Limits...
The maize AAA-type protein SKD1 confers enhanced salt and drought stress tolerance in transgenic tobacco by interacting with Lyst-interacting protein 5.

Xia Z, Wei Y, Sun K, Wu J, Wang Y, Wu K - PLoS ONE (2013)

Bottom Line: ATPase associated with various cellular activities (AAA) proteins are important regulators involved in diverse cellular functions.Collectively, these data demonstrate that ZmSKD1could be involved in salt and drought stress responses and its over-expression enhances salt or drought stress tolerance possibly through interacting with LIP5 in tobacco.This study may facilitate our understandings of the biological roles of SKD1-mediated ESCRT pathway under stress conditions in higher plants and accelerate genetic improvement of crop plants tolerant to environmental stresses.

View Article: PubMed Central - PubMed

Affiliation: College of Life Science, Henan Agricultural University, Zhengzhou, PR China. xiazl08@163.com

ABSTRACT
ATPase associated with various cellular activities (AAA) proteins are important regulators involved in diverse cellular functions. To date, the molecular mechanisms of AAA proteins involved in response to salt and drought stresses in plants are largely unknown. In this study, a putative SKD1 (suppressor of K(+) transport growth defect 1) ortholog from Zea mays (ZmSKD1), which encodes a putative AAA protein, was isolated. The transcript levels of ZmSKD1 were higher in aerial tissues and were markedly up-regulated by salt or drought stress. Over-expression of ZmSKD1 in tobacco plants enhanced their tolerances not only to salt but to drought. Moreover, reactive oxygen species accumulations in ZmSKD1 transgenic lines were relative less than those in wild-type plants during salt or PEG-induced water stress. The interaction between ZmSKD1 and NtLIP5 (Lyst-Interacting Protein 5 homolog from Nicotiana tabacum) was confirmed by both yeast two-hybrid and immuno-precipitation assays; moreover, the α-helix-rich domain in the C-terminus of ZmSKD1 was identified to be required for its interaction with NtLIP5 using truncation mutations. Collectively, these data demonstrate that ZmSKD1could be involved in salt and drought stress responses and its over-expression enhances salt or drought stress tolerance possibly through interacting with LIP5 in tobacco. This study may facilitate our understandings of the biological roles of SKD1-mediated ESCRT pathway under stress conditions in higher plants and accelerate genetic improvement of crop plants tolerant to environmental stresses.

Show MeSH

Related in: MedlinePlus

Dynamic changes of ROS in wild-type and ZmSKD1-overexpressing tobacco plants in response to salt or PEG-induced water stress.A H2O2 production in leaves of wild-type and OE (OE-7) plants treated with distilled water(control), 200 mM NaCl or 15% PEG-6000 solutions, respectively, was visualized by staining with 3, 3′-diaminobenzidine (DAB). Plants were treated for 1, 2, 6, and 12 h, and were subsequently stained with DAB as described in the experimental procedures. B Relative H2O2 levels were quantified in leaves of wild-type and OE (OE-7) plants exposed to 200 mM NaCl or 15% PEG-6000 for 1, 2, 6, and 12 h. Error bars indicate SE (n = 6). **t-test, with P<0.01; *t-test, with P<0.05. C O2− production in leaves of wild-type and OE (OE-7) plants treated with 200 mM NaCl or 15% PEG-6000 solutions, respectively, was visualized by staining with nitroblue tetrazolium (NBT). Plants were treated for 1, 2, 6, and 12 h, and were subsequently stained with NBT as described in the experimental procedures. D Relative O2− levels were quantified in leaves of wild-type and OE (OE-7) plants exposed to 200 mM NaCl or 15% PEG-6000 for 1, 2, 6, and 12 h. Error bars indicate SE (n = 6). **t-test, with P<0.01; *t-test, with P<0.05.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3722157&req=5

pone-0069787-g005: Dynamic changes of ROS in wild-type and ZmSKD1-overexpressing tobacco plants in response to salt or PEG-induced water stress.A H2O2 production in leaves of wild-type and OE (OE-7) plants treated with distilled water(control), 200 mM NaCl or 15% PEG-6000 solutions, respectively, was visualized by staining with 3, 3′-diaminobenzidine (DAB). Plants were treated for 1, 2, 6, and 12 h, and were subsequently stained with DAB as described in the experimental procedures. B Relative H2O2 levels were quantified in leaves of wild-type and OE (OE-7) plants exposed to 200 mM NaCl or 15% PEG-6000 for 1, 2, 6, and 12 h. Error bars indicate SE (n = 6). **t-test, with P<0.01; *t-test, with P<0.05. C O2− production in leaves of wild-type and OE (OE-7) plants treated with 200 mM NaCl or 15% PEG-6000 solutions, respectively, was visualized by staining with nitroblue tetrazolium (NBT). Plants were treated for 1, 2, 6, and 12 h, and were subsequently stained with NBT as described in the experimental procedures. D Relative O2− levels were quantified in leaves of wild-type and OE (OE-7) plants exposed to 200 mM NaCl or 15% PEG-6000 for 1, 2, 6, and 12 h. Error bars indicate SE (n = 6). **t-test, with P<0.01; *t-test, with P<0.05.

Mentions: Drought or salt stress may induce production of reactive oxygen species (ROS), which cause oxidative damage to plant cells, or act as signaling molecules for regulating development and various physiological responses [15]. This promoted us to examine cellular levels of two prominent ROS (O2− and H2O2) accumulations in OE and WT plants during salt or PEG-induced water stress. Histochemical detection of H2O2 accumulation was performed with DAB staining in NaCl or PEG-treated leaves along with controls from OE and WT plants within 12 h. As shown in Fig. 5A, both OE and WT leaves showed more DAB staining at 1 h of NaCl or PEG treatment; after 2 h, marked difference in DAB staining intensity was observed between OE and WT leaves. WT showed more DAB staining at 2 h of salt or drought stress, and remained higher levels till 12 h, whereas OE showed less staining after 2 h; moreover, the contrasting staining was much clear between the OE and WT leaves at 6 h, and till the end of the stresses (Fig. 5A). No significant differences in DAB staining intensity were observed between WT and OE lines within 12 h of distilled water-treated controls (Fig. 5A). The fluctuation in H2O2 level, especially in wild-type plants treated with PEG and stained with DAB was also noted (Fig. 5A). Quantitative determination of H2O2 accumulation at 1, 2, 6 and 12 h of salt or drought stress further demonstrated that compared to their corresponding controls, WT increased significantly in DAB staining intensity at 2, 6, or 12 h of both stresses, while the OE plants had no significant changes (Fig. 5B).


The maize AAA-type protein SKD1 confers enhanced salt and drought stress tolerance in transgenic tobacco by interacting with Lyst-interacting protein 5.

Xia Z, Wei Y, Sun K, Wu J, Wang Y, Wu K - PLoS ONE (2013)

Dynamic changes of ROS in wild-type and ZmSKD1-overexpressing tobacco plants in response to salt or PEG-induced water stress.A H2O2 production in leaves of wild-type and OE (OE-7) plants treated with distilled water(control), 200 mM NaCl or 15% PEG-6000 solutions, respectively, was visualized by staining with 3, 3′-diaminobenzidine (DAB). Plants were treated for 1, 2, 6, and 12 h, and were subsequently stained with DAB as described in the experimental procedures. B Relative H2O2 levels were quantified in leaves of wild-type and OE (OE-7) plants exposed to 200 mM NaCl or 15% PEG-6000 for 1, 2, 6, and 12 h. Error bars indicate SE (n = 6). **t-test, with P<0.01; *t-test, with P<0.05. C O2− production in leaves of wild-type and OE (OE-7) plants treated with 200 mM NaCl or 15% PEG-6000 solutions, respectively, was visualized by staining with nitroblue tetrazolium (NBT). Plants were treated for 1, 2, 6, and 12 h, and were subsequently stained with NBT as described in the experimental procedures. D Relative O2− levels were quantified in leaves of wild-type and OE (OE-7) plants exposed to 200 mM NaCl or 15% PEG-6000 for 1, 2, 6, and 12 h. Error bars indicate SE (n = 6). **t-test, with P<0.01; *t-test, with P<0.05.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3722157&req=5

pone-0069787-g005: Dynamic changes of ROS in wild-type and ZmSKD1-overexpressing tobacco plants in response to salt or PEG-induced water stress.A H2O2 production in leaves of wild-type and OE (OE-7) plants treated with distilled water(control), 200 mM NaCl or 15% PEG-6000 solutions, respectively, was visualized by staining with 3, 3′-diaminobenzidine (DAB). Plants were treated for 1, 2, 6, and 12 h, and were subsequently stained with DAB as described in the experimental procedures. B Relative H2O2 levels were quantified in leaves of wild-type and OE (OE-7) plants exposed to 200 mM NaCl or 15% PEG-6000 for 1, 2, 6, and 12 h. Error bars indicate SE (n = 6). **t-test, with P<0.01; *t-test, with P<0.05. C O2− production in leaves of wild-type and OE (OE-7) plants treated with 200 mM NaCl or 15% PEG-6000 solutions, respectively, was visualized by staining with nitroblue tetrazolium (NBT). Plants were treated for 1, 2, 6, and 12 h, and were subsequently stained with NBT as described in the experimental procedures. D Relative O2− levels were quantified in leaves of wild-type and OE (OE-7) plants exposed to 200 mM NaCl or 15% PEG-6000 for 1, 2, 6, and 12 h. Error bars indicate SE (n = 6). **t-test, with P<0.01; *t-test, with P<0.05.
Mentions: Drought or salt stress may induce production of reactive oxygen species (ROS), which cause oxidative damage to plant cells, or act as signaling molecules for regulating development and various physiological responses [15]. This promoted us to examine cellular levels of two prominent ROS (O2− and H2O2) accumulations in OE and WT plants during salt or PEG-induced water stress. Histochemical detection of H2O2 accumulation was performed with DAB staining in NaCl or PEG-treated leaves along with controls from OE and WT plants within 12 h. As shown in Fig. 5A, both OE and WT leaves showed more DAB staining at 1 h of NaCl or PEG treatment; after 2 h, marked difference in DAB staining intensity was observed between OE and WT leaves. WT showed more DAB staining at 2 h of salt or drought stress, and remained higher levels till 12 h, whereas OE showed less staining after 2 h; moreover, the contrasting staining was much clear between the OE and WT leaves at 6 h, and till the end of the stresses (Fig. 5A). No significant differences in DAB staining intensity were observed between WT and OE lines within 12 h of distilled water-treated controls (Fig. 5A). The fluctuation in H2O2 level, especially in wild-type plants treated with PEG and stained with DAB was also noted (Fig. 5A). Quantitative determination of H2O2 accumulation at 1, 2, 6 and 12 h of salt or drought stress further demonstrated that compared to their corresponding controls, WT increased significantly in DAB staining intensity at 2, 6, or 12 h of both stresses, while the OE plants had no significant changes (Fig. 5B).

Bottom Line: ATPase associated with various cellular activities (AAA) proteins are important regulators involved in diverse cellular functions.Collectively, these data demonstrate that ZmSKD1could be involved in salt and drought stress responses and its over-expression enhances salt or drought stress tolerance possibly through interacting with LIP5 in tobacco.This study may facilitate our understandings of the biological roles of SKD1-mediated ESCRT pathway under stress conditions in higher plants and accelerate genetic improvement of crop plants tolerant to environmental stresses.

View Article: PubMed Central - PubMed

Affiliation: College of Life Science, Henan Agricultural University, Zhengzhou, PR China. xiazl08@163.com

ABSTRACT
ATPase associated with various cellular activities (AAA) proteins are important regulators involved in diverse cellular functions. To date, the molecular mechanisms of AAA proteins involved in response to salt and drought stresses in plants are largely unknown. In this study, a putative SKD1 (suppressor of K(+) transport growth defect 1) ortholog from Zea mays (ZmSKD1), which encodes a putative AAA protein, was isolated. The transcript levels of ZmSKD1 were higher in aerial tissues and were markedly up-regulated by salt or drought stress. Over-expression of ZmSKD1 in tobacco plants enhanced their tolerances not only to salt but to drought. Moreover, reactive oxygen species accumulations in ZmSKD1 transgenic lines were relative less than those in wild-type plants during salt or PEG-induced water stress. The interaction between ZmSKD1 and NtLIP5 (Lyst-Interacting Protein 5 homolog from Nicotiana tabacum) was confirmed by both yeast two-hybrid and immuno-precipitation assays; moreover, the α-helix-rich domain in the C-terminus of ZmSKD1 was identified to be required for its interaction with NtLIP5 using truncation mutations. Collectively, these data demonstrate that ZmSKD1could be involved in salt and drought stress responses and its over-expression enhances salt or drought stress tolerance possibly through interacting with LIP5 in tobacco. This study may facilitate our understandings of the biological roles of SKD1-mediated ESCRT pathway under stress conditions in higher plants and accelerate genetic improvement of crop plants tolerant to environmental stresses.

Show MeSH
Related in: MedlinePlus