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Original Article 
JVS. 2019; 6(1): 12-23


Detection and molecular identification of phytoplasma associated with Gazania in Egypt

shimaa mohamed gad, ahmed Abd El aziz Kheder, mohamed ahmed awad.

Abstract
Abstract
Background: Phytoplasmas, known as mycoplasma-like organisms (MLOs) cause diseases in several commercial ornamental plants, leads to serious economic losses all over the world. During 2017-2018, phytoplasma disease showing phyllody, yellowing, proliferation, virescence and little leaf symptoms was observed on gazania and other ornamental plants in Giza, Egypt.
Methods: Phytoplasma disease was detected and isolated from naturally infected gazania plants during surveys in flower nurseries and open filed in Giza governorate, it was transmitted from naturally infected Gazania to healthy periwinkle and other ornamental plants by dodder (Cuscuta reflexa), and insects (Empoasca decipiens). Diene's stain was used to detect and differentiate the phloem tissues of leaf sections from infected gazania and periwinkle plants. Transmission Electron Microscopy (TEM) observed the presence of phytoplasma in the sieve tubes and parenchyma cells of leaf midribs in infected plants. DNA extracted from symptomatic samples was used as a template in nested polymerase chain reaction (PCR) using universal primers pairs P1/P7 and R16F2n/R16R2. Sequencing and phylogenetic analysis were performed to identify the detected phytoplasmas.
Results: Phytoplasma was transmitted successfully from naturally infected gazania to healthy ornamental plants by dodder and insect. Light microscopy (LM) and transmission electron microscopy (TEM) revealed that, phytoplasma-like bodies were detected inside phloem, sieve tubes and parenchyma cells of leaf midribs tissues in infected plants and ranging from 200 to 400nm in diameters. The 16SrRNA gene from phytoplasma was amplified by nested-PCR assay and direct sequenced using specific primer pairs. Phylogenetic tree was calculated based on obtained sequences data.
Conclusion: The phytoplasma associated with gazania exhibiting phyllody, yellowing, proliferation, virescence and little leaf symptoms was confirmed by the results of LM and TEM observations and nested-PCR testing. Based on direct sequence date, phylogeny analysis, the associated phytoplasma was classified as related to 16SrII group.

Key words: Key words: Phytoplasma, Gazania, Light Microscopy, Electron microscopy, nested-PCR


 
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How to Cite this Article
Pubmed Style

gad sm, Kheder aAEa, awad ma, . Detection and molecular identification of phytoplasma associated with Gazania in Egypt. JVS. 2019; 6(1): 12-23.


Web Style

gad sm, Kheder aAEa, awad ma, . Detection and molecular identification of phytoplasma associated with Gazania in Egypt. http://www.j-virol-sci.com/?mno=28916 [Access: January 27, 2020].


AMA (American Medical Association) Style

gad sm, Kheder aAEa, awad ma, . Detection and molecular identification of phytoplasma associated with Gazania in Egypt. JVS. 2019; 6(1): 12-23.



Vancouver/ICMJE Style

gad sm, Kheder aAEa, awad ma, . Detection and molecular identification of phytoplasma associated with Gazania in Egypt. JVS. (2019), [cited January 27, 2020]; 6(1): 12-23.



Harvard Style

gad, s. m., Kheder, a. A. E. a., awad, m. a. & (2019) Detection and molecular identification of phytoplasma associated with Gazania in Egypt. JVS, 6 (1), 12-23.



Turabian Style

gad, shimaa mohamed, ahmed Abd El aziz Kheder, mohamed ahmed awad, and . 2019. Detection and molecular identification of phytoplasma associated with Gazania in Egypt. Journal of Virological Sciences, 6 (1), 12-23.



Chicago Style

gad, shimaa mohamed, ahmed Abd El aziz Kheder, mohamed ahmed awad, and . "Detection and molecular identification of phytoplasma associated with Gazania in Egypt." Journal of Virological Sciences 6 (2019), 12-23.



MLA (The Modern Language Association) Style

gad, shimaa mohamed, ahmed Abd El aziz Kheder, mohamed ahmed awad, and . "Detection and molecular identification of phytoplasma associated with Gazania in Egypt." Journal of Virological Sciences 6.1 (2019), 12-23. Print.



APA (American Psychological Association) Style

gad, s. m., Kheder, a. A. E. a., awad, m. a. & (2019) Detection and molecular identification of phytoplasma associated with Gazania in Egypt. Journal of Virological Sciences, 6 (1), 12-23.








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