title: Characterization of polypeptides accumulated in the latex cytosol of rubber trees affected by the tapping panel dryness syndrome.
abstract: The tapping panel dryness (TPD) syndrome of rubber is characterized by the reduction or ultimately total cessation of latex flow upon tapping, due to physiological disorders in the bark tissue. The protein pattern in the cytoplasm from healthy and TPD tree latex cells was compared by electrophoresis. Two polypeptides (P15 and P22) of 15 and 22 kDa, respectively, were found to accumulate in the cytosol of the TPD-affected trees, whereas a 29 kDa polypeptide (P29) appeared de novo. P15 and P22 were identified as REF (Hev b1) and SRPP (Hev b3), respectively, two proteins proposed to be involved in rubber biosynthesis. P29 appeared to be a new member of the patatin-like protein family. Specific molecular probes were designed for a detailed characterization of REF and SRPP gene expression and RFLP mapping. This allowed the demonstration that REF and SRPP display very similar expression profiles. They are highly over-expressed by the tapping-induced metabolic activation, although not by wounding per se, or ethylene or ABA. In addition to this similarity in gene expression, they were found to share one common locus in the genome. No significant difference in REF and SRPP gene expression was observed between healthy and TPD trees, indicating that their TPD-related accumulation in the cytosol was not transcriptionally regulated. Western blot analysis demonstrated that osmotic lysis of the sedimentable organelles (lutoids) in vitro caused the release of REF and SRPP from the rubber particle membrane into the cytosol. A mechanism of cellular delocalization as a consequence of the lutoids instability is proposed to explain REF and SRPP accumulation in the cytosol of TPD trees.
authors.affiliation: Department of Biotechnology, Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand.
authors.forename: Unchera
authors.initials: U
authors.lastname: Sookmark
authors.forename: Valérie
authors.initials: V
authors.lastname: Pujade-Renaud
authors.forename: Hervé
authors.initials: H
authors.lastname: Chrestin
authors.forename: Régis
authors.initials: R
authors.lastname: Lacote
authors.forename: Chinda
authors.initials: C
authors.lastname: Naiyanetr
authors.forename: Marc
authors.initials: M
authors.lastname: Seguin
authors.forename: Phayao
authors.initials: P
authors.lastname: Romruensukharom
authors.forename: Jarunya
authors.initials: J
authors.lastname: Narangajavana
mesh_terms: D000485:Allergens / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D015152:Blotting, Northern; D015153:Blotting, Western; D002874:Chromosome Mapping; D003600:Cytosol / Q000378:metabolism; D028482:Hevea / Q000235:genetics / Q000502:physiology*; D007840:Latex / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism*; D010935:Plant Diseases; D010940:Plant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013314:Stress, Mechanical
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D007840:Latex; D010455:Peptides; D010940:Plant Proteins; C119716:SRPP protein, Hevea brasiliensis; C061477:REF protein, Hevea brasiliensis