Detection and
molecular characterization of phytoplasma associated with Hibiscus
Witches’-Broom in Egypt
Samah A. Mokbel
Department
of Virus and Phytoplasma Research, Plant Pathology Research Institute, Agricultural
Research Center., Egypt. Dr.SamahMokbel@gmail.com
Abstract
Hibiscus
rosa-sinensisis a valuable ornamental species widely planted in Egypt. The
Phytoplasma associated witches’-Broom has been detected on symptomatic hibiscus
plants. Samples of hibiscus leaves were collected from El Giza, Alexandria,
Qlubia, El-Fayom, and El-Mansoura governorates and analyzed for phytoplasma
infection. The collected hibiscus plants showed characteristic symptoms for
Phytoplasma associated witches’-Broom disease, which is characterized by
excessive axillary branching, abnormally small leaves, and deformed flowers.
Dienes’ stain was used for detection of witches’ broom infection midribs of the
symptomatic hibiscus plant. The phloem of infected tissues showed scattered
area stained bright blue. Molecular detection utilizing nested and direct PCR
as well as DNA sequencing was used for the diagnosis of the witches’ Broom
infection. Total DNA was isolated from leaf tissues of infected hibiscus plants.
Nested polymerase chain reaction (PCR) was performed using the universal -
phytoplasma specific primers; P1/P7, R16F2n/R16R2. Witches’ broom specific
primers SR1/ SR2, at the spacer region (SR), were used for the direct PCR.
Amplicons of expected size characteristic for phytoplasma associated witches’
broom were obtained from infected hibiscus samples. DNA sequencing and
phylogenetic analysis of the 16S rRNA gene fragment from the hibiscus witches'
broom phytoplasma showed 100% homology with the same region of the hibiscus
witches' broom strain that isolated in Brazil (Phytoplasma Brasiliense). The
DNA sequence was submitted for the gene bank as the first report of the
Brasiliense phytoplasma associated witches’ broom affecting hibiscus in Egypt.
Key
words: Hibiscus, Dienes’ stain, 16SRNA, phytoplasma, PCR, Sequencing.
How to cite this article:
Mokbel, A.S., El-Attar, A.K. and Farag, A.G. (2013). Detection and molecular characterization of phytoplasma associated with Hibiscus Witches’-Broom in Egypt. Egyptian J. of Virology. 10:162-171.
Mokbel, A.S., El-Attar, A.K. and Farag, A.G. (2013). Detection and molecular characterization of phytoplasma associated with Hibiscus Witches’-Broom in Egypt. Egyptian J. of Virology. 10:162-171.
