Molecular Biology Commons^™

Myosin Xi Is Essential For Tip Growth In Physcomitrella Patens, L Vidali, Gm Burkart, Rc Augustine, E Kerdavid, E Tuzel, M Bezanilla

Magdalena Bezanilla

Class XI myosins are plant specific and responsible for cytoplasmic streaming. Because of the large number of myosin XI genes in angiosperms, it has been difficult to determine their precise role, particularly with respect to tip growth. The moss Physcomitrella patens provides an ideal system to study myosin XI function. P. patens has only two myosin XI genes, and these genes encode proteins that are 94% identical to each other. To determine their role in tip growth, we used RNA interference to specifically silence each myosin XI gene using 5′ untranslated region sequences. We discovered that the two myosin XI …

Rapid Screening For Temperature-Sensitive Alleles In Plants, L Vidali, Rc Augustine, Sn Fay, P Franco, Ka Pattavina, M Bezanilla

Magdalena Bezanilla

We developed a simple and fast method to identify temperature-sensitive alleles of essential plant genes. We used primary and tertiary structure information to identify residues in the core of the protein of interest. These residues were mutated and tested for temperature sensitivity, taking advantage of the exceptionally rapid 1-week complementation assay in the moss Physcomitrella patens. As test molecules, we selected the actin-binding proteins profilin and actin-depolymerizing factor, because they are essential and their loss-of-function phenotype can be fully rescued. Screening a small number of candidate mutants, we successfully identified temperature-sensitive alleles of both profilin and actin-depolymerizing factor. Plants harboring …

Lifeact-Megfp Reveals A Dynamic Apical F-Actin Network In Tip Growing Plant Cells, L Vidali, Cm Rounds, Pk Hepler, M Bezanilla

Magdalena Bezanilla

Background

Actin is essential for tip growth in plants. However, imaging actin in live plant cells has heretofore presented challenges. In previous studies, fluorescent probes derived from actin-binding proteins often alter growth, cause actin bundling and fail to resolve actin microfilaments.

Methodology/Principal Findings

In this report we use Lifeact-mEGFP, an actin probe that does not affect the dynamics of actin, to visualize actin in the moss Physcomitrella patens and pollen tubes from Lilium formosanum and Nicotiana tobaccum. Lifeact-mEGFP robustly labels actin microfilaments, particularly in the apex, in both moss protonemata and pollen tubes. Lifeact-mEGFP also labels filamentous actin structures in …

Profilin Is Essential For Tip Growth In The Moss Physcomitrella Patens, L Vidali, Rc Augustine, Kp Kleinman, M Bezanilla

Magdalena Bezanilla

The actin cytoskeleton is critical for tip growth in plants. Profilin is the main monomer actin binding protein in plant cells. The moss Physcomitrella patens has three profilin genes, which are monophyletic, suggesting a single ancestor for plant profilins. Here, we used RNA interference (RNAi) to determine the loss-of-function phenotype of profilin. Reduction of profilin leads to a complete loss of tip growth and a partial inhibition of cell division, resulting in plants with small rounded cells and fewer cells. We silenced all profilins by targeting their 3′ untranslated region sequences, enabling complementation analyses by expression of profilin coding sequences. …

The Role Of Arp2/3 And Scar/Wave Complexes In Polar Growth Of The Apical Cell Of The Moss Physcomitrella, Rs Quatrano, Pf Perroud, P Harries, M Bezanilla, Ah Pan, P Klueh, D Cove

Magdalena Bezanilla

No abstract provided.