title: A stress responsive alternative splicing mechanism in Citrus clementina leaves.
abstract: Chitinases are often considered pathogenesis-related proteins since their activity can be induced by viral infections, fungal and bacterial cell wall components, and also by more general sources of stress such as wounding, salicylic acid, ethylene, auxins and cytokinins. In the present study, comparative proteomic analysis showed the defense-related acidic chitinase II to be specifically induced in Citrus clementina leaves infested by the two-spotted spider mite Tetranychus urticae or treated with MeJA. In parallel, changes in the mRNA profiles of two partially homologous chitinase forms were shown by RT-PCR. In particular, the appearance of an additional cDNA chitinase fragment in T. urticae-infested and MeJA-treated leaves was observed. This finding may indicate a specific regulatory mechanism of chitinase expression. We report evidence for alternative splicing in T. urticae-infested C. clementina, where a premature stop codon after the first 135 amino acids was introduced. We observed inducible chitinase activity after MeJA treatment, indicative of a rapid plant response to infestation. This work provides the first evidence of chitinase alternative splicing in C. clementina. In addition, the presence of the dual-band pattern for chitinase cDNA by RT-PCR may represent a suitable predictive marker for early diagnosis of plant biotic stress.
authors.affiliation: Institute of Clinical Physiology, National Council of Research, Via Moruzzi 1, 56124 Pisa, Italy. rdc@ifc.cnr.it
authors.forename: Renata
authors.initials: R
authors.lastname: Del Carratore
authors.forename: Eleonora
authors.initials: E
authors.lastname: Magaldi
authors.forename: Alessandra
authors.initials: A
authors.lastname: Podda
authors.forename: Pascale
authors.initials: P
authors.lastname: Beffy
authors.forename: Quirico
authors.initials: Q
authors.lastname: Migheli
authors.forename: Bianca Elena
authors.initials: BE
authors.lastname: Maserti
mesh_terms: D000085:Acetates / Q000494:pharmacology; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002688:Chitinases / Q000235:genetics / Q000378:metabolism*; D002957:Citrus / Q000187:drug effects / Q000201:enzymology* / Q000469:parasitology; D003517:Cyclopentanes / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D054883:Oxylipins / Q000494:pharmacology; D018515:Plant Leaves / Q000187:drug effects / Q000201:enzymology* / Q000469:parasitology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000187:drug effects* / Q000378:metabolism; D013312:Stress, Physiological; D040102:Tetranychidae / Q000502:physiology
chemical_list: D000085:Acetates; D003517:Cyclopentanes; D054883:Oxylipins; D010940:Plant Proteins; D012333:RNA, Messenger; C072239:methyl jasmonate; D002688:Chitinases