New Zealand Journal of Crop and Horticultural Science

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Peroxidase as a biochemical marker of maturity

levels in potato (Solanum tuberosum) cultivars
grown under short days

S. K. Sandhu , R. S. Marwaha & S. K. Pandey

To cite this article: S. K. Sandhu , R. S. Marwaha & S. K. Pandey (2007) Peroxidase as a

biochemical marker of maturity levels in potato (Solanum�tuberosum) cultivars grown under
short days, New Zealand Journal of Crop and Horticultural Science, 35:1, 171-175, DOI:
10.1080/01140670709510181

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New Zealand Journal of Crop and Horticultural Science, 2007, Vol. 35: 171-175
0014-0671/07/3501-0171 © The Royal Society of New Zealand 2007
Peroxidase as a biochemical marker of maturity levels in potato
(Solanum tuberosum) cultivars grown under short days
S. K. SANDHU*
R. S. MARWAHA
Central Potato Research Station
Jalandhar 144003, India
S. K. PANDEY
Central Potato Research Institute
Shimla 171001, India
*Present address: Department of Plant Breeding,
Genetics & Biotech, Punjab Agricultural University,
Ludhiana 141001, India, email: surinderksandhu@
yahoo.com
Abstract Peroxidase activity was determined in
the leaves of early (70-80 days maturity), medium
(90-100 days), and late maturing (100-120 days)
potato (Solanum tuberosum) germplasm accessions
at 30, 50, and 70 days after planting. Enzyme activity
was lowest in early maturing cultures, whereas late
maturing cultures exhibited the maximum activities
with medium maturing ones showing intermediate
values at all three stages of crop growth. Peak
enzyme activity was observed at the 30-day stage,
irrespective of the maturity group, and declined
thereafter. Mean peroxidase activity in the leaves
at the pre-tuber initiation stage (30 days) was 25.3,
-1 -1
40.3, and 74.0 ΔA min g fresh weight, in early,
medium, and late maturing cultures, respectively.
The large differences in the enzyme activities of
(at the 30-day stage) can be used as a biochemical
marker for identifying cultivars belonging to different
maturity groups.
Keywords peroxidase activity; germplasm
accessions; varieties; maturity levels; leaves; days
after planting; potato
H05117; Online publication date 12 April 2007
Received 6 October 2005; accepted 25 August 2006
171
INTRODUCTION
Peroxidases are widely distributed in plant tissues
and are of immense physiological interest because of
their association with numerous catalytic functions.
Peroxidase has been implicated in the oxidation
of indole-3-acetic acid (Stonier et al. 1979),
hydoxylation of proline (Ridge & Osborne 1970),
wound healing (Kawashima & Uritani 1963), water
stress (Krishnamurthy 2003), disease resistance
(Katoch et al. 2003), shelf life (Vijayalakshmi &
Bangarusamy 2003), seed quality (Yadav et al.
2003), and zinc tolerance (Singh et al. 2005) in
legumes, root, rhizome, and fruit crops. In potato
(Solanum tuberosum L.), it has been associated with
keeping quality and resistance against rotting during
storage (Tripathi et al. 1974), salt tolerance (Rahnama
& Ebrahimzadeh 2005), and has been used as a
biochemical marker to determine physiological age
of tubers (Frydecka-Mazurczyk & Zgorska 1993).
Potato possesses a wide range of adaptation
and climate flexibility. In Europe and the United
States, it is grown under long day conditions (c. 14-h
photoperiod) and the crop has a duration of 120-
180 days. In the Indian plains, it is grown under
short day conditions (9.75-11.5-h photoperiod)
during the winter where early to medium maturing
potato varieties are essentially required to fit the
multiple cropping sequences to increase agricultural
production in the country (Shekhawat et al. 1999). As
a result, the varieties and technological requirements
of potato cultivation in India are quite different
from those of the temperate world. The Central
Potato Research Institute, Shimla has a germplasm
collection of c. 1200 accessions of S. tuberosum ssp.
tuberosum. If a reliable biochemical marker such as
peroxidase activity is identified in the leaves at the
pre-tuber initiation stage for differentiating early,
medium, or late maturity cultivars, this will be of
immense help to screen the germplasm accessions
at an early stage. This will assist potato breeders in
selecting the desired parental lines for hybridisation
without waiting for the crop to mature and will
also help to determine the maturity levels of the
172 new Zealand Journal of Crop and horticultural Science, 2007, Vol. 35

progenies. Keeping this objective in mind, the activity
of peroxidase was determined in the leaves at three
stages of crop growth in potato cultures belonging to
three different maturity groups to establish if there
exists a relationship between enzyme activity and
maturity levels.
MATERIAL AND METHODS
Planting of germplasm material and sampling
Ten potato germplasm accessions viz., aura, CIP
382284, Craigs Defiance, Early Gem, Kinga, LT-7,
Rushmore, universal netherlands, 25/40, 27/40,
and three Indian commercial varieties viz., Kufri
alankar, Kufri Chandramukhi, and Kufri Lauvkar
belonging to the early maturity group (70-80 days);
10 accessions viz., aBZ 69-1, atol, BR 63-75, Cea
69-1, Meise, Perricholi, Pirola, Red skin, TS-4,
3392 (1), and three Indian commercial varieties
viz., Kufri Jyoti, Kufri Lalima, and Kufri Sutlej
belonging to the medium maturity group (90-100
days) ; and an equal number of germplasm accessions
viz., ackersegen, andinita, Belle d e Locronay,
Claudia, Kennebec, Marvia, Muruta, Ruta, Sissay,
TM-1, and three Indian commercial varieties viz.,
Kufri Badshah, Kufri dewa, and Kufri Sindhuri
belonging to the late maturity group (100-120
days) were grown in sandy loam soil at the Central
Potato Research Station, Jalandhar, located in the
north-western Indian plains (75°40 e, 31°21 n
250m a.s.l.) during the autumn from October to
January under short days with decreasing day length
(11.5-9.75h) and temperature (min. 5-18°C; max.
17-33°C). The germplasm accessions belonging
to S. tuberosum ssp. tuberosum, under the present
study were collected from different countries and
maintained vegetatively under field conditions. The
maturity levels of these germplasm accessions after
introduction to India were recorded at the Central
Potato Research Institute, Shimla (Kufri farm,
c. 2530m a.s.l.) under long day conditions (100-
110 days for early, 110-135 days for medium, and
135-160 days for late maturing ones). The Institute
has allocated its own numbers, designated with CP,
to the accessions received from different countries
of the world (Gaur et al. 1984). The germplasm
accessions undertaken in the present study have been
mentioned with their original country names as well
as with CP numbers in the tables.
The trial was laid during 2000-01 and 2001-02
in a randomised block design with three replications
and the planting was done on 1 October in both
years. Twelve plants of each genotype in a single row
planted at inter- and intra-row distances of 60 and
20 cm, respectively, represented each replication. all
the recommended cultural practices were followed.
The crop was grown under irrigated conditions

Table 1 Peroxidase activity (Da min–1 g–1 fresh weight) in the leaves of early maturing potato (Solanum tuberosum)
germplasm accessions and Indian commercial varieties at different stages of growth.

Germplasm accession Growth stage (days after planting)

*
Genotype name CPno. 30 50 70 Mean
aura CP 1700 21.2 22.8 30.8 24.9
CIP 382284 CP3155 22.8 18.4 18.8 20.0
Craigs Defiance CP 1680 26.0 19.6 34.0 26.5
early Gem CP 1747 36.4 20.8 25.2 27.5
Kinga CP3188 22.4 16.8 20.0 19.7
LT-7 CP 2176 33.6 14.4 24.0 24.0
Rushmore CP 1763 20.4 16.4 22.4 19.7
universal netherlands CP2023 20.0 13.6 29.0 20.9
25/40 CP3191 28.8 18.8 17.2 21.6
27/40 CP3192 27.6 11.6 20.2 19.8
Indian varieties
Kufri alankar CP2137 20.4 8.4 16.0 14.9
Kufri Chandramukhi CP2141 19.6 10.8 14.4 14.9
Kufri Lauvkar CP2150 29.6 13.2 15.6 19.5
Mean _ 25.3 15.8 22.1
LSd (0.05) Stage (S) Genotype (G) S×G
0.91 1.9 3.28
*numbers allotted by the Institute.
Sandhu et al.—Peroxidase as marker of maturity levels in potato 173

and intercultural operations like manual weeding,
spraying against weeds, and preventive spraying
against late blight (Phytophthora infestans) were
applied on the schedule dates. The early, medium,
and late maturing cultures were dehaulmed on 75,
90, and 110 days after planting, respectively, and
were harvested after 20 days of curing in the soil.
For the determination of peroxidase activity, the 4th
leaf from the top of the plants was sampled at the
30, 50, and 70 day stage after planting.
Preparation of crude enzyme extract and assay
Five gram leaf samples were macerated in 45 ml
of 0.1M cold phosphate buffer (ph 6.5) in a pre-
chilled mortar and pestle. The slurry was filtered
through four layers of muslin cloth and the filtrate
was centrifuged at 12 000g for 15 min at 4°C. The
resulting supernatant was used as enzyme extract.
Peroxidase activity was measured by a slightly
modified procedure of Malik & Singh (1980).
The reaction mixture contained 3.5 ml of 0.1M
phosphate buffer, 0.2 ml of o-dianisidine (2.93 mg/
ml in methanol), 0.1 ml enzyme extract, and 0.2
ml of 0.2M hydrogen peroxide in a total volume of
4.0 ml. The reaction was monitored by the increase
in absorbance (a) at 430 nm in a Spectronic 21 d
spectrophotometer at 15-s intervals for 3 min. The
enzyme activity was expressed as Damin–1 g–1 fresh
weight (Fw). Three replications for each estimation
were taken and the data were pooled over 2 years and
statistically analysed (Gomez & Gomez 1984).
RESULTS AND DISCUSSION
Peroxidase activity during different
stages of crop growth and development
Mean peroxidase activity was maximum in the leaves
at the 30-day stage in all the germplasm accessions
and cultivars, irrespective of the maturity group
(Tables 1, 2, 3). The activity declined significantly
at 50 days and 70 days after planting. The average
reduction in activity at 50 and 70 days was c. 38%
and 13% in early, 39% and 21% in medium, and 44%
and 40% in late maturing germplasm and cultivars,
respectively, when compared at 30 days. Similar
results showing maximum activities of peroxidase,
amylase, and catalase during the first half of the
growth period followed by a decrease towards the
maturation stage was reported in cotton (Vodogreeva
1978). huang & Zhang (2000) also reported that
catalase and peroxidase activities were maximum
in soybean at 25 days after flowering and rapidly
decreased afterwards. Our results showed that the
peroxidase activity was very high in the leaves before
the tuber initiation stage which fell significantly
during the active bulking stage of the tubers. In early
maturing cultures, the tuber initiation starts between

Table 2 Peroxidase activity (Da min–1 g–1 fresh weight) in the leaves of medium maturing potato (Solanum tuberosum)
germplasm accessions and Indian commercial varieties at different stages of growth.

Germplasm accession Growth stage (days after planting)

*
Genotype name CPno. 30 50 70 Mean
aBZ 69-1 CP 2377 39.2 27.6 36.0 34.3
atol CP3180 49.2 32.8 39.6 40.5
BR 63-75 CP2164 48.8 24.8 30.4 34.7
Cea 69-1 CP2386 38.4 23.6 30.0 30.7
Meise CP 1458 36.4 22.8 26.4 28.5
Perricholi CP2292 40.0 23.6 35.2 32.9
Pirola CP 2427 40.8 30.4 33.2 34.8
Red Skin CP2042 35.2 27.6 32.4 31.7
TS-4 CP3197 48.8 24.0 38.4 37.1
3392 (1) CP1515 34.0 16.8 21.6 24.1
Indian varieties
Kufri Jyoti CP2144 44.0 21.6 30.4 32.0
Kufri Lalima CP2149 35.6 23.2 28.4 29.1
Kufri Sutlej - 34.0 21.6 29.6 28.4
Mean _ 40.3 24.6 31.7
LSd (0.05) Stage (S) Genotype (G) S×G
0.79 1.65 2.86
*numbers allotted by the Institute.
174 New Zealand Journal of Crop and Horticultural Science, 2007, Vol. 35

Table 3 Peroxidase activity (AA min^g^ 1 fresh weight) in the leaves of late maturing potato (Solarium tuberosum)
germplasm accessions and Indian commercial varieties at different stages of growth.

Germplasm accession Growth stage (days after planting)

Genotype name *CPno. 30 50 70 Mean
Ackersegen CP 1636 81.6 41.2 52.0 58.3
Andinita CP3195 87.2 35.6 41.2 54.7
Belle De Locronay CP 1702 67.6 38.0 39.6 48.4
Claudia CP 1704 71.2 40.4 43.6 51.7
Kennebec CP2124 74.8 42.4 52.0 56.4
Mar via CP 2406 68.0 52.0 59.2 59.7
Muruta CP3147 81.6 42.4 52.8 58.9
Ruta CP3158 69.2 34.8 41.6 48.5
Sissay CP3189 82.4 44.4 41.2 56.0
TM-1 CP 2374 70.4 42.4 45.2 52.7
Indian varieties
Kufri Badshah CP2138 64.8 34.4 38.4 45.9
Kufri Dewa CP 2142 72.8 49.2 31.6 51.2
Kufri Sindhuri CP2158 70.4 42.0 38.5 50.3
Mean — 74.0 41.5 44.4
LSD (0.05) Stage (S) Genotype (G) SxG
1.41 2.91 5.08
'Numbers allotted by the Institute.

25 and 30 days, in medium maturing between 35
and 40 days, and in late maturing between 40 and
45 days after planting, which is followed by the
bulking stage. Presence of high peroxidase activity in
the leaves before tuber initiation may be associated
with a high rate of accumulation of metabolites
and an active metabolic pool in the leaves because
of high photosynthetic activity and absence of
translocation towards the sink at this stage. It is
reported that developing tubers are a dominant sink
for carbohydrates (Moorby 1968) and compete with
haulms for the available assimilates under tuberising
conditions (Minhas et al. 2002).
As can be seen in the tables, peroxidase activity
was maximum in the leaves of late maturing cultures
at all the three stages of growth, whereas the early
maturing showed minimum activity, with medium
maturing ones exhibiting intermediate levels.
Vodogreeva (1978) also reported lower activities
of peroxidase and catalase in the plants of early
maturing cotton cultivars than in late-maturing ones.
Among early maturing cultures, average peroxidase
activity was minimum in Kufri Alankar and Kufri
Chandramukhi, whereas it was maximum in Early
Gem but was at par with Craigs Defiance. Out of
medium maturing cultures, the activity was lowest in
3392(1) and highest in Atol, whereas Kufri Badshah
and Marvia showed minimum and maximum
activities, respectively, among late maturing cultures.
Marvia showed maximum activity among the late
maturing group, but it was at par with Muruta and
Ackersegen. The large differences in the activities
of enzyme at the 30-day stage in the leaves can be
used as a biochemical marker for differentiating
early, medium, and late maturing varieties. The
method for the estimation of peroxidase is simple,
quick, and reliable and will be a useful tool for
screening germplasm accessions for different
maturity levels.
ACKNOWLEDGMENTS
We are thankful to Dr S. M. Paul Khurana, former
Director, Central Potato Research Institute, Shimla and
Dr G. S. Kang, Head of the Central Potato Research
Station, Jalandhar for their keen interest and for providing
necessary facilities. Thanks also to Mr J. S. Jassal and Mr
Raj Kumar for help in chemical and statistical analyses,
respectively.
REFERENCES
Frydecka-Mazurczyk A, Zgorska K 1993. Biochemical
markers indicating the physiological age of potato
tubers. Ziemniaka 43: 147-161.
Sandhu et al. —Peroxidase as marker of maturity levels in potato
Gaur PC, Mishra PC, Nayar, NM 1984. Catalogue of
potato germplasm collection group tuberosum.
Central Potato Research Institute, Shimla, India.
Technical Bulletin No. 13.
Gomez KA, Gomez AA 1984. Statistical procedures for
agricultural research. New York, John Wiley &
Sons. 110 p.
Huang Fu-J, Zhang G 2000. Physiological and biochemical
changes during seed filling in relation to leaf
senescence in soybean. Biología Plantarum 43:
545-548.
Katoch R, Mann APS, Sohal BS 2003. Salicylic acid as
elicitor for the induction of systematic acquired
resistance in garden pea (Pisum sativum L.).
Journal of Plant Biology 30: 365-369.
Kawashima N, Uritani I 1963. Occurrence of peroxidases
in sweet potato infected by the black root
fungus. Agricultural Biology and Chemistry 27:
409-414.
Krishnamurthy KS 2003. Physiological and biochemical
changes during water stress in ginger. Journal of
Plant Biology 30: 313-318.
Malik CP, Singh MB 1980. Plant enzymology and histo-
enzymology. New Delhi, Kalyani Publishers.
53 p.
Minhas JS, Rai VK, Saini HS 2002. Importance of invertase
during tuberisation in potato plants. Proceedings
of Global Conference on Potato, Indian Potato
Association, New Delhi. Pp. 820-825.
Moorby J 1968. The influence of carbohydrates and
mineral nutrient supply on the growth of potato
tubers. Annals of Botany 32: 57-68.
175
Rahnama H, Ebrahimzadeh H 2005. The effect of NaCl on
antioxidant enzyme activities in potato seedlings.
Biología Plantarum 49: 93-97.
Ridge I, Osborne DJ 1970. Hydroxyproline and peroxidase
in cell walls of Pisum sativum; regulation by
ethylene. Journal of Experimental Botany 21:
583-587.
Shekhawat GS, Gaur PC, Pandey SK. 1999. Managing
biodiversity in potato and associated crops. Journal
of Indian Potato Association 26: 119-125.
Singh RA, Singh AP, Roy, NK, Singh AK 2005. Pigment
concentration and activity of antioxidant enzymes
in zinc tolerant and susceptible chickpea genotypes
subjected to zinc stress. Indian Journal of Plant
Physiology 10: 48-53.
Stonier T, Stasinos S, Reddy KBS 1979. The masking of
peroxidase catalysed oxidation of IAA in Vigna.
Phytochemistry 18: 25-30.
Tripathi RK, Verma MN, Gupta VK 1974. Peroxidase and
isoenzymes in relation to resistance in potatoes
against rotting. Indian Journal of Experimental
Biology 12: 591-592.
Vijayalakshmi D, Bangarusamy U 2003. Effect of certain
post-harvest treatments on shelf life, physiological
loss in weight and oxidative enzyme activities in
sapota fruits cv. Co 1 and Co 2. Journal of Plant
Biology 30: 313-318.
Vodogreeva LG 1978. Enzyme activity during ontogenesis
of cotton cultivars of different maturity.
Referativnyi Zhurnal 55: 486-491.
Yadav S, Bhatia VS, Guruprasad KN 2003. Role of
peroxidase and catalase enzymes in deterioration
of soyabean seeds due to field weathering. Indian
Journal of Plant Physiology 8: 195-200.