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Measuring Germination Percentage in Wheat
小麦发芽率的测定   

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Abstract

Knowledge of the viability of seeds is a prerequisite for establishing the seeding rate in crop production and for germination-related trait evaluation in crop plants. This method explains a simple procedure to establish germination percentage in wheat seeds.

Materials and Reagents

  1. Wheat seeds
  2. Sterile deionized water
  3. Filter paper (Whatman No.1)
  4. Permanent marker
  5. Lab wipes and gloves
  6. 5% (w/v) Sodium hypochlorite

Equipment

  1. Forceps
  2. Laminar flow hood
  3. Timer
  4. 70% Alcohol and flame
  5. Metal forceps
  6. Ruler
  7. Petri dishes (11 cm diameter)

Procedure

  1. Obtain a representative sample (i.e. seeds of a particular variety or cultivar) of your wheat seeds. Random selection within this sample is required to avoid any bias. The rest of the procedure is done under Laminar flow hood.
  2. Wipe the working area with alcohol and alcohol/flame sterilize the forceps.
  3. Spread fresh Whatman paper 1 on the petri dish and moisten until thoroughly damp (~2 ml water is added in our case). Avoid standing water above the Whatman paper 1.
  4. The seeds are placed in petri dishes, covered with disinfecting (5% (w/v) sodium hypochlorite) for 15 min, stirred, drained, and washed four times with sterile deionized water.
  5. Gently place the seeds spread out in the petri dish with a sterile forceps and record time. Use four replicates of five seeds per dish. It is advisable to have extra sets in case of microbial contamination.
  6. Place the petri dishes with seeds in a dark place with stable room temperature (~73 °F).
  7. Seeds are considered to be germinated when radicle has emerged approximately ≥ 2 mm (Figure 1). Germination percentage is recorded every 24 h for 6 days. Keep an eye for contamination. Do not let the whatman 1 paper dry out (Periodically inspect the moisture level with the help of a timer. In our case every 12 h, ~1 ml sterile water was added under aseptic conditions to each of the petri dishes).


    Figure 1. Germinated wheat seeds at the 4th day of germination initiation

  8. Germination rate is estimated by using the following formula:
    Germination Percentage = seeds germinated/total seeds x 100

Acknowledgments

This protocol is adapted from Manmathan et al. (2013).

References

  1. Maynard, Donald N. and George J. Hochmuth. 1997. Knotts Handbook for Vegetable Growers, 4th Edition.
  2. Manmathan, H., Shaner, D., Snelling, J., Tisserat, N. and Lapitan, N. (2013). Virus-induced gene silencing of Arabidopsis thaliana gene homologues in wheat identifies genes conferring improved drought tolerance. J Exp Bot 64(5): 1381-1392.
  3. New York: John Wiley and Sons, Inc. Yaklich, R.W., Editor. (1985). Rules for Testing Seeds, J Seed Technol. Lansing, Michigan: Association of Official Seed Analysts. Vol. 6, No. 2.

简介

种子生存力的知识是在作物生产中确定接种速率和在作物植物中进行发芽相关性状评价的先决条件。 该方法解释了在小麦种子中建立发芽率的简单程序

材料和试剂

  1. 小麦种子
  2. 无菌去离子水
  3. 滤纸(Whatman No.1)
  4. 永久标记
  5. 实验室擦拭布和手套
  6. 5%(w/v)次氯酸钠

设备

  1. 镊子
  2. 层流罩
  3. 计时器
  4. 70%酒精和火焰
  5. 金属钳
  6. 标尺
  7. 培养皿(直径11厘米)

程序

  1. 获取您的小麦种子的代表性样品(即特定品种或品种的种子)。 需要在该样本内随机选择以避免任何偏差。 其余步骤在层流净化罩下进行
  2. 用酒精和酒精擦拭工作区域,对镊子进行火焰消毒
  3. 传播新鲜的Whatman纸1在培养皿,并润湿,直到彻底潮湿(在我们的情况下,约2毫升水)。避免在Whatman纸1上面积水。
  4. 将种子置于培养皿中,用消毒(5%(w/v)次氯酸钠)覆盖15分钟,搅拌,排干,并用无菌去离子水洗涤四次。
  5. 用无菌镊子轻轻地将种子散布在培养皿中,并记录时间。每碟使用四个重复五个种子。建议在微生物污染的情况下增加一套。
  6. 将培养皿中的种子放在黑暗的地方,室温稳定(约73°F)
  7. 当胚根出现约≥2mm时,种子被认为是发芽的(图1)。每24小时记录6天的萌发百分比。注意污染。不要让Whatman 1纸干燥(在定时器的帮助下定期检查水分含量。在我们的情况下,每12小时,在无菌条件下向每个培养皿中加入〜1ml无菌水)。


    图1.在发芽开始的第4天发芽的小麦种子

  8. 发芽率使用以下公式估算:
    发芽百分比=种子发芽/总种子×100

致谢

该协议改编自Manmathan等人(2013)。

参考文献

  1. Maynard,Donald N.和George J. Hochmuth。 1997. Knotts Handbook for Vegetable Growers,4 th Edition。
  2. Manmathan,H.,Shaner,D.,Snelling,J.,Tisserat,N.和Lapitan,N。(2013)。 小麦中拟南芥基因同源物的病毒诱导基因沉默鉴定基因 赋予改善的耐旱性。 64(5):1381-1392。
  3. New York:John Wiley and Sons,Inc.Yaklich,R.W.,Editor。 (1985)。 测试种子的规则 J Seed Technol 。 兰辛,密歇根州:官方种子分析师协会。 Vol。 6,No.2。
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Copyright: © 2013 The Authors; exclusive licensee Bio-protocol LLC.
引用:Manmathan, H. and Lapitan, N. L. (2013). Measuring Germination Percentage in Wheat. Bio-protocol 3(16): e866. DOI: 10.21769/BioProtoc.866.
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