Mycoplasma/Phytoplama (PPLO) Diseases of Plants: Citrus greening, Rice yellow

dwarf: Little leaf of Brinjal, Sandal Spike.

There are some diseases which until 1967 were attributed to viruses, although no viral
particles had been found in the affected plants. Even so, given the observation and
detection methods available at the time, this was the most probable hypothesis.In 1967
it was conclusively shown that they were nothing to do with viruses.With the coming
of the electron microscope it was possible to identify some polymorphic structures
which were present in the phloem of the sick plants and not in that of healthy ones.
These structures had certain similarities to prokaryotic micro-organisms known as
mycoplasmas, which had long been recognized as agents of infection in animals or as
saprophytes on mucus.

These mycoplamsas are similar to bacteria except that they have no true cell wall. They
are generally assigned an intermediate place between bacteria and viruses.
Mycoplasmas are Eubacteria of the class Mollicutes, a group of organisms
phylogenetically related to Gram-positive bacteria. Their more characteristic features
reside in the small size of their genomes, the low guanine (G) plus cytosine (C) content
of their genomic DNA and the lack of a cell wall.

Plant pathogenic mycoplasmas are responsible for several hundred diseases and belong
to two groups: the phytoplasmas and the spiroplasmas. The phytoplasmas (previously
called MLOs, for mycoplasma like organisms) were discovered first; they are
pleiomorphic, and have so far resisted in vitro cultivation. Phytoplasmas represent the
largest group of plant pathogenic Mollicutes. Only three plant pathogenic spiroplasmas
are known today. Spiroplasma citri, the agent of citrus stubborn was discovered and
cultured in 1970 and shown to be helical and motile. S. kunkelii is the causal agent of
corn stunt. S. phoeniceum, responsible for periwinkle yellows, was discovered in Syria.
There are many other spiroplasmas associated with insects and ticks. Plant pathogenic
mycoplasmas are restricted to the phloem sieve tubes in which circulates the
photosynthetically-enriched sap, the food for many phloem-feeding insects (aphids,
leafhoppers, psyllids, etc.). Interestingly, phytopathogenic mycoplasmas are very
specifically transmitted by leafhoppers or psyllid species.

1. Citrus Greening

Citrus greening disease is a major cause of crop and tree loss in many parts of Asia and
Africa. The disease occurs in most citrus growing areas in the states of Jammu,
Kashmir, Punjab, P, Rajasthan and Madhya Pradesh.Before it was identified as one
disease, it became known by various names: yellow shoot (huanglungbin) in China;
likubin (decline) Taiwan; dieback in India; leaf mottle in the Philippines; vein phloem.
Losses due to greening are not easy to assess. Sometimes only sectors of a tree are
affected and losses are small, but in other cases the entire tree is infected and crop loss
is total. The disease is associated with tristeza virus.

In India and appears to be even more dangerous than tristeza virus. Some cases they
jointly caused citrus die back disease.

Symptoms

• Trees affected are generally stunted, have a sparce yellow foliage, and show
twig dieback.

• Sometimes, at early stages of infection, symptoms are seen only on one part
of the canopy, hence the name of the disease in China (yellow shoot).

• Due to shortening of internode the branches give a bushy appearance.

 • The most characteristic symptom is leaf mottle (normal-green patches on a
pale green background).

• This symptom is conspicuous on sweet orange leaves, but less prominent on

mandarin leaves.

• Leaf mottle is not specific of greening, as stubborn-affected leaves show

similar symptoms but it is the best foliar symptom for the diagnosis of greening.

• Zinc-deficiency-like symptoms are commonly associated with greening,

resulting in its confusion with nutritional problems.

• Scattered green areas are visible in yellow area of leaf lamina.

• Yellow area surrounded by midrib on one side and by lateral one on other sides.

• Leaf size is reduce and become thicker and more or less erect.

• Fruits on severely affected trees are small, lopsided and poorly colored, remaining dull
green, hence the name greening is given to the disease in South Africa

• Occasionally, color development on affected fruits starts at the peduncular

end rather than at the stylar end as it is the case in normal fruits.

• This symptom is referred to as color inversion or "red nose"! Variable

numbers of aborted seeds are present in fruits of affected sweet orange and mandarin
trees.
 Causal organism

• Lafleche and Bove were the first to show, by electron microscopy, that a
microorganism, not a virus, was present in the phloem sieve tubes of greening-affected
trees. They first thought that it was a mycoplasma-like organism (MLO) as MLOs
were known since 1967 to infect phloem of numerous plants. However in 1972, the
disease was reported to be caused by a phloem inhabiting fastidious bacterium, in
which cell wall appears more as a second membrane than a cell wall. The bacterium is
Gram negative , non motile , rigid, measuring 1.0-2.0 X 0.2-0.5 µm. The cell occurs in
mature sieve elements, irregularly distributed among vascular bundles. They are
sensitive to different antibiotics, relatively more to penicillin.

Transmission

Lean and Oberholzer demonstrated that greening disease was graft-transmissible, they
also established that the disease was naturally transmitted by the African citrus psylla,
Trioxa erytreae and Diaphorina citri, more prevalent in northernn and central India.
After acquiring agent, the vector remains infective for whole of its life. Incubation
period of bacterium is 8-12 days.

Control and Disease Management

• Physical Methods includes Heat Therapy, Heat therapy has long been used in
the field of plant disease management because high temperatures can cause cell lysis
and thus kill bacteria

• Healthy graft from disease free should be used for planting

• Using the seedlings which are acquired from diseased trees or polluted
breeding bases is responsible for the crazy spread of citrus greening.
• Stem tips culture: plant apex tissues containing little or no germs and viruses
are the ideal material to obtain pathogen-free seedlings

• So, building a citrus pathogen-free breeding system is important for

containing the transmission of HLB

• Diseased plant should be removed from the orchards .

• New plants should be raised only from indexed stock.

• Insect vectors should be eradicated by using inceticides like diazinon

(002%), endrin (0.02%), or parathion (0.02%)/

In Punjab full recovery of the affected trees could be achieved by spraying them with
a mixture of Bavistin and Ledermycin (500ppm each) six times at 10 days interval.

Since antibiotics were discovered, they have made a significant contribution to the
control of bacterial diseases in plants (Stockwell and Duffy, 2012).

Tetracycline has been widely used to treat HLB in various countries as early as the
1970s (Su and Chang, 1974). However, it was later replaced by other antibiotics, such
as penicillin, streptomycin, and terramycin, according to the poor results of
assessment for tetracycline

2. Rice yellow dwarf

Yellow dwarf disease of rice has been reported in most of the rice growing countries of
the East and South East Asia. In some areas of Japan and India, the disease has been
observed to cause serious damage to the crop. In recent years with continuous
cropping, with the release of varieties which are comparatively photo-insensitive, the
rice yellow dwarf disease is tending to attain economic importance in India.
 Symptoms
• The infected plants are stunted and have yellowish green to whitish green leaves.
There is excessive tillering and leaves became soft and droop slightly.
• Root growth is also reduced significantly.
• Chlorosis occurs on the leaves occasionally even spreading to the leaf sheaths.
• Streaks may also form parallel to the leaf veins.
• If plants are infected early they usually die before maturity, and even if they do
survive no panicles are produced or only a small number with no grains.
Causal organism
• Caused by a phytoplasma (rice yellow dwarf phytoplasma designated as a novel
taxon, ‘Candidatus Phytoplasma oryzae’).
• These are generally Observed in phloem tubes of yellow dwarf infected rice plants.
• These are pleomorphic bodies measured from 80-800 mm, devoid of cell walls and
bound by unit membrane.

Transmission
• The Mycoplasma Like Organism is transmitted by grasshopper Nephotettix
virescens and N. nigropictus with a latent period of 25-30 days.It survives on several
grass weeds.This virus can multiply in the body of a vector and can be transmitted
from a mother insect to her progeny by means of transvarial transmission.The insect
can acquire the pathogen by feeding on diseased plant by half an hour and require long
incubation period (20 days) to transmit the phytoplasma. The insects remain infected
until they die. A combination of low temperature and high humidity favours the
leafhopper population resulting in higher disease incidence.
Control and Disease Management
• Deep ploughing during summer and burning of stubbles.
• Rice varieties like IR62 and IR64 are resistant to the disease.
• The management practices followed for Rice Tungro disease may be adopted for this
disease also.
• Avoiding early-planted rice will prevent an increase in vector density and proportion of
infected vectors.
• Planting fallow rice fields with non-vector hosts, ploughing fallow paddy fields, and
late-planting, synchronous planting, or avoiding an over-lap of early- and late-planted
rice crops.
• Field sanitation
Seed treatment with Carbofuran and subsequent soaking in Carbofuran solution (75%
WP) for 36 hours prior to transplanting (by effectively controlling the vector)
• Spraying Dimecron (0.03%), Carbofuran (0.04%), Dimethoate (0.025%) and Methyl
Dometin (0.025%) Spraying of ch lorotetracycline at 100 ppm

3. Little leaf of Brinjal

Brinjal is the most common, popular and widely grown vegetable crop of both tropics
and subtropics of the world. Little leaf of brinjal or eggplant (Solanum melongena L.)
is one of the most serious diseases of brinjal in the areas of its cultivation. Occured in
almost all the states of India. This graft transmissible disease was first reported from
Coimbatore (Thomas and Krishnaswami, 1939).There is hardly any variety of Brinjal
which is resistance to this disease. Little leaf of brinjal is known to cause heavy
economic losses (more than 90%) in India.
Symptoms
• The main symptom of the disease is the production of very short leaves by
affected plant.
 • The petioles are so much reduced in size that leaves appear sticking to the stem.
Such leaves are narrow, soft, smooth and yellowish in colour.
• Newly formed leaves are further reduced in size.
• The internodes are shortened and at the same time large number of axillary buds
are stimulated to grow into short branches with small leaves.
• This gives whole plant a bushy appearance. Usually such plant unable to form
flowers. Fruiting is very rare.

Causal Organism

Verma et al (1969) showed MLO, spherical to ovoid(40- 3000nm)structures, without

cell wall, present in sieve tubes of the phloem cells of diseased plants, which lacking
cell wall. The disease is thus shown to be caused by Mycoplasma. The disease is
caused by a plant pathogenic mollicute, Phytoplasma, discovered in 1967 by Japnese
scientists (earlier known as mycoplasma like organism or MLO).

Transmission

In nature the disease is transmitted by insect Hishimonus phycitis, which belongs to the
group of leafhoppers. Bitter gourd, Ground gourd, Water melon, Menthi, Sunhemp and
Carrot are good hosts of the vectors, while Beet, Castor, Soyabean, Chenopodium are
suitable hosts for oviposition.

Control and Disease Management

 Spray insecticides- Ajaneyalu and Ramkrishnan (1969,1972b) and Verma et al. (1973)
reported tetracycline therapy at 1000 ppm3 times at 7 days interval to be effective in
controlling this disease.
 Spraying with ledermycin at 500 ppm has shown beneficial effects because it
suppresses the symptoms.
 Removal and destruction of weeds and infected plants -- Early destruction of infected
plants, uproot and burn infected plants and weeds also to control the disease.
 Spray insecticides to manage and control vector.
 Use of disease resistant varieties– as BB-7, BWR-12, Pant Ritu Raj& H8.
 Adopting sanitary measures including the eradication of susceptible volunteer crop
plants from a previous planting can reduce the damage.
 Use of barriers of trap crops and early removal and destruction of infected plants is
also recommended.
 The sowing time can be adjusted to avoid the main flights of the beet leafhopper.
 Spraying Malathion (2ml/litre of water) starting with the appearance of the leaf
hoppers controls their population

4. Sandal Spike
Sandal (Santalum album L.) is a semi parasitic, perennial tree belonging to the family
Santalaceae. It possesses a highly valuable wood, known for its scented oil and
carving. The oil found in the root and heartwood, is acknowledged as one of the most
precious perfumery items from antiquity down to the modern times. Sandalwood spike
disease (SSD) is likely to be the most destructive of known diseases to infect Indian
sandalwood, Santalum album L., which is valued for its wood and scented essential oil
derived from the heartwood.Sandal spike disease was first observed in the Reserve 51
of Marayoor forest range of Munnar forest division during June 1980. More than 50
per cent of the sandal trees in this reserve had been found to be affected with the
 disease. Spike disease of sandal is considered to be the most serious yellows-type
disease of forest trees known in the world today (Seliskar and Wilson, 1981).
• The disease was earlier known to be graft-transmissible (Coleman, 1917), caused by an
insect-borne virus (Anon., 1955). Recently. after the discovery of association of
Mycoplasma-like organisms (MLO) with some yellows-type of diseases (Doi et al.,
1967; Ishiie et al., 1967). mycoplasmal etiology of spike disease was confirmed by
Dijkstra and Ie (1969), Hull et al. (1969) and Varma et al. (1969) through electron
microscopic studies.
Symptoms
• The occurrence of spike disease was first reported by McCarthy from Coorg in 1899
(Barber, 1903).
• The characteristic symptoms are an extreme reduction in size of leaves and internodes,
accompanied by stiffering of leaves.
• In advance stage , the whole shoots appear like a spike type of inflorescence due to
progressive reduction in size of leaves and internodes.
• The shoot appear as spike , bearing four rows of fine bristles.
• Spike plants do not bears flower occasionally.
• Spiked trees usually die within one or two years, depending upon their age and the
prevalling environment condition.
• Beside reduction in size , the leaves may even become chlorotic, sometimes brwonish
followed by defoliation.
• The root become shrivelled and gradually disinegrate.
Causal organism
• It was earlier thought cause by virus, however in 1969, a group of workers observed
mycoplasma like bodies in the sieve tubes.
• Sensitivity of the causal agent to tetracyclines further confirmed the mycoplasma
etiology of the sandal spike disease.
• From TEM studeis , these bodies appears mostly round, and 40-750 nm in diameter.
• A few of these may also be avoid to ellipsoidal in shape.
• They do not have rigid cell wall and unit membrane aorund cytoplasm is 10 to 12 nm.
• MLO ( Mycoplasma like organisms) is the causal organism. MLO size in the host
ranges from 60-750 nm. The common form is ellipsoid, measuring 180-220 x 250-300
nm.
• The MLO is reported to infect a large number of plant species such as Eucalyptus
grandis, Vinca rosea, Zizyphus oenoplea, Dodonea viscosa
Transmission
• Vector transmissionn was not confirmed in spike disease, but recent studies also did
not give any conclutive evidence of vector transmission.
• The disease is transmitted through root contacts and insect vectors.
• In 1984 Indian researcher, suggested the trasnmisiion through an inscet Redarator
bimaculatus. But not yet confirmed.
• The disease can also be transmitted through Vinca rosea and vice versa through
dodder. The most common insect Vectors are Moonia alhimaculata and Nephotettix
virescence.

Disease Management
No specific method to control the disease has so far been developed. However, it is
claimed that planting of Mysore gum trees (ahybriod of Eucalyptus tereticornis) at a
distance of 10-20 metres from the Sandal trees keeps the latter free of infection.
• Spraying of chemical and antibiotic.
• The disease repairs after varying period of time.
• Thus , the most effective methods to eliminate SSD from sandalwood tissue are
likely to be the use of tissue culture to create disease-free superior genotypes.