Within leaf variation is the largest source of variation in agroinfiltration of Nicotiana benthamiana

Hany Bashandy, Salla Jalkanen, Teemu Heikki Teeri

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Abstract
Background: Transient gene expression utilizing syringe agroinfiltration offers a simple and efficient technique for
different transgenic applications. Leaves of Nicotiana benthamiana show reliable and high transformation efficiency,
but in quantitative assays also a certain degree of variation. We used a nested design in our agroinfiltration experiments
to dissect the sources of this variation.
Results: An intron containing firefly luciferase gene was used as a reporter for agroinfiltration. A number of 6 week
old tobacco plants were infiltrated for their top leaves, several samples were punched from the leaves after 2 days of
transient expression, and protein extracts from the samples were repeatedly measured for luciferase activity. Interestingly,
most of the variation was due to differences between the sampling spots in the leaves, the next important
source being the different leaves on each plant. Variation between similar experiments, between plants and between
repetitive measurements of the extracts could be easily minimized.
Conclusions: Efforts and expenditure of agroinfiltration experiments can be optimized when sources of variation are
known. In summary, infiltrate more plants but less leaves, sample more positions on the leaf but run only few technical
replicates.
Original languageEnglish
Article number47
JournalPlant Methods
Volume11
Number of pages7
ISSN1746-4811
DOIs
Publication statusPublished - Oct 2015
MoE publication typeA1 Journal article-refereed

Fields of Science

  • 1183 Plant biology, microbiology, virology
  • 414 Agricultural biotechnology

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