Characterization of Actin Filament Dynamics during Mitosis in Wheat Protoplasts under UV-B Radiation

Chen, Huize; Han, Rong

doi:10.1038/srep20115

Cited by 14 publications

(11 citation statements)

References 35 publications

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“…One of the many applications of the protoplasts technology is to establish a transient expression system which can be used to study high-throughput analysis and functional characterization of genes. To establish, efficient expression system through protoplasts there is requirement of high-quality protoplasts [24].…”

Section: Discussionmentioning

confidence: 99%

Optimization of Isolation and Culture of Protoplasts in Alfalfa (<i>Medicago sativa</i>) Cultivar Regen-SY

Sangra¹,

Shahin²,

Dhir³

2019

AJPS

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Alfalfa (Medicago sativa) is an important forage crop belonging to the Fabaceae family. It is cultivated across the world for fodder and originated in Asia. Alfalfa cultivar Regen-SY was used in this study which is a hybrid of first-generation self-parents from Regen-S (M. sativa) and Regen-Y (Medicago falcata) research cultivars. The main objective of the study was to optimize conditions for the isolation and liquid culture of alfalfa Regen-SY protoplasts. Several factors like enzyme combination, incubation time, plant age, centrifugation speed and shaker speed affecting protoplast isolation and culture were optimized in the study. The yield and viability of the protoplasts was determined by using hemocytometer and Fluorescein diacetate (FDA) staining respectively. Results showed that factors like enzyme combination, incubation time, plant age, centrifugation speed and Mannitol concentration significantly (p ≤ 0.05) affect protoplast yield and viability whereas shaker speed didn't result in any significant difference in the yield and viability of protoplasts. Using optimum conditions protoplasts were cultured in the liquid medium and microcalli formation was achieved after five weeks of the culture. The protocol established in this study will assist researchers in the isolation and culture of protoplasts in alfalfa and will accelerate the research processes like protoplast fusion and genetic engineering.

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Section: Discussionmentioning

confidence: 99%

Optimization of Isolation and Culture of Protoplasts in Alfalfa (<i>Medicago sativa</i>) Cultivar Regen-SY

Sangra¹,

Shahin²,

Dhir³

2019

AJPS

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“…Previous results showed that UV-B promotes actin filament bundle formation in wheat protoplasts and induces abnormal chromosome movement [ 26 ]. Recently, it was reported that light was channeled through the stem to the roots [ 27 ], and UV-B signals could be transmitted to the roots through plant endodermis tissues [ 8 ].…”

Section: Introductionmentioning

confidence: 99%

Actin filaments mediated root growth inhibition by changing their distribution under UV-B and hydrogen peroxide exposure in Arabidopsis

Wang²,

Chen

et al. 2020

Biol Res

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Background UV-B signaling in plants is mediated by UVR8, which interacts with transcriptional factors to induce root morphogenesis. However, research on the downstream molecules of UVR8 signaling in roots is still scarce. As a wide range of functional cytoskeletons, how actin filaments respond to UV-B-induced root morphogenesis has not been reported. The aim of this study was to investigate the effect of actin filaments on root morphogenesis under UV-B and hydrogen peroxide exposure in Arabidopsis. Results A Lifeact-Venus fusion protein was used to stain actin filaments in Arabidopsis. The results showed that UV-B inhibited hypocotyl and root elongation and caused an increase in H2O2 content only in the root but not in the hypocotyl. Additionally, the actin filaments in hypocotyls diffused under UV-B exposure but were gathered in a bundle under the control conditions in either Lifeact-Venus or uvr8 plants. Exogenous H2O2 inhibited root elongation in a dose-dependent manner. The actin filaments changed their distribution from filamentous to punctate in the root tips and mature regions at a lower concentration of H2O2 but aggregated into thick bundles with an abnormal orientation at H2O2 concentrations up to 2 mM. In the root elongation zone, the actin filament arrangement changed from lateral to longitudinal after exposure to H2O2. Actin filaments in the root tip and elongation zone were depolymerized into puncta under UV-B exposure, which showed the same tendency as the low-concentration treatments. The actin filaments were hardly filamentous in the maturation zone. The dynamics of actin filaments in the uvr8 group under UV-B exposure were close to those of the control group. Conclusions The results indicate that UV-B inhibited Arabidopsis hypocotyl elongation by reorganizing actin filaments from bundles to a loose arrangement, which was not related to H2O2. UV-B disrupted the dynamics of actin filaments by changing the H2O2 level in Arabidopsis roots. All these results provide an experimental basis for investigating the interaction of UV-B signaling with the cytoskeleton.

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“…In wheat protoplasts, UV-B irradiation disturbs the actin cytoskeleton, which forms foci at different stages of the cell cycle. Those effects are accompanied by the formation of apoptotic bodies (Chen and Han, 2016). UV-B-induced programmed cell death has also been reported for BY2 cells (Lytvyn et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

UV-B Induces Chloroplast Movements in a Phototropin-Dependent Manner

et al. 2019

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We examined the impact of UV-B irradiation on chloroplast movements in Arabidopsis leaves. Directional chloroplast movements induced by blue light have been described in multiple plant species. In weak light, chloroplasts accumulate at periclinal cell walls to increase light capture. In strong light, chloroplasts exhibit the avoidance response, as they move towards anticlinal walls to protect the photosynthetic apparatus from light-induced damage. In Arabidopsis, chloroplast movements are triggered by phototropins, phot1 and phot2, which are known as blue/UV-A photoreceptors. We found that irradiation with UV-B of 3.3 µmol·m−2·s−1 induced chloroplast accumulation in wild-type plants. UV-B-triggered accumulation was dependent on the presence of phototropins, especially phot1, but not on UVR8 (the canonical UV-B photoreceptor). Irradiation with strong UV-B of 20 µmol·m−2·s−1 did not induce substantial chloroplast relocations in wild-type leaves. However, in the jac1 mutant, which is defective in chloroplast accumulation, strong UV-B elicited chloroplast avoidance. This indicated that UV-B can also activate signaling to the avoidance response. To assess the possibility of indirect effects of UV-B on chloroplast movements, we examined the impact of UV-B on the actin cytoskeleton, which serves as the motile system for chloroplast movements. While irradiation with UV-B of 3.3 µmol·m−2·s−1 did not affect the actin cytoskeleton, strong UV-B disrupted its structure as shown using an Arabidopsis line expressing Lifeact-green fluorescent protein (GFP). In wild-type plants, pretreatment with strong UV-B attenuated chloroplast responses triggered by subsequent blue light irradiation, further indicating that this UV-B intensity also indirectly affects chloroplast movements. Taken together, our results suggest that the effect of UV-B on chloroplast movement is twofold: it directly induces phototropin-mediated movements; however, at higher intensities, it attenuates the movements in a nonspecific manner.

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Characterization of Actin Filament Dynamics during Mitosis in Wheat Protoplasts under UV-B Radiation

Cited by 14 publications

References 35 publications

Optimization of Isolation and Culture of Protoplasts in Alfalfa (<i>Medicago sativa</i>) Cultivar Regen-SY

Optimization of Isolation and Culture of Protoplasts in Alfalfa (<i>Medicago sativa</i>) Cultivar Regen-SY

Actin filaments mediated root growth inhibition by changing their distribution under UV-B and hydrogen peroxide exposure in Arabidopsis

UV-B Induces Chloroplast Movements in a Phototropin-Dependent Manner

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Characterization of Actin Filament Dynamics during Mitosis in Wheat Protoplasts under UV-B Radiation

Cited by 14 publications

References 35 publications

Optimization of Isolation and Culture of Protoplasts in Alfalfa (&lt;i&gt;Medicago sativa&lt;/i&gt;) Cultivar Regen-SY

Optimization of Isolation and Culture of Protoplasts in Alfalfa (&lt;i&gt;Medicago sativa&lt;/i&gt;) Cultivar Regen-SY

Actin filaments mediated root growth inhibition by changing their distribution under UV-B and hydrogen peroxide exposure in Arabidopsis

UV-B Induces Chloroplast Movements in a Phototropin-Dependent Manner

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Optimization of Isolation and Culture of Protoplasts in Alfalfa (<i>Medicago sativa</i>) Cultivar Regen-SY

Optimization of Isolation and Culture of Protoplasts in Alfalfa (<i>Medicago sativa</i>) Cultivar Regen-SY