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Molecular Characterization of Stolbur Phytoplasma Associated with Pea Plants in Poland
Author(s) -
Zwolińska Agnieszka,
Krawczyk Krzysztof,
Pospieszny Henryk
Publication year - 2012
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/j.1439-0434.2012.01903.x
Subject(s) - phytoplasma , biology , pisum , restriction fragment length polymorphism , sativum , chlorosis , nested polymerase chain reaction , 16s ribosomal rna , cultivar , shoot , botany , polymerase chain reaction , restriction enzyme , horticulture , ribosomal rna , gene , genetics
Pea plants ( Pisum sativum ) showing symptoms of stunting, shoot proliferation and leaf chlorosis were observed in 2008 during routine greenhouse cultivation of garden pea cultivars from commercially obtained seeds. The disease incidence occurred in over 25% of grown plants. To confirm phytoplasma infection, fresh tissue samples, from symptomatic and asymptomatic peas, were collected, and total DNA was extracted, using a modified CTAB method. Nested‐PCR assay was carried out with specific phytoplasma 16S rDNA primers: P1/P7 followed by R16F2n/R16R2. The product, of expected size 1.2 kb, was restricted with 6 different endonucleases, and on the basis of obtained RFLP profile, phytoplasma was identified as a 16Sr XII‐A ribosomal subgroup member. For further differentiation, the nucleotide sequence of the tuf gene encoding a transcription factor was analysed. This is the first report of phytoplasma affecting pea plants.