Peroxidase as a Biochemical Marker of Maturity Levels in Potato Solanum Tuberosum Cultivars Grown Under Short Days

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  Full Terms & Conditions of access and use can be found at New Zealand Journal of Crop and Horticultural Science ISSN: 0114-0671 (Print) 1175-8783 (Online) Journal homepage: Peroxidase as a biochemical marker of maturitylevels in potato ( Solanum tuberosum ) cultivarsgrown 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 abiochemical 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 To link to this article: Published online: 19 Feb 2010.Submit your article to this journal Article views: 568Citing articles: 1 View citing articles  New Zealand Journal of  Crop  and  Horticultural  Science,  2007,  Vol. 35:  171󰀭1750014󰀭0671/07/3501󰀭0171 © The Royal Society of New Zealand 2007171  eroxidase  s biochemical  marker  of  maturity  levels in  potato  Solanum  tuberosum)  cultivars  grown  under  short  d ys S. K. SANDHU*R. S. MARWAHA Central  Potato Research Station Jalandhar  144003, IndiaS. K. PANDEY Central  Potato Research InstituteShimla 171001, India *Present  address: Department of Plant Breeding, Genetics  Biotech, Punjab Agricultural University, Ludhiana  141001, India, email: surinderksandhu@  bstract  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 activitywas lowest in early maturing cultures, whereas latematuring cultures exhibited the maximum activitieswith medium maturing ones showing intermediatevalues at all three stages of crop growth. Peakenzyme activity was observed at the 30󰀭day stage,irrespective of the maturity group, and declinedthereafter. Mean peroxidase activity in the leaves at  the pre󰀭tuber initiation stage (30 days) was 25.3, 40.3,  and 74.0 ∆A  min 󰀭1  g 󰀭1  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 biochemicalmarker for identifying cultivars belonging to different maturity  groups. Keywords  peroxidase activity; germplasmaccessions; varieties; maturity  levels;  leaves; daysafter planting; potato H05117;  Online publication  date  12  April  2007 Received  6  October  2005;  accepted  25  August  2006 INTRODU TION 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 oxidationof indole󰀭3󰀭acetic acid (Stonier et al. 1979),hydoxylation of proline (Ridge Osborne 1970),wound healing (Kawashima Uritani 1963), waterstress (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) inlegumes, root, rhizome, and fruit crops. In potato  Solanum  tuberosum  L.), it has been associated withkeeping quality and resistance against rotting duringstorage (Tripathi et al. 1974), salt tolerance (Rahnama Ebrahimzadeh 2005), and has been used as abiochemical marker to determine physiological ageof tubers (Frydecka󰀭Mazurczyk Zgorska 1993). Potato  possesses  a wide range of adaptation and  climate  flexibility.  In Europe and the UnitedStates, 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 undershort day conditions (9.75󰀭11.5󰀭h photoperiod)during the winter where early to medium maturing potato  varieties are essentially required to fit themultiple cropping sequences to increase agricultural production  in the country (Shekhawat et al. 1999). As a  result, the varieties and technological requirementsof potato cultivation in India are quite differentfrom those of the temperate world. The Central Potato  Research Institute, Shimla has a germplasmcollection of c. 1200 accessions of  S.  tuberosum  ssp. tuberosum.  If a reliable biochemical marker such asperoxidase activity is identified in the leaves at the pre󰀭tuber  initiation stage for differentiating early, medium,  or late maturity cultivars, this  will  be ofimmense help to screen the germplasm accessions at  an early stage. This  will assist  potato breeders inselecting the desired parental lines for hybridisationwithout waiting for the crop to mature and  will also help to determine the maturity  levels  of the  172new Zealand Journal of Crop and horticultural Science, 2007, Vol. 35progenies. Keeping this objective in mind, the activityof peroxidase was determined in the leaves at threestages of crop growth in potato cultures belonging tothree different maturity groups to establish if thereexists a relationship between enzyme activity andmaturity levels. MATERIAL AND METHODSPlanting of germplasm material and sampling Ten potato germplasm accessions viz., aura, CIP382284, Craigs Defiance, Early Gem, Kinga, LT-7,Rushmore, universal netherlands, 25/40, 27/40,and three Indian commercial varieties viz., Kufrialankar, Kufri Chandramukhi, and Kufri Lauvkarbelonging 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 varietiesviz., Kufri Jyoti, Kufri Lalima, and Kufri Sutlejbelonging to the medium maturity group (90-100days)   and an equal number of germplasm accessionsviz., ackersegen, andinita, Belle de Locronay,Claudia, Kennebec, Marvia, Muruta, Ruta, Sissay, TM-1,  and three Indian commercial varieties viz.,Kufri Badshah, Kufri dewa, and Kufri Sindhuribelonging to the late maturity group (100-120days) were grown in sandy loam soil at the CentralPotato Research Station, Jalandhar, located in thenorth-western Indian plains (75°40 e, 31°21 n250m  a.s.l.)  during the autumn from October toJanuary under short days with decreasing day length(11.5-9.75h) and temperature (min. 5-18°C; max.17-33°C). The germplasm accessions belongingto  S. tuberosum  ssp.  tuberosum,  under the presentstudy were collected from different countries andmaintained vegetatively under field conditions. Thematurity levels of these germplasm accessions afterintroduction to India were recorded at the CentralPotato Research Institute, Shimla (Kufri farm,c. 2530m  a.s.l.)  under long day conditions (100-110 days for early, 110-135 days for medium, and135-160 days for late maturing ones). The Institutehas allocated its own numbers, designated with CP,to the accessions received from different countriesof the world (Gaur et al. 1984). The germplasmaccessions undertaken in the present study have beenmentioned with their srcinal country names as wellas with CP numbers in the tables.The trial was laid during 2000-01 and 2001-02in a randomised block design with three replicationsand the planting was done on 1 October in bothyears. Twelve plants of each genotype in a single rowplanted at inter- and intra-row distances of 60 and20 cm, respectively, represented each replication. allthe recommended cultural practices were followed.The crop was grown under irrigated conditions Table 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.GermplasmGenotype nameauraCIP 382284Craigs Defianceearly GemKingaLT-7Rushmoreuniversal netherlands25/4027/40 Indian varieties Kufri alankarKufri ChandramukhiKufri LauvkarMeanLSd (0.05)accession * CPno.CP 1700CP3155CP 1680CP 1747CP3188CP 2176CP 1763CP2023CP3191CP3192CP2137CP2141CP2150_3021.222.826.036.422.433.620.420.028.827.620.419.629.625.3Stage (S)0.91Growth stage (days after planting)5022.818.419.620.816.814.416.413.618.811.68.410.813.215.8Genotype (G)1.97030.818.834. S×G3.28Mean24.920.026.527.5 19.724.019.720.921.619.814.914.919.5*numbers allotted by the Institute.  Sandhu et al.—Peroxidase as marker of maturity levels in potato173and intercultural operations like manual weeding,spraying against weeds, and preventive sprayingagainst late blight  Phytophthora infestans)  wereapplied on the schedule dates. The early, medium,and late maturing cultures were dehaulmed on 75, 90,  and 110 days after planting, respectively, andwere harvested after 20 days of curing in the soil.For the determination of peroxidase activity, the 4thleaf 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 mlof 0.1M cold phosphate buffer (ph 6.5) in a pre-chilled mortar and pestle. The slurry was filteredthrough four layers of muslin cloth and the filtratewas centrifuged at 12 000g for 15 min at 4°C. Theresulting supernatant was used as enzyme extract.Peroxidase activity was measured by a slightlymodified procedure of Malik & Singh (1980).The reaction mixture contained 3.5 ml of 0.1Mphosphate buffer, 0.2 ml of o-dianisidine (2.93 mg/ml in methanol), 0.1 ml enzyme extract, and 0.2ml of 0.2M hydrogen peroxide in a total volume of4.0 ml. The reaction was monitored by the increasein absorbance (a) at 430 nm in a Spectronic 21 dspectrophotometer at 15-s intervals for 3 min. Theenzyme activity was expressed as Damin– 1  g –1  freshweight (Fw). Three replications for each estimationwere taken and the data were pooled over 2 years andstatistically analysed (Gomez & Gomez 1984). RESULTS AND DISCUSSIONPeroxidase activity during differentstages of crop growth and development Mean peroxidase activity was maximum in the leavesat the 30-day stage in all the germplasm accessionsand cultivars, irrespective of the maturity group(Tables 1, 2, 3). The activity declined significantlyat 50 days and 70 days after planting. The averagereduction 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. Similarresults showing maximum activities of peroxidase,amylase, and catalase during the first half of thegrowth period followed by a decrease towards thematuration stage was reported in cotton (Vodogreeva1978). huang & Zhang (2000) also reported thatcatalase and peroxidase activities were maximumin soybean at 25 days after flowering and rapidlydecreased afterwards. Our results showed that theperoxidase activity was very high in the leaves beforethe tuber initiation stage which fell significantlyduring the active bulking stage of the tubers. In earlymaturing 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. GermplasmGenotype name aBZ 69-1 atol BR  63-75 Cea 69-1 MeisePerricholiPirola Red  Skin TS-4 3392  (1)  ndian varieties Kufri JyotiKufri LalimaKufri SutlejMean LSd  0.05)accession * CPno.CP  2377CP3180CP2164CP2386 CP 1458CP2292CP  2427 CP2042 CP3197 CP1515 CP2144CP2149 -_3039.249.248.838.436.440.040.835.248.834.044.035.634.040.3 Stage  (S)0.79Growth  stage days after planting) 5027.632.824.823.622.823.630.427.624.016.821.623.221.624.6 Genotype  (G)1.657036.039.630.430.026.435.233.232.438.421.630.428.429.631.7S×G2.86 Mean 34.340.534.730.728.532.934.831.737.*numbers  allotted  by the Institute.
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