CN102018969B - Device for disinfecting surfaces of explants of plants - Google Patents
Device for disinfecting surfaces of explants of plants Download PDFInfo
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Abstract
一种生物技术领域的植物外植体表面消毒装置，包括：储液瓶、氯气发生器、盐酸滴入管、储气瓶和灭菌瓶，其中：第一储液瓶与氯气发生器相连接，盐酸滴入管的输出端与氯气发生器相连，氯气发生器的输出端与灭菌瓶相连，灭菌瓶的底部与储气瓶的输入端相连，储气瓶的输出端与第二储液瓶相连接，第一储液瓶和第二储液瓶的底部相连通。本发明装置密闭，确保氯气不外泄；一次生成氯气后可以重复使用，节约成本；装置内氯气的量保持不变，不同的植物材料可以通过试验确定合适的消毒时间，便于把消毒条件准确量化。本发明安全环保，简单有效，操作方便，造价便宜，适用于大部分植物的外植体接种前表面消毒，具有很大的应用价值。
A plant explant surface disinfection device in the field of biotechnology, comprising: a liquid storage bottle, a chlorine gas generator, a hydrochloric acid drip tube, a gas storage bottle and a sterilization bottle, wherein: the first liquid storage bottle is connected to the chlorine gas generator , the output end of the hydrochloric acid dropping tube is connected to the chlorine generator, the output end of the chlorine gas generator is connected to the sterilizing bottle, the bottom of the sterilizing bottle is connected to the input end of the gas storage bottle, and the output end of the gas storage bottle is connected to the second storage bottle. The liquid bottles are connected, and the bottoms of the first liquid storage bottle and the second liquid storage bottle are connected. The device of the present invention is airtight to ensure that the chlorine gas does not leak out; once the chlorine gas is generated, it can be reused to save costs; the amount of chlorine gas in the device remains unchanged, and different plant materials can be tested to determine the appropriate disinfection time, which is convenient for accurately quantifying the disinfection conditions . The invention is safe and environment-friendly, simple and effective, convenient to operate and cheap in cost, suitable for surface disinfection of most plant explants before inoculation, and has great application value.
Description
技术领域technical field
本发明涉及的是一种生物技术领域的装置，具体是一种植物外植体表面消毒装置。The invention relates to a device in the field of biotechnology, in particular to a plant explant surface disinfection device.
背景技术Background technique
植物组织培养技术，是植物细胞工程学、遗传学、植物生理学、生物化学与分子生物学研究的重要基本技术。植物组织培养技术不仅仅用于快速繁殖，在单倍体育种、种质保存、生理学研究和基因转化等方面也有着广泛的用途。在植物组织培养中始终贯穿着一个无菌的概念，一个植物外植体在接种之前，必须先通过表面消毒然后才能获得无菌材料。Plant tissue culture technology is an important basic technology for plant cell engineering, genetics, plant physiology, biochemistry and molecular biology research. Plant tissue culture technology is not only used for rapid propagation, but also has a wide range of applications in haploid breeding, germplasm preservation, physiological research and gene transformation. There is always a concept of sterility in plant tissue culture. Before inoculation, a plant explant must be sterilized on the surface before obtaining sterile materials.
传统的用于植物外植体接种前消毒的途径，主要是把外植体用一种或几种消毒溶液浸泡，然后再用无菌水洗去残留的有毒离子。这种方法的缺点是：1.操作繁琐；2.有毒成分渗透到植物组织内部后，对后续培养造成不利影响；3.使用过的有毒废液污染环境。The traditional approach for disinfection of plant explants before inoculation is mainly to soak the explants with one or several disinfection solutions, and then wash away residual toxic ions with sterile water. The disadvantages of this method are: 1. The operation is cumbersome; 2. After the toxic component penetrates into the plant tissue, it will adversely affect the subsequent cultivation; 3. The used toxic waste liquid pollutes the environment.
氯气消毒是近年来发明的一种有效的外植体消毒方法，消毒效果好，操作方便。经过对现有技术的检索发现，刘海坤，卫志明2002年在《植物生理学通讯》上发表的“一种大豆成熟种子的消毒方法”一文中，采用如图1所示的氯气消毒装置，消毒效果比较理想，但该装置容易把氯气泄露在周围环境中，长期使用会引起慢性中毒，重者致死致残。Chlorine disinfection is an effective explant disinfection method invented in recent years. It has good disinfection effect and is easy to operate. After searching the prior art, it was found that in the article "A Disinfection Method for Mature Soybean Seeds" published by Liu Haikun and Wei Zhiming in "Plant Physiology Letters" in 2002, a chlorine gas disinfection device as shown in Figure 1 was used to disinfect The effect is relatively ideal, but the device is easy to leak chlorine gas into the surrounding environment, and long-term use will cause chronic poisoning, and severe cases may cause death or disability.
发明内容Contents of the invention
本发明针对现有技术存在的上述不足，提供一种植物外植体表面消毒装置，通过液泵使氯气在储气瓶和盛灭菌材料的容器中往返转移，从而达到表面消毒的目的。The present invention aims at the above-mentioned deficiencies in the prior art, and provides a device for disinfecting the surface of plant explants, which uses a liquid pump to transfer chlorine gas back and forth between the gas storage bottle and the container containing the sterilizing material, thereby achieving the purpose of surface disinfection.
本发明是通过以下技术方案实现的，本发明包括：若干个储液瓶、氯气发生器、盐酸滴入管、储气瓶和灭菌瓶，其中：第一储液瓶与氯气发生器相连接，盐酸滴入管的输出端与氯气发生器相连，氯气发生器的输出端与灭菌瓶相连，灭菌瓶的底部与储气瓶的输入端相连，储气瓶的输出端与第二储液瓶相连接，第一储液瓶和第二储液瓶的底部相连通。The present invention is achieved through the following technical solutions, and the present invention includes: several liquid storage bottles, chlorine gas generators, hydrochloric acid drip tubes, gas storage bottles and sterilization bottles, wherein: the first liquid storage bottle is connected with the chlorine gas generator , the output end of the hydrochloric acid dropping tube is connected to the chlorine gas generator, the output end of the chlorine gas generator is connected to the sterilizing bottle, the bottom of the sterilizing bottle is connected to the input end of the gas storage bottle, and the output end of the gas storage bottle is connected to the second storage bottle. The liquid bottles are connected, and the bottoms of the first liquid storage bottle and the second liquid storage bottle are connected.
所述的氯气发生器上设有次氯酸钠入口，所述的盐酸滴入管上设有控速开关和体积刻度，该盐酸滴入管的底部与氯气发生器的瓶底接触。The chlorine gas generator is provided with a sodium hypochlorite inlet, and the hydrochloric acid dripping tube is provided with a speed control switch and a volume scale, and the bottom of the hydrochloric acid dripping tube is in contact with the bottle bottom of the chlorine gas generator.
所述的第一储液瓶和第二储液瓶的体积相等，所述的灭菌瓶和储气瓶的体积相等，所述的储液瓶的体积为储气瓶体积的两倍。The volumes of the first liquid storage bottle and the second liquid storage bottle are equal, the volumes of the sterilization bottle and the gas storage bottle are equal, and the volume of the liquid storage bottle is twice the volume of the gas storage bottle.
所述的第一储液瓶和第二储液瓶之间设有液泵，该液泵为带有双向开关的普通液体泵。A liquid pump is arranged between the first liquid storage bottle and the second liquid storage bottle, and the liquid pump is an ordinary liquid pump with a bidirectional switch.
所述的氯气发生器和灭菌瓶之间、灭菌瓶与储气瓶之间均设有开关夹。Switch clips are provided between the chlorine generator and the sterilizing bottle, and between the sterilizing bottle and the gas storage bottle.
本发明工作时，先将该装置连接好置于超净工作台上，待灭菌材料放在灭菌材料存放瓶中，并把开关夹和第二储液瓶的盖子取下来，用饱和氯化钠溶液装满第二储液瓶。迅速往氯气发生器中加入一定体积的次氯酸钠，然后按比例加入一定体积的浓盐酸。顺序为先加次氯酸钠，再加浓盐酸。加好后迅速加盖密封，同时打开液泵到最大流速，使第二储液瓶中的一半体积的液体在1分钟之内转移到储液瓶中，从而使得生成的氯气转移到灭菌材料存放瓶中。最后关闭液泵，夹上开关夹，开始计时。When the present invention works, first connect the device and place it on the ultra-clean workbench, put the sterilized material in the sterilized material storage bottle, and take off the switch clip and the cover of the second liquid storage bottle, and use saturated chlorine Sodium NaCl solution fills the second reservoir bottle. Quickly add a certain volume of sodium hypochlorite to the chlorine generator, and then add a certain volume of concentrated hydrochloric acid in proportion. The sequence is to add sodium hypochlorite first, then concentrated hydrochloric acid. After filling, quickly cover and seal, and at the same time turn on the liquid pump to the maximum flow rate, so that half the volume of the liquid in the second liquid storage bottle is transferred to the liquid storage bottle within 1 minute, so that the generated chlorine gas is transferred to the sterilized material Store in bottle. Finally, turn off the liquid pump, clamp the switch clip, and start timing.
灭菌结束后，打开开关夹。打开液泵，把第二储液瓶中剩余的一半液体全部抽到第一储液瓶中，从而使氯气发生器中氯气全部转移到储气瓶中。夹上开关夹，取下待灭菌材料瓶，消毒完成。After sterilization, open the switch clip. Turn on the liquid pump, and pump all the remaining half of the liquid in the second liquid storage bottle into the first liquid storage bottle, so that all the chlorine gas in the chlorine gas generator is transferred to the gas storage bottle. Clamp the switch clamp, remove the material bottle to be sterilized, and the disinfection is completed.
下次使用时，不需要再生成氯气，只要把液泵开到相反的方向，使得储气瓶中的氯气转移至待灭菌材料瓶，消毒结束后再把氯气转移到氯气发生器中即可。In the next use, there is no need to regenerate chlorine gas, just turn the liquid pump to the opposite direction, so that the chlorine gas in the gas storage bottle is transferred to the material bottle to be sterilized, and then transfer the chlorine gas to the chlorine gas generator after disinfection .
与已有装置相比，本发明装置密闭，确保氯气不外泄；一次生成氯气后可以重复使用，节约成本；由于装置内氯气的量保持不变，不同的植物材料可以通过试验确定合适的消毒时间，便于把消毒条件准确量化。Compared with the existing device, the device of the present invention is airtight to ensure that the chlorine gas does not leak out; the chlorine gas can be reused after being generated once, and the cost is saved; since the amount of chlorine gas in the device remains unchanged, different plant materials can be tested to determine the appropriate disinfection time, it is convenient to accurately quantify the disinfection conditions.
附图说明Description of drawings
图1为现有技术结构示意图。Fig. 1 is a schematic structural diagram of the prior art.
图2为本发明的结构示意图。Fig. 2 is a structural schematic diagram of the present invention.
具体实施方式Detailed ways
下面对本发明的实施例作详细说明，本实施例在以本发明技术方案为前提下进行实施，给出了详细的实施方式和具体的操作过程，但本发明的保护范围不限于下述的实施例。The embodiments of the present invention are described in detail below. This embodiment is implemented on the premise of the technical solution of the present invention, and detailed implementation methods and specific operating procedures are provided, but the protection scope of the present invention is not limited to the following implementation example.
如图2所示，本实施例包括：第一储液瓶1、氯气发生器2、盐酸滴入管3、储气瓶4、灭菌瓶5和第二储液瓶6，其中：第一储液瓶1与氯气发生器2相连接，盐酸滴入管3的输出端与氯气发生器2相连，氯气发生器2的输出端与灭菌瓶5相连，灭菌瓶5的底部与储气瓶4的输入端相连，储气瓶4的输出端与第二储液瓶6相连接，第一储液瓶1和第二储液瓶6的底部相连通。As shown in Figure 2, the present embodiment includes: a first liquid storage bottle 1, a chlorine gas generator 2, a hydrochloric acid dripping tube 3, a gas storage bottle 4, a sterilization bottle 5 and a second liquid storage bottle 6, wherein: the first The liquid storage bottle 1 is connected with the chlorine gas generator 2, the output end of the hydrochloric acid dripping tube 3 is connected with the chlorine gas generator 2, the output end of the chlorine gas generator 2 is connected with the sterilizing bottle 5, and the bottom of the sterilizing bottle 5 is connected with the gas storage The input end of the bottle 4 is connected, the output end of the gas storage bottle 4 is connected with the second liquid storage bottle 6 , and the bottom of the first liquid storage bottle 1 and the second liquid storage bottle 6 are connected.
所述的氯气发生器2上设有次氯酸钠入口7，所述的盐酸滴入管3上设有控速开关8和体积刻度，该盐酸滴入管3的底部与氯气发生器2的瓶底接触。Described chlorine gas generator 2 is provided with sodium hypochlorite inlet 7, and described hydrochloric acid dripping tube 3 is provided with speed control switch 8 and volume scale, and the bottom of this hydrochloric acid dripping tube 3 contacts with the bottle bottom of chlorine gas generator 2 .
所述的第一储液瓶1和第二储液瓶6的体积相等，所述的灭菌瓶5和储气瓶4的体积相等，所述的储液瓶的体积为储气瓶4体积的两倍。The volumes of the first liquid storage bottle 1 and the second liquid storage bottle 6 are equal, the volumes of the sterilization bottle 5 and the gas storage bottle 4 are equal, and the volume of the described liquid storage bottle is the volume of the gas storage bottle 4 twice as much.
所述的第一储液瓶1和第二储液瓶6之间设有液泵9，该液泵9为带有双向开关的普通液体泵。A liquid pump 9 is provided between the first liquid storage bottle 1 and the second liquid storage bottle 6, and the liquid pump 9 is an ordinary liquid pump with a bidirectional switch.
所述的氯气发生器2和灭菌瓶5之间、灭菌瓶5与储气瓶4之间均设有开关夹。Between the chlorine generator 2 and the sterilizing bottle 5 , and between the sterilizing bottle 5 and the gas storage bottle 4 , switch clips are arranged.
本实施例用氯气消毒装置对大豆种子进行接种前表面消毒，设置第一储液瓶1与第二储液瓶6的体积为300毫升，氯气发生器2与储气瓶4的体积为150毫升。待灭菌大豆100粒。所加新开瓶次氯酸钠溶液75毫升，浓盐酸25毫升。灭菌时间为1小时。灭菌结束后，全部接种MS固体培养基，无一例污染，发芽率达100%。In this embodiment, a chlorine gas disinfection device is used to sterilize the surface of soybean seeds before inoculation. The volume of the first liquid storage bottle 1 and the second liquid storage bottle 6 is set to be 300 milliliters, and the volume of the chlorine gas generator 2 and the gas storage bottle 4 is 150 milliliters. . 100 soybeans to be sterilized. Add 75 ml of newly opened sodium hypochlorite solution and 25 ml of concentrated hydrochloric acid. The sterilization time is 1 hour. After the sterilization, all the MS solid media were inoculated, without any contamination, and the germination rate reached 100%.
本实施例中使用的装置，适用于大部分植物的外植体接种前表面消毒，具有很大的应用价值。The device used in this example is suitable for surface disinfection of explants of most plants before inoculation, and has great application value.
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| CN102293155B (en) * | 2011-07-20 | 2013-10-30 | 天津大学 | Convenient explant sterilization method by cup method |
| CN104026013B (en) * | 2014-06-17 | 2015-07-08 | 西北农林科技大学 | A plant tissue surface aseptic disinfection device |
| CN104737910A (en) * | 2015-03-06 | 2015-07-01 | 中南民族大学 | Easy seed sterilization method and device |
| CN106119076A (en) * | 2016-06-29 | 2016-11-16 | 无锡南理工科技发展有限公司 | A kind of crops training spore suspension preparation facilities and method |
| CN108552060A (en) * | 2018-07-19 | 2018-09-21 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | The sterilizing methods of sweet potato tissue cultures explant |
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| CN201131939Y (en) * | 2007-12-19 | 2008-10-15 | 吕延利 | Environmentally friendly miniature medical ethylene oxide sterilizer |
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| JP2004215930A (en) * | 2003-01-15 | 2004-08-05 | Kitano Takahiro | Method and device for sterilizing and cleaning endoscope |
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| JP特开2004-215930A 2004.08.05 |
| 刘海坤，卫志明.一种大豆成熟种子的消毒方法.《植物生理学通讯》.2002,第38卷(第3期),第260-261页. * |
| 循环概念在有毒气体实验中的运用;黄无违;《实验教学与仪器》;20030630;第20卷(第6期);第25页左栏第1-20行、图1-2 * |
| 黄无违.循环概念在有毒气体实验中的运用.《实验教学与仪器》.2003,第20卷(第6期), |
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