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Plasma membrane receptor-like kinase leaf panicle 2 acts downstream of the DROUGHT AND SALT TOLERANCE transcription factor to regulate drought sensitivity in rice.

Wu F, Sheng P, Tan J, Chen X, Lu G, Ma W, Heng Y, Lin Q, Zhu S, Wang J, Wang J, Guo X, Zhang X, Lei C, Wan J - J. Exp. Bot. (2014)

Bottom Line: A leucine-rich repeat (LRR)-RLK gene named Leaf Panicle 2 (LP2) was previously found to be strongly expressed in leaves and other photosynthetic tissues, but its function remains unclear.In addition, LP2 was identified as a functional kinase localized to the plasma membrane and interacted with the drought-responsive aquaporin proteins OsPIP1; 1, OsPIP1; 3, and OsPIP2; 3.Thus, the findings provided evidence that the LRR-RLK LP2, transcriptionally regulated by the drought-related transcription factor DST, served as a negative regulator in drought response.

View Article: PubMed Central - PubMed

Affiliation: National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China.

No MeSH data available.


Related in: MedlinePlus

LP2 is transcriptionally regulated by DST via direct binding to the promoter. (A) Schematic diagram of the promoter regions of LP2. A black line represents the promoter region of LP2; the black box on the line represents the putative DST-binding site. Upper numbers indicate relative distances from the ATG initiation codon shown as +1 (scale bars=200bp). (B) Yeast one-hybrid assays showing that DST activates the LacZ reporter gene driven by the LP2 promoter containing the putative DST-binding motif, but not LacZ reporter genes driven by the LP2 promoter without the binding motif, or with mutations in the binding motif. Red bars in the construct of LP2p-1-m and LP2p-1d-m indicate the positions of mutations. AD, activation domain. (C) ChIP-qPCR assays showing that LP2 promoter fragments containing the putative DST-binding site are specifically enriched. Four pairs of primers were used for the ChIP-qPCR experiment (means ±SD, n=3). Immunoprecipitation with a pre-immune (Pre.) serum was used as the negative control. (D) Comparison of transcript abundance of LP2 in the wild type (ZH11) and dst mutant by qRT–PCR.
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Figure 6: LP2 is transcriptionally regulated by DST via direct binding to the promoter. (A) Schematic diagram of the promoter regions of LP2. A black line represents the promoter region of LP2; the black box on the line represents the putative DST-binding site. Upper numbers indicate relative distances from the ATG initiation codon shown as +1 (scale bars=200bp). (B) Yeast one-hybrid assays showing that DST activates the LacZ reporter gene driven by the LP2 promoter containing the putative DST-binding motif, but not LacZ reporter genes driven by the LP2 promoter without the binding motif, or with mutations in the binding motif. Red bars in the construct of LP2p-1-m and LP2p-1d-m indicate the positions of mutations. AD, activation domain. (C) ChIP-qPCR assays showing that LP2 promoter fragments containing the putative DST-binding site are specifically enriched. Four pairs of primers were used for the ChIP-qPCR experiment (means ±SD, n=3). Immunoprecipitation with a pre-immune (Pre.) serum was used as the negative control. (D) Comparison of transcript abundance of LP2 in the wild type (ZH11) and dst mutant by qRT–PCR.

Mentions: Analysis of the promoter sequence of LP2 revealed a DST-binding element (DBE) at position –670 to –661 (Huang et al., 2009; Fig. 6A). DST is a zinc finger transcription factor that negatively regulates stomatal closure by direct modulation of genes related to H2O2 homeostasis (Huang et al., 2009). To test the possibility that DST directly binds to the DBE in the LP2 promoter, a yeast one-hybrid assay was performed. The results showed that DST protein binds directly to the promoter sequence of LP2 (–2126 to –1) (Fig. 6B). To determine whether the predicted DBE is functional, several truncation derivatives of the –2126 to –1 promoter sequence were subjected to transactivity assays. As shown in Fig. 6B, only the fragment including the DBE interacted with DST in yeast. When the DBE was deleted or mutated, there was no binding (Fig. 6B). A chromatin immunoprecipitation-quantitative real-time PCR (ChIP-qPCR) assay was further applied to investigate the interaction between DST and the LP2 promoter. The results indicated a significant enrichment of DST binding to the DBE of the LP2 promoter, in contrast to its negligible binding to other positions (Fig. 6C). The expression level of LP2 in the dst mutant was also assayed and, as shown in Fig. 6D, expression of LP2 was greatly decreased in the mutant. These results indicated that DST bound to the DBE at position –670 to –661 of the LP2 promoter to regulate LP2 expression directly.


Plasma membrane receptor-like kinase leaf panicle 2 acts downstream of the DROUGHT AND SALT TOLERANCE transcription factor to regulate drought sensitivity in rice.

Wu F, Sheng P, Tan J, Chen X, Lu G, Ma W, Heng Y, Lin Q, Zhu S, Wang J, Wang J, Guo X, Zhang X, Lei C, Wan J - J. Exp. Bot. (2014)

LP2 is transcriptionally regulated by DST via direct binding to the promoter. (A) Schematic diagram of the promoter regions of LP2. A black line represents the promoter region of LP2; the black box on the line represents the putative DST-binding site. Upper numbers indicate relative distances from the ATG initiation codon shown as +1 (scale bars=200bp). (B) Yeast one-hybrid assays showing that DST activates the LacZ reporter gene driven by the LP2 promoter containing the putative DST-binding motif, but not LacZ reporter genes driven by the LP2 promoter without the binding motif, or with mutations in the binding motif. Red bars in the construct of LP2p-1-m and LP2p-1d-m indicate the positions of mutations. AD, activation domain. (C) ChIP-qPCR assays showing that LP2 promoter fragments containing the putative DST-binding site are specifically enriched. Four pairs of primers were used for the ChIP-qPCR experiment (means ±SD, n=3). Immunoprecipitation with a pre-immune (Pre.) serum was used as the negative control. (D) Comparison of transcript abundance of LP2 in the wild type (ZH11) and dst mutant by qRT–PCR.
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Figure 6: LP2 is transcriptionally regulated by DST via direct binding to the promoter. (A) Schematic diagram of the promoter regions of LP2. A black line represents the promoter region of LP2; the black box on the line represents the putative DST-binding site. Upper numbers indicate relative distances from the ATG initiation codon shown as +1 (scale bars=200bp). (B) Yeast one-hybrid assays showing that DST activates the LacZ reporter gene driven by the LP2 promoter containing the putative DST-binding motif, but not LacZ reporter genes driven by the LP2 promoter without the binding motif, or with mutations in the binding motif. Red bars in the construct of LP2p-1-m and LP2p-1d-m indicate the positions of mutations. AD, activation domain. (C) ChIP-qPCR assays showing that LP2 promoter fragments containing the putative DST-binding site are specifically enriched. Four pairs of primers were used for the ChIP-qPCR experiment (means ±SD, n=3). Immunoprecipitation with a pre-immune (Pre.) serum was used as the negative control. (D) Comparison of transcript abundance of LP2 in the wild type (ZH11) and dst mutant by qRT–PCR.
Mentions: Analysis of the promoter sequence of LP2 revealed a DST-binding element (DBE) at position –670 to –661 (Huang et al., 2009; Fig. 6A). DST is a zinc finger transcription factor that negatively regulates stomatal closure by direct modulation of genes related to H2O2 homeostasis (Huang et al., 2009). To test the possibility that DST directly binds to the DBE in the LP2 promoter, a yeast one-hybrid assay was performed. The results showed that DST protein binds directly to the promoter sequence of LP2 (–2126 to –1) (Fig. 6B). To determine whether the predicted DBE is functional, several truncation derivatives of the –2126 to –1 promoter sequence were subjected to transactivity assays. As shown in Fig. 6B, only the fragment including the DBE interacted with DST in yeast. When the DBE was deleted or mutated, there was no binding (Fig. 6B). A chromatin immunoprecipitation-quantitative real-time PCR (ChIP-qPCR) assay was further applied to investigate the interaction between DST and the LP2 promoter. The results indicated a significant enrichment of DST binding to the DBE of the LP2 promoter, in contrast to its negligible binding to other positions (Fig. 6C). The expression level of LP2 in the dst mutant was also assayed and, as shown in Fig. 6D, expression of LP2 was greatly decreased in the mutant. These results indicated that DST bound to the DBE at position –670 to –661 of the LP2 promoter to regulate LP2 expression directly.

Bottom Line: A leucine-rich repeat (LRR)-RLK gene named Leaf Panicle 2 (LP2) was previously found to be strongly expressed in leaves and other photosynthetic tissues, but its function remains unclear.In addition, LP2 was identified as a functional kinase localized to the plasma membrane and interacted with the drought-responsive aquaporin proteins OsPIP1; 1, OsPIP1; 3, and OsPIP2; 3.Thus, the findings provided evidence that the LRR-RLK LP2, transcriptionally regulated by the drought-related transcription factor DST, served as a negative regulator in drought response.

View Article: PubMed Central - PubMed

Affiliation: National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China.

No MeSH data available.


Related in: MedlinePlus