The plant growth process is accompanied by dynamic changes in the microtubules,and the rearrangement of microtubules is regulated by diverse microtubule-associated proteins(MAPs).Plant MAP65s have been exhaustively ch...The plant growth process is accompanied by dynamic changes in the microtubules,and the rearrangement of microtubules is regulated by diverse microtubule-associated proteins(MAPs).Plant MAP65s have been exhaustively characterized in some species,but the information about MAP65 family members in tomato(Solanum lycopersicum)is limited.In this study,nine SlMAP65 family genes were identified in the tomato genome.Then a systematic analysis that considered the physio-chemical properties,evolution,conserved motifs,domains,gene structure,and cis-regulatory elements of SlMAP65 family members was conducted.The family member Sl MAP65-1,which had the highest expression,was knocked out by CRISPR/Cas9.The tomato fruit of slmap65-1 loss of function lines showed an elongated morphology,and the data indicated that SlMAP65-1 is involved in fruit morphogenesis at the early fruit development stage.These results provide new insights for fruit morphogenesis-related research and future functional studies of the SlMAP65 family members in tomato.展开更多
[Objective] This study aimed to test the characteristics of MAP65 in wheat mesophyll cells by co-sedimentation experiments. [Method] We used wheat Linyou 2018 as the experimental material. Western blotting analysis wa...[Objective] This study aimed to test the characteristics of MAP65 in wheat mesophyll cells by co-sedimentation experiments. [Method] We used wheat Linyou 2018 as the experimental material. Western blotting analysis was carried out to de- tect the existing of 65 kDa MAP in the wheat mesophyll cells; spectrophotometery and SDS-PAGE were adopted to analyze the role of MAP65 in microtubules poly- merization in vitro. [Result] MAP65 indeed existed in wheat mesophyll cells. Co-sedi- mentation experiments showed that MAP65 can bind to microtubules, with the basic characteristics of the microtubule-associated proteins. Turbidimetric experiments showed that microtubule polymerization depended on the concentration of MAP65. Low concentrations of MAP65 promoted microtubule polymerization while high con- centrations of MAP65 inhibited microtubule polymerization. [Conclusion] This study laid significant basis for further research on physiological function of MAP65 protein.展开更多
Cytokinesis, the partitioning of the cytoplasm following nuclear division, requires extensive coordination between membrane trafficking and cytoskeletal dynamics. In plants, the onset of cytokinesis is characterized b...Cytokinesis, the partitioning of the cytoplasm following nuclear division, requires extensive coordination between membrane trafficking and cytoskeletal dynamics. In plants, the onset of cytokinesis is characterized by the assembly of a bipolar microtubule array, the phragmoplast, and of a transient membrane compartment, the cell plate. Little is known about the coordination between membrane deposition at the cell plate and the dynamics of phragmoplast microtubules. In this study, we monitor the localization dynamics of microtubule and membrane markers throughout cytokinesis. Our spatiotemporal resolution is consistent with the general view that microtubule dynamics drive membrane movements. Nonetheless, we provide evidence for active sorting at the cell plate and show that this is, at least in part, mediated by the TRAPPII tethering complex. We also characterize phragmoplast microtubule organization and cell plate formation in a suite of cytokinesis-defective mutants. Of four mutant lines with defects in phragmoplast microtubule organization, only morl microtubule-associated mutants exhibited aberrant cell plates. Conversely, the mutants with the strongest impairment in phragmoplast microtubule reorganization are keule alleles, which have a primary defect in membrane fusion. Our findings identify the SEC1/Munc18 protein KEULE as a central regulatory node in the coordination of membrane and microtubule dynamics during plant cytokinesis.展开更多
基金supported by the National Natural Sciences Foundation of China(32302623 and 32172639)。
文摘The plant growth process is accompanied by dynamic changes in the microtubules,and the rearrangement of microtubules is regulated by diverse microtubule-associated proteins(MAPs).Plant MAP65s have been exhaustively characterized in some species,but the information about MAP65 family members in tomato(Solanum lycopersicum)is limited.In this study,nine SlMAP65 family genes were identified in the tomato genome.Then a systematic analysis that considered the physio-chemical properties,evolution,conserved motifs,domains,gene structure,and cis-regulatory elements of SlMAP65 family members was conducted.The family member Sl MAP65-1,which had the highest expression,was knocked out by CRISPR/Cas9.The tomato fruit of slmap65-1 loss of function lines showed an elongated morphology,and the data indicated that SlMAP65-1 is involved in fruit morphogenesis at the early fruit development stage.These results provide new insights for fruit morphogenesis-related research and future functional studies of the SlMAP65 family members in tomato.
基金Supported by the National Science Foundation of China (30671061)the Natural Science of Shanxi Province (2008011059-1)~~
文摘[Objective] This study aimed to test the characteristics of MAP65 in wheat mesophyll cells by co-sedimentation experiments. [Method] We used wheat Linyou 2018 as the experimental material. Western blotting analysis was carried out to de- tect the existing of 65 kDa MAP in the wheat mesophyll cells; spectrophotometery and SDS-PAGE were adopted to analyze the role of MAP65 in microtubules poly- merization in vitro. [Result] MAP65 indeed existed in wheat mesophyll cells. Co-sedi- mentation experiments showed that MAP65 can bind to microtubules, with the basic characteristics of the microtubule-associated proteins. Turbidimetric experiments showed that microtubule polymerization depended on the concentration of MAP65. Low concentrations of MAP65 promoted microtubule polymerization while high con- centrations of MAP65 inhibited microtubule polymerization. [Conclusion] This study laid significant basis for further research on physiological function of MAP65 protein.
文摘Cytokinesis, the partitioning of the cytoplasm following nuclear division, requires extensive coordination between membrane trafficking and cytoskeletal dynamics. In plants, the onset of cytokinesis is characterized by the assembly of a bipolar microtubule array, the phragmoplast, and of a transient membrane compartment, the cell plate. Little is known about the coordination between membrane deposition at the cell plate and the dynamics of phragmoplast microtubules. In this study, we monitor the localization dynamics of microtubule and membrane markers throughout cytokinesis. Our spatiotemporal resolution is consistent with the general view that microtubule dynamics drive membrane movements. Nonetheless, we provide evidence for active sorting at the cell plate and show that this is, at least in part, mediated by the TRAPPII tethering complex. We also characterize phragmoplast microtubule organization and cell plate formation in a suite of cytokinesis-defective mutants. Of four mutant lines with defects in phragmoplast microtubule organization, only morl microtubule-associated mutants exhibited aberrant cell plates. Conversely, the mutants with the strongest impairment in phragmoplast microtubule reorganization are keule alleles, which have a primary defect in membrane fusion. Our findings identify the SEC1/Munc18 protein KEULE as a central regulatory node in the coordination of membrane and microtubule dynamics during plant cytokinesis.
