Genet Resour Crop Evol

https://doi.org/10.1007/s10722-020-00900-z (0123456789().,-volV)
(0123456789().,-volV)

RESEARCH ARTICLE

Parasitic interactions of Orobanche with selected Nicotiana

species and identification of effective resistant genotypes
G. Swarnalatha . K. Sarala . K. Prabhakara Rao . K. Baghyalakshmi .
K. R. S. Sambasiva Rao . J. Poorna Bindu

Received: 2 May 2019 / Accepted: 6 February 2020

Ó Springer Nature B.V. 2020

Abstract Tobacco is an important commercial crop
in India and other parts of the world. Recently,
Orobanche cernua Loefl. (broomrape), a total root
parasite is causing considerable yield losses to tobacco
crop cultivated in India. Control strategies focusing
largely on agronomic practices are not successful in
achieving its control. Unfortunately there is no tobacco
variety having good levels of resistance to O. cernua is
available for managing its menace. Due to lack of the
availability of O. cernua resistance even in germplasm
G. Swarnalatha
Acharya Nagarjuna University, Nagarjunanagar, Guntur,
Andhra Pradesh 522510, India
e-mail: g.swarnalatha85@gmail.com
K. Sarala (&)
K. Prabhakara Rao
J. Poorna Bindu
ICAR-Central Tobacco Research Institute, Rajahmundry,
Andhra Pradesh 533 105, India
e-mail: ksarala@rediffmail.com
K. Prabhakara Rao
e-mail: prabhakarabt@yahoo.co.in
J. Poorna Bindu
e-mail: j.poornabindu@gmail.com
of cultivated tobacco species, a study was conducted
with seven selected species of Nicotiana viz., N.
repanda Lehm., N. benthamiana-repanda, N. neso-
phila, N. umbratica-nesophila, N. stocktonii, N. pan-
iculata L. and N. tabacum L. cv Siri for identifying
Orobanche resistance for further utilization in trans-
ferring resistance factors to cultivated tobacco. Emer-
gence of higher number of Orobanche and reduction in
plant height and weight in N. paniculata, N. repanda,
and N. tobacum cv. Siri indicated they are susceptible.
Higher reduction in plant height and weight with less
number of Orobache shoots N. stocktonii proved to be a
susceptible species. With relatively low reduction in
plant height and weight under O. cernua treatment, N.
benthamiana-repanda and N. nesophila can be con-
sidered as tolerant species. N. umbratica-nesophila
with less number of Orobanche spikes, higher plant
height and dry weights under both control and treated
conditions can be considered as a resistant species and
can be used in breeding resistant cultivars. Resis-
tant/tolerant species either increased or maintained
their nutrient uptake abilities under parasite infestation
thereby maintained their growth.

K. Baghyalakshmi
Keywords Nutrient uptake
Orobanche
Resistant

ICAR-Central Institute of Cotton Research, Coimbatore,
Tamil Nadu 641 003, India Tobacco
Wild species
e-mail: kauverk@gmail.com

K. R. S. Sambasiva Rao List of abbreviations

Mizoram University (a Central University), Aizawl, N Nitrogen
Mizoram 796004, India P Potassium
e-mail: krssrao@yahoo.com

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K Phosphorus
PH Plant height
C Control
T Treated
SL Strigolactone
Introduction
Tobacco belongs to the family Solonaceae, the genus
Nicotiana and is well- known for its narcotic proper-
ties. There are above 70 species in Nicotiana family
and the two of them are cultivated, N. tabacum L. and
N. rustica L.. Every year the tobacco production is
expected to be around 6.7 million tons throughout the
world. Tobacco is an important commercial crop of
India with about 800 m kg of production and adds
approx Rs. 20,000 Cr. to the national exchequer by
way of excise duty, and approx. Rs. 5000 Cr. by way of
foreign exchange every year. Around 0.25% of India’s
cultivated land is used for tobacco production. It is
also life line of millions of people including farmers,
farm labourers and others involved in its trade. The
production of good quality tobacco leaves are highly
accentuated in its cultivation. Owing to its flourishing
biomass, it is being subjected to various biotic stresses
like insects, diseases and parasitic weeds which
radically reduce the yield and quality. Among these
stresses, the parasitic weed Orobanche is posing a
major challenge in tobacco cultivation in India and
other countries in the world.
Orobanche commonly known as broomrape is one of
the root parasites on tobacco which belongs to Oroban-
chaceae family (Gevezova et al. 2012). Broomrapes are
holo-parasites which are of the most difficult weeds to be
controlled and can even cause total crop failure
(Baghyalakshmi et al. 2019). Broomrapes are con-
strained to obtain their nutrition by feeding off other
plants using the haustorium, a unique structure through
which the parasites divert the water and nutrients from
the host (Parker and Riches 1993). Among various
species of broomrape, about 20 species are considered as
harmful parasitic weeds. Broomrape members found to
infest important crops, including O. cumana on sun-
flower, O. crenata and O. foetida on legumes, O. cernua
on tobacco, tomato and potato, O. ramosa on oilseed
rape and tobacco, and O. aegyptiaca Pers. on tomato and
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Genet Resour Crop Evol
tobacco. Yield loss due to Orobanche in various crops
ranges from 20 to 100% depending on the intensity of
infestation (Barker et al. 1996). Orobanche infestation
has been observed consistently for the last few years
(Reddy et al. 2018) in India and its incidence was very
high in tobacco grown in various parts of the country
during 2018–2019 resulting in loss of yield and leaf
quality. Studies indicated that the species infesting
tobacco is Orobanche cernua Loefl. and causing crop
losses vary from 20 to 50% depending on the time of
infection and the availability of soil moisture (CABI
2019; Krishnamurthy et al. 1977). Molecular character-
isation of Orobanche collected from different tobacco
areas also confirmed that the O. cernua is infecting FCV,
burley and bidi tobacco crops in Andhra Pradesh
(Swarnalatha et al. 2019).
In view of Orobanche ability to produce a large
number of seed/plant, very small sized seed, easy seed
dispersal, longevity of seed in the soil, below ground
establishment and damage to the host even before its
emergence, wide host range etc. making its complete
eradication a difficult proposition. A wide variety of
management approaches, like hand weeding, use of
selective herbicides, biological control, suicidal ger-
mination, and soil treatments by fumigation and solar
heating have been explored (Krishnamurthy 1994;
Krishnamurthy et al. 1976), but none have been found
to be sufficiently effective and affordable. As success
through agronomic practices has been marginal, an
attractive approach for tackling the broomrape prob-
lem is the selection and use of resistant or tolerant
varieties. But, unfortunately there is no tobacco
variety having good levels of resistance to Orobanche
cernua is currently available for managing this
menace (Nagarajan and Reddy 2001; Sarala 2019).
Hence, developing Orobanche resistance tobacco
varieties became one of the priority areas in tackling the
Orobanche menace in tobacco cultivation. However,
known source of resistance to O. cernua that can be
utilized in breeding resistant cultivars is not reported in
germplasm of cultivated tobacco species. In view of this,
it was proposed to evaluate wild Nicotiana species for O.
cernua resistance. The resistance species, if any, thus
identified can be utilized as a source for developing O.
cernua resistant tobacco varieties.
Genet Resour Crop Evol
Materials and methods
The experiment was conducted during 2016–17 at
ICAR-Central Tobacco Research Institute, Rajah-
mundry using O. cernua seed and seven selected
Nicotiana species accessions available in the germ-
plasm of Indian Council of Agricultural Research-
Central Tobacco Research Institute (ICAR-CTRI),
Rajahmundry, based on the earlier work carried out at
the institute.
Orobanche germination: The O. cernua seed col-
lected from dried spikes from FCV tobacco fields at
Katheru Farm were used in screening the Nicotiana
species. Before infecting the soil with Orobanche
seeds, the viability of the parasite seeds was studied in
three replicated petriplates placed with two layers of
filter paper. On the bottom layer Orobanche seeds
were spread uniformly and on the top layer the green
gram (Vigna mungo) seeds were placed. Enough
moisture was given for the green gram seeds to
germinate. After 3 days the bottom layer with
Orobanche seeds was viewed using the microscope
(10 9 magnification). The Orobanche seed showing
germtube formation under the microscope are treated
as viable. Seed germination percentage estimated as
percent viable seed out of total seed sown and mean of
three replications calculated.
Screening of Nicotiana species: Five species viz.,
N. repanda Lehm., N. benthamiana-repanda, N.ne-
sophila, N.umbratica-nesophila, and N. stocktonii that
are found to record tolerant reaction to Orobanche
under field conditions at ICAR-CTRI were screened
under artificial conditions along with N. paniculata L.,
the susceptible species and a cultivated tobacco
variety, N. tabacum cv Siri. Seeds of all the species
were grown in pots (10 cm 9 15 cm) containing a
mixture of soil, sand and compost. The seedlings were
raised in greenhouse and were transplanted on 45th
day in earthen pots of dimension 30 cm 9 25 cm. The
experiment was conducted in Completely Random-
ized Block Design (CRBD) under two different
treatments (control and Orobanche infestation). The
soil of the each of the Orobanche treated pot was
mixed with about 3 g of O. cernua seed. Care was
taken to ensure that the controlled pots were not
infested with parasitic seeds. Nicotiana seedlings were
planted and thinned to two per pot after establishment.
The pots were watered every day and all the recom-
mended cultural practices were followed to maintain
healthy plants. Observations were recorded on the
emergence of first spike and number and weight of
spikes of Orobanche per plant (including below
ground). Final root and shoot biomass along with the
nutrient status (as mentioned below) in both control
and treated conditions were recorded. Percentage
reduction in plant height and weight in treatment
compared to control was estimated.
NPK estimation: The plant samples collected from
both control and infested pots were washed thoroughly
to remove adhering dirt and soil particles. The samples
were initially air dried and later dried in an oven at
60 °C to constant weight. The oven dry weight was
recorded and computed as g/plant. The oven dried
samples were then powdered in a wiley mill. Total
nitrogen was estimated in the powdered samples by
Micro Kjeldahl digestion and distillation method,
phosphorus by triple acid digestion with vanado-
molybdate yellow colour method and potassium by
triple acid digestion and flame photometry (Jackson
1973). The per cent concentration of the nutrients was
multiplied by respective dry matter content to estimate
the nutrient uptake (g/plant). Percent reduction nutri-
ent uptake under treatment over control was estimated
in each genotype. Percent of total nutrient uptake of
Orobanche developed on each genotype was estimated
using the formula: Nutrient uptake by Orobanche/
Combined nutrient uptake by tobacco and Orobanche
9 100.
Statistical analysis: Analysis of variance of the data
was carried out using the design of experiments
software developed by Upendra et al. (2004) at the
Institute. Graphic representation of plant and Oro-
banche characters, reduction percentages over control
are depicted in graphs using Microsoft office excel.
Results
The seed viability of the Orobanche cernua seeds
collected from Katheru Farm was tested using green
gram exudates method. The germination tube from
Orobanche found to form between 3 and 5 days. The
germination percent of parasite seeds was found to
have above 85. After confirming germination, O.
cernua seed was incorporated into the pots used for
screening Nicotiana species.
The Nicotiana species tested differed in their
response to O. cernua treatment with respect to
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Fig. 1 Uprooted Nicotiana species grown under control and Orobanche infection (First row total plants and second row roots)
Table 1 Days taken for the S. no Name of the genotype
emergence of Orobanche
Cernua in Nicotiana species 1 N. paniculata
2 N. repanda
3 N. benthamiana-epanda
4 N. nesophila
5 N. stocktonii
6 N. umbratica-nesophila
7 N. tobacum cv. siri
emergence of broom rape spikes (Fig. 1) and plant
characters. Emergence of O. Cernua spikes in differ-
ent species in the treated plots observed between 40
and 80 days (Table 1) and produced its above ground
shoot. Orobanche emergence observed first in N.
paniculata (40 days) followed by N. tabacum cv. Siri
(43 days). The Orobanche shoots emerged at 45 days
in N. stocktonii and N. nesophila. Delayed emergence
of Orobanche observed N. repanda (80 days), N.
benthamiana-repanda (70 days) and N. umbratica-
nesophila (65 days) compared to all the genotypes.
Less number of Orobanche spikes observed in N.
benthamiana-repanda (2 No.), N. umbratica-ne-
sophila (3 No.) and N. stocktonii (8 No.) (Table 2).
Significantly higher number of Orobanche spikes
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Genet Resour Crop Evol
Mean no. of days taken for the first spike emergence
40
80
70
45
45
65
43
recorded in N. nesophila (20 No.), N. repanda (15
No.), N. paniculata (14 No.) and N. tobacum cv. Siri
(12 No.). Dry weight of Orobanche shoots was
significantly higher in N. tobacum cv. Siri (15 g) and
N. nesophila. (11 g). Lowest weight observed in N.
benthamiana-repanda (2 g). Though, N. repanda
recorded higher number of Orobanche, total weight
of the shoots was relatively less (4.30 g).
Except in N. umbratica–nesophila, plant height and
weights (roots and shoot) decreased under treated
condition compared to control (Table 2). Contrary, N.
umbratica-nesophila recorded increase in plant height
and dry weight of the shoots under infestation
compared to control (Table 2; Fig. 2). Dry weight of
shoots, roots and total plant was higher in the N.
Genet Resour Crop Evol

Table 2 Mean performance of Nicotiana species studied under control (C) and O. cernua infestation (T)
Species Species plant characters Orobanche per plant
*
PH (cm) Dry weight of Dry weigh of Total dry weight No. of Dry weight
shoots (g) roots (g) of plants(g) shoots of shoots(g)
C T C T C T C T C T C T

N.paniculata 37.30 10.00 21.00 8.00 7.00 4.30 28.00 12.30 – 14.00** – 5.67
N.repanda 16.30 15.66 19.00 12.00 8.30 5.03 27.30 15.03 – 15.00** – 4.30
N.benthamiana– 46.60** 42.20** 23.00 19.00 7.30 4.63 30.30 23.63 – 2.00 – 2.00
repanda
N.stocktonii 42.60 15.33 25.00 11.00 8.60 3.63 33.60 14.63 – 8.00 – 4.00
N.nesophila 43.00 39.20** 27.00 26.00** 10.00 5.33 37.00 31.33** – 20.00** – 11.00**
N. umbratica – 50.33** 51.60** 35.00** 41.00** 11.60** 9.63** 46.60** 50.63** – 3.00 – 2.00
nesophila
N.tabacum (siri) 51.33** 17.66 70.00** 35.00** 19.00** 7.63** 89.00** 42.63** – 12.00* – 15.00**
GM 41.07 27.38 31.43 21.72 10.26 5.74 41.69 27.70 – 10.57 – 6.28
CD@5% 2.19 0.89 1.83 1.14 0.45 0.32 2.33 1.15 – 2.14 – 1.01
CD@1% 3.09 1.26 2.58 1.60 0.64 0.45 3.28 1.62 – 3.01 – 1.42
CV 3.01 1.84 3.29 2.96 2.49 3.13 3.15 2.40 – 11.40 – 9.05
SEM 0.71 0.29 0.60 0.37 0.15 0.10 0.76 0.38 – 0.70 – 2.45
*PH Plant height; GM General Mean; CD Critical Difference; CV Coefficient of variation; SEM Error mean square
** Significantly superior over General Mean (GM)

Fig. 2 Plant height (PH) and total plant weight (TPD) of Nicotiana species under control (C) and O. cernua infestation (T)

tabacum cv. Siri under control condition compared to condition. N. umbratica-nesophila and N. nesophila
other species (Fig. 2). Though plant height was higher recorded significantly higher plant height and dry
in Siri than other species under controlled condition, weights under both control and treated conditions than
marked reduction (66%) observed under treated mean.

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Fig. 3 Reduction (%) in plant height (PH) and total plant weight (TPW) in Nicotiana species due to Orobanche treatment compared to
control
The plant height found to be significantly higher
than mean in N. benthamiana-repanda (42.2 cm)
under Orobanche infection. Plant height of N. repanda
under treatment was comparable to control.
The reduction in plant height and total plant weight
was higher in N. paniculata, N. tabacum cv. Siri and N.
stocktonii (Fig. 3). Except in N. paniculata, flowering
observed in all the Nicotiana sp. under broom rape
treatment. Though the number of Orobanche spikes
emerged is less, marked reduction in plant height and
total dry weight was observed in N. stocktonii. N.
nesophila and N. benthamiana-repanda recorded
lower reduction in plant height and dry weight under
Orobanche treatment. The reduction in plant height
was less compared to plant weight in N. repanda. N.
umbratica-nesophila recorded increase in plant dry
weight under Orobanche treatment.
Nutrition uptake
Except in N. umbratica nesophila, all the species
recorded higher total nutrient uptake under control
than treated condition (Table 3; Fig. 4). When com-
pared to tobacco under control, combined total
nutrient uptake of tobacco and Orobanche was more
or less same in N. paniculata and N. repanda, less in N.
benthamiana repanda, N. stocktonii and N. tabacum
(Siri) and high in N. nesophila and N. umbratica
nesophila (Fig. 4). Nutrient uptake by Orobanche
infecting derivatives of two species viz., N. umbratica
nesophila and N. benthamiana repanda was lower
compared to one species.
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Genet Resour Crop Evol
The highest nutrition uptake (N,P&K) was
observed under controlled condition in the cultivated
genotype, Siri (1.14 N, 0.06 P and 1.12 K g/plant)
with maximum plant height and total dry weight
(Table 3). Whereas all the wild species recorded
medium nutrient uptake ranging from 0.3 to 0.65 g per
plant. Among the species, N & P uptake was higher in
N. umbratica nesophila and K in N. nesophila under
treated condition (Table 3). Total nutrient uptake was
higher in N. nesophila and N. umbratica-nesophila
under Orobanche infestation followed by N. tabacum
cv. Siri (Fig. 4). The nutrient uptake (g/plant) was
found to be highest (0.96 N; 0.05 P and 0.35 K) in N.
umbratica nesophila under Orobanche infestation,
which was even higher when compared to its con-
trolled condition (0.51 N; 0.04 P and 0.31 K).
Reduction per cent in nutrition uptake was higher in
Siri followed by N. stocktonii, whereas the reduction
per cent was negative in N. umbratica nesophila. Siri
recorded 53% less N and 63% less P uptake under
infestation with reduced total plant weight in the
infested condition. The N uptake was significantly
higher than mean in the species; N. benthamiana-
repanda, N. nesophila, N. umbratica-nesophila and N.
tobacum cv. Siri under Orobanche infestation. Similar
trend was also found in P & K uptake.
The nutrient uptake of N, P & K by species under
stress was found to be reduced compared to that of
controlled condition in N. stocktonii and N.tabacum
(siri) (Fig. 5, 6, 7) and N. repanda maintained same
under both the conditions. N uptake in N.benthami-
ana- repanda found to be comparable under both
conditions and P & K uptake reduced under
 Genet Resour Crop Evol

Table 3 NPK uptake by Nicotiana species and Orobanche spike under control and O. cernua treatment
Species Condition Nutrient uptake (g/Plant) Uptake by Orobanche (g/plant)
N P K N P K
Mean R% Mean R% Mean R% Mean % of total uptake Mean % of total uptake Mean % of total uptake

N. paniculata C 0.3 3.33 0.03 66.67 0.3 80 – – – – – –

T 0.29 0.01 0.06 0.087 23.08 0.01 50 0.153 71.83
N. repanda C 0.29 20.69 0.02 50 0.24 25 – – – – – –
T 0.23 0.01 0.18 0.075 24.59 0.01 44.44 0.057 24.05
N.benthamiana- C 0.58 5.17 0.04a 50 0.42 33.33 – – – – – –
repanda a
T 0.55 0.02a 0.28a 0.034 5.82 0 0 0.034 10.83
N. stocktonii C 0.57 50.88 0.04a 50 0.43 37.21 – – – – – –
T 0.28 0.02a 0.27 0.065 18.84 0.01 33.33 0.076 21.97
a
N. nesophila C 0.65 13.85 0.04a 25 0.51a 11.76 – – – – – –
T 0.56a 0.03a 0.45a 0.178 24.12 0.016 34.78 0.203 31.09
a
N.umbratica nesophila C 0.51 - 88.24 0.04 - 25 0.31 - 12.90 – – – – – –
T 0.96a 0.05a 0.35a 0.039 3.90 0 0 0 0
a a
N.tabacum (siri) C 1.14 52.63 0.06 50 1.12a 63.39 – – – – – –
T 0.54a 0.03a 0.41a 0.275 33.74 0 0 0 0
Mean ± SE C 0.577 ± 0.081 0.0385 ± 0.003 0.475 ± 0.085
T 0.487 ± 0.072 0.0242 ± 0.004 0.285 ± 0.039
a
N Nitrogen; P Phosphorus; K Potash; C Control; T Orobanche treated; R % Reduction percent over control; SE Standard error

123
 Genet Resour Crop Evol

Fig. 4 Total nutrient uptake in Nicotiana species and O. cernua under control and treatment

Fig. 5 N uptake by tobacco

genotypes in controlled
condition and combined
uptake by tobacco and
Orobanche in treated
condition (g/plant)

Fig. 6 P uptake by tobacco

genotypes in controlled
condition and combined
uptake by tobacco and
Orobanche in treated
condition (g/plant)

infestation. Nutrient uptake under Orobanche infec- N uptake in Orobanche ranged from 0.034 to
tion found to be higher in N. nesophila and N. 0.275 g/plant, P uptake from 0 to 0.016 g/plant and K
umbratica nesophila (Fig. 8). uptake from 0 to 0.203 g/plant. In general, K uptake

123
Genet Resour Crop Evol

Fig. 7 K uptake by tobacco

genotypes in controlled
condition and combined
uptake by tobacco and
Orobanche in treated
condition (g/plant)

Fig. 8 Cumulative uptake of different nutrients in Nicotiana species and Orobanche emerged under broomrape infestation

Fig. 9 Nutrient uptake in Orobanche infecting different Nicotiana species

percentage (out of total uptake) was higher (0–72%) Lowest uptake of nutrients recorded in N. umbratica
followed by P and N. Orobanche recorded higher N nesophila (Fig. 9).
uptake in Siri and K in N. nesophila. Total nutrient
uptake was highest in Orobanche infesting N.
nesophila followed by N. tabacum and N. paniculata.

123

Discussion
The emergence of parasite shoots observed at the end
of grand growth period in different genotypes. Studies
show that germination of parasitic weed seeds is
promoted by strigolactone (SL) a signaling compound
secreted by the host root (Butler 1995; Umehara et al.
2015). These SLs are produced during the flower
initiation period which inhibits lateral branching habit
of the plants, thereby inducing production of floral
buds (Umehara et al. 2015).
N. paniculata proved to be susceptible with very
early emergence of higher number of Orobanche
spikes with more weight and marked reduction in plant
height and plant weight without flowering under
treated condition. Emergence of higher number of
Orobanche spikes and reduction in plant height and
weight in N. repanda, and N. tobacum cv. Siri
indicated they are predisposed to Orobanche infesta-
tion and are susceptible. However, N. tabacum cv. Siri
recorded higher dry weight of shoots, roots and total
plant under control condition compared to other
species indicating its higher yielding nature. N.
benthamiana-repanda, N. umbratica-nesophila and
N. Stocktonii recorded few Orobanche spikes indicat-
ing they are not favorable for parasitic emergence.
Though, number of Orobache shoots are less in N.
stocktonii, the resultant reduction in plant height and
weight are more making it a susceptible species. N.
benthamiana-repanda with few number of O. cernua
shoots and relatively low reduction in plant height and
weight may be considered as tolerant species. Though,
N. nesophila recoded higher number of Orobanche
shoots, the reduction in plant height and weight are not
much indicating it as a tolerant species. Differential
response of Nicotiana sp. to O. cernua treatment
indicated that genetic differences among the species.
Increase in plant dry weight of N. umbratica-nesophila
under O. cernua infestation compared to control may
be due to its ability to cope with the competition
created by the parasite to the host as an extra sink in the
initial stages with increasing the metabolism activity
of the plant. N. umbratica-nesophila with less number
of Orobanche spikes, higher plant height and dry
weights under both control and treated conditions can
be considered as a resistant species and can be used in
breeding resistant cultivars.
Most of the species recorded reduction in plant
height, weight and total nutrient uptake under treated
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Genet Resour Crop Evol
condition compared to control. The germinated O.
cernua compete with the host and depend totally on
the host for all nutrition, drawing sugars and nitrogen
compounds from the phloem and also drawing most of
their water from the host xylem. The parasite becomes
an active sink, comparable to an actively growing part
of the host plant itself, such that effects on the host are
generally proportional to the biomass of the parasite.
Thus the mass of the parasite is reflected in a very
similar loss in mass of the host crop (Hibberd et al.
1998 1999). In general, the type and extent of the
impact are determined by four factors: (1) parasite
size, (2) the parasite’s metabolic activity and growth
rate, (3) the degree of dependency on the host for
resources, and (4) the host’s stage of development
(Hamamouch 2004). Graves (1995) also reported that
Orobanche-induced yield reductions are not primarily
due to competition for water, but rather due to
carbohydrate loss to the parasite. It further reduces
the water uptake capacity of the host roots. Yield loss
in crops ranges from 20 to 100% depending on the
intensity of Orobanche infestation (Barker et al.
1996).
The current study further indicated that the species
N. nesophila and N. umbratica nesophila which were
showing tolerance to the Orobanche infection could
uptake more nutrients in order to compensate the loss
due to weed infestation. The combined total nutrient
uptake of tobacco and Orobanche was either same or
less in susceptible species, N. paniculata, N. repanda,
N. benthamiana repanda, N. stocktonii and N.
tabacum (Siri) indicating their inability to compensate
the nutrient loses to the weed. The N uptake was much
diverted to Orobanche in the susceptible entries when
compared to the tolerant species. N being the impor-
tant nutrition to the tobacco plants, loss of N to
Orobanche will have an adverse effect on the quality
and yield of the tobacco leaves. The germinated
Orobanche compete with the host for water and
metabolites (Graves 1995), this is evident from the
loss of nutrients in the susceptible host. In the current
study, the K uptake percentage (out of total uptake) of
Orobanche found to be higher (0–72%) followed by P
and N indicating the major role of K and P in
Orobanche nutrition. Ernst (1986) observed that in
tobacco infected with O. ramosa, the leaf potassium
concentration of infected plants was reduced by 60%
and phosphorus concentration was reduced by more
than 50% in roots. These changes in the nutrient
Genet Resour Crop Evol
availability of infected plants could be the principal
reason for a reduction in host growth and yield.
Siri being the cultivated variety recorded higher
nutrients showing that the cultivated species respond
well to fertilizer application. However, Siri recorded
53% less N and 63% less P uptake under infestation
with reduced plant weight and total plant weight in the
infested condition indicating possible yield reduction
under field condition. Krishnamurthy et al. (1977)
reported yield losses of 24–52% in tobacco cultivated
in India with additional loss due to reduction in leaf
quality with Orobanche infestation. It is found that the
Orobanche infesting Siri had the highest nitrogen
accumulation content and the lowest accumulation
was observed in N.benthamiana- repanda. In most of
the Orobanche spikes, it was found that the N
accumulation was higher compared to K and the least
was P content. Similarly, the parasite spikes emerging
from the resistant species were found to accumulate
very less nutrient content
Control strategies designed for non-parasitic weeds
such as cultural and chemical methods do not neces-
sarily achieve the required level of control for
broomrape due to its mixed traits as weeds and root
parasite. Hence, host plant resistance is the best option
for the control of this parasitic weed. The present study
confirmed that the two species namely, N.bethamiana-
repanda and N. nesophila were found to be tolerant
and N. umbratica nesophila resistant to parasite
infestation. These species after further investigations
can be exploited for developing Orobanche resistant
tobacco cultivars. Tobacco sources resistant to O.
ramosa and O. aegyptiaca were identified in France
(Julio et al. 2018) and Iran (Porkabiri et al. 2019),
respectively. Wild forms of Nicotiana varies in their
genome composition as there is a large variation in
basic number viz., x = 6, 9, 10, 12, 16, 18, 19, 20, 21,
22, 23, 32 indicating that the genes responsible for the
resistance could be present in only few species (Knapp
et al. 2004). Similarly, there may be few genes in the
wild type of tobacco which provides resistance to the
parasite which would have been lost during domesti-
cation process making the cultivated high yielding
lines more prone to the parasite attack.
In sunflower, six wild species namely Helianthus
divaricatus, H. grosseserratus, H. divaricatus, H.
mutelii, H. grosseserratus and H. tuberosus (Pérez-
Vich et al. 2002) are reported to be the sources of
Orobanche resistance genes. About 7 genes/QTL
regions were identified to confer resistance towards
this parasitic weed (Alonso et al. 1996), (Skoric et al.
2010), (Rodrı́guez-Ojeda et al. 2001) in sunflower. In
sunflower the gene Or5 resistance towards Orobanche
race E has been mapped by Tang et al. (2003) using
SSR markers and mapped on LG3. RNA-sequence
analysis and physical mapping of O. aegyptiaca
resistance in tobacco entry, Wika found to be
controlled by a recessive gene (14 m bp deletion) on
chromosome number 14 (Julio et al. 2018).
Conclusions
Screening of Nicotiana sp. against O. cernua indicated
that species differ in their response to broomrape
infestation differently. The present study confirmed
that the two species namely, N.bethamiana-repanda
and N. nesophila were tolerant and N. umbratica
nesophila resistant to parasite infestation. The Oro-
banche tolerant or resistant species found to demon-
strate their ability to compensate nutrient loss to the
parasite while maintaining growth and metabolism.
These species after further investigations can be
exploited for transferring the resistant trait or genes
to cultivated species either by conventional or modern
biotech tools.
Acknowledgements Authors profoundly acknowledge the
Competent authority at Indian Council of Agricultural
Research-Central Tobacco Research Institute, Rajahmundry
for providing encouragement in carrying out the research work.
Funding The research work presented in the manuscript is not
funded by any external agency and it is the work carried out
using in-house resources only.
Compliance with ethical standards
Conflict of interest We also declare that there is no conflict of
interest
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