Plant Disease "First Look" paper http://dx.doi.org/10.

1094/PDIS-05-15-0534-PDN posted 07/27/2015

This paper has been peer reviewed and accepted for publication but has not yet been copyedited or proofread. The final published version may differ.

Page 1 of 3

Disease Note

Candidatus Phytoplasma aurantifolia (16SrII group) associated with Witches

Broom disease of Bamboo (Dendrocalamus strictus) in India

Amit Yadav , Vipool Thorat and Yogesh Shouche

Microbial Culture Collection, National Centre for Cell Science, Pashan, Pune 411021,

India

25329034, Fax: +91-20-25329001.

Keywords:

Corresponding author: Amit Yadav, e-mail: amityadav@nccs.res.in; Phone: +91-20-

Candidatus

Phytoplasma

aurantifolia,

16SrII

group,

Bamboo,

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Dendrocalamus strictus

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Solid or Calcutta bamboo (Dendrocalamus strictus Nees., Poaceae) is found in the

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tropical regions of Indian subcontinent, Southeast Asia and China. D. strictus is

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semelparous and has a flowering cycle of 20 to 65 years. It is one of the toughest

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bamboos in India and extensively used in making pulp, rayon, paper mills and

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construction of light-weight houses as well as furniture, making it a commercially

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significant plant species. Its young shoots are used as food, leaves as fodder and it is

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also used in traditional medicine (Nadgauda et al. 1993). Leaf samples from

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symptomatic bamboo plants were collected from five geographically separated

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plantations from Maharashtra state during September 2014 to January 2015.

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Observed symptoms were typical witches broom phenotype with severe

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proliferative branching at nodal region and reduction in leaf size, as described earlier

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by Suryanarayana et. al. (2009) which were suggestive of a phytoplasma infection

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(Figure 1). To confirm the presence of phytoplasma, 1 g of leaf tissue from each of

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the symptomatic and asymptomatic plants were used for total DNA extraction using

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CTAB method. The phytoplasma 16S rRNA gene was detected in all 16 collected

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symptomatic D. strictus samples using nested PCR with phytoplasma universal

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primers P1 and P7 followed by primers R16F2n/R16R2 (Smart et. al. 1996) on

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agarose gel. No amplification was observed in template DNA isolated from

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asymptomatic plants. PCR fragments generated using R16F2n/1492R primers were

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sequenced directly using six different primers viz. R16F2n, R16R2, 343R, 704F, 907R,

Plant Disease "First Look" paper http://dx.doi.org/10.1094/PDIS-05-15-0534-PDN posted 07/27/2015

This paper has been peer reviewed and accepted for publication but has not yet been copyedited or proofread. The final published version may differ.

Page 2 of 3

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and 1492R. The obtained 16S rRNA gene sequence (GenBank accession number:

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LN811707, 1337 bp) showed 99.4 % homology with a strain WBDL belonging to

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Peanut Witches broom group phytoplasma; (Candidatus Phytoplasma aurantifolia,

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U15442) found in Catharanthus roseus (Asteraceae) when compared using the

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EzTaxon 16S rRNA database. The virtual RFLP pattern (Zhao et al. 2009) derived from

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the 16S rRNA sequence (LN811707) was found most similar to the reference pattern

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of the 16Sr group II, subgroup C (AJ293216), with a pattern similarity coefficient of

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0.99. To our knowledge, this is the only report of Ca. Phytoplasma aurantifolia

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associated with witches broom disease of D. strictus from India in addition to Ca.

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Phytoplasma asteris (AB242433) associated with Phyllostachys nigra (black bamboo)

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reported from Korea (Jung et. al., 2006). While Suryanayana et. al. (2009) reported

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symptoms of witches broom disease on D. strictus from Karnataka, India; it was not

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clear whether observed symptoms were related to phytoplasma infection.

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Additionally, there was no evidence to establish the species of the disease causing

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organism. D. strictus being an economically important plant, its susceptibility to

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phytoplasma diseases, especially involving vegetative propagation, can cause

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significant negative effect on its yield and therefore socio-economic status of the

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population engaged in its cultivation. Diagnostic accreditation of bamboo species for

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phytoplasma diseases and management of phytoplasma insect vectors will help

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reduce its exposure to witches broom disease.

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References: (1) R. Nadgauda et al., (1993). Tree Physiol., 13: 401- 408. (2) Smart, C.

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et al., (1996). Appl. and Environ. Microbiol., 62: 2988-2993. (3) Jung H. et. al. (2006).

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J. Gen. Plant Pathol. 72 (4): 261- 263. (4) Suryanarayana V. et. al. Ind. Phytoplathol.,

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62: 245- 248. (5) Zhao Y. et al. (2009). Int. J. Syst. Evol. Microbiol., 59: 2582-2593.

Plant Disease "First Look" paper http://dx.doi.org/10.1094/PDIS-05-15-0534-PDN posted 07/27/2015

This paper has been peer reviewed and accepted for publication but has not yet been copyedited or proofread. The final published version may differ.

Page 3 of 3

Figure 1: Infected Dendrocalamus strictus plant showing typical witches broom symptoms associated with
Candidatus Phytoplasma aurantifolia at Pune, India. Proliferative branches on plant (A), single branch (B)
and node (C) with profuse tillering.
388x121mm (150 x 150 DPI)