搜索

1-MCP (1-methylcyclopropene) Treatment Protocol for Fruit or Vegetables
水果或蔬菜的1-MCP(1-甲基环丙烯)处理实验方案   

下载 PDF 引用 收藏 提问与回复 分享您的反馈 Cited by

本文章节

Abstract

1-MCP (1-methylcyclopropene) is a simple synthetic hydrocarbon molecule that interacts with the ethylene receptor and inhibits the response of fruit or plant to ethylene. 1-MCP has opened new opportunities in handling harvested crops and serves as a powerful tool to learn about plant response to ethylene (Watkins and Miller, 2006). 1-MCP is manufactured by Agrofresh and known by its commercial name SmartfreshSM.

Keywords: Ripening inhibition(成熟抑制), Ethylene(乙烯), Fruit(水果), Postharvest(采后), Storage(存储)

Background

Application of 1-MCP can serve as a powerful tool to examine the response of the plant to ethylene. The commercial production of 1-MCP by Agrofresh allows simple practice of this treatment, especially postharvest. The application of 1-MCP to plants, fruit or vegetable was mentioned in many articles but was not described in detail. This was the motivation to provide the following protocol. In this protocol tomatoes served as candidates for the treatment.

Materials and Reagents

  1. Hand gloves (latex or nitrile)
  2. Rubber septa (for the volumetric flask) (Sigma-Aldrich, catalog number: Z553964 )
  3. Syringe 10 ml (OMG, catalog number: OMG-W-10M )
  4. Needle 0.60 x 30 mm (Pic, catalog number: 03.070140.300.800 )
  5. Weighing papers for the 1-MCP powder (Whatman, catalog number: 28414662 )
  6. Freshly harvested tomatoes (any type and origin)
  7. 1-MCP powder (Agrofresh Inc.) containing 0.14% 1-MCP as active ingredient
  8. Tap water

Equipment

  1. 30 L airtight HDPE plastic barrel wide mouth and cover with lid equipped with flexible latex hose or rubber septa (Chen Samuel chemicals, catalog number: 0100230 )
  2. Brand volumetric flask 1,000 ml (Sigma-Aldrich, catalog number: Z326828 )
  3. Benchtop weight scales with 0.01 g resolution (Vibra, catalog number: AJ-3200CE )

Procedure

  1. Preparation of 1-MCP gas stock (1,000 ppm or 0.1%)
    1. Gently pour 0.82 g of 1-MCP powder into the 1,000 ml volumetric flask and close tightly with the rubber septa.
    2. Inject a volume of 50 ml of water into the closed flask (5 times, 10 ml of water) and dissolve the powder to release the 1-MCP gas (Figure 1A).
    3. Turn the volumetric flask upside down to check that the gas volume is 1,000 ml otherwise add water up to the marked line (Figure 1B).


      Figure 1. Preparation of 1-MCP gas stock and application to the tomato fruit. A. Injection of water into the volumetric flask through the rubber septa; B. Volumetric flask filled with water up to the marked line to allow accurate volume of gas; C. Injection of 1-MCP gas through flexible latex hose into the closed barrel with tomatoes inside (Pictures: Dan Gamrasni).

  2. 1-MCP treatment.
    1. Place the tomatoes (It is recommended not to exceed over one third of the barrel volume) inside the barrel and close the lid tightly. You can use wrapped plastic or other solution to prevent any gas leakage. The tomatoes and the barrel should be placed in a 20-25 °C room.
    2. To apply 1 ppm of 1-MCP: injection of 30 ml of 1,000 ppm 1-MCP in 30 L barrel will give a concentration of 1 ppm 1-MCP to the tomatoes.

      C1 x V1 = C2 x V2; 1,000 (ppm) x V1 (ml) = 1 (ppm) x 30,000 (ml)

      Where, C1 = the concentration of 1-MCP gas stock in the volumetric flask (ppm); V1 = the unknown volume of the 1-MCP gas stock sample that should inject into the Barrel (ml); C2 = the wanted concentration of 1-MCP in the barrel (ppm); V2 = the volume of the barrel (ml).
    3. The application of 1-MCP: Inject 10 ml of water into the volumetric flask (1-MCP) through the septa, draw up 10 ml of 1-MCP gas and inject immediately into the closed barrel through the flexible latex hose or the septa (Figure 1C).
    4. Repeat step 2c two more times until injecting a total of 30 ml 1-MCP gas into the barrel.
    5. Incubate for 24 h.
    6. Open the barrel and air the tomatoes for at least 2 h to allow the tomatoes to adjust to the atmospheric environment. It is possible to measure ethylene levels and/or carbon dioxide in the barrel before opening.
    7. As a control, untreated tomatoes should be placed in similar conditions without 1-MCP or any other gas treatment (Figure 2).


      Figure 2. The effect of 1-MCP treatment on the color change of tomatoes (Lycopersicon esculentum cv. Ailsa Craig) harvested at different ripening stages (mature green, breaker or early orange) 48 h after the end of the treatment (Picture: Dario Breitel)

Notes

  1. The volume of the tomatoes should not exceed one third of the barrel to prevent CO2 accumulation if quicklime is not available, otherwise use bigger barrel (200 L) or do the treatment to smaller samples of tomatoes in few barrels.
  2. To prevent carbon dioxide accumulation due to fruit respiration in the closed barrel, it is possible to add 0.5 kg of quicklime (known as burnt lime) that will spontaneously react with CO2 from the air (you can keep this powder in any paper bag).
  3. It is possible to use the 1-MCP stock solution up to 2 weeks keeping the volumetric flask upside down at low temperature (4 °C).

Acknowledgments

The protocol was adapted from Gamrasni et al. (2010), and was similarly applied in Breitel et al. (2016).
We would like to thank RIMI and Agrofresh Inc. for the supply of 1-MCP.

References

  1. Breitel, D. A., Chappell-Maor, L., Meir, S., Panizel, I., Puig, C. P., Hao, Y., Yifhar, T., Yasuor, H., Zouine, M., Bouzayen, M., Granell Richart, A., Rogachev, I. and Aharoni, A. (2016). AUXIN RESPONSE FACTOR 2 intersects hormonal signals in the regulation of tomato fruit ripening. PLoS Genet 12(3): e1005903.
  2. Gamrasni, D., Ben-Arie, R. and Goldway, M. (2010). 1-Methylcyclopropene (1-MCP) application to Spadona pears at different stages of ripening to maximize fruit quality after storage. Postharvest Biol Technol 58(2):104-112.
  3. Watkins, C. B. and Miller, W. B. (2006). A summary of physiological processes or disorders in fruits, vegetables and ornamental products that are delayed or decreased, increased, or unaffected by application of 1-methylcyclopropene (1-MCP).

简介

1-MCP(1-甲基环丙烯)是与乙烯受体相互作用并抑制果实或植物对乙烯的反应的简单合成烃分子。 1-MCP已经为处理收获的作物开辟了新的机会,并且是了解植物对乙烯的反应的有力工具(Watkins和Miller,2006)。 1-MCP由Agrofresh制造,以商业名称Smartfresh SM 而闻名。

背景 1-MCP的应用可以作为检测工厂对乙烯的响应的有力工具。 Agrofresh的1-MCP的商业化生产允许这种处理的简单实践,特别是收获后。 许多文章中提到了1-MCP对植物,水果或蔬菜的应用,但没有详细描述。 这是提供以下协议的动机。 在这个协议中,西红柿作为治疗的候选人。

关键字:成熟抑制, 乙烯, 水果, 采后, 存储

材料和试剂

  1. 手套(乳胶或腈)
  2. 橡胶隔垫(用于容量瓶)(Sigma-Aldrich,目录号:Z553964)
  3. 注射器10ml(OMG,目录号:OMG-W-10M)
  4. 针0.60 x 30 mm(Pic,目录号:03.070140.300.800)
  5. 1-MCP粉末称重纸(Whatman,目录号:28414662)
  6. 新鲜收获的西红柿(任何类型和来源)
  7. 含有0.14%1-MCP作为活性成分的1-MCP粉末(Agrofresh Inc.)
  8. 自来水

设备

  1. 30L密封HDPE塑料桶广口盖和盖子配有柔性乳胶软管或橡胶隔垫(陈塞缪尔化学品,目录号:0100230)
  2. 品牌容量瓶1000ml(Sigma-Aldrich,目录号:Z326828)
  3. 台秤重量为0.01g分辨率(Vibra,目录号:AJ-3200CE)

程序

  1. 1-MCP气体原料的制备(1,000ppm或0.1%)
    1. 轻轻地将0.82g 1-MCP粉末倒入1,000ml容量瓶中,并用橡胶隔片紧密地紧贴。
    2. 将50ml体积的水注入封闭的烧瓶中(5次,10ml水),并溶解粉末以释放1-MCP气体(图1A)。
    3. 将容量瓶倒置倒置,以检查气体体积为1,000 ml,否则将水加至标线(图1B)。


      图1. 1-MCP气体原料的制备和番茄果实的应用。 A.通过橡胶隔片将水注入容量瓶中; B.容量瓶中装满水,直到标记线,以允许精确的气体体积; C.通过柔性乳胶软管将1-MCP气体注射到带有西红柿的封闭桶中(图片:Dan Gamrasni)。

  2. 1-MCP治疗。
    1. 在桶内放置西红柿(建议不要超过桶体积的三分之一以上),并将盖子紧紧关闭。您可以使用包裹的塑料或其他溶液来防止任何气体泄漏。西红柿和桶应放在20-25°C的房间。
    2. 应用1ppm的1-MCP:在30L桶中注入30ml的1,000ppm的1-MCP,将给番茄1ppm的1-MCP浓度。

      C1×V1 = C2×V2; 1,000(ppm)×V1(ml)= 1(ppm)×30,000(ml)

      其中C1 =容量瓶中1-MCP气体的浓度(ppm); V1 =应注入桶(ml)的1-MCP气体样品的未知体积; C2 =桶中1-MCP的需要浓度(ppm); V2 =桶的体积(ml)。
    3. 1-MCP的应用:通过隔垫将10ml水注入容量瓶(1-MCP)中,抽出10ml的1-MCP气体,并通过柔性胶乳软管或隔垫立即注入封闭的筒体(图1C)
    4. 重复步骤2c两次,直到将30ml 1-MCP气体注入桶中
    5. 孵育24小时。
    6. 打开桶并将番茄放入至少2小时,以使番茄适应大气环境。在打开之前可以测量桶中的乙烯水平和/或二氧化碳。
    7. 作为对照,未经处理的番茄应处于类似的条件下,无需1-MCP或任何其他气体处理(图2)。


      图2. 1-MCP处理对番茄颜色变化的影响( 番茄红霉素 cv。Ailsa Craig)不同的成熟阶段(成熟的绿色,破碎或早期的橙色)治疗结束后48 h (图片:Dario Breitel)

笔记

  1. 如果生石灰不可用,番茄的体积不应超过桶的三分之一,以防止二氧化碳积累,否则使用较大的桶(200升),或少量处理较小的番茄样本桶。
  2. 为了防止封闭桶内水果呼吸引起的二氧化碳积聚,可以添加0.5公斤的生石灰(称为生石灰),这些生石灰会与空气中的CO 2自发反应(您可以保持这一点任何纸袋中的粉末)
  3. 可以使用1-MCP储存溶液长达2周,使容量瓶在低温(4°C)下倒置。

致谢

该协议改编自Gamrasni等人。 (2010),并且类似地应用于Breitel等人。 (2016)。
我们要感谢RIMI和Agrofresh Inc.为1-MCP的供应。

参考

  1. Breitel,DA,Chappell-Maor,L.,Meir,S.,Panizel,I.,Puig,CP,Hao,Y.,Yifhar,T.,Yasuor,H.,Zouine,M.,Bouzayen, Granair Richart,A.,Rogachev,I。和Aharoni,A.(2016)。  AUXIN RESPONSE FACTOR 2在激素信号调节番茄果实成熟中相交。 PLoS Genet 12(3):e1005903。
  2. Gamrasni,D.,Ben-Arie,R.和Goldway,M.(2010)。< a class ="ke-insertfile"href ="http://www.sciencedirect.com/science/article/pii/在不同成熟阶段的梨子中的1-甲基环丙烯(1-MCP)应用,以使存储后的果实质量达到最大化。采收后生物技术 58(2):104-112。
  3. Watkins,CB和Miller,WB(2006)。生理概要延迟或减少,增加或不受施用1-甲基环丙烯(1-MCP)影响的水果,蔬菜和观赏产品中的过程或障碍。
  • English
  • 中文翻译
免责声明 × 为了向广大用户提供经翻译的内容,www.bio-protocol.org 采用人工翻译与计算机翻译结合的技术翻译了本文章。基于计算机的翻译质量再高,也不及 100% 的人工翻译的质量。为此,我们始终建议用户参考原始英文版本。 Bio-protocol., LLC对翻译版本的准确性不承担任何责任。
Copyright: © 2017 The Authors; exclusive licensee Bio-protocol LLC.
引用:Dan, G., Martin, G., Yosi, S., Dario, B. and Asaph, A. (2017). 1-MCP (1-methylcyclopropene) Treatment Protocol for Fruit or Vegetables. Bio-protocol 7(10): e2278. DOI: 10.21769/BioProtoc.2278.
提问与回复

(提问前,请先登录)bio-protocol作为媒介平台,会将您的问题转发给作者,并将作者的回复发送至您的邮箱(在bio-protocol注册时所用的邮箱)。为了作者与用户间沟通流畅(作者能准确理解您所遇到的问题并给与正确的建议),我们鼓励用户用图片或者视频的形式来说明遇到的问题。由于本平台用Youtube储存、播放视频,作者需要google 账户来上传视频。

当遇到任务问题时,强烈推荐您提交相关数据(如截屏或视频)。由于Bio-protocol使用Youtube存储、播放视频,如需上传视频,您可能需要一个谷歌账号。