝酚钠、黄腐酸和改性甘露聚糖，二次搅拌(转速为160r/min，时间为25min)后高速剪切(转速为3500r/min，时间为8min)，得到抗菌保水凝胶。
对比例1
同实施例1，区别仅在于，改变裙带菜提取物的加入步骤，具体如下：
将水苏糖加入水(水苏糖和水的质量/体积比为1g:50mL)中加热(85℃)溶解，保持温度不变，然后加入聚乳酸和明胶，进行一次搅拌(转速为100r/min，时间为30min)后加入裙带菜提取物、水杨酸、吲哚丁酸、复硝酚钠、黄腐酸和改性甘露聚糖，二次搅拌(转速为180r/min，时间为20min)后高速剪切(转速为3600r/min，时间为6min)，得到抗菌保水凝胶。
对比例2
同实施例1，区别仅在于，将改性甘露聚糖替换成质量比为1：5的谷氨酸和甘露聚糖。
对比例3
同实施例1，区别仅在于，抗菌保水凝胶的制备方法如下：
将水苏糖加入水(水苏糖和水的质量/体积比为1g:50mL)中加热(85℃)溶解，保持温度不变，然后加入聚乳酸、裙带菜提取物、明胶、水杨酸、吲哚丁酸、复硝酚钠、黄腐酸和改性甘露聚糖进行一次搅拌(转速为100r/min，时间为30min)，接着进行二次搅拌(转速为180r/min，时间为20min)后高速剪切(转速为3600r/min，时间为6min)，得到抗菌保水凝胶。
对比例4
同实施例1，区别仅在于，将水杨酸替换成等质量份数的改性甘露聚糖。
效果例1
茶树嫁接：
(1)砧木的选择：选择5年树龄、立地条件好、根系发达、生长健壮的茶树上直径为2.0厘米左右、生长健壮的茎干作砧木。
(2)接穗的选择：选择云抗14号或普景1号作接穗，接穗应选择生长健壮、无病虫害、茎皮呈红棕色、芽比较饱满的一年生木质化枝条，枝条当天采摘，当天嫁接。
(3)嫁接时期：2月份左右。
(4)嫁接过程
处理组：
A.剪砧木：砧木离地面12cm左右处剪断准备用于嫁接，其余茎干齐根剪平。
B.切砧木：用嫁接刀从砧木断面带部分木质部的部位纵切一刀，切口深度2.5cm左右。
C.选取的接穗保留一个芽，上端在距芽0.5厘米处剪断，在其下部左右各削一刀，形成楔形，接穗削面长度与砧木切口深度相同。
D.接合：将削好的接穗插入砧木切口，使接穗的形成层与砧木的形成层对齐，最后在嫁接接口处涂覆10g本发明实施例及对比例制备的抗菌保水凝胶，静置2min左右后用纱布包裹(绑缚)。
E.进行常规的嫁接后管理，不喷施任何杀菌剂。
对照组：仅步骤(D)的处理不同：将削好的接穗插入砧木切口，使接穗的形成层与砧木的形成层对齐，最后用塑料条将嫁接口进行绑缚。
嫁接(每个处理嫁接50株)完成后观察并记录处理组和对照组完全愈合的时间，并统计计算嫁接接口的发霉率(嫁接1个月后统计)以及嫁接的成活率(嫁接6个月后统计)。
结果见表1和表2。
发霉率＝发霉株数/嫁接株数×100％；成活率＝成活株数/嫁接株数×100％。
表1接穗为云抗14号
分组 愈合所需时间(d) 发霉率(％) 成活率(％)
实施例1 35 4 96
实施例2 37 6 94
实施例3 36 4 94
对比例1 42 10 88
对比例2 47 24 72
对比例3 44 18 80
对比例4 39 8 90
对照组 92 34 58
表2接穗为普景1号
分组 愈合所需时间(d) 发霉率(％) 成活率(％)
实施例1 32 2 98
实施例2 34 4 94
实施例3 32 2 96
对比例1 37 8 90
对比例2 43 20 76
对比例3 39 16 84
对比例4 36 8 88
对照组 83 32 61
从表1和表2中可以看出，在嫁接接口处涂覆本发明实施例制备的抗菌保水凝胶可以降低接口的愈合时间和发霉率，提高茶树嫁接的成活率。
以上所述的实施例仅是对本发明的优选方式进行描述，并非对本发明的范围进行限定，在不脱离本发明设计精神的前提下，本领域普通技术人员对本发明的技术方案做出的各种变形和改进，均应落入本发明权利要求书确定的保护范围内。

Claims (10)

1.一种抗菌保水凝胶，其特征在于，包括以下质量份数的原料：水苏糖20～30份、聚乳酸12～15份、裙带菜提取物8～10份、明胶10～13份、水杨酸1.5～2.5份、吲哚丁酸0.5～0.8份、复硝酚钠1～2份、黄腐酸2～3份和改性甘露聚糖4～5份。
2.根据权利要求1所述的抗菌保水凝胶，其特征在于，所述裙带菜提取物的制备方法，包括以下步骤：
(1)将裙带菜孢子叶粉碎后加水混合均匀，调节pH至5.5～6.5，然后加入复合酶保温酶解后收集上清液；
(2)将上清液浓缩后用乙醇溶液沉淀，得到所述裙带菜提取物。
3.根据权利要求2所述的抗菌保水凝胶，其特征在于，所述裙带菜孢子叶和水的质量/体积比为1g:(8～10)mL；所述复合酶的添加量为裙带菜孢子叶质量的2.0～2.5％；所述乙醇溶液的浓度为95vol.％。
4.根据权利要求3所述的抗菌保水凝胶，其特征在于，所述保温酶解的温度为45～55℃，时间为2.5～3.5h；所述复合酶，包括以下质量份数的组分：纤维素酶2～3份、胰蛋白酶1～2份和木瓜蛋白酶1～2份。
5.根据权利要求2所述的抗菌保水凝胶，其特征在于，所述改性甘露聚糖的制备方法，包括以下步骤：
将谷氨酸、甘露聚糖和水混合均匀，加热回流反应后浓缩、干燥，得到所述改性甘露聚糖。
6.根据权利要求5所述的抗菌保水凝胶，其特征在于，所述加热回流反应的温度为80～90℃，时间为8～10h。
7.一种权利要求1～6任一项所述的抗菌保水凝胶的制备方法，其特征在于，包括以下步骤：
将水苏糖加入水中加热溶解，然后加入聚乳酸、裙带菜提取物和明胶，进行一次搅拌后加入水杨酸、吲哚丁酸、复硝酚钠、黄腐酸和改性甘露聚糖，二次搅拌后高速剪切，得到所述抗菌保水凝胶。
8.根据权利要求7所述的制备方法，其特征在于，所述水苏糖和水的质量/体积比为1g:(45～55)mL；加热溶解的温度为80～85℃；所述一次搅拌的转速为80～100r/min，时间为30～40min；所述二次搅拌的转速为150～180r/min，时间为20～30min；所述高速剪切的转速为3500～3800r/min，时间为5～8min。
9.一种权利要求1～6任一项所述的抗菌保水凝胶在茶树嫁接中的应用。
10.根据权利要求9所述的应用，其特征在于，所述应用的方法，具体包括：将所述抗菌保水凝胶涂敷在嫁接接口处，静置2～3min，然后包裹纱布。
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