RALF, a 5-kDa ubiquitous polypeptide in plants, arrests root growth and development

Pearce, Gregory; Moura, Daniel S.; Stratmann, Johannes W.; Ryan, Clarence A.

doi:10.1073/pnas.201416998

Cited by 389 publications

(511 citation statements)

References 21 publications

(16 reference statements)

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“…Several peptides have been characterized recently that have roles in wounding (systemin; Pearce et al, 1991), self-incompatibility (SCR; Schopfer et al, 1999), stomatal patterning (EPF1; Hara et al, 2007), cellular proliferation and expansion (PSY1; Amano et al, 2007), abscission (IDA; Butenko et al, 2003), pollen formation (TPD1; Yang et al, 2003), root development (RALF; Pearce et al, 2001), shoot meristem development (CLAVATA3; Fletcher et al, 1999), and innate immunity (AtPep1; Huffaker et al, 2006). In the case of systemin, PSY1, SCR, and CLAVATA3, the corresponding peptide receptors have been identified, suggesting a general model of hormone activity via ligand-receptor pairing (Matsubayashi et al, 2001).…”

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Blufensin1Negatively Impacts Basal Defense in Response to Barley Powdery Mildew

2008

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Plants have evolved complex regulatory mechanisms to control the defense response against microbial attack. Both temporal and spatial gene expression are tightly regulated in response to pathogen ingress, modulating both positive and negative control of defense. BLUFENSIN1 (BLN1), a small peptide belonging to a novel family of proteins in barley (Hordeum vulgare), is highly induced by attack from the obligate biotrophic fungus Blumeria graminis f. sp. hordei (Bgh), casual agent of powdery mildew disease. Computational interrogation of the Bln1 gene family determined that members reside solely in the BEP clade of the Poaceae family, specifically, barley, rice (Oryza sativa), and wheat (Triticum aestivum). Barley stripe mosaic virus-induced gene silencing of Bln1 enhanced plant resistance in compatible interactions, regardless of the presence or absence of functional Mla coiled-coil, nucleotide-binding site, Leu-rich repeat alleles, indicating that BLN1 can function in an R-gene-independent manner. Likewise, transient overexpression of Bln1 significantly increased accessibility toward virulent Bgh. Moreover, silencing in plants harboring the Mlo susceptibility factor decreased accessibility to Bgh, suggesting that BLN1 functions in parallel with or upstream of MLO to modulate penetration resistance. Collectively, these data suggest that the grass-specific Bln1 negatively impacts basal defense against Bgh.

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Blufensin1Negatively Impacts Basal Defense in Response to Barley Powdery Mildew

2008

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“…Moura, J. Stratmann, C.A. Ryan [2001] Proc Natl Acad Sci USA 98: 12843-12847). Two unique poplar RALF cDNAs (PtdRALF1 and PtdRALF2) were isolated from a poplar cDNA library and used to study RALF expression in poplar saplings and cultured poplar cells.…”

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confidence: 99%

“…Therefore, the rapid pH changes observed in cell culture are essential components of signal transduction pathways. Furthermore, other plant peptides have been shown to trigger cell culture alkalinization: A recent report described the purification of a novel peptide from tobacco (Nicotiana tabacum) that causes rapid culture medium alkalinization (Pearce et al, 2001). However, this peptide, named the rapid alkalinization factor (RALF), does not induce defense-signaling pathways, but inhibits root growth when present in the surrounding medium.…”

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confidence: 99%

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Rapid Alkalinization Factors in Poplar Cell Cultures. Peptide Isolation, cDNA Cloning, and Differential Expression in Leaves and Methyl Jasmonate-Treated Cells

Haruta

Constabel

2003

Plant Physiology

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A family of peptides inducing rapid pH alkalinization in hybrid poplar (Populus trichocarpa ϫ Populus deltoides) cell culture medium was isolated from hybrid poplar leaves. Five related approximately 5-kD peptides were purified by highperformance liquid chromatography and analyzed by matrix-assisted laser desorption ionization-mass spectrometry. The N-terminal sequence of one of the isolated peptides was very similar to a previously characterized peptide from tobacco (Nicotiana tabacum), rapid alkalinization factor (RALF), which causes a rapid increase in culture medium pH when added to tobacco cell cultures (G. Pearce, D.S. Moura, J. Stratmann, C.A. Ryan [2001] Proc Natl Acad Sci USA 98: 12843-12847). Two unique poplar RALF cDNAs (PtdRALF1 and PtdRALF2) were isolated from a poplar cDNA library and used to study RALF expression in poplar saplings and cultured poplar cells. Both genes were found to be expressed constitutively in poplar saplings and cultured cells. However, PtdRALF2 was expressed in leaves at very low levels, and its expression in suspension culture cells was transiently suppressed by methyl jasmonate (MeJa). Although the function of these novel peptides remains enigmatic, our experiments suggest their role may be developmental rather than stress related. Overall, our study confirms the presence of active RALF peptides in other plants, and provides new data on the complexity of the RALF gene family in poplar.An essential feature of all plant cells is the electrochemical proton gradient across the plasma membrane, generated by the plasma membrane H ϩ -ATPase, which uses ATP to pump H ϩ outside the cell. This H ϩ gradient is important for many physiological processes including ion uptake, solute transport, and cell wall growth (Sanders and Bethke, 2000). Moreover, transient changes in extracellular or intracellular concentrations of H ϩ , and the accompanying plasma membrane depolarization or hyperpolarization, are implicated in the rapid responses of cells to environmental stimuli; for example, changes in turgor, gravity, and pathogen attack (Blumwald et al., 1998;Felix et al., 1999Felix et al., , 2000Johannes et al., 2001). A common observation is the rapid alkalinization of the extracellular solution, which can be conveniently observed in suspension cell cultures; as a consequence, stress-induced culture medium alkalinization has become a useful tool for monitoring the rapid events that accompany stress signal transduction (Felix et al., 1993; Blumwald et al., 1998;Schaller and Oecking, 1999). Depending on the system, these rapid increases in medium pH could potentially be caused by several mechanisms, including activation of K ϩ /H ϩ antiporters, H ϩ /solute cotransporters, and other ion channels, as well as an inhibition of the plasma membrane H ϩ -ATPase (Mathieu et al., 1993).The response of cells to plant pathogens and pathogen-derived signal molecules called elicitors has been extensively studied using changes in culture medium pH. For example, glycoproteins, peptides, sterols, lipo-chitoo...

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“…An alkalinization assay was developed in the Ryan laboratory that has been a key to the purification of the defense peptides HypSys and AtPeps, as well as the peptide Rapid Alkalinization Factor (Pearce et al, 2001b), which is involved in developmental processes. This alkalinization assay has tremendous potential to unlock the nature of peptide signaling in any plant species that can be maintained in liquid culture.…”

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GmPep914, an Eight-Amino Acid Peptide Isolated from Soybean Leaves, Activates Defense-Related Genes

et al. 2011

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Only a handful of endogenous peptide defense signals have been isolated from plants. Herein, we report a novel peptide from soybean (Glycine max) leaves that is capable of alkalinizing the media of soybean suspension cells, a response that is generally associated with defense peptides. The peptide, DHPRGGNY, was synthesized and found to be active at 0.25 nM and requiring only 5 to 10 min to obtain a maximal pH change. The peptide is located on the carboxy-terminal end of a 52-amino acid precursor protein (Glyma12g00990) deduced from the soybean genome project. A search of the soybean databank revealed a homolog (Glyma09g36370) that contained a similar peptide, DLPRGGNY, which was synthesized and shown to have identical activity. The peptides, designated GmPep914 (DHPRGGNY) and GmPep890 (DLPRGGNY), were capable of inducing the expression of both Glyma12g00990 (GmPROPEP914) and Glyma09g36370 (GmPROPEP890) in cultured soybean suspension cells within 1 h. Both peptides induced the expression of defense genes, including CYP93A1, a cytochrome P450 gene involved in phytoalexin synthesis, chitinaseb1-1, a chitinase involved in pathogen defense, and Glycine max chalcone synthase1 (Gmachs1), chalcone synthase, involved in phytoalexin production. Both GmPROPEP914 and GmPROPEP890 were highly expressed in the roots, relative to the aerial portions of the plant. However, treatment of the aerial portion of soybean plants with hormones involved in elicitation of defense responses revealed a significant increase in expression levels of GmPROPEP914 and GmPROPEP890. A search of gene databases revealed homologous sequences in other members of the Fabales and also in the closely related Cucurbitales but not in any other order of plants.

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RALF, a 5-kDa ubiquitous polypeptide in plants, arrests root growth and development

Cited by 389 publications

References 21 publications

Blufensin1Negatively Impacts Basal Defense in Response to Barley Powdery Mildew

Blufensin1Negatively Impacts Basal Defense in Response to Barley Powdery Mildew

Rapid Alkalinization Factors in Poplar Cell Cultures. Peptide Isolation, cDNA Cloning, and Differential Expression in Leaves and Methyl Jasmonate-Treated Cells

GmPep914, an Eight-Amino Acid Peptide Isolated from Soybean Leaves, Activates Defense-Related Genes

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