When confronted with inorganic phosphate (Pi) starvation, plants activate an array of adaptive responses to sustain their growth. These responses, in a large extent, are controlled at the transcriptional level. Arabidopsis (Arabidopsis thaliana) PHOSPHATE RESPONSE1 (PHR1) and its close homolog PHR1-like 1 (PHL1) belong to a 15-member family of MYB-CC transcription factors and are regarded as the key components of the central regulatory system controlling plant transcriptional responses to Pi starvation. The knockout of PHR1 and PHL1, however, causes only a partial loss of the transcription of Pi starvation-induced genes, suggesting the existence of other key components in this regulatory system. In this work, we used the transcription of a Pi starvation-induced acid phosphatase, AtPAP10, to study the molecular mechanism underlying plant transcriptional responses to Pi starvation. We first identified a DNA sequence on the AtPAP10 promoter that is critical for the transcription of AtPAP10. We then demonstrated that PHL2 and PHL3, two other members of the MYB-CC family, specifically bind to this DNA sequence and activate the transcription of AtPAP10. Unlike PHR1 and PHL1, the transcription and protein accumulation of PHL2 and PHL3 are upregulated by Pi starvation. RNA-sequencing analyses indicated that the transcription of most Pi starvation-induced genes is impaired in the phl2 mutant, indicating that PHL2 is also a key component of the central regulatory system. Finally, we showed that PHL2, and perhaps also PHL3, acts redundantly with PHR1 to regulate plant transcriptional response to Pi starvation.

Background Masson pine (Pinus massoniana) is primarily present in subtropical and tropical areas of China, which are severely deficient in inorganic phosphate (Pi). Although some studies identified transcriptomic and proteomic responses to Pi deficiency in Masson pine seedlings, different tissues, especially the roots, that exhibit primary responses to low-Pi stress, have not been well studied. To shed further light on the complex responses of Masson pine to Pi deficiency, a spatiotemporal experiment was performed to identify differentially expressed mRNAs and miRNAs under Pi-deficient conditions. Results Spatiotemporal analyses of 72 RNA sequencing libraries provided a comprehensive overview of the dynamic responses of Masson pine to low-Pi stress. Differentially expressed gene analysis revealed several high-affinity phosphate transporters and nitrate transporters, reflecting the crosstalk between nitrate and Pi homeostasis in plants. The MYB family was the most abundant transcription factor family identified. miRNA differential expression analysis identified several families that were associated with Pi deficiency, such as miR399. In addition, some other families, including novel miRNA families in Masson pine, were dramatically changed in response to Pi starvation. GO and KEGG analyses of these mRNAs and targets of miRNAs indicated that metabolic processes were most enriched under Pi deficiency. Conclusions This study provided abundant spatiotemporal transcriptomic information to functionally dissect the response of Masson pine seedlings to Pi deficiency, which will aid in further elucidation of the biological regulatory mechanisms of pines in response to low-Pi stress.

No abstract

Plants have evolved an array of adaptive responses to cope with phosphate (Pi) starvation. These responses are mainly controlled at the transcriptional level. In Arabidopsis, PHR1, a member of the MYB-CC transcription factor family, is a key component of the central regulatory system controlling plant transcriptional responses to Pi starvation. Its homologs in the MYB-CC family, PHL1 (PHR1-LIKE 1), PHL2, and perhaps also PHL3, act redundantly with PHR1 to regulate plant Pi starvation responses. The functions of PHR1’s closest homolog in this family, PHL4, however, have not been characterized due to the lack of its null mutant. In this work, we generated two phl4 null mutants using the CRISPR/Cas9 technique and investigated the functions of PHL4 in plant responses to Pi starvation. The results indicated that the major developmental, physiological, and molecular responses of the phl4 mutants to Pi starvation did not significantly differ from those of the wild type. By comparing the phenotypes of the phr1 single mutant and phr1phl1 and phr1phl4 double mutants, we found that PHL4 also acts redundantly with PHR1 to regulate plant Pi responses, but that its effects are weaker than those of PHL1. We also found that the overexpression of PHL4 suppresses plant development under both Pi-sufficient and -deficient conditions. Taken together, the results indicate that PHL4 has only a minor role in the regulation of plant responses to Pi starvation and is a negative regulator of plant development.

We previously demonstrated that epidermal growth factor (EGF) induces a several-fold increase in ornithine decarboxylase (ODC) activity and the steady-state level of ODC mRNA in cultured SV40-transformed human keratinocytes (1). Pretreatment of cell cultures with ultraviolet B (UVB) radiation resulted in a reduction of EGF-induced ODC activity. To determine whether UVB inhibits the accumulation of ODC mRNA by EGF, cells were pretreated with 20 mJ/cm2 UVB or sham-irradiated and then incubated with 100 ng/ml EGF. Northern blot analysis revealed that UVB irradiation entirely blocked the EGF induction of ODC mRNA. Since the binding of EGF to its plasma membrane receptor is the first step in initiating a biological response, the effect of UVB on EGF binding was evaluated. UVB treatment of cultured keratinocytes resulted in an immediate and dose-dependent reduction of EGF binding. Scatchard analysis revealed that the reduction of EGF binding was due to a 52% decrease in the number of available receptors, from 6.2 x 10(4)/cell to 3.0 x 10(4)/cell. However, UVB decreased the EGF-binding affinity very little (Kd = 0.60 nM in control and Kd = 0.75 nM in UVB-treated Z114 cells). In addition, UVB did not alter the rate of EGF internalization. These data suggest that UVB blocks the signal transduction pathway of EGF that is involved in regulation of ODC gene expression. Immunoblot analysis of extracts from irradiated cells showed that UVB induced tyrosine phosphorylation of EGFR and that the quantity of EGFR protein was unaffected by UVB treatment. Phosphorylation of EGFR may be responsible for decreased binding of EGF to its receptor.