Plumeria rust caused by Coleosporium plumeriae on frangipani trees in Sumatra, Indonesia

Abstract

Powdery rust pustules of bright yellow-orange color were observed forming on both mature and young leaves of red frangipani (Plumeria rubra) in the province of Riau, Sumatra, Indonesia. Based on morphological characteristics and DNA sequence of the internal transcribed spacer region (ITS), the rust fungus was identified as Coleosporium plumeriae, only urediniospores were found on the infected leave. Inoculation trials confirmed the pathogenicity of C. plumeria, fulfilling the Koch’s postulates. This is the first report and characterization of plumeria rust caused by C. plumeria on frangipani on the island of Sumatra, Indonesia.

Full text

Plumeria rust caused by Coleosporium plumeriae on frangipani
trees in Sumatra, Indonesia
Leonardo Sarno Soares Oliveira
1
&Eko Sulistyono
2
&Pantun David Mangatas Lbn Gaol
1
&Tisha Melia
2
&Alvaro Durán
1
Received: 21 July 2019 /Accepted: 25 October 2019
#Australasian Plant Pathology Society Inc. 2019
Abstract
Powdery rust pustules of bright yellow-orange color were observed forming on both mature and young leaves of red frangipani
(Plumeria rubra) in the province of Riau, Sumatra, Indonesia. Based on morphological characteristics and DNA sequence of the
internal transcribed spacer region (ITS), the rust fungus was identified as Coleosporium plumeriae,onlyurediniosporeswere
found on the infected leave. Inoculation trials confirmed the pathogenicity of C. plumeria, fulfilling the Koch’s postulates. This is
the first report and characterization of plumeria rust caused by C. plumeria on frangipani on the island of Sumatra, Indonesia.
Keywords Rust disease .Flowering plants .Urediniospores .Temple tree
Plumeria spp. are flowering plants of the family
Apocynaceae, with center of origin in West Indies and
Central America, but widely cultivated in other tropical and
sub-tropical areas, especially in Asian countries and the
Pacific Islands (Nelson 2009). In Indonesia, Plumeria spp.
are commonly found in urban landscapes or as potted plants,
especially in the island of Bali. Their flowers are associated
with the Balinese culture and the plants are locally named as
frangipani, temple tree, Bkamboja^or Bjepun^.
Kobayashi et al. (1994)reportedP. rubra plants (red fran-
gipani) found suffering from a serious rust disease caused by
Coleosporium plumeriae, and this is considered the first offi-
cial report of this disease in Indonesia. The disease was sub-
sequently reported in the same plant species in West Java
(Kakishima et al. 2017). Coleosporium plumeriae Pat.
(Patouillard 1902) belongs to the Family Coleosporiaceae
(Pucciniales). It was originally reported on P. alba plants in
Santo Domingo, West Indies, in 1852 as Uredo domingensis
Berk. (Arthur 1918). This disease is now known to have a
worldwide distribution. The reason for its unexpected and
rapid expansion from Central America to Pacific Islands re-
mains unclear but has been hypothesized to be related to
human-mediated introductions and also due to climate chang-
es related to El Niño and La Niña events in the Pacific Ocean
(Kakishima et al. 2017).
P. rubra has been widely planted in the Indonesian territory
but there are no official reports of plumeria rust from Sumatra,
the largest island that is located entirely in Indonesia. The aim
of this study was thus to report and characterize the morphol-
ogy, pathogenicity and DNA profile of Coleosporium
plumeriae from Riau, Sumatra, Indonesia.
Numerous powdery rust pustules (uredinia) were observed
forming on the abaxial surface of both mature and young
leaves (Fig. 1). No symptoms were observed on flowers,
stems or branches. As the disease progressed, coalescent pus-
tules could be observed on the upper leaf surface as sunken,
angular, yellowish flecks or even necrotic lesions. In case of
severe infection, curly and distorted leaves were observed
followed by defoliation.
In September 2018, infected leaves of P. rubra showing
rust symptoms were collected from urban trees located in
Pangkalan Kerinci, Riau, Indonesia (coordinates: 0° 24′0″
N, 101° 51′0″E). A total of sixteen samples were analyzed
at the RGE Plant Pathology Laboratory (Pangkalan Kerinci).
With the aid of a soft brush, urediniospores were collected
from infected leaves and stored in glycerol 15% at −80 °C.
Additionally, a representative specimen was selected and de-
posited in the culture collection of Bogor Agricultural
University (IPB/LIPI -Accession number: 243234). For exam-
ination of morphological features, uredinia were mounted in
*Leonardo Sarno Soares Oliveira
leonardo_oliveira@aprilasia.com
1
Plant Health Program, Research and Development, Asia Pacific
Resources International Holdings Ltd. (APRIL), Pangkalan
Kerinci, Riau 28300, Indonesia
2
Molecular Biology and Genomics Program, Research and
Development, Asia Pacific Resources International Holdings Ltd.
(APRIL), Pangkalan Kerinci, Riau 28300, Indonesia
Australasian Plant Disease Notes (2019) 14:34
https://doi.org/10.1007/s13314-019-0366-1

cotton blue or distilled water and fungal structures were ob-
served and measured using a light microscope. Microscopic
examination of infected leaves revealed the presence of
urediniospores (Fig. 1c) that were a bright yellow-orange col-
or, ellipsoidal or globose, sometimes angular, echinulate, and
measured 16.3–42.2 × 16.4–29.8 μm, matching with ranges
reported by Traquair and Kokko (1980). Teliospores were
not observed in any of the samples collected.
For molecular identification, DNA was extracted from
urediniospores of four samples (A1-CARCP01, A2-
CARCP02, A3-CARCP03 and A4-CARCP04) each obtained
from single pustules, following the protocols described by
Doyle and Doyle (1987). Amplification of ITS region was
performed using ITS1/ITS4 primer (White et al. 1990). PCR
amplification was performed according to protocols described
by Langrell et al. (2008). Purified DNA was then sent for
sequencing (Macrogen Inc., South Korea). Sequences were
desposited in GenBank (accession number: MK788144,
MK788146, MK788147 and MK788148), with an average
of sequence length about 700 bp. BLASTn searches in
NCBI GenBank showed 100% identity to sequence of
Coleosporium plumeriae from the neighboring country,
Malaysia (GenBank accession number: MF769628).
Multiple sequence analysis (MSA) was performed with the
four isolates from the present study, and compared against
other nucleotide sequences from GenBank. Austropuccinia
psidii (EF210142) was used as an outgroup taxon (Table 1)
based on ClustalW (Thompson et al. 1994). Phylogenetic
analyses were performed in MEGA6 (Tamura et al. 2013)
using the maximum-likelihood method with additional 1000
Fig. 1 Symptoms and morphology of plumeria rust caused by Coleosporium plumeriae in Plumeria rubra.aand b–Rust symptoms; c–
Urediniospores. Scale bar = 10 μm
Table 1 Details of Colesoporium
plumeriae isolates from Plumeria
rubra and other rust fungi used in
the phylogenetic analyses in this
study
Species Isolate code Host Locality ITS-rDNA Accession
number
Coleosporium
plumeriae
A1-CARCP01 Plumeria rubra Riau, Indonesia MK788144
a
Coleosporium
plumeriae
A2-CARCP02 Plumeria rubra Riau, Indonesia MK788146
a
Coleosporium
plumeriae
A3-CARCP03 Plumeria rubra Riau, Indonesia MK788147
a
Coleosporium
plumeriae
A4-CARCP04 Plumeria rubra Riau, Indonesia MK788148
a
Coleosporium
plumeriae
MCA3480 Plumeria sp. Sabah, Malaysia MF769628
Coleosporium
solidaginis
MCA3473 Solidago sp. Solidago, North
America
MF769632
Coleosporium
campanulae
IT7AG Campanula
rapuncoloides
Turin, Italy KY296542
Coleosporium
zanthoxyli
KUS-F29608 Zanthoxylum
planispinum
South Korea MH465095
Coleosporium
euodiae
–Tetradium
glabrifolium
P. R. China KP017557
Coleosporium
cimicifugatum
–Cimicifuga sp. P. R. China KP017559
Austropuccinia psidii UFV-9 Eucalyptus sp. Tasmania,
Australia
EF210142
a
Submitted in this study
34 Page 2 of 4 Australasian Plant Dis. Notes (2019) 14:34

bootstrap replications to increase accuracy on phylogenetic
tree topology. Phylogenetic analyses showed that four isolates
grouped on the same clade of Coleosporium plumeriae,with
high bootstrap support of 99% (Fig. 2).
Inoculations were performed on young plants of P. rubra.
Three month-old plants were transplanted into 3 L capacity
polybags containing a mixture of cocopeat (80%) and carbon-
ized rice husk (20%) and kept under growth chamber with
controlled temperature and humidity (25 °C / 80%). Two
months later, plants were inoculated by spraying an inoculum
suspension at 2 × 10
4
spores/mL in water containing 1%
Tween 20. Plants were covered with black plastic bags to
ensure darkness for the first 24 h. The plastic bags were then
removed and plants were kept for additional 72 h in the same
growth chamber with photoperiod of 12 h. Inoculated plants
were then moved to open area until observation of first symp-
toms. To complete the requirements of Koch’s postulate,
urediniospores were collected from inoculated plants and mor-
phological features were compared with those of the inocu-
lum. Typical rust symptoms were observed on inoculated
P. rubra plants while non-inoculated plants used as control
remained asymptomatic throughout the entire experiment.
Urediniospores collected from inoculated plants had the same
morphological features and DNA sequences as those original-
ly collected from naturally infected plants, fulfilling the
Koch’s postulates for obligate pathogens (biotrophic).
Observations of natural infections as well as inoculations
conducted under controlled conditions showed that P. rubra is
hightly susceptible to C. plumeriae isolates collected in
Pangkalan Kerinci, Riau, Sumatra. This pathogen has a very
specific host range, affecting species of the genus Plumeria,
but has also been reported on Catharanthus roseus, also in the
Apocynaceaee (Holcomb and Aime 2010). Host reactions
among Plumeria spp. and hybrids range from highly suscep-
tible to highly resistant (Nelson 2009).
Other than genetic resistance, integrated practices can be
applied to reduce new infections and consequently minimize
the disease impact. In this case, removal and elimination in-
fected leaves from underneath the canopy early in the season
combined with pruning of infected leaves still attached to the
branches are important practices to reduce the source of inoc-
ulum. Infected fallen leaves may serve as inoculum source
once the spores can survive on leaf debris and then spread
by the wind. Additionally, hyperparasites have been reported
infecting urediniospores of C. plumeriae (Manimohan and
Mannethody 2011) and potential biocontrol could function
as an alternative component of integrated management.
Chemical control is also an alternative (Nelson 2009). To
avoid the spread of C. plumeriae into disease free areas, re-
striction of the movement of infected plant material and quar-
antine measures should be considered.
Acknowledgements We thank Asia Pacific Resources International
Holdings Ltd. (APRIL) for financial assistance that made this study pos-
sible. We also thank Dr. MJ Wingfield whose suggestions helped improve
the submitted manuscript.
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