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International Journal of Trend in Scientific Research and Development (IJTSRD)
Volume 6 Issue 6, September-October 2022 Available Online: www.ijtsrd.com e-ISSN: 2456 – 6470
@ IJTSRD | Unique Paper ID – IJTSRD51977 | Volume – 6 | Issue – 6 | September-October 2022 Page 867
Various Plant Diseases Caused by Xanthomonas Species
Dr. Dilip Kumar Sharma
Director, Vardhman Mahaveer Open University (VMOU), Kota, Rajasthan, India
ABSTRACT
Xanthomonas is a Gram-negative, aerobic, short rod-shaped
bacterium belonging to the family Pseudomonadaceae. This genus
includes several pathovars, which are mainly plant pathogens.
Extracellular proteases, and Type II secretion systems chiefly help
these bacteria in colonizing the host plant. It produces a characteristic
yellow pigment, xanthomonadin, which often is used as a
chemotaxonomic and diagnostic marker of this genus. Xanthan gum
produced by Xanthomonas is commercially important. It is an
important additive in the food industry. Recently, Xanthomonas was
also observed to undergo programmed cell death (PCD), the
morphological and biochemical features of which closely resemble
eukaryotic PCD. The determination of the genus Xanthomonas and
its species is relatively easy, however, the characterization of X.
campestris pathovars poses problems. An unambiguous identification
of the pathovars of X. campestris can be of great use in plant
pathology. The X. campestris pathovars that are defined by the host
or disease symptoms are difficult to identify by other phenotypic
characteristics. X. campestris group is the largest of all and causes
diseases in many plant species. It is, therefore, classified into
pathovars differentiated by the host reaction. However, application of
the newer techniques of classification has been useful. A relationship
of nutritional properties, host specificity and DNA homology groups
has been observed. Genetic diversity among the strains of different
pathovars of X. campestris has also been studied for a number of
pathovars. It is believed that the variability could be more
pronounced in the regions where the host plant originated. Ribosomal
RNA and DNA probes could be useful tools for the epidemiological
studies and in following the genetic evolution of the strains.
How to cite this paper: Dr. Dilip Kumar
Sharma "Various Plant Diseases Caused
by Xanthomonas Species" Published in
International
Journal of Trend in
Scientific Research
and Development
(ijtsrd), ISSN:
2456-6470,
Volume-6 | Issue-6,
October 2022,
pp.867-875, URL:
www.ijtsrd.com/papers/ijtsrd51977.pdf
Copyright © 2022 by author (s) and
International Journal of Trend in
Scientific Research and Development
Journal. This is an
Open Access article
distributed under the
terms of the Creative Commons
Attribution License (CC BY 4.0)
(http://creativecommons.org/licenses/by/4.0)
KEYWORDS: diseases, plants,
Xanthomonas, species, bacteria,
pathovars, identification
INTRODUCTION
Members of this genus are short Gram-negative rods
of linear shape, which are generally 0.4–0.7 µm wide
and 0.8–2 µm long. The bacteria exist singularly or in
pairs, and they are motile due to a single flagellum
(∼1.7–3.0 µm long). These bacteria have a GC
content of 63.3–69.7 mol.%. They are catalase
positive, urease, oxidase negative (or weakly
positive), nondenitrifying or nitrate reducers, and they
can produce H2S but not indole or acetoin. Their
growth is inhibited by 6% NaCl; 30% glucose; 0.01%
lead acetate, methyl green, or thionin; and by 0.1%
(and usually by 0.02%) triphenyl tetrazolium
chloride. Proteins are readily digested by these
bacteria and some species are able to hydrolyze
cellulose, pectin, starch, and Tween 80. They are
chemoorganotrophic, able to use various
carbohydrates and salts of organic acids as their sole
carbon source, and strictly aerobic (as they have
respiratory metabolism with oxygen as the terminal
electron acceptor)[1,2]
The main fatty acids found in cells of this genus are
9-methyl decanoic acid (C11:0 iso), 3-hydroxy-9-
methyl decanoic acid (C11:0 iso 3OH), and 3-
hydroxy-11-methyl dodecanoic acid (C13:0 iso 3OH).
This composition of fatty acids serves as a useful
criterion to differentiate Xanthomonas from other
bacteria. The optimal growth temperature for
Xanthomonas is 20–30°C depending on the species,
with the minimum temperature for growth being
>4°C and the maximum being 27.5–39°C. These
bacteria ideally grow at pH 6.5–7.5, but anything less
than pH 4.5 inhibits growth.
IJTSRD51977
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Xanthomonas metabolize glucose using the Entner–
Doudoroff pathway in conjunction with the
tricarboxylic acid (TCA) cycle pathway. The pentose
phosphate pathway may also be used but this only
accounts for a small portion (8%–16%) of the total
glucose consumed. The glyoxylate cycle may also be
used for substrate catabolism and energy production.
Most Xanthomonas species can grow in chemically
defined medium containing minerals, ammonium,
nitrogen, a suitable carbon source (i.e., glucose), and
amino acids (usually glutamate or methionine). They
also show satisfactory growth on nutrient agar (peptic
digest of animal tissue, sodium chloride, beef extract,
yeast extract, and agar at 5, 5, 1.5, 1.5, and 15 g/L,
respectively) with or without yeast extract
supplementation, GYCA medium (glucose, yeast
extract, CaCO3, and agar at 10, 5, 30, and 20 g/L,
respectively), GPPYA medium (glucose, proteose
peptone, yeast extract, and agar at 10, 5, 5, and 20
g/L, respectively), and YM agar (glucose, peptone,
yeast extract, and agar at 20, 5, 3, and 17 g/L,
respectively). Other culture media containing starch
may be used for the general growth of Xanthomonas.
These are usually fortified with antimicrobials, such
as cephalexin, kasugamycin, chlorothalonil,
gentamycin, brilliant cresyl blue, methyl green, and
methyl violet to make them selective for
Xanthomonas; Features that distinguish Xanthomonas
from other related bacteria [3,4] (i.e., Pseudomonas
and Enterobacteriaceae species) include their ability
to hydrolyze starch and their independence from
using asparagine as a source of carbon and nitrogen.
Culture media for xanthan production have been
widely studied and these may have different
characteristics to those intended for bacterial growth.
Xanthomonas strains capable of causing spoilage of
salad vegetables and fruits can grow on pectate agar
media commonly used for isolation of soft-rotting
Pseudomonas and Erwinia. Xanthomonas strains,
however, can be readily differentiated from
pseudomonads based on their ability to produce
yellow-pigmented xanthomonadin and mucoid
xantham gum. Like other xanthomonads, soft-rotting
Xanthomonas are unable to grow in the minimum
medium without the addition of organic supplements.
The xanthomonads are sensitive to those
antimicrobials incorporated in Pseudomonas selective
agars, for example triphenyl tetrazolium chloride and
others. For isolation of plant-pathogenic
Xanthomonas, a number of antimicrobials such as
cycloheximide methyl green and vancomycin are
frequently used. A number of selective agar media for
isolation of a specific species or pathovars of
Xanthomonas are available. It has not been
determined if these selective agars are suitable for
isolation of soft-rotting strains of xanthomonads.[5,6]
Several Xanthomonas species and pathovars attack
each of the cultivated cereals and wild grasses, and
some of them cause severe losses to their respective
hosts. The most common bacterial diseases of these
crops are bacterial stripe, bacterial blight, stripe, or
streak of several cereals
(X. campestris pv. translucens), bacterial leaf blight
of rice (X. oryzae pv. oryzae) bacterial leaf streak of
rice and leaf scald of sugarcane (X. albilineans). The
symptoms appear on leaf blades and sheaths as small,
linear, water-soaked areas that soon elongate and
coalesce into irregular, narrow, yellowish, or
brownish stripes. Droplets of white exudate are
common on the stripes. Severe infections cause leaves
to turn yellow and die from the tip downward, they
also retard spike elongation and cause blighting.
Small lesions form on the kernels as well. The
diseases develop mainly in rainy, damp weather.
Bacteria overwinter on the seed and in crop residue
and are spread by rain, direct contact, and insects. The
main control measures are use of disease-free or
treated seed and crop rotation.[7,8]
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Symptoms due to Xanthomonas
Discussion
Bacterial leaf spot-The disease is caused by Xanthomonas eleusinae Rangaswami, Prasad, Eswaran. Linear spots
are seen on both upper and lower surfaces of the leaf blade spreading along the veins. The spots measure 2−4
mm long, but often extend up to 25 mm or more. In the beginning, spots are light yellowish brown, but soon
become dark brown. At the advanced stage, the leaf splits along the streak giving a shredded appearance. All the
leaves, including the tender shoots, in a plant are affected. The bacterium mainly affects the leaves, but at times
characteristic streaks may be found on the peduncle.
Bacterial blight-Bacterial blight is caused by Xanthomonas oryzaepv. oryzae (Xoo). The symptoms of
bacterial blight can be seen during the seedling stage in the form of grayish-green rolled leaves which turn
yellow as the disease progresses. As the disease progresses further, the yellow color changes to straw-colored
wilt leaves and ultimately the death of whole seedlings. Bacterial blight mostly occurs in tropical and temperate
regions of the world, especially in irrigated and rainfed areas when strong winds are coupled with heavy rains.
Blight is the most destructive rice disease in Asia and resulted in a historic epidemic in India during the late
1970s[9,10]
Bacterial leaf blight (BLB), caused by Xanthomonas oryzae pv. oryzae (Xoo), is found in both tropical and
temperate regions. BLB also occurs in Australia, Africa, Latin America, the Caribbean and the USA. Yield
losses of 10%–50% from BLB have been reported. Outbreaks of BLB are most common during the monsoon
season in South-East Asia and India. Rice was introduced for cultivation into the USA (North Carolina) more
than 200 years ago and has been cultivated in other parts of the USA for over 100 years. Although many rice
diseases have either been introduced or developed on rice during the history of its cultivation in the USA, Xoo
has not established in the USA. The climates of rice-producing areas in the USA and USA rice cultivation
practices are not conducive to the long-term survival or spread of Xoo. For these reasons, Xoo is of low risk to
US agriculture.
BLB is efficiently controlled by the use of resistant rice cultivars. However, because Xoo has the capacity to
express effectors that suppress some host defence responses, often this resistance is eventually overcome.
Resistance genes of the non-RD pattern recognition receptor class typically confer long-lasting resistance
because they recognize conserved microbial signatures, which, when mutated, cripple the virulence of the
pathogen. Control of the disease with copper compounds, antibiotics and other chemicals has not proven to be
effective. [11,12]
Xanthomonas oryzae pv. oryzae is a rod-shaped, Gram-negative bacterium. It produces a yellow soluble
pigment, called xanthomonadin, and extracellular polysaccharide (EPS). EPS is important in the protection of
bacteria from desiccation and for the attenuation of wind- and rain-borne dispersal. Xoo is disseminated by
irrigation water systems, splashing or wind-blown rain, as well as by contaminated rice stubble from the
previous crop season, which is the most important source of primary inoculum. Xoo infects the rice leaf typically
through hydathodes at the leaf tip, broken trichomes, leaf margins and wounds in the leaves or roots, multiplies
in the intercellular spaces and enters into xylem vessels. Within a few days of infection, bacterial cells and EPS
fill the xylem vessels and ooze out from the hydathodes, and form beads of exudate on the leaf surface, a
characteristic sign of the disease and a source of secondary inoculum.
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The genus Xanthomonas currently consists of 20 species including axonopodis X. Six distinct genomic groups
have been defined within axonopodis X., with many pathovars causing economically important diseases on
different host plants of agronomic significance.
Cassava (Manihot esculenta) is the staple food of nearly 600 million people in the world's tropical regions.
Xanthomonas axonopodis pv. manihotis (Xam) is the causal agent of CBB, a major disease, endemic in tropical
and subtropical areas. This foliar and vascular disease severely affects cassava production worldwide. Losses of
between 12% and 100% affect both yield and planting material. Over recent years, a significant recurrence of the
disease has been reported in different regions in Africa and Asia. Xam induces a wide combination of symptoms,
including angular leaf lesions, blight, wilt, stem exudates and stem canker. Host resistance is still the most
effective way to control this disease.[13,14] However, no breeding strategy is being developed for the control of
CBB disease. Only two cassava CBB resistance genes have been identified so far (C. Lopez, personal
communication, Universidad Nacional, Bogota, Colombia). Plant defence responses to Xam have been well
characterized. Genomic tools for cassava, such as a large expressed sequence tag (EST) database and a cassava
microarray, have been developed and used for Xam–plant expression studies.
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Bacterial blight of cotton is a disease affecting the cotton plant resulting from infection by Xanthomonas
axonopodis pathovar malvacearum (Xcm) a Gram negative, motile rod-shaped, non-spore-forming bacterium
with a single polar flagellum. The bacteria can affect the cotton plant during all growth stages, infecting stems,
leaves, bracts and bolls. It causes seedling blight, leaf spot, blackarm (on stem and petioles), black vein and boll
rot. On cotyledons small, green, water-soaked rounded (or irregular) spots form which turn brown. Cotyledons
can be distorted if the infection is intense. Black and elongated lesions can girdle the hypocotyls and kill
seedlings. On the leaves, scattered small dark-green, water-soaked, areolate spots, form measuring 1–2 mm on
the lower surface, which appear translucent against transmitted light. The spots increase in diameter to 5 mm,
become angular (due to leaf veination), brown and later turn dark brown to blackish, becoming visible on the
upper surface. On susceptible cultivars numerous spots can occur, causing chlorosis, necrosis and distortion, and
eventually defoliation.[1]
the black arm symptom is characterized by dark brown to black lesions which may
coalesce and the extended necrosis then girdles the stem with blockage of the vascular system at infected sites
which hinder the movement of water and nutrients though the plant system. Drooping of leaves is associated
with this hindrance of water and nutrient movement. Cracking of the stem and gummosis is also observed in
infected plants.[15,16] The resultant breaking of the stem which hang typically as a dry black twig is referred to
as the ‘black arm’. The pathogen also reportedly causes blackening of the veins and veinlets, giving a typical
‘blighting’ appearance.
Xanthomonas campestris pv. vesicatoria is a bacterium that causes bacterial leaf spot (BLS) on peppers and
tomatoes. It is a gram-negative and rod-shaped. Xanthomonas campestris pv. vesicatoria survives on tomato and
pepper plants, seeds, and debris from infected plants as it cannot live in the soil for more than a few weeks. The
bacterium can also be found in association with wheat roots and some weed species which are both considered
sources of inoculum as well as diseased tomato and pepper plants. In cold climates, Xanthomonas campestris pv.
vesicatoria infection is mostly caused by contaminated seed material, both on and inside of seeds. If it survives
on seeds, it will infect the cotyledons of the growing plant as it emerges from the seed coat. Internally infected
seeds will produce diseased plants from the point of germination. Systemic symptoms such as wilting,
yellowing, and dwarfing are not typical of plants infected at the point of germination. However, foliage loss can
happen when localized symptoms on leaves become severe. If the bacterium survives on debris, it may infect
healthy plants through stomata as well as wounds on leaves and fruit. It is spread by direct contact of plants with
debris, human movement of the bacteria from debris to plants, and can easily travel from debris to healthy plants
through saturated soils via water movement. Once infected, plants begin to develop lesions on the leaves as well
as fruit, becoming inoculum sources for further infection.[17,18]
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Results
Contaminated seeds, weeds, infected plant debris are the main route of transmission. Infection starts with
epiphytic stage – i.e. bacteria grow on the aerial tissues of plant host (leaf, fruit, etc) followed by endophytic
stage when bacteria enter and colonise host tissues through wounds or natural openings. When population of
bacteria increases it re-emerges to the surface and is transmitted mainly by wind, rain or through seeds or
agricultural machinery, while animal and insect vectors seems to play minor role.
Xanthomonas uses surface polysacharides, adhesion proteins and type IV pili to attach to the surface and can
form biofilms to sustain abiotic stresses (UV, drought, etc). Xanthomonas produce xanthomonadins - yellow
pigments that protect from radiation caused from natural light. Resistance to UV is mostly conferred by genes
related to oxidative stress and DNA repair. Response to light is important in pathogenicity of these bacteria and
regulates surface attachment and production of biofilm.
Xanthomonas possess almost all known secretion systems (types I to VI) that play different roles in the life and
disease cycle, with type III secretion system (T3SS) being the key factor of pathogenicity. Typically,
Xanthomonas T3SS injects a cocktail of 20-30 effector proteins that interfere with plant immune system and
various host cellular processes. Many of the effectors are presumably redundant as individual deletions of
effector genes does not impair virulence, however mutations in T3SS apparatus has strong effect. Secretion of
the effectors is coordinated with expression of other virulence factors via shared regulatory networks. The
effector repertoire has been proposed to be a determinant of host specificity. Xanthomonas actively kill other
bacterial using type IV secretion system and defend itself from amoeba using type VI secretion system. [19]
To prevent infections, limiting the introduction of the bacteria is key. Some resistant cultivars of certain plant
species are available as this may be the most economical means for controlling this disease. For chemical
control, preventative applications are best to reduce the potential for bacterial development. Copper-containing
products offer some protection along with field-grade antibiotics such as oxytetracycline, which is labeled for
use on some food crops in the United States. Curative applications of chemical pesticides may slow or reduce the
spread of the bacterium, but will not cure already diseased plants. It is important to consult chemical pesticide
labels when attempting to control bacterial diseases, as different Xanthomonas species can have different
responses to these applications. Over-reliance on chemical control methods can also result in the selection of
resistant isolates, so these applications should be considered a last resort.
Potential use of bacteriophages is also considered, however major limiting factors are their sensitivity to
environmental conditions and in particular to UV radiation. Plant beneficial microorganisms or attenuated strains
of Xanthomonas are being tested as a biocontrol reasoning that they could compete by occupying the same niche
and even eradicate pathogenic strain. Generation of plant species resistant to Xanthomonas is another potential
strategy.
Xanthomonas species produce an edible polysaccharide called xanthan gum that has a wide range of industrial
uses, including foods, petroleum products, and cosmetics. Xanthan also plays role in the disease cycle of
Xanthomonas. In particular, xanthan gum is one of the main components of biofilm matrix. Biofilms help these
bacteria sustain abiotic stresses on the leaf surface. Genes for Xanthan gum biosynthesis comprise the gum
operon (gumB-gymM) coding for 12 enzymes. Xanthan production by Xanthomonas spp. that thrive in vascular
plant systems might block the water flow of the plant and as a result cause wilting.
Xanthomonas translucens on SPA media. Notice the yellow pigment
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Xanthomonas spp. life cycle
Conclusions
Phytopathogenic bacteria are economically important
because they affect crop yields and threaten the
livelihoods of farmers worldwide. The genus
Xanthomonas is particularly significant because it is
associated with some plant diseases that cause
tremendous loss in yields of globally essential crops.
Current management practices are ineffective,
unsustainable and harmful to natural ecosystems.
Bacteriophage (phage) biocontrol for plant disease
management has been of particular interest from the
early nineteenth century to date. Xanthomonas phage
research for plant disease management continues to
demonstrate promising results under laboratory and
field conditions.[20,21] AgriPhage has developed
phage products for the control of Xanthomonas
campestris pv. vesicatoria and Xanthomonas citri
subsp. citri. These are causative agents for tomato,
pepper spot and speck disease as well as citrus canker
disease.Phage-mediated biocontrol is becoming a
viable option because phages occur naturally and are
safe for disease control and management. Thorough
knowledge of biological characteristics of
Xanthomonas phages is vital for developing effective
biocontrol products. Several Xanthomonas phages are
evaluated for their potential as biocontrol agents
against Xanthomonas species. So far, most of these
belong to order Caudovirales and are lytic to a broad
range of host strains. They are isolated from diverse
ecosystems and distributed across the globe
depending on the presence of the pathogen they
infect. Their structural integrity and functionality in in
vitro conditions is maintained under optimal growth
and storage conditions. Pathogenesis of Xanthomonas
phages in bacteria induce molecular alterations that
may have regulatory functions important during their
life cycle. Although few studies have focused on this
aspect of biology, more research is needed to
understand their life cycle.
From their first discovery in filtrates to applications
as phage/pathogen suspensions, or in combination
with other antimicrobials or with UV-protectants or
as cocktail/monophage treatments, phages have
proved to be promising alternatives to agrochemicals
and antibiotics. They can reduce disease severity or
inhibit bacteria growth in diverse field settings. So
far, two Xanthomonas phage-based biocontrol
products are commercially available for plant disease
control. As the transition into commercial products
continues, more studies are needed to tap into the
many unexploited potentials of Xanthomonas phages
for a range of Xanthomonas related plant
diseases.[22]
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PLANT DISEASE BACTERIA

  • 1. International Journal of Trend in Scientific Research and Development (IJTSRD) Volume 6 Issue 6, September-October 2022 Available Online: www.ijtsrd.com e-ISSN: 2456 – 6470 @ IJTSRD | Unique Paper ID – IJTSRD51977 | Volume – 6 | Issue – 6 | September-October 2022 Page 867 Various Plant Diseases Caused by Xanthomonas Species Dr. Dilip Kumar Sharma Director, Vardhman Mahaveer Open University (VMOU), Kota, Rajasthan, India ABSTRACT Xanthomonas is a Gram-negative, aerobic, short rod-shaped bacterium belonging to the family Pseudomonadaceae. This genus includes several pathovars, which are mainly plant pathogens. Extracellular proteases, and Type II secretion systems chiefly help these bacteria in colonizing the host plant. It produces a characteristic yellow pigment, xanthomonadin, which often is used as a chemotaxonomic and diagnostic marker of this genus. Xanthan gum produced by Xanthomonas is commercially important. It is an important additive in the food industry. Recently, Xanthomonas was also observed to undergo programmed cell death (PCD), the morphological and biochemical features of which closely resemble eukaryotic PCD. The determination of the genus Xanthomonas and its species is relatively easy, however, the characterization of X. campestris pathovars poses problems. An unambiguous identification of the pathovars of X. campestris can be of great use in plant pathology. The X. campestris pathovars that are defined by the host or disease symptoms are difficult to identify by other phenotypic characteristics. X. campestris group is the largest of all and causes diseases in many plant species. It is, therefore, classified into pathovars differentiated by the host reaction. However, application of the newer techniques of classification has been useful. A relationship of nutritional properties, host specificity and DNA homology groups has been observed. Genetic diversity among the strains of different pathovars of X. campestris has also been studied for a number of pathovars. It is believed that the variability could be more pronounced in the regions where the host plant originated. Ribosomal RNA and DNA probes could be useful tools for the epidemiological studies and in following the genetic evolution of the strains. How to cite this paper: Dr. Dilip Kumar Sharma "Various Plant Diseases Caused by Xanthomonas Species" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-6 | Issue-6, October 2022, pp.867-875, URL: www.ijtsrd.com/papers/ijtsrd51977.pdf Copyright © 2022 by author (s) and International Journal of Trend in Scientific Research and Development Journal. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0) (http://creativecommons.org/licenses/by/4.0) KEYWORDS: diseases, plants, Xanthomonas, species, bacteria, pathovars, identification INTRODUCTION Members of this genus are short Gram-negative rods of linear shape, which are generally 0.4–0.7 µm wide and 0.8–2 µm long. The bacteria exist singularly or in pairs, and they are motile due to a single flagellum (∼1.7–3.0 µm long). These bacteria have a GC content of 63.3–69.7 mol.%. They are catalase positive, urease, oxidase negative (or weakly positive), nondenitrifying or nitrate reducers, and they can produce H2S but not indole or acetoin. Their growth is inhibited by 6% NaCl; 30% glucose; 0.01% lead acetate, methyl green, or thionin; and by 0.1% (and usually by 0.02%) triphenyl tetrazolium chloride. Proteins are readily digested by these bacteria and some species are able to hydrolyze cellulose, pectin, starch, and Tween 80. They are chemoorganotrophic, able to use various carbohydrates and salts of organic acids as their sole carbon source, and strictly aerobic (as they have respiratory metabolism with oxygen as the terminal electron acceptor)[1,2] The main fatty acids found in cells of this genus are 9-methyl decanoic acid (C11:0 iso), 3-hydroxy-9- methyl decanoic acid (C11:0 iso 3OH), and 3- hydroxy-11-methyl dodecanoic acid (C13:0 iso 3OH). This composition of fatty acids serves as a useful criterion to differentiate Xanthomonas from other bacteria. The optimal growth temperature for Xanthomonas is 20–30°C depending on the species, with the minimum temperature for growth being >4°C and the maximum being 27.5–39°C. These bacteria ideally grow at pH 6.5–7.5, but anything less than pH 4.5 inhibits growth. IJTSRD51977
  • 2. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD51977 | Volume – 6 | Issue – 6 | September-October 2022 Page 868 Xanthomonas metabolize glucose using the Entner– Doudoroff pathway in conjunction with the tricarboxylic acid (TCA) cycle pathway. The pentose phosphate pathway may also be used but this only accounts for a small portion (8%–16%) of the total glucose consumed. The glyoxylate cycle may also be used for substrate catabolism and energy production. Most Xanthomonas species can grow in chemically defined medium containing minerals, ammonium, nitrogen, a suitable carbon source (i.e., glucose), and amino acids (usually glutamate or methionine). They also show satisfactory growth on nutrient agar (peptic digest of animal tissue, sodium chloride, beef extract, yeast extract, and agar at 5, 5, 1.5, 1.5, and 15 g/L, respectively) with or without yeast extract supplementation, GYCA medium (glucose, yeast extract, CaCO3, and agar at 10, 5, 30, and 20 g/L, respectively), GPPYA medium (glucose, proteose peptone, yeast extract, and agar at 10, 5, 5, and 20 g/L, respectively), and YM agar (glucose, peptone, yeast extract, and agar at 20, 5, 3, and 17 g/L, respectively). Other culture media containing starch may be used for the general growth of Xanthomonas. These are usually fortified with antimicrobials, such as cephalexin, kasugamycin, chlorothalonil, gentamycin, brilliant cresyl blue, methyl green, and methyl violet to make them selective for Xanthomonas; Features that distinguish Xanthomonas from other related bacteria [3,4] (i.e., Pseudomonas and Enterobacteriaceae species) include their ability to hydrolyze starch and their independence from using asparagine as a source of carbon and nitrogen. Culture media for xanthan production have been widely studied and these may have different characteristics to those intended for bacterial growth. Xanthomonas strains capable of causing spoilage of salad vegetables and fruits can grow on pectate agar media commonly used for isolation of soft-rotting Pseudomonas and Erwinia. Xanthomonas strains, however, can be readily differentiated from pseudomonads based on their ability to produce yellow-pigmented xanthomonadin and mucoid xantham gum. Like other xanthomonads, soft-rotting Xanthomonas are unable to grow in the minimum medium without the addition of organic supplements. The xanthomonads are sensitive to those antimicrobials incorporated in Pseudomonas selective agars, for example triphenyl tetrazolium chloride and others. For isolation of plant-pathogenic Xanthomonas, a number of antimicrobials such as cycloheximide methyl green and vancomycin are frequently used. A number of selective agar media for isolation of a specific species or pathovars of Xanthomonas are available. It has not been determined if these selective agars are suitable for isolation of soft-rotting strains of xanthomonads.[5,6] Several Xanthomonas species and pathovars attack each of the cultivated cereals and wild grasses, and some of them cause severe losses to their respective hosts. The most common bacterial diseases of these crops are bacterial stripe, bacterial blight, stripe, or streak of several cereals (X. campestris pv. translucens), bacterial leaf blight of rice (X. oryzae pv. oryzae) bacterial leaf streak of rice and leaf scald of sugarcane (X. albilineans). The symptoms appear on leaf blades and sheaths as small, linear, water-soaked areas that soon elongate and coalesce into irregular, narrow, yellowish, or brownish stripes. Droplets of white exudate are common on the stripes. Severe infections cause leaves to turn yellow and die from the tip downward, they also retard spike elongation and cause blighting. Small lesions form on the kernels as well. The diseases develop mainly in rainy, damp weather. Bacteria overwinter on the seed and in crop residue and are spread by rain, direct contact, and insects. The main control measures are use of disease-free or treated seed and crop rotation.[7,8]
  • 3. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD51977 | Volume – 6 | Issue – 6 | September-October 2022 Page 869 Symptoms due to Xanthomonas Discussion Bacterial leaf spot-The disease is caused by Xanthomonas eleusinae Rangaswami, Prasad, Eswaran. Linear spots are seen on both upper and lower surfaces of the leaf blade spreading along the veins. The spots measure 2−4 mm long, but often extend up to 25 mm or more. In the beginning, spots are light yellowish brown, but soon become dark brown. At the advanced stage, the leaf splits along the streak giving a shredded appearance. All the leaves, including the tender shoots, in a plant are affected. The bacterium mainly affects the leaves, but at times characteristic streaks may be found on the peduncle. Bacterial blight-Bacterial blight is caused by Xanthomonas oryzaepv. oryzae (Xoo). The symptoms of bacterial blight can be seen during the seedling stage in the form of grayish-green rolled leaves which turn yellow as the disease progresses. As the disease progresses further, the yellow color changes to straw-colored wilt leaves and ultimately the death of whole seedlings. Bacterial blight mostly occurs in tropical and temperate regions of the world, especially in irrigated and rainfed areas when strong winds are coupled with heavy rains. Blight is the most destructive rice disease in Asia and resulted in a historic epidemic in India during the late 1970s[9,10] Bacterial leaf blight (BLB), caused by Xanthomonas oryzae pv. oryzae (Xoo), is found in both tropical and temperate regions. BLB also occurs in Australia, Africa, Latin America, the Caribbean and the USA. Yield losses of 10%–50% from BLB have been reported. Outbreaks of BLB are most common during the monsoon season in South-East Asia and India. Rice was introduced for cultivation into the USA (North Carolina) more than 200 years ago and has been cultivated in other parts of the USA for over 100 years. Although many rice diseases have either been introduced or developed on rice during the history of its cultivation in the USA, Xoo has not established in the USA. The climates of rice-producing areas in the USA and USA rice cultivation practices are not conducive to the long-term survival or spread of Xoo. For these reasons, Xoo is of low risk to US agriculture. BLB is efficiently controlled by the use of resistant rice cultivars. However, because Xoo has the capacity to express effectors that suppress some host defence responses, often this resistance is eventually overcome. Resistance genes of the non-RD pattern recognition receptor class typically confer long-lasting resistance because they recognize conserved microbial signatures, which, when mutated, cripple the virulence of the pathogen. Control of the disease with copper compounds, antibiotics and other chemicals has not proven to be effective. [11,12] Xanthomonas oryzae pv. oryzae is a rod-shaped, Gram-negative bacterium. It produces a yellow soluble pigment, called xanthomonadin, and extracellular polysaccharide (EPS). EPS is important in the protection of bacteria from desiccation and for the attenuation of wind- and rain-borne dispersal. Xoo is disseminated by irrigation water systems, splashing or wind-blown rain, as well as by contaminated rice stubble from the previous crop season, which is the most important source of primary inoculum. Xoo infects the rice leaf typically through hydathodes at the leaf tip, broken trichomes, leaf margins and wounds in the leaves or roots, multiplies in the intercellular spaces and enters into xylem vessels. Within a few days of infection, bacterial cells and EPS fill the xylem vessels and ooze out from the hydathodes, and form beads of exudate on the leaf surface, a characteristic sign of the disease and a source of secondary inoculum.
  • 4. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD51977 | Volume – 6 | Issue – 6 | September-October 2022 Page 870 The genus Xanthomonas currently consists of 20 species including axonopodis X. Six distinct genomic groups have been defined within axonopodis X., with many pathovars causing economically important diseases on different host plants of agronomic significance. Cassava (Manihot esculenta) is the staple food of nearly 600 million people in the world's tropical regions. Xanthomonas axonopodis pv. manihotis (Xam) is the causal agent of CBB, a major disease, endemic in tropical and subtropical areas. This foliar and vascular disease severely affects cassava production worldwide. Losses of between 12% and 100% affect both yield and planting material. Over recent years, a significant recurrence of the disease has been reported in different regions in Africa and Asia. Xam induces a wide combination of symptoms, including angular leaf lesions, blight, wilt, stem exudates and stem canker. Host resistance is still the most effective way to control this disease.[13,14] However, no breeding strategy is being developed for the control of CBB disease. Only two cassava CBB resistance genes have been identified so far (C. Lopez, personal communication, Universidad Nacional, Bogota, Colombia). Plant defence responses to Xam have been well characterized. Genomic tools for cassava, such as a large expressed sequence tag (EST) database and a cassava microarray, have been developed and used for Xam–plant expression studies.
  • 5. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD51977 | Volume – 6 | Issue – 6 | September-October 2022 Page 871 Bacterial blight of cotton is a disease affecting the cotton plant resulting from infection by Xanthomonas axonopodis pathovar malvacearum (Xcm) a Gram negative, motile rod-shaped, non-spore-forming bacterium with a single polar flagellum. The bacteria can affect the cotton plant during all growth stages, infecting stems, leaves, bracts and bolls. It causes seedling blight, leaf spot, blackarm (on stem and petioles), black vein and boll rot. On cotyledons small, green, water-soaked rounded (or irregular) spots form which turn brown. Cotyledons can be distorted if the infection is intense. Black and elongated lesions can girdle the hypocotyls and kill seedlings. On the leaves, scattered small dark-green, water-soaked, areolate spots, form measuring 1–2 mm on the lower surface, which appear translucent against transmitted light. The spots increase in diameter to 5 mm, become angular (due to leaf veination), brown and later turn dark brown to blackish, becoming visible on the upper surface. On susceptible cultivars numerous spots can occur, causing chlorosis, necrosis and distortion, and eventually defoliation.[1] the black arm symptom is characterized by dark brown to black lesions which may coalesce and the extended necrosis then girdles the stem with blockage of the vascular system at infected sites which hinder the movement of water and nutrients though the plant system. Drooping of leaves is associated with this hindrance of water and nutrient movement. Cracking of the stem and gummosis is also observed in infected plants.[15,16] The resultant breaking of the stem which hang typically as a dry black twig is referred to as the ‘black arm’. The pathogen also reportedly causes blackening of the veins and veinlets, giving a typical ‘blighting’ appearance. Xanthomonas campestris pv. vesicatoria is a bacterium that causes bacterial leaf spot (BLS) on peppers and tomatoes. It is a gram-negative and rod-shaped. Xanthomonas campestris pv. vesicatoria survives on tomato and pepper plants, seeds, and debris from infected plants as it cannot live in the soil for more than a few weeks. The bacterium can also be found in association with wheat roots and some weed species which are both considered sources of inoculum as well as diseased tomato and pepper plants. In cold climates, Xanthomonas campestris pv. vesicatoria infection is mostly caused by contaminated seed material, both on and inside of seeds. If it survives on seeds, it will infect the cotyledons of the growing plant as it emerges from the seed coat. Internally infected seeds will produce diseased plants from the point of germination. Systemic symptoms such as wilting, yellowing, and dwarfing are not typical of plants infected at the point of germination. However, foliage loss can happen when localized symptoms on leaves become severe. If the bacterium survives on debris, it may infect healthy plants through stomata as well as wounds on leaves and fruit. It is spread by direct contact of plants with debris, human movement of the bacteria from debris to plants, and can easily travel from debris to healthy plants through saturated soils via water movement. Once infected, plants begin to develop lesions on the leaves as well as fruit, becoming inoculum sources for further infection.[17,18]
  • 6. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD51977 | Volume – 6 | Issue – 6 | September-October 2022 Page 872 Results Contaminated seeds, weeds, infected plant debris are the main route of transmission. Infection starts with epiphytic stage – i.e. bacteria grow on the aerial tissues of plant host (leaf, fruit, etc) followed by endophytic stage when bacteria enter and colonise host tissues through wounds or natural openings. When population of bacteria increases it re-emerges to the surface and is transmitted mainly by wind, rain or through seeds or agricultural machinery, while animal and insect vectors seems to play minor role. Xanthomonas uses surface polysacharides, adhesion proteins and type IV pili to attach to the surface and can form biofilms to sustain abiotic stresses (UV, drought, etc). Xanthomonas produce xanthomonadins - yellow pigments that protect from radiation caused from natural light. Resistance to UV is mostly conferred by genes related to oxidative stress and DNA repair. Response to light is important in pathogenicity of these bacteria and regulates surface attachment and production of biofilm. Xanthomonas possess almost all known secretion systems (types I to VI) that play different roles in the life and disease cycle, with type III secretion system (T3SS) being the key factor of pathogenicity. Typically, Xanthomonas T3SS injects a cocktail of 20-30 effector proteins that interfere with plant immune system and various host cellular processes. Many of the effectors are presumably redundant as individual deletions of effector genes does not impair virulence, however mutations in T3SS apparatus has strong effect. Secretion of the effectors is coordinated with expression of other virulence factors via shared regulatory networks. The effector repertoire has been proposed to be a determinant of host specificity. Xanthomonas actively kill other bacterial using type IV secretion system and defend itself from amoeba using type VI secretion system. [19] To prevent infections, limiting the introduction of the bacteria is key. Some resistant cultivars of certain plant species are available as this may be the most economical means for controlling this disease. For chemical control, preventative applications are best to reduce the potential for bacterial development. Copper-containing products offer some protection along with field-grade antibiotics such as oxytetracycline, which is labeled for use on some food crops in the United States. Curative applications of chemical pesticides may slow or reduce the spread of the bacterium, but will not cure already diseased plants. It is important to consult chemical pesticide labels when attempting to control bacterial diseases, as different Xanthomonas species can have different responses to these applications. Over-reliance on chemical control methods can also result in the selection of resistant isolates, so these applications should be considered a last resort. Potential use of bacteriophages is also considered, however major limiting factors are their sensitivity to environmental conditions and in particular to UV radiation. Plant beneficial microorganisms or attenuated strains of Xanthomonas are being tested as a biocontrol reasoning that they could compete by occupying the same niche and even eradicate pathogenic strain. Generation of plant species resistant to Xanthomonas is another potential strategy. Xanthomonas species produce an edible polysaccharide called xanthan gum that has a wide range of industrial uses, including foods, petroleum products, and cosmetics. Xanthan also plays role in the disease cycle of Xanthomonas. In particular, xanthan gum is one of the main components of biofilm matrix. Biofilms help these bacteria sustain abiotic stresses on the leaf surface. Genes for Xanthan gum biosynthesis comprise the gum operon (gumB-gymM) coding for 12 enzymes. Xanthan production by Xanthomonas spp. that thrive in vascular plant systems might block the water flow of the plant and as a result cause wilting. Xanthomonas translucens on SPA media. Notice the yellow pigment
  • 7. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD51977 | Volume – 6 | Issue – 6 | September-October 2022 Page 873 Xanthomonas spp. life cycle Conclusions Phytopathogenic bacteria are economically important because they affect crop yields and threaten the livelihoods of farmers worldwide. The genus Xanthomonas is particularly significant because it is associated with some plant diseases that cause tremendous loss in yields of globally essential crops. Current management practices are ineffective, unsustainable and harmful to natural ecosystems. Bacteriophage (phage) biocontrol for plant disease management has been of particular interest from the early nineteenth century to date. Xanthomonas phage research for plant disease management continues to demonstrate promising results under laboratory and field conditions.[20,21] AgriPhage has developed phage products for the control of Xanthomonas campestris pv. vesicatoria and Xanthomonas citri subsp. citri. These are causative agents for tomato, pepper spot and speck disease as well as citrus canker disease.Phage-mediated biocontrol is becoming a viable option because phages occur naturally and are safe for disease control and management. Thorough knowledge of biological characteristics of Xanthomonas phages is vital for developing effective biocontrol products. Several Xanthomonas phages are evaluated for their potential as biocontrol agents against Xanthomonas species. So far, most of these belong to order Caudovirales and are lytic to a broad range of host strains. They are isolated from diverse ecosystems and distributed across the globe depending on the presence of the pathogen they infect. Their structural integrity and functionality in in vitro conditions is maintained under optimal growth and storage conditions. Pathogenesis of Xanthomonas phages in bacteria induce molecular alterations that may have regulatory functions important during their life cycle. Although few studies have focused on this aspect of biology, more research is needed to understand their life cycle. From their first discovery in filtrates to applications as phage/pathogen suspensions, or in combination with other antimicrobials or with UV-protectants or as cocktail/monophage treatments, phages have proved to be promising alternatives to agrochemicals and antibiotics. They can reduce disease severity or inhibit bacteria growth in diverse field settings. So far, two Xanthomonas phage-based biocontrol products are commercially available for plant disease control. As the transition into commercial products continues, more studies are needed to tap into the many unexploited potentials of Xanthomonas phages for a range of Xanthomonas related plant diseases.[22]
  • 8. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD51977 | Volume – 6 | Issue – 6 | September-October 2022 Page 874 References [1] An SQ, Potnis N, Dow M, Vorhölter FJ, He YQ, Becker A, et al. (October 2019). "Mechanistic insights into host adaptation, virulence and epidemiology of the phytopathogen Xanthomonas". FEMS Microbiology Reviews. 44 (1): 1–32. doi:10.1093/femsre/fuz024. PMC 8042644. PMID 31578554. [2] Doidge EM (1921). "A tomato canker". Annals of Applied Biology. 7 (4): 407–30. doi:10.1111/j.1744-7348.1921.tb05528.x. [3] Dowson WJ (1939). "On the systematic position and generic names of the Gram- negative bacterial plant pathogens". Zentralblatt für Bakteriologie, Parasitenkunde, Infektionskrankheiten und Hygiene: 177–193. [4] Young JM, Dye DW, Bradbury JF, Panagopoulos CG, Robbs CF (1978). "A proposed nomenclature and classification for plant pathogenic bacteria". New Zealand Journal of Agricultural Research. 21 (1): 153– 177. doi:10.1080/00288233.1978.10427397. ISSN 0028-8233. [5] Stall RE, Beaulieu C, Egel DS (1994). "Two genetically diverse groups of strains are included in Xanthomonas campestris pv. vesicatoria". Int J Syst Bacteriol. 44: 47–53. doi:10.1099/00207713-44-1-47. [6] Vauterin L, Swings J, Kersters K, Gillis M, Mew TW, Schroth MN, et al. (1990). "Towards an improved taxonomy of Xanthomonas". Int J Syst Bacteriol. 40 (3): 312–316. doi:10.1099/00207713-40-3-312. [7] Rademaker JL, Louws FJ, Schultz MH, Rossbach U, Vauterin L, Swings J, de Bruijn FJ (September 2005). "A comprehensive species to strain taxonomic framework for xanthomonas". Phytopathology. 95 (9): 1098–111. doi:10.1094/phyto-95-1098. PMID 18943308. [8] Vauterin L, Hoste B, Kersters K, Swings J (1995). "Reclassification of Xanthomonas". Int J Syst Evol Microbiol. 45 (3): 472–489. doi:10.1099/00207713-45-3-472. [9] Ah-You N, Gagnevin L, Grimont PA, Brisse S, Nesme X, Chiroleu F, et al. (February 2009). "Polyphasic characterization of xanthomonads pathogenic to members of the Anacardiaceae and their relatedness to species of Xanthomonas". International Journal of Systematic and Evolutionary Microbiology. 59 (Pt 2): 306–18. doi:10.1099/ijs.0.65453-0. PMID 19196770. [10] Young JM, Wilkie JP, Park DS, Watson DR (2010). "New Zealand strains of plant pathogenic bacteria classified by multi-locus sequence analysis; proposal of Xanthomonas dyei sp. nov". Plant Pathol. 59 (2): 270–281. doi:10.1111/j.1365-3059.2009.02210.x. [11] Rodriguez-R LM, Grajales A, Arrieta-Ortiz ML, Salazar C, Restrepo S, Bernal A (March 2012). "Genomes-based phylogeny of the genus Xanthomonas". BMC Microbiology. 12: 43. doi:10.1186/1471-2180-12-43. PMC 3359215. PMID 22443110. [12] Büttner D, Bonas U (March 2010). "Regulation and secretion of Xanthomonas virulence factors". FEMS Microbiology Reviews. 34 (2): 107–33. doi:10.1111/j.1574- 6976.2009.00192.x. PMID 19925633. [13] Studholme, David J.; Wicker, Emmanuel; Abrare, Sadik Muzemil; Aspin, Andrew; Bogdanove, Adam; Broders, Kirk; Dubrow, Zoe; Grant, Murray; Jones, Jeffrey B.; Karamura, Georgina; Lang, Jillian; Leach, Jan; Mahuku, George; Nakato, Gloria Valentine; Coutinho, Teresa; Smith, Julian; Bull, Carolee T. (2020). "Transfer of Xanthomonas campestris pv. arecae and X. campestris pv. musacearum to X. vasicola (Vauterin) as X. vasicola pv. arecae comb. nov. and X. vasicola pv. musacearum comb. nov. and Description of X. vasicola pv. vasculorum pv. nov". Phytopathology. American Phytopathological Society. 110 (6): 1153–1160. doi:10.1094/phyto-03-19-0098-le. ISSN 0031- 949X. PMID 31922946. [14] Schaad NW, Jones JB, Chun W (2001). "Laboratory Guide for Identification of Plant Pathogenic Bacteria 3rd Ed": 175–199. [15] Boch J, Bonas U (September 2010). "Xanthomonas AvrBs3 family-type III effectors: discovery and function". Annual Review of Phytopathology. 48: 419–36. doi:10.1146/annurev-phyto-080508-081936. PMID 19400638. [16] Ritchie DF (2000). "Bacterial spot of pepper and tomato". The Plant Health Instructor. doi:10.1094/PHI-I-2000-1027-01. [17] Mew TW, Alvarez AM, Leach JE, Swings J (1993). "Focus on bacterial blight of rice". Plant Disease. 77: 5–13. doi:10.1094/pd-77- 0005.
  • 9. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD51977 | Volume – 6 | Issue – 6 | September-October 2022 Page 875 [18] Hajri A, Brin C, Hunault G, Lardeux F, Lemaire C, Manceau C, et al. (August 2009). Yang CH (ed.). "A "repertoire for repertoire" hypothesis: repertoires of type three effectors are candidate determinants of host specificityin Xanthomonas". PLOS ONE. 4 (8): e6632. Bibcode:2009PLoSO...4.6632H. doi:10.1371/journal.pone.0006632. PMC 2722093. PMID 19680562. [19] Souza DP, Oka GU, Alvarez-Martinez CE, Bisson-Filho AW, Dunger G, Hobeika L, et al. (March 2015). "Bacterial killing via a type IV secretion system". Nature Communications. 6 (1): 6453. Bibcode: 2015NatCo...6.6453S. doi:10.1038/ncomms7453. PMID 25743609. [20] Bayer-Santos E, Lima LD, Ceseti LM, Ratagami CY, de Santana ES, da Silva AM, et al. (April 2018). "Xanthomonas citri T6SS mediates resistance to Dictyostelium predation and is regulated by an ECF σ factor and cognate Ser/Thr kinase". Environmental Microbiology. 20 (4): 1562–1575. doi:10.1111/1462-2920.14085. PMID 29488354. S2CID 3579518. [21] Ellis SD, Boehm MJ, Coplin D (2008). "Bacterial Diseases of Plants". Ohio State University Fact Sheet. [22] Koebnik R (7 August 2012), The Xanthomonas Resource