American International Journal of Research in Formal, Applied & Natural Sciences
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Efficacy of aqueous plant extracts on the seed quality of pea (Pisum sativum L.) during storage Kiran Rana, K. C. Sharma and H. S. Kanwar1 Department of Entomology, 1 Department of Seed Science and Technology Dr. Y S Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh-173 230, India Abstract: Studies were carried out to evaluate the efficacy of aqueous plant extracts on seed deterioration and bruchid infestation in pea (Pisum sativum L.) cv. P-89 during storage. The seeds were treated with aqueous extracts of leaves of worm, s wood (Artemisia roxburghii L and A. annua L.), mint (Mentha longifolia L ; M. spicata L.), marigold (Tagetus erecta L.) and drupes of dharek (Melia azedarach L.) against the pulse beetle, Callosobruchus chinensis L. (Coleoptera: Bruchidae) at four different concentrations (1%, 2%, 4% and 6%) and evaluated along with control. All the quality parameters showed significant differences due to seed treatment with botanicals. The results revealed that seed treatment with aqueous extract of M. spicata recorded significantly higher germination (82.1%), seed vigour index-length (1077), seed vigour index-mass (1395) at the end of six months of storage period with nil percentage of insect infestation. This treatment was found on par with aqueous extract of M. longifolia (81.6%, 922.25, and 1364, respectively) compared to control (65.5%, 606.63 and 1044, respectively). All the botanicals showed nil percentage of insect infestation as compared to control (17%) after six months of storage. Key words: Pea, plant aqueous extracts, germination, seed vigour, storage I. Introduction Storage of seed up to next sowing season is an essential part of seed industry. In general, legumes are more susceptible to storage pests and pea is no exception. Because of its high protein content, pea seed is attacked by a number of insect pests in storage, which causes considerable damage to the seed and deteriorate the quality of seed. Among the important storage pests, the pulse beetle, Callosobruchus chinensis L. is a cosmopolitan pest causing great losses to stored legumes [1]. The infested seeds lose viability and vigour thus affecting germination to a greater extent. In order to get maximum germination of seed with synthetic insecticides is the only alternative, however the descriptive use of chemicals and their residual toxicity adversely affects the non-target animals including human beings besides affecting the seed quality. Hence, the safe and feasible approach is the treatment of seeds with botanicals which are safe, economical, ecofriendly and nonharmful to seed, animal and human beings. It has been proved beyond doubt that mixing the seed with botanicals prevents multiplication of beetles because of their repellent or antifeedent property and smoothen the seed surface so that beetle cannot proliferate on the seed coat. In view of this the present studies were carried out to evaluate the effect of seed treatment with some selected botanicals on bruchid infestation and storability of pea. II. Material and Methods A storage experiment was conducted at Seed Technology and Production Centre (STPC), Dr. YS Parmar University of Horticultural & Forestry, Nauni during 2011- 2012 to study the effect of seed treatment with some botanicals on storability of pea. Pea (Pisum sativum L.) cv. P-89 seeds were procured from seed store of STPC of the University having 8.5 per cent moisture content and 86 per cent germination. The seeds were treated with aqueous extracts of leaves of worm, s wood (Artemisia roxburghii L and A. annua L.), mint (Mentha longifolia L; M. spicata L.), marigold (Tagetus erecta L.), drupes of dharek (Melia azedarach L.) at 1%, 2%, 4% and 6% /100g of seed with five replications in each treatment and an untreated control. Aqueous extracts were prepared as per the method of Gahukar [2]. All the treated and untreated seeds were separately kept in plastic jars of 250 cc capacity and stored under ambient conditions (25-300C temperature and 70-80% relative humidity). Seeds were retrieved from storage
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at periodic interval of 2 months from the start of seed storage up to six months and tested for standard germination, seeding length, seed vigour index - length, seedling dry weight and seed vigour index-mass. The germination test was conducted with 50 seeds from each treatment in five replications using Between Paper (B.P.) method as per ISTA Rules [3]. The root length and shoot length of ten normal seedlings were measured on the 6th day. The seedling vigour index-length (SVI-L) was calculated by adopting the method suggested by Abdul-Baki and Anderson [4], SVI-L = Germination (%) x seedling length (cm). Dry weight of seedlings was measured by taking the ten normal seedlings used for root and shoot length. The seedlings were kept in blotting paper and dried in hot air oven maintained at 75o C for 24hours. The dried seedlings were cooled in a desicator for 60 minutes, then seedlings were weighed in an electronic balance and the weight was expressed in mg as dry weight of ten seedlings. Seedling vigour index-mass (SVI-M) was calculated as SVI-M = Germination (%) x seedling dry weight (mg). The data were analyzed statistically following completely randomized design (CRD) after proper transformations using SPSS computer programme. III. Results and Discussions Aqueous extracts of all the plant species except Artimesia spp. significantly improved the seed germination of treated seeds as compared to the untreated control seeds during storage. All the treatments prevented the bruchid infestation to nil (0 per cent) as compared to control where 17 per cent infestation was recorded after six months of storage. Significantly higher germination was observed in M. spicata (89%) followed by M. longifolia (88.93%) irrespective of months and both were equally effective in maintaining seed germination percentage (Table 1).This was followed by seed germination in T. erecta (71.72%) and M. azedarach (71.17%) and both were at par with one another but superior over control. In the present study, A. annua (68.77%) and A. roxburghii (64.36%) gave less reduction in germination as compared to control (69.93%). Ziborkere reported some loss of viability of cowpea seeds treated with chilli powder against C. maculates [5]. Table 1: Effect of aqueous plant extracts and storage period on seed germination of pea seeds Treatment
*Mean seed germination (%) of treated pea seeds in the indicated months and concentration 2nd Month 4th Month 6th Month 6%
4%
2%
1%
Mean
6%
4%
2%
1%
Mean
6%
4%
2%
1%
Mean
overallMean
Artemesia annua
72.00 (58.12)
74.80 (59.89)
75.20 (60.16)
78.00 (62.04)
75.00 (60.05)
68.80 (56.11)
72.00 (58.07)
72.00 (58.13)
74.40 (59.63)
71.80 (57.99)
53.20 (46.84)
58.80 (50.08)
62.00 (51.95)
64.00 (53.14)
59.50 (50.51)
68.77 (56.18)
Artemesia roxburghii
67.00 (55.08)
64.80 (53.65)
66.80 (54.84)
70.00 (56.80)
67.20 (55.09)
62.80 (52.43)
63.60 (52.94)
65.60 (54.10)
67.20 (55.07)
64.80 (53.64)
57.60 (49.38)
60.00 (50.79)
62.00 (51.97)
64.80 (53.62)
61.10 (51.44)
64.37 (53.39)
Melia azedarach
72.40 (58.37)
78.00 (62.09)
78.40 (62.24)
80.00 (63.49)
77.20 (61.57)
70.80 (57.38)
73.60 (59.22)
74.40 (59.75)
76.80 (61.33)
73.90 (59.42)
62.00 (51.98)
60.00 (50.79)
62.40 (52.22)
65.20 (53.86)
62.40 (52.21)
71.17 (57.74)
Mentha longifolia
89.20 (70.86)
93.60 (75.50)
94.80 (76.95)
95.60 (77.93)
93.30 (75.61)
88.00 (69.78)
92.00 (73.62)
93.20 (74.92)
94.40 (76.43)
91.90 (73.69)
80.80 (64.04)
81.20 (64.39)
82.00 (64.92)
82.40 (65.23)
81.60 (64.65)
88.93 (71.22)
Mentha spicata
91.20 (72.77)
93.20 (75.00)
93.60 (75.95)
94.40 (76.43)
93.10 (74.94)
89.60 (71.23)
92.40 (74.02)
92.40 (74.02)
92.80 (74.52)
91.80 (73.46)
78.40 (62.34)
81.60 (64.82)
83.60 (66.27)
84.80 (67.31)
82.10 (65.19)
89.00 (71.19)
Tagetes erecta
71.60 (57.83)
76.40 (62.48)
78.00 (62.12)
82.00 (64.96)
77.00 (61.84)
66.00 (54.37)
69.60 (56.71)
73.20 (58.93)
78.00 (62.10)
71.70 (50.03)
59.00 (50.19)
62.80 (52.43)
68.00 (55.57)
76.00 (60.78)
66.45 (54.74)
71.72 (58.21)
Control
75.60 (60.47)
76.00 (60.70)
74.00 (59.37)
74.80 (59.95)
75.10 (60.12)
71.20 (57.58)
70.40 (57.41)
68.40 (55.80)
66.80 (54.84)
69.20 (56.32)
68.40 (55.80)
67.20 (55.07)
64.00 (53.14)
62.40 (52.19)
65.50 (54.05)
69.93 (56.83)
Mean
77.03 (61.93)
79.54 (64.19)
80.11 (64.48)
82.11 (65.94)
73.89 (59.84)
76.23 (61.66)
77.03 (62.24)
78.63 (63.42)
65.63 (54.37)
67.37 (55.48)
69.14 (56.58)
71.37 (58.02)
*Mean of five replications; Figures in parenthesis are arc sine transformed values Overall CD (p=0.05) Treatment: 1.05; Concentration: 0.79; Treatment x Concentration: 2.09
When effect of aqueous extracts was recorded in two, four and six months, it was found that after two months of storage, the aqueous extracts of M. longifolia (93.3%) and M. spicata (93.1%) were the best and at par with each other followed by M. azedarach (77.2%), T. erecta (77%) and A. annua (75%). The lowest seed germination was recorded in A. roxburghii (67.2%). Similarly after 4th month of seed treatment, M. longifolia gave significantly higher germination (91.9%) which was at par with M. spicata (91.8%) followed by M. azedarach (73.9%), A. annua
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(71.8%), T. erecta (71.7%) and A. roxburghii (64.80%). After 6th month of storage, the seed germination in aqueous extract of M. spicata and M. longifolia was recorded as 82.1 and 81.6 per cent, respectively, whereas in the untreated control the germination was 65.5 per cent revealing thereby that the aqueous extracts of M. spicata and M. longifolia were effective in maintaining seed quality of pea. It was observed that the seed germination was indirectly proportional to concentrations of plant aqueous extracts and also with time after seed treatment except in A. roxburghii during 2nd month (64.8 per cent germination at 4% and 67 per cent germination at 6%) and M. azedarach during 6th month (60 per cent germination at 4% and 62 per cent germination at 6%). George and Patel reported no adverse effect of 10% mint (M. spicata) dry powder on green gram [6]. Keita et al. reported that seeds treated with botanical extracts or oils did not loose their viability [7]. The studies in India and elsewhere carried out by various workers revealed that though various plant products were effective in reducing oviposition and seed damage by pulse beetle, the seed quality and germination were not affected [8], [9], [10], [11]. Singh et al. reported that seed treatment of pigeon pea with neem based bio-insecticides showed higher germination percentage [12]. Table 2: Effect of aqueous plant extracts and storage period on Seed Vigour Index-Length (SVI-L) of pea seeds *Mean SVI-L of treated pea seeds in the indicated months and concentration 2nd Month
4th Month
6th Month
Treatment 6%
4%
2%
1%
Mean
6%
4%
2%
1%
Mean
6%
4%
2%
1%
Mean overallMean
Artemesia annua
899.58 870.38 888.77 985.96 896.17 754.94 779.59 (28.96) (29.31) (29.82) (31.32) (29.90) (27.45) (27.92)
810.27 904.53 812.33 511.41 563.53 592.23 661.96 582.28 (28.46) (30.02) (28.46) (22.62) (23.73) (24.34) (25.70) (24.10)
763.60 (27.49)
Artemesia roxburghii
566.13 589.85 643.57 730.38 632.48 468.62 526.86 (23.79) (24.28) (25.37) (27.03) (25.12) (21.63) (22.95)
580.54 641.74 554.44 419.70 492.62 520.24 605.74 509.57 (24.08) (25.32) (23.50) (20.49) (22.19) (22.81) (24.58) (22.52)
565.50 (23.71)
735.55 761.72 Melia azedarach (27.11) (27.60) 1029.00 1110.00 Mentha (32.07) (33.32) longifolia Mentha spicata Tagetes erecta Control Mean
793.41 (28.17) 1152.00 (33.95)
817.62 (28.60) 1198.00 (34.61)
777.08 (27.87) 1122.00 (33.49)
690.98 (26.27) 995.71 (31.55)
601.83 707.66 711.86 (24.49) (26.59) (26.67) 1049.00 1103.00 1114.00 (32.28) (33.21) (33.38)
678.08 (26.00) 1065.00 (32.63)
567.45 (23.80) 875.66 (29.59)
456.62 (21.36) 897.93 (29.97)
470.41 (21.65) 954.84 (30.89)
506.44 (23.68) 960.56 (30.99)
513.73 (22.62) 922.25 (30.36)
656.30 (25.50) 1037.00 (32.16)
1300.00 1411.00 1503.00 1566.00 1445.00 1252.00 1323.00 1393.00 1413.00 1345.00 891.81 1036.00 1195.00 1185.00 1077.00 (35.99) (37.55) (38.75) (39.58) (37.97) (35.34) (36.37) (37.29) (37.58) (36.64) (29.82) (32.12) (34.54) (34.42) (33.73)
1289.00 (35.78)
915.57 911.23 988.86 1049.00 966.23 802.81 783.53 (30.25) (30.12) (31.43) (32.39) (31.05) (28.28) (27.94)
906.58 961.47 836.60 586.81 634.31 695.16 806.36 680.74 (30.08) (30.99) (29.32) (24.23) (25.18) (26.37) (28.38) (26.04)
836.85 (28.80)
859.98 (29.29) 892.06 (29.64)
693.77 (26.34) 884.97 (29.43)
737.00 (27.04)
877.58 (29.60) 933.08 (30.28)
808.89 (28.42) 968.33 (30.84)
876.16 855.40 742.84 765.50 (29.61) (29.23) (27.23) (27.66) 1032.00 815.47 832.73 (31.88) (28.25) (28.53)
793.71 748.95 555.14 546.47 619.35 (28.18) (27.35) (23.42) (23.34) (24.89) 934.37 629.71 661.15 721.07 (30.31) (24.85) (25.41) (26.50)
705.57 606.63 (26.56) (24.55) 783.71 (27.76)
*Mean of five replications Figures in parenthesis are √x+0.05 transformed values Overall CD (p=0.05) Treatment: 0.48; Concentration: 0.36; Concentration x Treatment: 0.96
Seed vigour is the sum total of all those properties of seeds which determine the potential level of performance and activity of a non-dormant seed during germination and seedling emergence [13]. It was observed that the seed vigour index-length was significantly influenced by aqueous extracts of all these plant species (Table 2). Overall highest seed vigour index-length (1289) was recorded in M. spicata followed by M. longifolia (1037), T. erecta (836.85), A. annua (736.60), M. azedarach (656.30) and the lowest in A. roxburghii (565.50) and all these were significantly different from one another. After 2 nd, 4th and 6th month of seed treatment with aqueous plant extracts, the maximum seed vigour index-length was observed in seeds treated with aqueous extract of M. spicata (1445, 1345 and 1077, respectively) followed by M. longifolia (1122, 1065 and 922.25, respectively) and T. erecta (966.23, 836.60 and 680.74, respectively) as compared to control (855.40, 748.95 and 606.63, respectively). The least seed vigour was recorded in A. roxburghii (632.48, 554.44, and 509.57, respectively) at the end of 2 nd, 4th and 6th month of seed storage. Seed vigour index-length was also observed to be the highest at lowest concentration (1%) of these plant extracts. The overall seed vigour index-mass (Table 3) was also higher (1861) in M. spicata treated seeds followed by M. longifolia (1671), T. erecta (1339), A. annua (1233), M. azedarach (1129). Minimum seed vigour index-mass was recorded when the seeds were treated with A. roxburghii (967.36) in comparison to control where it was 1305 and was significantly different. After 2nd month of seed treatment, the seed vigour index-mass was maximum in seeds
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treated with aqueous extract of M. spicata (2176) followed by M. longifolia (1953) similarly after 4th month also, these two extracts resulted in higher seed vigour index- mass i.e. 2011 and 1696, respectively. Besides, the treatment with T. erecta (1384) and A. annua (1326) was also found to be superior over control (1214). After 6th month of storage, both the extracts of M. spicata (1395) and M. longifolia (1364) recorded significantly higher seed vigour index- mass while T. erecta (1041) and control (1044) were at par with each other. All other treatments resulted in low seed vigour index (Table 3). Table 3: Effect of aqueous plant extracts and storage period on Seed Vigour Index-Mass (SVI-M) of pea seeds *Mean SVI-M of treated pea seeds in the indicated months and concentration nd
4th Month
2 Month Treatment
6%
1360.0 0 (36.83) 1065.0 Artemesia 0 roxburghii (32.61) 1163.0 Melia 0 azedarach (34.09) 1835.0 Mentha 0 longifolia (42.85) 2084.0 Mentha 0 spicata (45.62) 1402.0 Tagetes 0 erecta (37.43) 1671.0 0 Control (40.87) 1511.0 Mean 0 (38.61) Artemesia annua
4%
2%
1%
Mean
6%
4%
1504.0 0 (38.78) 1099.0 0 (33.12) 1357.0 0 (36.83) 1978.0 0 (44.48) 2094.0 0 (45.75) 1629.0 0 (40.12) 1594.0 0 (39.92) 1608.0 0 (39.86)
1488.0 0 (38.58) 1185.0 0 (34.42) 1402.0 0 (37.45) 1997.0 0 (44.68) 2203.0 0 (46.94) 1652.0 0 (40.62) 1657.0 0 (40.70) 1655.0 0 (40.48)
1552.0 0 (39.39) 1293.0 0 (35.96) 1473.0 0 (38.39) 2001.0 0 (44.74) 2321.0 0 (48.18) 1682.0 0 (40.99) 1710.0 0 (41.35) 1719.0 0 (41.28)
1476.0 0 (38.39) 1160.0 0 (34.02) 1349.0 0 (36.69) 1953.0 0 (44.18) 2176.0 0 (46.62) 1591.0 0 (39.79) 1658.0 0 (40.71)
1213.0 0 (34.76) 884.05 (29.72)
1373.0 0 (37.05) 930.26 (30.47)
1060.0 0 (32.54) 1600.0 0 (39.99) 1973.0 0 (44.37) 1181.0 0 (34.34) 1290.0 0 (35.83) 1315.0 0 (35.94)
1090.0 0 (32.98) 1673.0 0 (40.89) 1988.0 0 (44.58) 1360.0 0 (36.69) 1296.0 0 (35.96) 1387.0 0 (36.94)
2%
6th Month 1%
Mean
6%
4%
1337.0 1382.0 1326.0 761.65 906.72 0 0 (27.56) (30.10) 0 (36.56) (37.16) (36.38) 987.52 1069.0 967.72 571.62 779.31 (31.42) 0 (31.08) (23.90) (27.88) (32.70) 1195.0 1269.0 1153.0 863.52 790.03 0 0 (29.37) (28.11) 0 (34.54) (35.63) (33.92) 1759.0 1751.0 1696.0 1304.0 1232.0 0 0 0 0 0 (41.94) (41.84) (41.17) (36.10) (35.09) 2011.0 2073.0 2011.0 1345.0 1389.0 0 0 0 0 0 (44.83) (45.50) (44.82) (36.66) (37.24) 1473.0 1522.0 1384.0 913.00 864.11 0 0 (30.20) (29.40) 0 (38.27) (38.95) (37.06) 1134.0 1136.0 1214.0 1020.0 1136.0 0 0 0 0 0 (33.69) (33.70) (34.78) (31.92) (33.68) 1414.0 1458.0 968.35 1014.0 0 0 (30.82) 0 (37.32) (37.92) (31.94)
2%
1%
Mean overallMean
938.43 957.73 895.63 (30.63) (31.24) (29.88)
1233.00 (34.89)
842.45 903.01 774.10 (29.02) (30.05) (27.71)
967.36 (30.94)
897.73 983.18 883.61 (29.94) (31.35) (26.69)
1129.00 (35.43)
1451.0 0 (38.09) 1426.0 0 (37.75) 1032.0 0 (32.12) 1011.0 0 (31.81) 1086.0 0 (32.77)
1671.00 (40.75)
1471.0 0 (38.35) 1418.0 0 (37.62) 1355.0 0 (36.79) 1010.0 0 (31.78) 1159.0 0 (33.88)
1364.0 0 (36.91) 1395.0 0 (37.32) 1041.0 0 (32.13) 1044.0 0 (32.30)
1861.00 (42.92) 1339.00 (36.32) 1305.00 (35.93)
*Mean of five replications; Figures in parenthesis are √x+0.05 transformed values Overall CD (p=0.05) Treatment: 0.58; Concentration: 0.44; Concentration x Treatment: 1.17
Keshavulu and Krishnasamy recorded higher SVI-I and SVI-II with soybean coated with H. rosasinensis and polykote colours [14]. Mahesh Babu and Hunje reported higher germination, more seed length and more seeding weight with botanicals in soybean treated with botanicals [15]. These findings are more or less in agreement with the findings observed in the present studies. The beneficial effect of botanicals is known to occur due to induced reduction and lipid peroxidation and quantitative changes in biochemical activities and increased free sugars during germination [16]. The present studies thus revealed that there is a great scope of using botanicals like M. spicata, M. longifolia and T. erecta @1%/100g of seed in protecting and maintaining the viability of pea during storage under ambient conditions. IV. References [1] [2] [3] [4] [5] [6] [7]
[8]
C.Park, S.I. Kim and Y.J. Ahn, Insecticidal activity of asarones identified in Acorus gramineus rhizome against three coleopteran stored-product insects. Journal of Stored Products Research, 39(3), 2003, 333-342. R.T. Gahukar, Formulation of neem based products/pesticides. Pestology., 20, 1996: 44-45 Anonymous. International rules for Seed testing. Seed Science Technology, 27, 1999: 25-30. A.A. Abdul Baki and J.D. Anderson , Vigour determination in Soybean seed by multiple criteria. Crop Science, 13, 1973: 630-633. D. S. Zibokere, Insecticidal potency of red pepper (Capsicum annum) on pulse beetle infesting cowpea (Vigna unguiculata) seeds during storage. Indian Journal of Agricultural Sciences, 64, 1994: 727-728. V.George and J.R. Patel, Mint Mentha spicata-a promising botanical protectant for green gram against pulse beetle Callosobruchus analis. Indian Journal of Plant Protection, 20, 1992: 66-69. S.M. Keita, C. Vincent, J. P. Schmit, J.T. Arnason and A. Belanger, Efficacy of essential oil of Ocimum basilicum L. and O. gratissimum L. applied as an insecticidal fumigant and powder to control Callosobruchus maculatus (Fab.) (Coleoptera: Bruchidae). Journal of Stored Products Research, 37(4) , 2001, 339-349. N. Raja, S. Albert, S. Ignacimuthu and S.Dorn, Effect of plant volatile oils in protecting stored cowpea Vigna unguiculata (L.) walpers against C. maculatus (F.) (Coleoptera: Bruchidae) infestation. Journal of Stored Products Research, 37(2), 2001,127-132.
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Kiran Rana et al., American International Journal of Research in Formal, Applied & Natural Sciences, 6(1), March-May 2014, pp. 07-11
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