Movatterモバイル変換

[0]ホーム
URL:

Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
Thehttps:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

NIH NLM Logo
Log inShow account info
Access keysNCBI HomepageMyNCBI HomepageMain ContentMain Navigation
pubmed logo
Advanced Clipboard
User Guide

Full text links

Silverchair Information Systems full text link Silverchair Information Systems Free PMC article
Full text links

Actions

Share

.2012 Jan;24(1):163-77.
doi: 10.1105/tpc.111.093575. Epub 2012 Jan 31.

POM-POM2/cellulose synthase interacting1 is essential for the functional association of cellulose synthase and microtubules in Arabidopsis

Affiliations

POM-POM2/cellulose synthase interacting1 is essential for the functional association of cellulose synthase and microtubules in Arabidopsis

Martin Bringmann et al. Plant Cell.2012 Jan.

Abstract

In plants, regulation of cellulose synthesis is fundamental for morphogenesis and plant growth. Cellulose is synthesized at the plasma membrane, and the orientation of synthesis is guided by cortical microtubules; however, the guiding mechanism is currently unknown. We show that the conditional root elongation pom2 mutants are impaired in cell elongation, fertility, and microtubule-related functions. Map-based cloning of the POM-POM2 locus revealed that it is allelic to CELLULOSE SYNTHASE INTERACTING1 (CSI1). Fluorescently tagged POM2/CSI1s associated with both plasma membrane-located cellulose synthases (CESAs) and post-Golgi CESA-containing compartments. Interestingly, while CESA insertions coincided with cortical microtubules in the pom2/csi1 mutants, the microtubule-defined movement of the CESAs was significantly reduced in the mutant. We propose that POM2/CSI1 provides a scaffold between the CESAs and cortical microtubules that guide cellulose synthesis.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Positional Cloning ofPOM2 and Microtubule-Related Defects in the Mutant.(A) Six-day-old etiolatedpom2-4 hypocotyl displaying twisting of cell files (red outline).(B) Rosette leaves of 3-week-oldpom2-4 plants that spiral.(C) Microtubule organization in 3-d-old elongating hypocotyl cells of the wild type (wt; left) andpom2-4 (right). Microtubules are labeled by mCherry:TUA5.(D) Microtubule angle distribution in elongating hypocotyl cells in the wild type andpom2-4. Angles were measured against the growth axis. Seven cells from seven seedlings were used for the analysis.(E) Mappedpom2 mutations inPOM2 (At2g22125). Black line indicates introns and boxed areas exons. aa, amino acids. Bars = 100 μm in(A), 2 cm in(B), and 10 μm in(C).
Figure 2.
Figure 2.
Triplepom2/csi1 csi2 csi3 Mutant Phenotypes.(A) Graph displaying length of 5-d-old etiolated hypocotyls, 5-d-old light-grown seedling roots, and mature siliques from 7-week-old soil-grown plants of wild-type,pom2-7/csi1-5, andpom2-7/csi1-5 csi2 csi3 genotypes.(B) Seven-week-old wild-type,pom2-7/csi1-5, andpom2-7/csi1-5 csi2 csi3 triple mutants. Bar = 5 cm.(C) Five-day-old etiolated wild-type,pom2-7/csi1-5, andpom2-7/csi1-5 csi2 csi3 triple mutant seedlings. Bar = 5 mm.(D) Five-day-old light-grown wild-type,pom2-7/csi1-5, andpom2-7/csi1-5 csi2 csi3 triple mutant seedlings. Bar = 5 mm.(E) Mature siliques from 7-week-old wild-type,pom2-7/csi1-5, andpom2-7/csi1-5 csi2 csi3 triple mutant plants. Bar = 1 cm.(F) to(H) Environmental scanning electron microscopy images of pollen grains on anthers from wild-type,pom2-7/csi1-5, andpom2-7/csi1-5 csi2 csi3 triple mutant plants. Bar = 70 μm.(I) Twisting rosette leaves in 4-week-oldpom2-7/csi1-5 andpom2-7/csi1-5 csi2 csi3 triple mutant plants. Bar = 5 cm.
Figure 3.
Figure 3.
POM2/CSI1-Containing Compartments Can Merge and Bifurcate at the Cell Cortex.(A) Kymographs from 4-d-old etiolated seedlings expressing tdTomato:CESA6 and POM2:CFP. Kymographs display CESA and POM2/CSI movement at the plasma membrane in nontreated cells (left) and CESA and POM/CSI1 movement at the cell cortex in isoxaben-treated (100 nm for 8 h) cells (right).(B) and(C) Images from two different time series in 4-d-old etiolated seedlings expressing tdTomato:CESA6 and POM2:CFP. The images show a split of one compartment into two (for both the CESA and POM2/CSI1 fluorescent signal) at 5 s(B), and a merge of the signals at 5 s in another time series(C). Right panel shows kymographs depicting the split of the signal into two(Top) and the merge of two compartments into one(Bottom). Bars = 2 μm.(D) and(E) Surface plots displayed as heat maps. The mean intensity of the signals above background was measured for(B) and(C) and is displayed as relative values ±sd for all pixels along the indicated trace.
Figure 4.
Figure 4.
POM2/CSI1 Fluorescent Signals Associate with Microtubule Depolymerizing Ends.(A) Images from time series (seeSupplemental Movie 1 online) of cells in 4-d-old hypocotyls expressing mCherry:TUA5 and POM2:CFP treated with 100 nM isoxaben for 2 h. Yellow arrowheads indicate POM2/CSI1 fluorescent signal associated with a depolymerizing microtubule end. Right panel displays kymographs, which show that the POM2/CSI1 signal follows the microtubule retracting end.(B) Time series (seeSupplemental Movie 2 online) of POM2/CSI1 collecting other POM2/CSI1s. Yellow arrowheads indicate collecting POM2/CSI1 and red arrowheads collected POM2/CSI1s.(C) Kymograph displaying POM2/CSI1 bidirectional migration (left), accelerating and stopping POM2/CSI1 (middle), and collecting POM2/CSI1 (right).(D) Surface plot displayed as a heat map. The mean intensity of the signals above background was measured for the right kymograph in(C) and is displayed as relative values ±sd.(E) Kymograph showing bifurcation and merge of POM2/CSI1 signal that is associated with a retracting microtubule end. When the end crosses another microtubule, the POM2/CSI1 signal bifurcates (yellow arrowheads). Merging events of two POM2/CSI1 signals happen along retracting microtubules that contain laterally associated POM2/CSI1 (red arrowheads). Bars = 5 μm(A),(C), and(E) and 1 μM in(B).
Figure 5.
Figure 5.
POM2/CSI1 Is Necessary for Coalignment of CESAs and Microtubules at the Cell Cortex.(A) Map of CESA complex delivery sites in an elongatingpom2-4 mutant cell from a 4-d-old etiolated hypocotyl. The sites were mapped on a 10-min average projection of microtubule signal. Red plus symbols indicate CESA insertion sites.(B) Single image frame showing CESA insertions (green channel; white arrowheads) on microtubules (red channel) in apom2-4 mutant cell from a 4-d-old etiolated hypocotyl. Bars = 5 μm in(A) and(B).(C) Quantification of colocalization between GFP:CESA3 and mCherry:TUA5 trajectories in 500-s time averages using Pearson correlation (n = 9 cells from 9 seedlings). Error bars indicatesd. Asterisks indicate significance (**P value < 0.01).(D) and(E) Average projections of 53 frames in elongating control(D) andpom2-4(E) cells in 4-d-old etiolated hypocotyls expressing mCherry:TUA5 and GFP:CESA3. Brackets indicate CESA migration trajectories. Bars = 5 μm.(F) Time frames of individual plasma membrane–located CESAs in the wild type(Top) andpom2-4(Bottom) in relation to cortical microtubules. CESAs (arrowheads) track along microtubules in the wild type(Top), but the tracking of CESAs inpom2-4 is independent of the microtubule orientation(Bottom). Kymographs of the observations are supplied to the right of the respective time frames. The intensity of both channels was plotted along the cyan trace.(G) and(H) Degree of linearity of moving CESAs over time. The displacement of individual CESA particles (145 particles from nine seedlings; between 45 and 50 particles per treatment and line) was traced over 8 min (100 frames). The left panel shows examples of individual traces. x and y coordinates at different time points were plotted ([G], right panel) and fitted against a linear curve. Deviations from linearity result in a lowR2 value; ideal linear traces are represented with anR2 of 1. The distribution ofR2 values (right panel) shows a significant reduction in linear CESA movement inpom2/csi1 mutants and in oryzalin-treated cells (t test: the wild type (wt) versuspom2-4 and/or oryzalin treatment; P < 0.01,pom2 versus oryzalin treatment; P > 0.5).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Adhami F., Müller S., Hauser M.-T. (1999). Nonradioactive labeling of large DNA fragments for genome walking, RFLP and northern blot analysis. Biotechniques 27: 314–320 - PubMed
    1. Alonso J.M., et al. (2003). Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301: 653–657 - PubMed
    1. Ambrose J.C., Shoji T., Kotzer A.M., Pighin J.A., Wasteneys G.O. (2007). The Arabidopsis CLASP gene encodes a microtubule-associated protein involved in cell expansion and division. Plant Cell 19: 2763–2775 - PMC - PubMed
    1. Arioli T., et al. (1998). Molecular analysis of cellulose biosynthesis in Arabidopsis. Science 279: 717–720 - PubMed
    1. Azarov A.S., Tokarev B.I., Netchepurenko A.E. (1990). Effect of sodium chloride on pollen germination and pollen tube growth in vitro in Arabidopsis thaliana (L.) Heynh. Arabidopsis Inf. Serv. 27.http://www.arabidopsis.org/ais/1990/azaro-1990-aacdq.html

Publication types

MeSH terms

Substances

Related information

Grants and funding

LinkOut - more resources

Full text links
Silverchair Information Systems full text link Silverchair Information Systems Free PMC article
Cite
Send To

NCBI Literature Resources

MeSHPMCBookshelfDisclaimer

The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Unauthorized use of these marks is strictly prohibited.

[8]ページ先頭
©2009-2025 Movatter.jp