An Assignment on

Bacterial diseases of crops in Nepal

Submitted by: Submitted to:

Mr. Prem Pandey Mr. Hira kaji Manandhar, Phd
PLP-07M-2016 Department of plant pathology
Department of plant pathology AFU, Rampur, Chitwan
AFU, Rampur, Chitwan, Nepal Nepal.
2016
 Summary
Bacterial diseases are major bottlenecked problem for agricultural crops losses
in Nepal. Reviewing various articles and research papers related to bacterial
diseases in Nepal is to find actual status of bacterial diseases in Nepal and their
possible management. Cereals like rice, maize and wheat are affected by severe
bacterial diseases like bacterial leaf blight of rice (Xanthomonas oryzae pv.
oryzae ) accounting 5-60% yield in terai and mid-hills during hot and humid
periods, Bacterial leaf streaks of rice (Xanthomonas campestris pv. oryzicola )
causing yield loss from 1 to 17% depending on the cultivar and the climatic
conditions , bacterial Sheath brown rot (Pseudomonas fuscovaginae), Bacterial
stalk rot of maize (Erwinia chrysanthemi pv. Zeae ) extended to some pockets
of the mountains and Kathmandu valley, Bacterial leaf stripe of maize
(Xanthomonas rubrilineans ) its incidence has been extremely low in case of
Nepal, Stewart wilt of maize (Erwinia stewartii ) occur in warmer region
causing negligible loss. The bacterial diseases causing vegetables are bacterial
wilt of vegetables (Pseudomonas solanacearum) causing yield loss of about 5-
70% , Soft rot of vegetable is (Erwinia carotovora ), bacterial speck of tomato
(Pseudomonas syringae pv. tomato), Black rot caused by Xanthomonas
campestris pv. campestris causing 24.5% infection in local cauliflower and 39
% infection in mustard seeds, bacterial spot on tomato (Xanthomonas
campestris pv.vesicatoria ) reported 50-60% loss, brown rot of potato
(Pseudomonas solanacearum ), two bacterial diseases of soybean; bacterial
postule (Xanthomonas campestris pv glycines) and seed decay (Bacillus
subtilis) and bacterial diseases causing on fruits in Nepal are citrus greening
caused by Candidatus Liberibacter asiaticus, Citrus canker (Xanthomonas
citri), fire blight of pome (Eriwinia amylovora ) and olive knot caused by
Pseudomonas savastanoi pv. savastanoi . These are economically important
bacterial diseases in all kinds of crops causing negligible to 100% losses, so all
concern personnel should be contemplation for the effective management of
these diseases.
Bacterial diseases of major crops in Nepal
Introduction
Bacteria are unicellular prokaryotes differing from higher organisms
for genetic material. Six main genera of bacteria accounts for most of
the bacterial species inciting plant disease. Bacteria survives on crop
residues, seeds and volunteer plants. They spreads through water,
wind, mechanical means, insects and seeds. Infection takes place
through natural openings and wounds. Bacteria infect almost all of the
cultivated crops and causes significant loss of yield and quality.
Bacterial diseases are of the important diseases of cereals, fruits,
vegetables including crucifer, cucurbits and solanaceous crops
(potato, tomato, tobacco, brinjal, pepper and so on) causing
significant damages. Bacteria of genus Pseudomonas, Xanthomonas,
Ralstonia, Erwinia and Clavibacter are found to be associated with
different solanaceous crops .
Although the plant pathology section was established in 1954
under department of agriculture in Nepal and bacame functional,
research on bacetrial diseases was established in 1963 in bacterial
leaf blight of rice. The plant pathogenic bacteria are distributed
or confined between plains (terai region ) and valley where
tropical to sub tropical climate is found in Nepal. average loss
caused by bacteria in crops is about 5-60% . Almost all
pathogenic bacteria are prevalent in Nepal except Rhodococcus
i.e Xanthomonas, Pseudomonas, Eriwinia, Agrobacterium,
Clavibacter, Curtobacterium, Streptomyces etc. Later, some other
bacteria were reported in rice seeds, They are Pseudomonas avenae
(now Acidovorax avenae) (Shakya et al. 1985), and P. glumae (now
Burkholderia glumae) (Shakya 1989), Pseudomonas fuscovaginae
(Mortensen et al. 1992, Shakya & Manandhar 1992), P. marginalis
and Erwinia sp. (Mortensen et al. 1992). A nematode,
Aphelenchoides besseyi, has been frequently isolated from rice seeds.
Major bacterial diseases in Nepal
1. Bacterial diseases of cereals

The Contribution of cereals on AGDP is 31.7 % and in GDP 10. 49%

(MoAD, 2014). cereals are the major staple food for nepalese people
and nepalese farming system is cereals dependent agricultre
system.The major cereals crop which are grown in Nepal are Rice,
Maize, Wheat, Barley, Finger millet.Plant pathogenic bacteria causes
many serious diseases of plants throughout the world
(Vidhyasekaran 2002).

A. Bacterial leaf blight of rice

Bacterial Leaf Blight is caused by Xanthomonas oryzae pv. oryzae

(Xoo), is a devastating disease of rice worldwide. In rice, bacterial
leaf blight reduced 5-60% yield in terai and mid-hills during hot and
humid periods. It is one of the oldest and first noticed by the farmers
of Japan in 1884. The bacterium was renamed Pseudomonas oryzae
and later Xanthomonas oryzae (Ishiyama, 1922). Subsequently, its
incidence has been reported from different parts of Asia, northern
Australia, Africa and USA (Gnanamanickam et.al., 1999). In Nepal it
causes losses about 32.5% (Thapa, 1978) , 26% (Adhikari and Mew
1992).

In Nepal, it is considered to be a major disease of rice (IRRI 1979;

Adhikari and Mew, 1988). The disease was first described in Balaju
(Kathmandu valley) in 1968 (Khadka et al. 1968) and has since
been reported throughout the country (Adhikari and Shrestha
1989). Recently, the disease has spread northward and is
becoming a major problem of rice in the mid-hills (Manandhar
et al. 1987). Disease surveys in various rice-growing regions
showed that BB is present wherever rice is grown, with the
warmest regions being the most seriously affected. Disease is
favoured by the prolonged periods of heavy rainfall and high
temperatures from August to October in Nepal. Kresek syndrome is
very prominent when the temperature is between 28 and 32 °C.
Bacterial ooze can disperse with wind-driven rain. Seed- borne
studies have not yet been conducted.

Epidemiology
The bacterium is aerobic, Gram negative, Non spore forming, Rod
with size ranging from 0.55-0.75×1.35-2.17µm
with monotrichous polar flagellum. Bacterial colonies are circular,
convex with entire margins, whitish yellow to straw yellow colored
and opaque.
Clipping of tip of the seedling at the time of transplanting Heavy rain,
heavy dew, flooding, deep irrigation water Severe wind and
temperature of 25-300C with relative humidity above 70%.
Application of excessive nitrogen, especially late top dressing are the
favourable conditions which ultimately lead the bacterial leaf blight in
rice.

Disease cycle
The infected seeds as a source of inoculum may not be important
since the bacteria decrease rapidly and die in the course of seed
soaking. The pathogen survives in soil and in the infected stubbles,
hosts
Leersia, Plantago najor,Paspalum dictum and Cyanodon dactylon. 
The pathogen spreads through irrigation water and also through rain
storms.

Prevention and control

Seeds from infested areas is either prohibited or should come from
pathogen-free crops and be subject to further testing for the pathogen.
Other control measures include burn the stubbles, Use optimum dose
of fertilizers, Avoid clipping of tip of seedling at the time of
transplanting, Avoid flooded conditions, Remove alternate hosts,
Grow resistant cultivars : example, IR 20 and TKM 6, Spraying
copper-oxychloride and streptomycin solution completely inhibits the
growth of bacterium (Tagami and Mizukami, 1962) and
Spray Streptomycin sulphate and tetracycline combination 300g +
Copper oxychloride 1.25 Kg/ha.

B. Bacterial leaf streaks

Bacterial leaf streaks is caused by Xanthomonas campestris pv.
oryzicola reported a bacterial leaf stripe disease on rice in the
Philippines. it is widely distributed in tropical asia and west africa in
both lowland and upland rice growing areas. The estimates of yield
loss from the disease range from 1 to 17% depending on the cultivar
and the climatic conditions.
In Nepal bacterial leaf streaks diseases was first reported in 1970
(Manandhar, 1987) as minor disease of rice. It was reported from terai
and inner terai region of Nepal.
The bacterum survives largely on the infested seed and straw. all wild
species of the genus Oryza can be infected by X. oryzae pv. oryzicola
and may serve as reervoirs of the inoculum. No other host of the
bacterium are known. The bacterium may also be able to survive in
irrigation water.
The bacterium enters the host through stomatas or wounds and
multiplies in the parenchymatous tissue. baterial exudates from leaf
lesins are disseminated primarily by splashing and wind blown rain
and also by leaf contact and irrigation water. Disease development is
favoured by rain, high humidity, and high temperature(28-30oC).

Management
The disease can be controlled with the use of resistant cultivars and
treated seed. In practice, growers have relied on the resistant cultivars
for the disease control.

C. Bacterial brown stripes

The disease was reported on rice by Goto & Ohata (1961). The
bacterial stripe is caused by Pseudomonas syringae pv. Panici and
Pseudomonas avenae. it is found in Japan, Tiwan, Philippines and
occurs in upland nurseries and causes only little damage. it occurs in
upland and wetland nurseries as well as in nursery boxes. Although it
is widely distributed in the rice growing countries (Shakya et al,
1985), it does not cause much damage to production.
Bacterial brown stripe disease was reported in Nepal as minor disease
of rice crop since So far the diseases have not been observed as major
problems.
Disease cycle and Epidemology
P.avenae is seed borne, and P. syrinage pv. panici is likely seed
borne. Natural infection of Panicum miliaceum, Hordeum vulgare and
Setaria italica by P. syringae pv. panici has been reported.

Management
Dry heat treatment at 65OC for 6 days can eliminate the pathogen
from seeds (Zeigler and Alvarez, 1988). In nursery boxes, spraying of
Kasugamycin can control the pathogen.

D. Bacterial sheath rot

The bacterial Sheath brown rot is caused by Pseudomonas

fuscovaginae [formerly described as P. marginalis (P. fluorescens
biovar II)] is induced by chilling temperatures and the causal
organism, Pseudomonas fuscovaginae 1st reported from Hokkido,
Japan (Tanii et al., 1976) and in Nepal from Western
Nepal( Pradhanang and Ghimire, 1994). It is most important disease
in high altitude and high rainfall areas of Nepal. The incidence of
disease was high in Lumle and Chhomrong (1600-2000m). The
disease was also found in wheat during spring season in LARC.
Symptoms of bacterial sheath rot are confused with fungal sheath
rot caused by Sarocladium oryzae, which is prevalent in warmer
areas.
The disease is seedborne and was first reported from seeds in early
1990s (Mortensen et al. 1992, Shakya & Manandhar 1992). Bacterial
sheath brown rot is not a serious problem in widely adapted local
landraces (Chhomrong, Takmare, Kalopatle, Sinjali, Seto Bhakunde,
Darmali etc) while it could be a major problem in introduced
varieties(Sharma et al., 1997). An improved variety Macchapuchhre-
3, a progeny of Chhomrong, is resistant to the disease No chemicals
and other control measures have been tested against the disease.
Bacterial sheath rot is first reported in wheat in 1996 infecting in hilly
region of Nepal by NWRP as minor diseases of wheat.

E. Bacterial Stalk rot

Bacterial stalk rot of maize is Caused by Erwinia chrysanthemi pv.
Zeae It is extended to some pockets of the mountains and Kathmandu
valley but with very low rates of infection. The disease usually
appears at the time of tasseling. Intensity of infection increases as
weather conditions become warmer and humid (Manandhar, 1983). In
the terai, the rate of stalk rot infection is high because 50% of the
stalk rot is caused by corn borer damage and high percent of yield
losses incurred. In the terai area of northern India, and in southern
Nepal and the southern Philippines, this disease represents a serious
problem for maize production. It has been observed that a high
disease incidence is associated with the use of sewage water for
irrigation; it is particularly favoured by high temperatures (≥ 28°C)
and high relative humidity, which prevails in most maize-growing
areas 3-4 weeks after sowing.

F. Bacterial leaf strip of maize

This disease caused by Xanthomonas rubrilineans was first reported
by Manadhar in 1976. Its incidence has been extremely low. However
in 1981 a severe infection appeared in epidemic form at Rampur,
Chitwan (Manandhar, 1983).

G. Stewart wilt of maize

Stewart wilt of maize disease is caused by Erwinia stewartii (E.F.

Smith Dye). The disease required warmer temperature for the
infection so can be reported from a few places in the terai region of
Nepal. Because the incidence of the disease is very low, yield losses
attributed to it are negligible (Manandhar, 1983).

2. Bacterial diseases of vegetables

A. Bacterial wilt of vegetables

Bacterial wilt, caused by Pseudomonas solanacearum is one of the

most important and widespread diseases of crop plants in Nepal.
In particular, the disease incidence in potato and tomato fields
ranged from 5-70%. Research on bacterial wilt at the Institute of
Agriculture and Animal Science (IAAS), Chitwan, Nepal has been
initiated with the main objectives to characterize strains of
P.solanacearum by pathogenicity tests, biochemical tests, and
polymerase chain reaction (PCR), and to develop an integrated
approach to manage the disease. The results showed two
distinct populations of P. solanacearum in Nepal. The strains
that originated from high hills (900 to 1800 m) and collected from
potato were race 3 and biovar II. The strains that collected from
lowland tropics (less than 900 m) were race 1 and biovar III. Several
tomato cultivars were tested for resistance to P. solanacearum
in the greenhouse and field. Tomato cultivar CL 1131 was highly
resistant to the disease. A 3-year crop rotation to integrate resistant
and non-host crops and possibility of using antagonistic bacteria to
manage this disease are in progress.

Host crops: potato, tomato, tobacco, banana, eggplant, pepper

etc. The disease was widely spread in many seed production areas
of the eastern, central,and western regions. In particular, the disease
was reported from IIlam, Jhapa, Morang, Sunsari, Dhankuta,
Mahottari, Dhanusa, Sarlahi, Kathmandu, Bhaktapur,
Sindhupalchok, Dolkha, Makwanpur, Kaski, Palpa, Nuwakot,
Kavre, Rupendehi, Banke, and Kanchanpur districts in Nepal
(Shrestha 1977, 1978; Adhikari, 1993). Bacterial wilt incidence in
potato fields has been reported as high as 70% (Pradhanang et. ai,
1992; Shrestha, 1988), while disease incidence on tomato varied
from 10-60%, depending upon the variety, location, and cultural
practices (Adhikari et aI., 1992).

B. Soft rot of vegetables

Soft rot of vegetable is caused by Erwinia carotovora  worldwide on a

wide range of hosts including potato, carrot, and cabbage. During the
spring of 2009, a soft rot with a foul smell was noted in stored potato
tubers of different local cultivars, especially Rato Alu and Seto Alu, in
the Kathmandu District, central region of Nepal.
Appearance of tan, water-soaked areas with watery ooze was
observed on potato. The rotted tissues were white-to-cream colored.
Seven different potato fields, where the stored tubers originated,
were surveyed and 23 samples consisting of approximately three
symptomatic tubers were collected.

Pathogenecity test

Bacteria were successfully isolated from all diseased tissues on nutrient

agar supplemented with 5% sucrose and incubated at 26 ± 1°C. After
purification on tripticase soy agar medium, 17 isolates were identified
as E. carotovora by the following deterministic tests: all strains were
gram-negative rods; oxidase negative; facultatively anaerobic; able to
degrade pectate; sensitive to erythromycin; negative for phosphatase;
unable to produce acid from α-methyl-glucoside; and produced acid
from trehalose. Pathogenicity of the strains was evaluated by
depositing a bacterial suspension (106 CFU/ml) on potato slices (cv.
Monalisa) and incubating at 30 ± 1°C. A reference strain of E.
carotovora subsp. carotovora (NCPPB 2577) and sterile distilled water
were used, respectively, as positive and negative controls. All strains
caused soft rot within a week. Bacteria were reisolated from the slices
and were shown to be identical to the original strains according to the
above morphological, cultural, and biochemical tests. The finding of this
pathogen is of fundamental value since this crop represents one of the
economically important crops of Nepal. This pathogen has already been
reported in the countries of China and India (1) with whom Nepal
shares its boundaries. The pathogen may have been introduced to this
region of Nepal via seed potato tubers from other countries.
C. Bacterial speck of tomato

An important disease in many tomato-growing areas of the world is

bacterial speck of tomato caused by Pseudomonas syringae pv.
tomato (Okabe) Young, Dye and Wilkie (Pst). Bacterial speck of
tomato was first seen in the Kathmandu valley on the cultivar B.L.
(Lamichhane et al., 2009), a common rainy season tomato cultivar
grown in different parts of Nepal. Now the disease is wide spread
over commercial regions of tomato cultivation overall the country.
The Pst pathogen has two races, race 0 and race 1 (Habazar and
Rudolph, 1997). Race 0 is the most widespread in countries where
tomato is grown (Goode and Sasser, 1980; Varvaro and Guario, 1983)
and it has also been reported from Nepal (Lamichhane et al., 2009);
while race 1 is less widespread but occurs in several countries,
including Canada and Italy (Lawton and MacNeill, 1986; Buonaurio et
al, 1996).

D. Black rot of crucifers

Black rot caused by Xanthomonas campestris pv. campestris
(Pammel) Dowson (Xcc) is one of the economically important
diseases of brassicaceous vegetable crops of Nepal. Manandhar and
thapa , (1992 ) reported that the disease is major disease in terai and
inner terai where cauliflower and other crucifers are commercially
cultivated. In some of the locations more than 60% yield reduction in
cauliflower has been reported (Shrestha, 1990).
The disease is infectious on other crucifers members such as radish,
cabbage, turnip, broad leaf mustard etc. it is reported that black rot
disease caused up to 24.5% infection in local cauliflower and up to
39% in mustard seeds. it is seed borne disease. Once the bacterium is
invaded in soil through infected seed and established the disease, it is
very difficult to eliminate. After infecting the lower leaves it gets
premature dropping and the symptom may disappear for several
weeks.

E. Bacterial spot on tomato

Bacterial spot on tomato is caused by Xanthomonas campestris

pv.vesicatoria. it was first identified in 2009 as devastating disease
causing 50-60% losses on tomato in different commercial farms at
Indrapur, Banke district, Mid-western region. The symptoms were
dark, circular and water soaked lesions of 3 mm in diameter
observed on leaves of tomato plants ( Solanum lycopersicum pv.
Pusa Ruby). Yellow pigmented bacterial colonies were
consistentlyisolated from diseased tissues on YDC medium and
incubated at 26+ 10C . six isolates were identified as Xanthomonas
campestris pv. vesicatoria on the basis of morphological and
biochemical tests. All the isolates were gram negative , rod shaped,
motile, aerobic, oxidase negative, catalase positive and amylolytic
positive.

F. Brown rot of potato

The brown rot of potato is caused by gram negative bacteria
Pseudomonas solanacearum. The disease was first identified by
Puskar Nath in 1963. Later Sherestha (1997) found the causative
agent Pseudomonas solancearum by performing several physical test,
biochemical test and host range test. Mainly the brown rot of potato is
transmitted through infected tubers. The Resistant genotype of this
disease are not identified yet but some of the tolerant lines are MS
35-9&BR 63-65 ( Shrestha 1988).
Maximum tolerance limit for seed multiplication & for ware potato
production were 0.1% & 2% respectively (National Potato
Development Programme). Adopting cropping pattern of ‘potato-
paddy- potato’ can somehow reduce the disease in field.

G.Bacterial diseases of soybean

In soybean two bacterial diseases were reported in Nepal. First one is
Bacterial postule caused by Xanthomonas campestris pv glycines and
next one is soybean seed decay caused by Bacillus subtilis. Bacterial
postule disease is severe in plains, moderate in mid hills and low in
high hills of Nepal. Exotic varieties of soybean are less susceptible
compared to indigenous Varieties(Manandhar & Sinclair 1982). No
chemicals are recommended yet for the control of this disease in
Nepal.

3. Bacterial diseases of fruits

A. Citurs greening
Three species of gram negative bacterium namely Candidatus
Liberibacter asiaticus, Candidatus Liberibacter africanus and
Candidatus Liberibacter americanus are the casual organisms of HLB,
respectively prevailing in the continent of Asia, Africa and South
America. In Nepal, the disease was first reported from Pokhara in
1968 (Thrower 1968) as the main cause of citrus decline in the
country (Roistacher 1996, Regmi and Yadav 2007). It was noted that
citrus greening aes expressed earlier and clearly in lower hills than
mid hills. Trees planted in 1975 were severely infected by citrus
greening in Pokhara valley. About 55% of citrus trees in Pokhara
valley and 100% in Horticulture Research Station were symptomatic
to HLB in 1980s (Regmi 1982). The disease is caused by a phloem-
restricted, non cultured, Gram-negative bacterium transmitted by
citrus psyllid insect vector; non-curable once trees are affected. Since
HLB is very destructive disease and slowly invading new citrus
growing areas represent a major threat to the world citrus industry
including Nepal. The Asian strain, i.e in case of Nepal it is heat
tolerant and able to cause symptoms at temperatures greater than
30oC.

At the initial time of its infection it was considered as Zn deficiency /

toxicity. Mandarin (Suntala), sweet orange (Junar) and acid lime
(Kagati) are the major commercial citrus species grown in Nepal
(MoAD 2013). Since all citrus species are infected with HLB in varying
degree and the disease is already spread several parts of the country
(Bove 2006) understanding the complex nature of the disease and its
management approaches is very vital to save the citrus industry of
the country.
B. Citrus canker
The disease caused by bacterium Xanthomonas citri (Hasse) occurs in
large areas of the world. The disease is endemic in India, Japan and
other south-East Asian countries, from where it has spread to all other
citrus producing continents except Europe. Government of Nepal has
initiated Citrus (Lime) Mission Programme in Tehrathum, Bhojpur
and Dhankuta districts with an objective to substitute the import of
this fruits by increasing its production from 2038 B.S. No detail
research has been carried out in Nepal on citrus canker. A study was
carried out during 2005 to 2006 in Kavrepalanchowk district as it is
representative of most of the hilly districts of Nepal.
Regarding the pathogen of citrus canker, in the late 1980s, strains
associated with canker A were proposed as a new species,
Xanthomanas citri, whereas types B and C as well as other strains
causing citrus bacterial spot remained within X. campestris as
pathovars aurentifolii and citrumelo respectively (Gabriel et al. 1989).
Schaad et al. (2000) proposed a reclassification that places citrus
canker and citrus bacterial spot strains within Xanthomonas as a
species citri (A strains), aurantifolli (B and C strains) and citrumelo
(citrus bacterial spot strains).

C. Olive knot
In commercial olive orchards in Bissingkhel locality, Makwanpur
district, knots (0·5–3 cm in diameter) resembling those caused
by Pseudomonas savastanoi pv. savastanoi  on olive trees, were
observed on twigs and branches of 12-year-old olive plants cv.
Frantoio during 2006-2008. From knots, bacterial cells were isolated
on nutrient agar medium supplemented by 5% sucrose (NAS). The
isolates were positive for oxidase, pectinolytic and arginine
dihydrolase activity, tobacco hypersensitivity; production of
fluorescent pigments and of indole on KB and on MM media,
respectively. The characteristic knot symptoms were observed on all
inoculated plants within six months after inoculation. Bacterial
cultures with identical characteristics to the original strain used for
inoculations were reisolated from all olive knots that developed.

Other bacterial diseases in Nepal

Other several bacterial diseases have been reported in Nepal causing
mild to severe infection on various crops in Nepal. Fire blight of
pome fruits caused by Erwinia amylovora, ratoon stunting of
sugarcane caused by leifsonia xyli subsp. xyli , Crown gall caused by
Agrobacterium tumifaciens affecting deciduous fruits of Nepal.
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