Int. J. Life. Sci. Scienti. Res.

, 3(1): 808-816

JANUARY- 2017

Research Article (Open access)

Distribution of Leaf Tip Necrosis Genes and its

Association with Expression of Lesion Mimic
Genes and Resistance to Spot Blotch in Spring
Wheat
Shweta Singh1,2,*, V.K. Mishra2, R.N. Kharwar1, R. N. Ahirwar2, Smrati Sharma2,3, P.S. Yadav2, Ramesh Chand4
1
Department of Botany, Institute of Science, Banaras Hindu University, Varanasi, India
2
Department of Genetics and Plant Breeding, Institute of Agricultural Sciences, Banaras Hindu University Varanasi, India
3
Department of Biotechnology, J C Bose Institute of Life Sciences, Bundelkhand University, Jhansi India
4
Department of Mycology and Plant Pathology, Institute of Agricultural Sciences, Banaras Hindu University Varanasi,
India
*

Address for Correspondence: Shweta Singh, Research Scholar, Department of Botany, Institute of Science,
BHU, Varanasi, India
Received: 06 November 2016/Revised: 23 November 2016/Accepted: 16 December 2016

ABSTRACT- The adult plant resistance (APR) gene Lr34 and Lr46 of wheat is associated with leaf tip necrosis (Ltn),
and provide resistance against multiple diseases, viz. leaf rust, stripe rust, stem rust, powdery mildew and spot blotch.
APR gene Lr46 is also associated with Ltn. Lesion mimic (lm) mutants express hypersensitive responses in the absence of
pathogens and also confer resistance to biotrophic pathogens, including leaf rust. However, association between spot
blotch and Ltn is reported, but not with Lm and Ltn. Five hundred diverse lines of spring wheat including two hundred
and ninety four wami population including were screened for Lr34 (CsLv34, 150 bp), Lr46 (STS1BL2, 600bp) and three
lesion mimic genes, viz. lm (Xwmc85.1 and Xgwm264) on 1B, lm1 (Xbarc147 and Xwmc674) on 3BS, and lm2 (Xgwm513
and Xgwm149) on 4BL.Lr34alone was found in only one line, but in 31% of lines it was in combination with Lr46. In
contrast, Lr46 was present singly in 63% of lines. The Lm genes appeared to increase spot blotch severity whereas Lr34
alone or with Lr46 provided moderate to high level of resistance. Lm expression was absent when both Lr34 and Lr46
were present indicating their masking effect on Lm genes. The presence of Lr46 alone showed variable Lm expression.
This suggests that the presence of Lr34 + Lr46 and/or absence of Lm genes enhance resistance to spot blotch.
Key-words- Leaf tip necrosis (Ltn), Lesion mimics (Lm), Spot blotch, Phenotypic marker, Adult plant resistance (APR)
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INTRODUCTION
The leaf tip necrosis (Ltn) is most popular trait among the
wheat pathologists and breeders for its association with the
durable rust resistance in adult plants. The adult plant
resistance gene, Lr34 (formerly LrT2), was first described
in a wheat lines PI58548 and was later shown to be present
in a number of wheat cultivars [1-2]. Leaf tip necrosis is
closely associated with Lr34 gene [3] and is being used as its
phenotypic marker [4].
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DOI: 10.21276/ijlssr.2017.3.1.10

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Lr34 confers horizontal resistance or slow rusting

resistance although under appropriate conditions it may
confer resistance in seedlings to certain rust races. The
efficianacy of Lr34 mediated APR also varies between
different cultivar backgrounds and growth conditions. More
importantly Lr34 has been recognized as major component
of durable rust resistance as it can act synergistically with
other leaf rust resistance genes. Lr34 encodes a putative
adenosine triphosphatebinding cassette (ABC) transporter.
It belongs to the ABC subtype G (formerly PDR), subclass
of transporters [5]. It was reported that developmentally
regulated resistance is correlated with elevated Lr34
transcript levels in flag leaves of adult plants. There are
several allelic forms of the Lr34 gene that have been
identified in the wheat germplasm, with two alleles
occurring at a high frequency [6]. Resistance allele (Lr34res)
differs by only two amino acid changes from the other
allelic forms associated with susceptibility (Lr34sus) and so

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Int. J. Life. Sci. Scienti. Res., VOL 3, ISSUE 1

gain-of its function of resistance. So it is assumed that the

resistance conferred by Lr34res is the result of altered
activity or substrate specificity. The presence of Lr34 and
Lr46 in wheat is associated with leaf tip necrosis (Ltn), a
type of necrosis that forms predominantly in the tips of
wheat flag leaves, and is dependent on expression of
Lr34res as well as on genetic. The gene Lr34 was
postulated in few Indian bread wheat cultivars reported the
association of leaf tip necrosis with the spot blotch of wheat
[7]
.
Spot blotch caused by Bipolaris sorokiniana has emerged
an important problem in the last century in 9 million
hectare what of South East Asia. This pathogen affects the
crops of millions of small and marginal farmers of this
region and cause severe damage. Annual average yield
losses due to spot blotch are reported around 19.6 and 15.5
% in wheat [8]. There are many program have been initiated
to improve the wheat against spot blotch [9-10]. But not
much success can be achieved to reduce or minimize the
damage due to spot blotch. Due to complex problems of
pathogen and tropical environment of South Asia Breeding
progress is slow. Substantial gain to spot blotch resistance
as made by sleeting wheat lines with Ltn [7]. [11] reported the
association of Lr34 and Lr46 with the spot blotch
resistance. In present studied population genotyping with
SSR markers shows the predominance of Lr46 alone or in
combination of Lr34. This study was performed for
variable level of spot blotch severity to understand the role
of different gene combination in the spot blotch resistance.
Spot blotch spreading is very much influenced by
environmental condition like temperature and radiation. It
was reported that an increase in these two factors also
shows additive effect in the susceptibility. Terminal heat
stress increased severity and showed positive correlation in
South Asia [10,12-13]. Temperature is an important non living
factor that influenced the developmental growth stages of
plant. The total temperature required for plant growth
development can be calculated by accumulating degree
days between the high and low temperature thresholds
throughout the crop season [14]. Incidence and spread of
spot blotch are influenced by the growth stages, and
maturity duration of wheat [15]. Plants that spontaneously
express necrotic lesions during development or in response
to changes in the environment, without the presence of any
pathogen, are denoted as lesion mimics exhibiting either
constitutive or unregulated cell death like HR after
pathogen infection. So it can be hypothesized that Lm in
wheat facilitating the spot blotch development and its rapid
spread in the host due to readily available necrotic tissue
for colonization of the pathogen. If this is true then Lm
free/least Lm wheat lines expected to have low
susceptibility and Lm expression should also be suppressed
by Ltn like spot blotch so the objective of this study was to
establish association between Ltn that is a trait of
resistance, lm and spot blotch, phenotypic and genotypic
expression and relation of different Ltn genes and
distribution of different Ltn genes in present population.
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MATERIALS AND METHODS

Screening of Germplasm:
Five hundred diverse wheat germplasm including two
hundred ninety four wami germplasm were evaluated for
leaf rust caused by Puccinia recondite, spot blotch causal
organism Bipolaris sorokiniana and Lesion mimic at the
Research Farm of Banaras Hindu University, Varanasi
during crop season 2013-14, 2014-2015 and 2015-16.

Sowing and maintenance of crop under field and

polyhouse condition
Banaras Hindu University falls in the rice wheat cropping
system zone. The field was prepared by ploughed thrice
after the harvest of rice crop. Fertilizers NPK were applied
@ 60, 60 and 40 Kg/ha during land preparation. Five
hundred wheat germplasm were sown in the alpha lattice
design in two replication, 25 blocks and each block
carrying 20 genotypes. Seeds were sown manually keeping
row to row distance 25 cm and plant to plant 12 cm. The
first irrigation was given at crown root initiation 20 days
after sowing and 30 Kg Nitrogen was also given as top
dressing after 3 days of irrigation while the last dose of
nitrogen was given after flowering (GS45).

Scoring of leaf tip necrosis

Leaf tip necrosis was scored on flag leaf in the field at the
GS 47 to 73. Flag leaves showing necrosis from the tip and
extended to 5cm were counted as Ltn. Leaf tip necrosis was
not appeared in poly house therefore it was not scored.
Under poly house conditions genotypes were screened at
GS 47-73 before ear emergence. Leaves were scored in two
way, 1st it was scored as presence or absence of Ltn. 2nd
when it was present it was scored as the level of their
expression. The scale was used for it is 0 to 4 where 0= no
Ltn, 1=25%, 2=50%, 3=75% and 4 more than 75%.

Scoring of lesion mimic

The plants were observed from seedling to adult stage both
in field and polyhouse for the expression of Lm symptoms.
Lesion mimic was recorded from seedling (GS25) to adult
plant stage (GS60) in poly house. Lm phenotype was
scored on flag leaf in the field and poly house at the GS 60.
The distribution of Lm on leaf i.e. apical, middle and basal
part was also recorded. Leaves showing typical LM
expression was scored with modification of 1- 6 rating
scale of [16] in 1-9 scale. Where 0=no visible specks
1= 1-10%, 2=10.1 to 20%, 3= 20.1-30,4= 30.1-40,
5= 401-50, 6= 50.1-60, 7= 60.1-70, 8, 70.1-80,
9= above80%.

Isolation of pathogen
All the genotypes produced leaf tip necrosis were collected
for the presence of Bipolaris sorokiniana or other pathogen.
Three well development lesions were taken for the isolation
of pathogen. A piece of 1mm2 leaf tissue was taken out
from the one week old lesion. Isolation was performed by
the method of [17].

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Inoculation of the pathogen

A pure culture of the most aggressive isolate of Bipolaris
sorokiniana (NABM MAT 1; NCBIJN128877, Banaras
Hindu University, Varanasi, India) known to be highly
aggressive was used to create artificial epiphytotic [7]. The
isolate was multiplied on sorghum grain and a spore
suspension adjusted to 10-4 spores/ml of water using
haemocytometer was uniformly sprayed at GS 50 on
Zadoks scale during evening hours [18].

Disease assessment
Spot blotch score for each line was evaluated on five
randomly tagged plants in the field at three different growth
stages (GS) viz; GS 63 (beginning of anthesis to half
complete), GS 69 (anthesis complete) and GS 77 (late
milking) [18] following double digit scale (DD, 00-99) [19].
The first digit (D1) indicates vertical disease progress on
the plant and the second (D2) indicates severity measured
in diseased leaf area. For each score, the disease severity
percentage was based on the following formula: % severity
= (D1/9) (D2/9)100.
Area under the disease progress curve based on disease
severity over time at GS63, GS69 and GS77 was calculated
using the percent severity estimations corresponding to the
disease ratings, as outlined by:
AUDPC =

{(Y + Y
i

(i + 1)

) / 2} (t

(i + 1)

- t )]
i

Where, Y = disease level at time ti

i
t(i + 1) - ti = Time (days) between two disease scores
n = number of dates at which spot blotch was recorded

Use of molecular markers for the leaf rust

resistance genes
Molecular marker analysis
A total of 2 SSR and STS markers associated with Ltn and
10 SSR markers associated with lesion mimic mutants,
were used for screening of wheat germplasm viz;
CsLv34(7DS) and STS1BL2 (1BL), Xwmc 674 (3BS);
Xbarc1033 (3BS); Xbarc147 (3BS); Xgwm513 (4BL);
Xgwm149 (4BL); Yao et al., 2009 Xbarc181 (1B); Xbarc61
(1B); Xgwm131 (1B); Xwmc85.1 (1B); Xgwm264.1 (1B)
[20]
.

10 cycles were performed at 94C for 1min, 65-55C for

1min and 72C for 1min. After completion of one cycle, the
annealing temperature lowers down 1C. Remaining 30
cycles were performed at 94C for 30s, 55C for 30s and
72C for 30s with a final extension at 72C for 7 min. The
15l PCR products were mixed with 2l loading dye (98%
formamide, 0.3% of each bromophenol blue and xylene
cyanol, and 10mM of EDTA). Electrophoresis was carried
out in 2.5% agarose gel stained with EtBr and 1 x TAE gel
electrophoresis buffer, prepared from 50 x TAE (242g
Tris-base, 57.1ml Glacial acetic acid and 100ml 0.5M
EDTA for 1L), was used for electrophoresis, at 75 W for 2
hours. Gels picture was visualized under UV light by using
UVP gel doc.

RESULTS
Experiment in Field and Poly house conditions
Ltn were first observed on the leaves of plants. The
expression of Ltn in all plant of same genotypes was
uniform however it was un-uniform in few genotypes. The
appearance of Ltn was recorded at late GS (47 to 73). Lm
symptoms were first observed on the leaf at the time of
booting stage and scored.The genotypes with good
expression of Ltn show no or very less expression of lm
(Fig. 1). Appearance and distribution of Ltn under
poly-house was not like the field. Ltn in poly house was not
expressed. There were no plant in poly house expressed
Ltn.

Effect of Temperature on expression of Ltn

During the experiment it was observed that no expression
of Ltn was seen in poly house (DD 1107) however it was
clearly expressed in field (DD 840). So it can be predict
that Ltn and DD are negatively associated. At the time of
experiment maximum temperature in field was 280C and in
polyhouse it was above 400C. So it can also be predicted
that at high temperature the Ltn did not expressed itself. So
it is clear that gene of Ltn is thermolabile. Appearance and
distribution of Lm under poly-house was different than the
field. Under poly-house more than 70% genotypes
expressed Lm at early (GS 25-30) (35 days after showing).
The Lm severity was higher in polyhouse than the field it
may be due to absence of Ltn and presence of high
temperature.

DNA extraction, PCR amplification and gel

electrophoresis

Leaf tip necrosis and its association with spot

Plant gnomic DNA extracted from the leaves of 15 days old blotch
seedling by following the modified CTAB method [21].
Isolated DNA was kept at -20C for further use. Each
Polymerase chain reaction (PCR) was performed in 15l
reaction volume containing 10ng genomic DNA, 1.5l
10nM Tris-HCL (pH 8.8), 0.2l dNTPs (MBI Fermentas),
0.2l 2.5mM MgCl2 (MBI fermentas), 0.2l Taq
polymerase 5U/l (MBI fermentas) and 0.6l forward and
reverse primers. The sequences of each primer are listed in
Table 1. Amplification were performed in touchdown i.e.,
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In the field Ltn and spot blotch were recorded in some of

different way as previously reported. All those lines which
express Ltn very well were not resistant against spot blotch.
Some lines with Ltn were also susceptible for the disease,
but some of the Ltn lines shows less severity or no severity
of disease. The susceptible genotypes which have Ltn in
their phenotype show the higher AUDPC (Table 2).

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Int. J. Life. Sci. Scienti. Res., VOL 3, ISSUE 1

Genotyping for Ltn lines

Lm, Ltn and Spot Blotch in field

Five hundred diverse lines including two hundred ninety

four wami population of spring wheat were screened for
Lr34 (CsLv34), Lr46 (STS1BL2). Genotyping of Ltn lines
clearly showed the presence of two different genes i.e.Lr34,
and Lr46 (Fig. 2). The frequency of these genes alone and
in combinations is given in table 2.In the populationLr34
alone was found in only four line, but in 31% of lines it was
in combination with Lr46 (Table 1). In contrast, Lr46 was
present singly in 63% of lines. Lr34 alone or with Lr46
provided moderate to high levels of resistance.

The lines with Ltn which have less or no expression of

lesion mimic were resistance or moderately resistance
against spot blotch. Table 1, shows the distribution of Ltn
in wheat genotypes and Table 2 shows slow or no Lm
frequency on those genotypes where Ltn was present.

Leaf tip necrosis and its association with spot

blotch
The genotyping shows that Ltn with Lr46 does not shows
the resistance against disease but if the gene Lr34 is present
then they were shown good resistance against the disease.
Lr34 provides more resistance against spot blotch in
comparison to Lr46 (Fig 1, Table 4).

Ltn, Lm and DD
Lm and DD (degree days) were positively correlated
whereas Ltn and DD are negatively correlated. Lesion
mimic expression was recorded after 35 days of sowing in
the poly house when it received the total DD 1101.1.
Whereas on the same genotypes LM appeared after 74 days
under filed conditions with the DD 740 (Table 4.) but in
case of Ltn in polyhouse at the 1101DD it does not appear
and in field at 840 DD it was appeared very well (Fig 1,
Table 3). Ltn is thermolabile where lm is
thermostable.

Table 1. Distribution of LTN genes among 500 wheat genotypes

Ltn gene (S)

Number of genotypes

% Germplasm

Ltn gene (S)

Lr34

0.8

Lr34

Lr46

315

63

Lr46

Lr34+Lr46

155

31

Lr34+Lr46

Table 2. List of wheat genotypes without Lesion mimic expression in the field and poly house, status of Lm gene,
Ltn and yield related characters. (a- absent, p- present)
Entry

Genotype

DH

AUDPC

1
2

VORONA/GEN
PARA2//JUP/BJY/3/VEE/JUN/4/2*K
AUZ
KAUZ/RAYON
RABE/2*MO88
OASIS/SKAUZ//4*BCN
HUAYTU CIAT
TARACHI F 2000
TAURUM
CNDO/R143//ENTE/MEXI_2/3/A
EGILOPSSQUARROSA
(TAUS)/4/WEAVER/5/2*KAUZ
KETUPA*2/PASTOR
REH/HARE//2*BCN/3/CROC_1/
AE.SQUARROSA
(213)//PGO/4/HUITES
CROC_1/AE.SQUARROSA
(205)//BORL95/3/PASTOR
ATTILA/3*BCN*2//BAV92
TUKURU//BAV92/RAYON
WBLL1*2/4/YACO/PBW65/3/KA
UZ*2/TRAP//KAUZ
PAVON
KEA/BUC//FCT

87
86

3
4
5
6
7
8
9

10
11

12
13
14
15
16
17

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TKW

Lm
field

417.22
564.45

PLOT
YIEL
D
119.5
55

lm

lm
1

lm
2

Ltn

A
A

lm
PolyHouse
a
a

31.8
24.8

p
p

p
a

P
P

p
P

82
86
86.5
83
86.5
86.5
84

447.66
438.09
365.81
406.73
453.03
366.48
351.92

125.5
114.5
110
116.5
107.5
63
148

25
24.8
22.6
30.2
30.4
21.6
30

a
A
A
A
A
A
A

a
a
a
a
a
a
a

p
p
p
p
p
p
p

p
p
a
p
a
p
p

P
P
P
A
P
P
P

a
p
p
p
p
p
p

86.5
82

573.21
523.77

104
147

20.3
32.4

A
A

a
a

p
p

p
p

P
p

a
a

84

490.81

107

29.8

86
82
84

396.05
562.22
461.24

130
128
134

25.4
28.3
28.4

A
A
A

a
a
a

p
a
p

a
a
p

P
P
P

a
p
p

87
80

436.91
505.13

106.5
127.5

22.8
33.2

A
A

a
a

p
p

p
p

P
P

p
p

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18
19

20

21

22
23
24
25
26
27
28

29

30

31
32
33
34

35

36

37

38

39

40
41
42
43
44

45
46

PRL/SARA//TSI/VEE#5
CNDO/R143//ENTE/MEXI_2/3/A
EGILOPS
SQUARROSA
(TAUS)/4/WEAVER
CROC_1/AE.SQUARROSA
(205)//JUP/BJY/3/SKAUZ/4/KAU
Z
PASTOR//SITE/MO/3/CHEN/AEGILO
PS SQUARROSA (TAUS)//BCN
BARBET1
MILVUS2
ATTILA*2/PBW65
VOROBEY
MILAN/KAUZ//PRINIA/3/BAV92
PASTOR/DHARWAR DRY
MILAN/KAUZ/3/URES/JUN//KAUZ
/4/CROC_1/AE.SQUARROSA
(224)//OPATA
CNO79//PF70354/MUS/3/PASTO
R/4/CROC_1/AE.SQUARROSA
(224)//OPATA
CROC_1/AE.SQUARROSA
(224)//OPATA/3/BJY/COC//PRL/B
OW/4/BJY/COC//PRL/BOW
CHEN/AE.SQ//2*OPATA/3/BAV9
2/4/JARU
CHONTE
WHEAR//2*PRL/2*PASTOR
KAUZ//ALTAR
84/AOS/3/MILAN/KAUZ/4/HUIT
ES
CROC_1/AE.SQUARROSA
(205)//BORL95/3/PRL/SARA//TSI
/VEE#5/4/FRET2
CROC_1/AE.SQUARROSA
(205)//BORL95/3/PRL/SARA//TSI
/VEE#5/4/FRET2
ALTAR
84/AE.SQUARROSA
(219)//OPATA/3/WBLL1/FRET2//
PASTOR
YAV_3/SCO//JO69/CRA/3/YAV79
/4/AE.SQUARROSA
(498)/5/2*OPATA
YAV_3/SCO//JO69/CRA/3/YAV79
/4/AE.SQUARROSA
(498)/5/2*OPATA
CHEWINK
THB/KEA//PF85487/3/DUCULA/
4/WBLL1*2/TUKURU
WBLL1*2/3/SNI/TRAP#1//KAUZ
*3/TRAP/4/KACHU
PBW343*2/KUKUNA*2//WHEA
R
MILAN/KAUZ//PRINIA/3/BAV92/5/
TRAP#1/BOW//VEE#5/SARA/3/Z
HE JIANG 4/4/DUCULA
ASTREB/OAX93.10.1//SOKOLL
BACANORA T 88

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84
83.5

557.8
520.75

132
162

30.2
36.6

A
A

a
a

p
a

p
p

P
P

a
p

86

409.88

151.5

30.6

82

425.06

151.5

36.8

84
82
82
86.5
84
74
81.5

502.9
540.06
439.33
366.05
443.83
211.85
239.14

124.5
130.5
127
121.5
115.5
157.5
159

27.2
29.2
27.8
25.4
30.6
34.2
36.9

A
A
A
A
0
0
A

a
a
a
a
0
a

p
p
p
p
p
a
p

p
p
p
p
p
a
p

P
p
P
P
P
A
P

p
p
p
p
p
p
p

84

522.66

118.5

28.4

82

454.32

110.5

29.4

82

434.26

132

32

86.5
80
82

528.53
548.34
536.55

141.5
110
144

31.2
31.6
32

A
A
A

a
a
a

p
a
p

a
p
p

A
P
P

p
p
p

84

498.15

103.5

24.4

84

538.83

101.5

26.2

81.5

345.74

135

34

82

177.66

159

31

83.5

487.04

104

27.8

84
82

214.75
585.43

158.5
147

28.4
33.4

A
A

a
a

a
a

p
a

P
A

p
p

85.5

395.99

133.5

33.6

81.5

375.74

140

32.8

82

373.77

119

31

82
80

211.14
534.14

131
105.5

31.6
24.2

A
A

a
a

p
p

p
a

P
P

p
p

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47
48

BECARD/KACHU
C80.1/3*BATAVIA//2*WBLL1/5/
REH/HARE//2*BCN/3/CROC_1/
AE.SQUARROSA
(213)//PGO/4/HUITES
KFA/3/PFAU/WEAVER//BRAMB
LING/4/PFAU/WEAVER*2//BRA
MBLING
MILAN/KAUZ//PRINIA/3/BAV92
QUAIU #3//MILAN/AMSEL
TACUPETO
F2001/SAUAL/4/BABAX/LR42//
BABAX*2/3/KURUKU
TRCH//INQALAB
91*2/KUKUNA
CROC_1/AE.SQUARROSA
(205)//KAUZ/3/SASIA/4/TROST
ESDA//ALTAR
84/AE.SQUARROSA
(211)/3/ESDA/4/CHOIX/5/WAXW
ING

49

50
51
52

53
54
55

84
85.5

485.99
506.18

119.5
133.5

29
27.2

A
A

a
a

p
p

p
p

P
P

p
p

87

372.16

138

29.7

82
86.5
86.5

439.39
472.22
474.45

128
143.5
125

30.5
28.2
28.6

A
A
A

a
a
a

p
p
p

p
p
p

P
A
P

a
p
a

76

430.87

92.5

28.4

82

420.07

135

31.4

82

503.42

137

28.6

84

417.78

145

34.8

Table 3. Cumulative degree days in poly house and field

Field
Min-Max
Da
ys

Week
No.

Month and
Date

50
51
52
1
2
3
4
5

Dec 10-16
Dec 17-23
Dec 24-31
Jan1-7
Jan 8-14
Jan 15-21
Jan 22-28
Jan 29-Feb4

13.6-26
6.7-26
5-22.2
2.5-25
2.8-27
6.5-32.5
4-22.5
6.6-26

2
7
8
7
7
7
7
7

0
169.1
145.4
138.8
142.7
183.3
144.8
171

6
7
8
9

Feb 05-11
Feb 12-18
Feb 19-25
Feb 26Mar04
Mar 5-11
Mar 12-18

8-27.7
11.6-28
11.8-28
13.1-30

7
7
7
7

13.5-35
16.4-33.8

7
7

10
11

Max

Min

Digreedays

MinMax

0
73.5
58.8
50
39.9
82.4
41.3
63.7

18.3
51.3
22.1
24.4
21.3
62.85
23.05
47.35

0
0

177.6
179.1
176.2
199.6

79.5
93.8
92.3
98.8

58.55
66.45
64.25
79.2

219.1
232.5

101.7
128.6

90.4
110.55
740.05

Days

0
0
1

26-46
26-42
31-43
31-50
36-48

Poly House
Max
Min

Digree days

0
0
28
192
194
192
227
249

0
0
40
291
296
288
296
312

0
0
24
171.5
175
170
191.5
210.5

186
0
0
0

232
0
0
0

159
0
0
0

0
0

0
0

0
0
1101.5

Table 4. Correlation coefficient of Lesion mimics with Ltn and other yield related traits
Character
Ltn
DH
AUDPC
PlotYield

2013-14
-0.87
-0.070*
0.088**
-0.102**

2014-15
-0.99
-0.187**
0.141**
-0.116**

2015-16
-0.85
-0.099**
0.193**
-0.003

TKW

-0.085**

-0.075*

-0.050

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Int. J. Life. Sci. Scienti. Res., VOL 3, ISSUE 1

Fig 1. Association of lesion mimics with Ltn and spot blotch

Fig 2. Screening of Ltn genes in present studied population

Fig 3. Different pattern of expression of Ltn on wheat plant in the field

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Int. J. Life. Sci. Scienti. Res., VOL 3, ISSUE 1

DISCUSSION
Five hundred wheat accessions including two hundred
ninety four wami populations were screened for Ltn and
most of them possess Lr46genes alone or in combinations
with Lr34. However, the expression of Ltn varied in the
wheat genotypes. The Lr34 are known to reduce the leaf
rust severity by reducing uredinial size. Thus, the wheat
genotypes carrying both of Ltn i.e. Lr34 and Lr46 will
serve an improvement of wheat against spot blotch in the
warmer region of South Asia. There are several genotypes
in this experiment identified without expression of Ltn
although they had either the genes alone or its combination.
These genotypes need to be tested under diverse
environment to validate the result. Expression of Ltn
absent in poly house revealed the thermolabile nature of
gene Lr34 and Lr46 both. It may be possible that at high
temperature these genes could not able to translate their
code into protein. So that there is no translation results in
absence of Ltn protein expression. The Lr34 gene encodes
an ABC transporter and has provided wheat, durable and
broad spectrum resistance against multiple fungal
pathogens for over many years. In wheat, transcriptional
expression of Lr34 and Lr46 are highest in adult plants and
confers with increased resistance and Ltn affecting the last
emerging leaf. Ltn also is a type of PCD and the genes can
only express in appropriate environmental condition.
Temperature is the important abiotic controlling element
for Ltn gene expression. Plant growth stage is also a factor
for these two gene expression in wheat.Lr34 and lr46 in
this population shows additive effect against spot blotch
resistance. So it is the need of future breeding program to
develop the genotypes which carry both the genes for
acquired the effective level of resistance against spot blotch
and various diseases.
The increase in global heat or temperature also cause the
threatened of escape of Ltn gene expression due to its
thermolabile nature.

CONCLUSION
Present investigation confirms that alone Lr34 or Lr46 does
not provide desire level of resistance but both the genes in
conjugation provide a good level of resistance against
spot- blotch and mask the expression of lesion mimic. So in
South Asia region the variety having both the genes is more
useful for food safety and yield loss. Ltn inhibit the
expression of lesion mimic phenotypically.

ACKNOWLEDGMENT
The authors are thankful for the financial support to UGC
and CGIAR and CYMMIT for research material.

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How to cite this article:

Singh S, Mishra VK, Kharwar RN, Ahirwar RN, Sharma S, Yadav PS, Chand R: Distribution of Leaf Tip Necrosis Genes and
its Association with Expression of Lesion Mimic Genes and Resistance to Spot Blotch in Spring Wheat. Int. J. Life. Sci.
Scienti. Res., 2017; 3(1): 808-816. DOI:10.21276/ijlssr.2017.3.1.10
Source of Financial Support: UGC-CGIAR-CYMMIT, Conflict of interest: Nil

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