Using chemical genomics to study cell wall formation and cell growth in Arabidopsis thaliana and Penium margaritaceum.

The cell wall is directly involved in cell growth, and its ability to loosen and rearrange allows for cell expansion through the existing turgor pressure. Thus, information on cell wall deposition and rearrangement can provide insights into the overall plant growth. This chapter describes two methods that can be used to evaluate cell expansion (1) in the model plant Arabidopsis thaliana and (2) the model alga Penium margaritaceum. These methods are further used to screen for small molecules that induce cell growth phenotypic changes affecting cell wall. Identification of such small molecules is beneficial due to their posttranslational mechanism of action that can be controlled in a temporal and spatial manner. Chemical genomics has the ability to overcome issues of genetic redundancy and lethality, which can hinder traditional genetic methods. The identification of small molecules in these screens will provide useful information on plant cell wall biology and overall plant growth.

Constitutive expression of soybean ferritin cDNA in transgenic wheat and rice results in increased iron levels in vegetative tissues but not in seeds.

We used particle bombardment to produce transgenic wheat and rice plants expressing recombinant soybean ferritin, a protein that can store large amounts of iron. The cDNA sequence was isolated from soybean by RT-PCR and expressed using the constitutive maize ubiquitin-1 promoter. The presence of ferritin mRNA and protein was confirmed in the vegetative tissues and seeds of transgenic wheat and rice plants by northern and western blot analysis, respectively. The levels of ferritin mRNA were similar in the vegetative tissues of both species, but ferritin protein levels were higher in rice. Both ferritin mRNA and protein levels were lower in wheat and rice seeds. ICAP spectrometry showed that iron levels increased only in vegetative tissues of transgenic plants, and not in the seeds. These data indicate that recombinant ferritin expression under the control of the maize ubiquitin promoter significantly increases iron levels in vegetative tissues, but that the levels of recombinant ferritin in seeds are not sufficient to increase iron levels significantly over those in the seeds of non-transgenic plants.