测试拟南芥耐咀嚼昆虫的选择和非选择性检测

Biology
 

Summary

植物耐咀嚼昆虫食草动物可以在几个方面进行测试。在这里,我们展示了如何设置一个选择和一个没有选择的实验与模型植物拟南芥,以确定对有害物种菜青虫的阻力。

Cite this Article

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De Vos, M., Jander, G. Choice and No-Choice Assays for Testing the Resistance of A. thaliana to Chewing Insects. J. Vis. Exp. (15), e683, doi:10.3791/683 (2008).

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Abstract

白色的小菜粉蝶的幼虫是一种有害生物,农业的设置。这种毛虫饲料白菜家族中的植物,其中包括许多作物,如白菜,花椰菜,甘蓝等昆虫饲养白菜植物在温室的地方。至少有两个笼子里饲养菜青虫需要。和其他包含成人,蝴蝶的幼虫。为了研究植物激素和有毒的植物化学物质,在抵抗虫害的作用,我们展示了两个实验。首先,耐咀嚼昆虫菜青虫茉莉酸(JA - 往往在抗虫性植物激素)的作用的决心。 JA生产受损的野生型和突变型植物,可以比较卡特彼勒增长。这个实验被认为是“别无选择”,因为幼虫被迫生存在一个厂,合成或不足之处是司法机构政务长。第二,我们展示了一个实验,调查的硫代葡萄糖苷,这是用来作为产卵(产卵)信号的作用。在这里,我们使用WT和突变的拟南芥在“生死抉择”的实验中,女性的蝴蝶允许选择打下它们的蛋,无论是基因型的植物甙生产受损。这个视频演示实验装置的分析以及代表性的结果。

Protocol

无选择试验:

  1. 拟南芥植物的兴趣仔细括在一个塑料杯。在这种情况下:COL - 0(WT)和JA生产受损的突变。确保您限制损坏的叶子。对于我用画笔后面。
  2. 使用精细的画笔,从一个刚孵出的幼虫上的白菜殖民地。
  3. 小心地将拟南芥的幼虫。
  4. 在第4天,7,10的重量幼虫。提示:短日照的条件(8小时光照)增加营养生长(1幼虫能够履行其全生命周期的1株)下生长的植物。

选择实验:

  1. 选定的两家工厂,每个基因型之一。最好是植物生长在同放入锅内,以限制其他因素,如托盘的影响,等等。
  2. 放置在蝴蝶笼的植物。
  3. 选择肥沃的女性。我通常赶上一位女性,正积极铺设鸡蛋在饲养的​​殖民地。
  4. 选择两个24小时的植物,在笼中释放的女性。

提示1:为了消除影响学习,不重用随后选择测试的女性。

提示2:只能比较,也有类似的增长的表型的植物。或者,你可以从每个基因型安装一片叶子,一个锥形瓶中,并比较它们的吸引力女性。

关于本议定书和昆虫取食植物免疫力的更多信息, 请访问植物生物学年度评论。

Comments

6 Comments

  1. I thought you did an excellent job, nice and clear explainations, good transitions and summary.

    Reply
    Posted by: Anonymous
    June 14, 2008 - 3:06 PM
  2. 1. perhaps you could explain why you use  arabidopsis instead of cabbage. My assumption is that much work, including sequencing of the genome, had been done on the arabidopsis so you were able to add-remove genetic components you wanted to test for. ². how long is a generation of the cabbage butterfly? 3. Choice experiment. Why did you not use multiple larvae released into cage with the two types of arabidopsis? Some what confusing because there were two different growth stages and two different stimuli tested for in the same testing 4. Excellent job of editing and presenting. It fit nicely on the video   Jay

    Reply
    Posted by: Anonymous
    July 6, 2008 - 8:00 AM
  3. Dear Jay, Thank you for your comments. Let me answer your questions and give you some additional information on the life cycle of Pieris rapae. 1. Larvae of the small white butterfly will feed from almost all cruciferous plants. We use Arabidopsis thaliana because of its short generation time, small size and above all its sequenced genome and vast number of available mutants. These mutants allow us to unravel the defense responses during the host-insect interaction. As demonstrated in the video, we show that jasmonic acid plays a major role in the plant's defensive capacities towards Pieris rapae. Interestingly, we also show that glucosinolates, whose breakdown products are toxic to many generalist caterpillars, are used as by Pieris rapae female butterflies (specialist on cruciferous plants) as oviposition cues. We have recently published a paper in Plant Physiology (March ²008) on this topic. ². Pieris rapae life cycle. The development of Pieris rapae strongly depends on the temperature and ranges from 5-7 weeks. We rear our insects at ²3C. Eggs hatch in 3-7 days; Larvae have 5 larval stages (L1-L5; ~ 10-14 day); Pupae (7-14 days); Adults (~14-²0 days). Under our rearing conditions eggs hatch in 5 days, complete larval development until pupation takes 1² days, pupa take ~7 days to eclosion, adult life is ~² weeks. 3. Using the video we wanted to show different experimental set-ups for testing plant-insect interaction. A choice test (preference) will give you additional insights that are not necessarily displayed in a no-choice test (edibility). There are other possibilities for choice tests, including the one you mentioned. The results obtained with this approach, e.g. leaf area eaten, are hard to quantify and rather subjective. Therefore, we mostly use the no-choice set-up to determine the effect on caterpillar performance. Martin 

    Reply
    Posted by: Anonymous
    July 7, 2008 - 11:05 AM
  4. The demo was very clear to me, but the music was .........
    Maybe you should point out in the video, that these experiments have to repeated.

    Reply
    Posted by: Birgid B. S.
    July 7, 2009 - 2:13 AM
  5. I had a question regarding the increase of weight of the Caterpillar.well the readings are taken on 4,7,10 day.The coi1 plants if are consumed earlier ,how come the weight is increased,if no plant material is left till 10th day?

    Reply
    Posted by: Anonymous
    January 14, 2012 - 11:19 PM
  6. I love this vedio and may you tag this vedio to me

    Reply
    Posted by: Anonymous
    March 16, 2012 - 11:44 PM

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