Limits...
Cloning of Gossypium hirsutum sucrose non-fermenting 1-related protein kinase 2 gene (GhSnRK2) and its overexpression in transgenic Arabidopsis escalates drought and low temperature tolerance.

Bello B, Zhang X, Liu C, Yang Z, Yang Z, Wang Q, Zhao G, Li F - PLoS ONE (2014)

Bottom Line: Under normal and stress conditions, the expression levels of stress-inducible genes, such as AtRD29A, AtRD29B, AtP5CS1, AtABI3, AtCBF1, and AtABI5, were increased in the GhSnRK2-overexpressing plants compared to the wild-type plants.GhSnRK2 gene silencing alleviated drought tolerance in cotton plants, indicating that VIGS technique can certainly be used as an effective means to examine gene function by knocking down the expression of distinctly expressed genes.The results of this study suggested that the GhSnRK2 gene, when incorporated into Arabidopsis, functions in positive responses to drought stress and in low temperature tolerance.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Cotton Biology, Cotton Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.

ABSTRACT
The molecular mechanisms of stress tolerance and the use of modern genetics approaches for the improvement of drought stress tolerance have been major focuses of plant molecular biologists. In the present study, we cloned the Gossypium hirsutum sucrose non-fermenting 1-related protein kinase 2 (GhSnRK2) gene and investigated its functions in transgenic Arabidopsis. We further elucidated the function of this gene in transgenic cotton using virus-induced gene silencing (VIGS) techniques. We hypothesized that GhSnRK2 participates in the stress signaling pathway and elucidated its role in enhancing stress tolerance in plants via various stress-related pathways and stress-responsive genes. We determined that the subcellular localization of the GhSnRK2-green fluorescent protein (GFP) was localized in the nuclei and cytoplasm. In contrast to wild-type plants, transgenic plants overexpressing GhSnRK2 exhibited increased tolerance to drought, cold, abscisic acid and salt stresses, suggesting that GhSnRK2 acts as a positive regulator in response to cold and drought stresses. Plants overexpressing GhSnRK2 displayed evidence of reduced water loss, turgor regulation, elevated relative water content, biomass, and proline accumulation. qRT-PCR analysis of GhSnRK2 expression suggested that this gene may function in diverse tissues. Under normal and stress conditions, the expression levels of stress-inducible genes, such as AtRD29A, AtRD29B, AtP5CS1, AtABI3, AtCBF1, and AtABI5, were increased in the GhSnRK2-overexpressing plants compared to the wild-type plants. GhSnRK2 gene silencing alleviated drought tolerance in cotton plants, indicating that VIGS technique can certainly be used as an effective means to examine gene function by knocking down the expression of distinctly expressed genes. The results of this study suggested that the GhSnRK2 gene, when incorporated into Arabidopsis, functions in positive responses to drought stress and in low temperature tolerance.

Show MeSH

Related in: MedlinePlus

Seedling growth of the WT and GhSnRK2-overexpressing lines in response to NaCl and ABA treatment.(A) WT and GhSnRK2 transgenic plants subjected to different concentrations of NaCl. (B) Approximately one-week-old seedlings were transferred to 1/2MS medium supplemented with different concentrations of NaCl; the root length was measured after 7 days. Each of the three biological replicates consisted of 16 plants. The mean values were compared using Student's T-test (p<0.05). (C) The root growth of the WT and GhSnRK2 transgenic plants treated with different concentrations of ABA. (D) The plants were cultured vertically on MS-containing medium treated with different concentrations of ABA (0.5 µm, 1 µm, or 2 µm); the root length was measured after 7 days. Each of the three biological replicates consisted of 16 plants. Student's T-test was used to compare the mean values of the WT and transgenic lines. Asterisk denotes a significant difference (P<0.05).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4231032&req=5

pone-0112269-g005: Seedling growth of the WT and GhSnRK2-overexpressing lines in response to NaCl and ABA treatment.(A) WT and GhSnRK2 transgenic plants subjected to different concentrations of NaCl. (B) Approximately one-week-old seedlings were transferred to 1/2MS medium supplemented with different concentrations of NaCl; the root length was measured after 7 days. Each of the three biological replicates consisted of 16 plants. The mean values were compared using Student's T-test (p<0.05). (C) The root growth of the WT and GhSnRK2 transgenic plants treated with different concentrations of ABA. (D) The plants were cultured vertically on MS-containing medium treated with different concentrations of ABA (0.5 µm, 1 µm, or 2 µm); the root length was measured after 7 days. Each of the three biological replicates consisted of 16 plants. Student's T-test was used to compare the mean values of the WT and transgenic lines. Asterisk denotes a significant difference (P<0.05).

Mentions: To investigate the effect of GhSnRK2 overexpression in response to NaCl and exogenous ABA treatment, a root growth experiment was conducted to elucidate the physiological differences between the transgenic and WT plants. The transgenic plants overexpressing GhSnRK2 exhibited enhanced seedling growth under NaCl and ABA stress treatments, suggesting that GhSnRK2 may be involved in the oxidative stress response pathway. Seedlings of WT and GhSnRK2 transgenic lines grew normally in 0 mM NaCl. In 100 mM and 150 mMNaCl, the transgenic seedlings formed longer roots and displayed significantly larger growth than the WT seedlings. When the NaCl concentration was increased to 200 mm, the growth of the WT seedlings was completely inhibited, and the color of these seedlings was found to have turned brownish, whereas the transgenic seedlings remained green and continued to grow, although at a slower rate (n = 3) (Figure 5A, B).


Cloning of Gossypium hirsutum sucrose non-fermenting 1-related protein kinase 2 gene (GhSnRK2) and its overexpression in transgenic Arabidopsis escalates drought and low temperature tolerance.

Bello B, Zhang X, Liu C, Yang Z, Yang Z, Wang Q, Zhao G, Li F - PLoS ONE (2014)

Seedling growth of the WT and GhSnRK2-overexpressing lines in response to NaCl and ABA treatment.(A) WT and GhSnRK2 transgenic plants subjected to different concentrations of NaCl. (B) Approximately one-week-old seedlings were transferred to 1/2MS medium supplemented with different concentrations of NaCl; the root length was measured after 7 days. Each of the three biological replicates consisted of 16 plants. The mean values were compared using Student's T-test (p<0.05). (C) The root growth of the WT and GhSnRK2 transgenic plants treated with different concentrations of ABA. (D) The plants were cultured vertically on MS-containing medium treated with different concentrations of ABA (0.5 µm, 1 µm, or 2 µm); the root length was measured after 7 days. Each of the three biological replicates consisted of 16 plants. Student's T-test was used to compare the mean values of the WT and transgenic lines. Asterisk denotes a significant difference (P<0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4231032&req=5

pone-0112269-g005: Seedling growth of the WT and GhSnRK2-overexpressing lines in response to NaCl and ABA treatment.(A) WT and GhSnRK2 transgenic plants subjected to different concentrations of NaCl. (B) Approximately one-week-old seedlings were transferred to 1/2MS medium supplemented with different concentrations of NaCl; the root length was measured after 7 days. Each of the three biological replicates consisted of 16 plants. The mean values were compared using Student's T-test (p<0.05). (C) The root growth of the WT and GhSnRK2 transgenic plants treated with different concentrations of ABA. (D) The plants were cultured vertically on MS-containing medium treated with different concentrations of ABA (0.5 µm, 1 µm, or 2 µm); the root length was measured after 7 days. Each of the three biological replicates consisted of 16 plants. Student's T-test was used to compare the mean values of the WT and transgenic lines. Asterisk denotes a significant difference (P<0.05).
Mentions: To investigate the effect of GhSnRK2 overexpression in response to NaCl and exogenous ABA treatment, a root growth experiment was conducted to elucidate the physiological differences between the transgenic and WT plants. The transgenic plants overexpressing GhSnRK2 exhibited enhanced seedling growth under NaCl and ABA stress treatments, suggesting that GhSnRK2 may be involved in the oxidative stress response pathway. Seedlings of WT and GhSnRK2 transgenic lines grew normally in 0 mM NaCl. In 100 mM and 150 mMNaCl, the transgenic seedlings formed longer roots and displayed significantly larger growth than the WT seedlings. When the NaCl concentration was increased to 200 mm, the growth of the WT seedlings was completely inhibited, and the color of these seedlings was found to have turned brownish, whereas the transgenic seedlings remained green and continued to grow, although at a slower rate (n = 3) (Figure 5A, B).

Bottom Line: Under normal and stress conditions, the expression levels of stress-inducible genes, such as AtRD29A, AtRD29B, AtP5CS1, AtABI3, AtCBF1, and AtABI5, were increased in the GhSnRK2-overexpressing plants compared to the wild-type plants.GhSnRK2 gene silencing alleviated drought tolerance in cotton plants, indicating that VIGS technique can certainly be used as an effective means to examine gene function by knocking down the expression of distinctly expressed genes.The results of this study suggested that the GhSnRK2 gene, when incorporated into Arabidopsis, functions in positive responses to drought stress and in low temperature tolerance.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Cotton Biology, Cotton Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.

ABSTRACT
The molecular mechanisms of stress tolerance and the use of modern genetics approaches for the improvement of drought stress tolerance have been major focuses of plant molecular biologists. In the present study, we cloned the Gossypium hirsutum sucrose non-fermenting 1-related protein kinase 2 (GhSnRK2) gene and investigated its functions in transgenic Arabidopsis. We further elucidated the function of this gene in transgenic cotton using virus-induced gene silencing (VIGS) techniques. We hypothesized that GhSnRK2 participates in the stress signaling pathway and elucidated its role in enhancing stress tolerance in plants via various stress-related pathways and stress-responsive genes. We determined that the subcellular localization of the GhSnRK2-green fluorescent protein (GFP) was localized in the nuclei and cytoplasm. In contrast to wild-type plants, transgenic plants overexpressing GhSnRK2 exhibited increased tolerance to drought, cold, abscisic acid and salt stresses, suggesting that GhSnRK2 acts as a positive regulator in response to cold and drought stresses. Plants overexpressing GhSnRK2 displayed evidence of reduced water loss, turgor regulation, elevated relative water content, biomass, and proline accumulation. qRT-PCR analysis of GhSnRK2 expression suggested that this gene may function in diverse tissues. Under normal and stress conditions, the expression levels of stress-inducible genes, such as AtRD29A, AtRD29B, AtP5CS1, AtABI3, AtCBF1, and AtABI5, were increased in the GhSnRK2-overexpressing plants compared to the wild-type plants. GhSnRK2 gene silencing alleviated drought tolerance in cotton plants, indicating that VIGS technique can certainly be used as an effective means to examine gene function by knocking down the expression of distinctly expressed genes. The results of this study suggested that the GhSnRK2 gene, when incorporated into Arabidopsis, functions in positive responses to drought stress and in low temperature tolerance.

Show MeSH
Related in: MedlinePlus