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SCF E3 ligase PP2-B11 plays a positive role in response to salt stress in Arabidopsis.

Jia F, Wang C, Huang J, Yang G, Wu C, Zheng C - J. Exp. Bot. (2015)

Bottom Line: Isobaric tag for relative and absolute quantification analysis revealed that 4311 differentially expressed proteins were regulated by AtPP2-B11 under salt stress.Moreover, AtPP2-B11 influenced the expression of Na(+) homeostasis genes under salt stress, and the AtPP2-B11 overexpressing lines exhibited lower Na(+) accumulation.These results suggest that AtPP2-B11 functions as a positive regulator in response to salt stress in Arabidopsis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, PR China.

No MeSH data available.


AtPP2-B11 regulates the levels of AnnAt1 under salt stress conditions. (A) Transcript level of AnnAt1 in the OE11 transgenic and wild-type plants under normal and salt stress conditions. Two-week-old seedlings of the OE11 transgenic and wild-type plants were treated with water or 200mM NaCl for 6h. Total RNA was obtained and analysed by real-time RT-PCR. (B) Protein levels of AnnAt1 in AtPP2-B11 transgenic and wild-type plants under normal and salt stress conditions. Coomassie Brilliant Blue R250 staining was used to show protein loading levels. (C, D) AnnAt1 promoter activities (C) and relative LUC fluorescence (D) after co-expressing AtPP2-B11 in the leaves of 4-week-old N. benthamiana plants compared with the control. (E) Transcription levels of ABF2/3/4 in 2-week-old AtPP2-B11 transgenic and wild-type plants under normal (CK) or salt stress (NaCl) conditions. (This figure is available in colour at JXB online.)
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Figure 6: AtPP2-B11 regulates the levels of AnnAt1 under salt stress conditions. (A) Transcript level of AnnAt1 in the OE11 transgenic and wild-type plants under normal and salt stress conditions. Two-week-old seedlings of the OE11 transgenic and wild-type plants were treated with water or 200mM NaCl for 6h. Total RNA was obtained and analysed by real-time RT-PCR. (B) Protein levels of AnnAt1 in AtPP2-B11 transgenic and wild-type plants under normal and salt stress conditions. Coomassie Brilliant Blue R250 staining was used to show protein loading levels. (C, D) AnnAt1 promoter activities (C) and relative LUC fluorescence (D) after co-expressing AtPP2-B11 in the leaves of 4-week-old N. benthamiana plants compared with the control. (E) Transcription levels of ABF2/3/4 in 2-week-old AtPP2-B11 transgenic and wild-type plants under normal (CK) or salt stress (NaCl) conditions. (This figure is available in colour at JXB online.)

Mentions: Based on the results of iTRAQ analysis, we focused on the highly upregulated and downregulated proteins. Interestingly, among the eight Arabidopsis annexins, three members in the OE11 plants were remarkably increased compared with the wild-type plants upon salt stress. As shown in Table 1, AnnAt1, AnnAt2, and AnnAt3 were upregulated with fold changes of 2.576, 2.667, and 3.593, respectively. To confirm the relationship between AtPP2-B11 and the annexins, the transcript level of AnnAt1 was detected using real-time RT-PCR analysis, since AnnAt1 plays a dominant role among eight annexins in Arabidopsis. As shown in Fig. 6A, no significant difference in the AnnAt1 mRNA level was observed between the OE11 and wild-type plants under normal conditions. After 200mM NaCl treatment for 6h, the AnnAt1 mRNA expression was more induced in OE11 than in wild-type plants (Fig. 6A). Moreover, Western blot analysis showed that AnnAt1 was significantly induced in the AtPP2-B11 overexpressing lines but depressed in the R-5 line under salt stress conditions (Fig. 6B). To further confirm that AtPP2-B11 indeed upregulated the expression of AnnAt1, we used a LUC reporter assay to examine AnnAt1 promoter activity after co-expressing AtPP2-B11 in leaves of 4-week-old N. benthamiana plants, whereas co-expressing pA::LUC and 35S::GUS served as a negative control. Consistent with the real-time RT-PCR results (Fig. 6A), AnnAt1 promoter activity was induced in N. benthamiana epidermal cells when co-expressed with AtPP2-B11 in comparison with the control (Fig. 6C, D). These results suggested that AtPP2-B11 functions in the salt stress response probably by regulating the expression of AnnAt1.


SCF E3 ligase PP2-B11 plays a positive role in response to salt stress in Arabidopsis.

Jia F, Wang C, Huang J, Yang G, Wu C, Zheng C - J. Exp. Bot. (2015)

AtPP2-B11 regulates the levels of AnnAt1 under salt stress conditions. (A) Transcript level of AnnAt1 in the OE11 transgenic and wild-type plants under normal and salt stress conditions. Two-week-old seedlings of the OE11 transgenic and wild-type plants were treated with water or 200mM NaCl for 6h. Total RNA was obtained and analysed by real-time RT-PCR. (B) Protein levels of AnnAt1 in AtPP2-B11 transgenic and wild-type plants under normal and salt stress conditions. Coomassie Brilliant Blue R250 staining was used to show protein loading levels. (C, D) AnnAt1 promoter activities (C) and relative LUC fluorescence (D) after co-expressing AtPP2-B11 in the leaves of 4-week-old N. benthamiana plants compared with the control. (E) Transcription levels of ABF2/3/4 in 2-week-old AtPP2-B11 transgenic and wild-type plants under normal (CK) or salt stress (NaCl) conditions. (This figure is available in colour at JXB online.)
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Figure 6: AtPP2-B11 regulates the levels of AnnAt1 under salt stress conditions. (A) Transcript level of AnnAt1 in the OE11 transgenic and wild-type plants under normal and salt stress conditions. Two-week-old seedlings of the OE11 transgenic and wild-type plants were treated with water or 200mM NaCl for 6h. Total RNA was obtained and analysed by real-time RT-PCR. (B) Protein levels of AnnAt1 in AtPP2-B11 transgenic and wild-type plants under normal and salt stress conditions. Coomassie Brilliant Blue R250 staining was used to show protein loading levels. (C, D) AnnAt1 promoter activities (C) and relative LUC fluorescence (D) after co-expressing AtPP2-B11 in the leaves of 4-week-old N. benthamiana plants compared with the control. (E) Transcription levels of ABF2/3/4 in 2-week-old AtPP2-B11 transgenic and wild-type plants under normal (CK) or salt stress (NaCl) conditions. (This figure is available in colour at JXB online.)
Mentions: Based on the results of iTRAQ analysis, we focused on the highly upregulated and downregulated proteins. Interestingly, among the eight Arabidopsis annexins, three members in the OE11 plants were remarkably increased compared with the wild-type plants upon salt stress. As shown in Table 1, AnnAt1, AnnAt2, and AnnAt3 were upregulated with fold changes of 2.576, 2.667, and 3.593, respectively. To confirm the relationship between AtPP2-B11 and the annexins, the transcript level of AnnAt1 was detected using real-time RT-PCR analysis, since AnnAt1 plays a dominant role among eight annexins in Arabidopsis. As shown in Fig. 6A, no significant difference in the AnnAt1 mRNA level was observed between the OE11 and wild-type plants under normal conditions. After 200mM NaCl treatment for 6h, the AnnAt1 mRNA expression was more induced in OE11 than in wild-type plants (Fig. 6A). Moreover, Western blot analysis showed that AnnAt1 was significantly induced in the AtPP2-B11 overexpressing lines but depressed in the R-5 line under salt stress conditions (Fig. 6B). To further confirm that AtPP2-B11 indeed upregulated the expression of AnnAt1, we used a LUC reporter assay to examine AnnAt1 promoter activity after co-expressing AtPP2-B11 in leaves of 4-week-old N. benthamiana plants, whereas co-expressing pA::LUC and 35S::GUS served as a negative control. Consistent with the real-time RT-PCR results (Fig. 6A), AnnAt1 promoter activity was induced in N. benthamiana epidermal cells when co-expressed with AtPP2-B11 in comparison with the control (Fig. 6C, D). These results suggested that AtPP2-B11 functions in the salt stress response probably by regulating the expression of AnnAt1.

Bottom Line: Isobaric tag for relative and absolute quantification analysis revealed that 4311 differentially expressed proteins were regulated by AtPP2-B11 under salt stress.Moreover, AtPP2-B11 influenced the expression of Na(+) homeostasis genes under salt stress, and the AtPP2-B11 overexpressing lines exhibited lower Na(+) accumulation.These results suggest that AtPP2-B11 functions as a positive regulator in response to salt stress in Arabidopsis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, PR China.

No MeSH data available.