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Int.J.Curr.Microbiol.App.Sci (2020) 9(7): 3484-3499 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 9 Number 7 (2020) Journal homepage: http://www.ijcmas.com Original Research Article https://doi.org/10.20546/ijcmas.2020.907.407 Effect of Seed Priming with Botanicals on Plant Growth and Seed Yield of Lentil (Lens culinaris M.) Deepak Chand Bhateshwar, Deepti Prabha, Deepak Jangid and Mohammad Salman* Department of Seed Science and Technology, Hemvati Nandan Bahuguna Garhwal University, Srinagar (Garhwal), Uttarakhand (A Central University), India *Corresponding author ABSTRACT Keywords Lentil, Seed Priming, Botanicals, Neem leaf extract, Castor oil, Ginger extract, Onion extract, Garlic extract etc Article Info Accepted: 22 June 2020 Available Online: 10 July 2020 A field experiment was conducted during rabi season, 2018-19 at the Research Farm, Department of Seed Science and Technology, Chauras Campus, H. N. B. Garhwal University, Srinagar (Garhwal), Uttarakhand, India to study the effect of seed priming with botanicals on plant growth and seed yield of lentil (Lens culinaris M.). The experiment was laid out into Randomized Block Design and replicated three times. The field experiment consisted of six treatments viz., seed priming with neem leaf extract (50%), castor oil (20%), ginger extract (50%), onion extract (50%) and garlic extract (50%) and control. The seed priming with 50% extract of garlic inhibits germination due to the presence of Allicin. The maximum germination percentage (90.80%), number of branches per plant (5.20), seeds per pod (1.87), 1000 seeds weight (20.24 g), yield per plant (3.51 g) and yield per ha (9.94 g) were observed when seeds were primed with neem leaf extracts (50%). Whereas, maximum plant height at 60 DAS (12.80 cm) and number of pods per plant (174.73) were observed when seeds were primed with onion extract (50%). Introduction Pulses are the edible seeds of plants in the legume family and grown in major area in India. It contributes a significant role in Indian agriculture due to their adaptability in various climatic conditions. In Indian meal, the pulses are used as a source of vegetable protein. Pulses grow in pods and come in a variety of shapes, sizes and colors. The United Nations Food and Agriculture Organization (FAO) recognizes 11 types of pulses: dry beans, dry broad beans, dry peas, Bambara beans, pigeon peas, chickpeas, cow peas, lentils, vetches, lupins and pulses nes. Pulses are healthy, nutritious and easy to cook with. Growing pulses also promotes sustainable agriculture, as pulse crops help decrease greenhouse gases, increase soil health, and use less water than other crops. Importantly, seed priming has shown its effectiveness in improving seed germination, seedling growth and crop stand against the 3484 Int.J.Curr.Microbiol.App.Sci (2020) 9(7): 3484-3499 negative impacts brought about by stress in the field. Seed priming is a commercially used technique for improving seed germination and vigour. It involves imbibitions of seeds in water under controlled conditions to initiate early events of germination, followed by drying the seed back to its initial moisture content (Varier et al., 2010).Treatment with water, salt solution, certain hormones, organic and inorganic chemicals, and pesticides, etc. (Vasileva and Ilieva, 2007) are primarily practiced in seed priming. Also, pre-soaking, hardening, hormonal priming, hydropriming, halopriming, Osmoconditioning, and ascorbate priming are some common priming techniques to improve speed and synchrony of seed germination. Botanical priming is new endeavor and many botanical extracts have been studied for their effect on seed and seedling parameters. In recent years, the use of local botanicals has gained much importance, mainly among researchers, because of its high benefits in plant growth, yield and seed quality attributes. The neem tree, Azadirachtinindica, a source of several insecticidal alkaloids is a subtropical tree native to the arid areas of Asia and Africa (Saha et al., 2006). Azadirachtin is the main pesticidal component of neem. Neem products are naturally available materials, cheaper, and also safe for beneficial organisms. Seed treatment with neem leaf extract and their use in agriculture is increasing because of beneficial effects on plants. Some other botanicals like garlic extract, onion extracts, castor oil etc. can also be used for seed treatment and various effect of them can be seen on growth and yield of a plant. Botanical seed treatment is extracted from naturally occurring sources based on botanical ingredients. It has synergistic effect on early and uniform seed germination and enhances tolerance to pest and disease during early crop stage. Botanical extracts been reported to possess antifungal activities against seed borne fungies (Suratuzzaman et al., 1994; Ashrafuzzaman and Hossain, 1992; Hossain and Schlosser 1993). Botanicals have been found to be effective for reduction of population of Fusarium associate with seed (Bowers and Locke, 2000). Available literature indicate that plant extracts and plant essential oils possess effective antimicrobial principles against food and grain storage fungi (Mishra and Dubey, 1994). Materials and Methods Climate In the experimental site except during the rainy season, rests of the months are usually dry, with exception of occasional showers during winter or early spring. Srinagar Garhwal region comes under sub-tropical climate, with both extremes in the temperature i.e., winter and summer. The monthly meteorological data on various weather parameters were recorded at meteorological observatory of Research Farm, Department of Seed Science and Technology, Chauras Campus, H. N. B. Garhwal University, Srinagar, Garhwal, Uttarakhand (India), during the whole experimental period in 2018-19. The data for the climatic features of the experimental site have been recorded during the period of experimental trail i.e, November-March and is presented in the following figure.1. Meteorological data The Research farm falls under humid and subtropical climatic condition which exhibits dry summer and rigorous winter with occasional dense fog from mid-November to midFebruary. In winter, there is less rainfall comparison to summer season. The 3485 Int.J.Curr.Microbiol.App.Sci (2020) 9(7): 3484-3499 temperature reaches 45 ºC on some days from May-June and 2 ºC in December-January. The maximum temperature range between 13ºC26ºC and minimum between 5 ºC-20 ºC. The maximum precipitation during experiment is 16.3 mm mostly received during December and minimum during November. Fertilizer application 15 t/ha FYM were applied in the field and then cross ploughing was done with the help of tractor so that FYM could be mixed thoroughly. Preparation of botanical extracts The data of temperature, relative humidity and rainfall during the experiment was obtained from the meteorological observatory of Horticulture Research Centre, Department of Horticulture, Chauras Campus, Srinagar Uttarakhand has been presented in appendix I and depicted in fig.1. Soil condition In order to find out the soil texture, response and fertility status, a composite soil sample representing the distinct areas of the experimental site was drawn from the experimental plot from 0-20 cm depth before transplanting the crop. Prepared and processed composite soil sample was analyzed at departmental laboratory and the results thus obtained have been presented in the table Materials During the course of experiment, Lentil Cv. PL-8was used. This variety is developed by G B Pant University of Agriculture and Technology, Pantnagar, Uttarakhand. The fruits of this variety are green and long tender. It is long duration crop. This variety is high fruit weight and good bearing. Preparation of stock solution The stock solution of botanical extracts was prepared by grinding the 25 g of botanical with adding 25 ml water in it. By this, 1:1 solution of all the botanicals was prepared. In case of neem, leaves were used, while rhizomes in ginger, bulbs in onion and cloves in garlic were used to prepare stock solution. Method of priming The 50% solution of botanicals was prepared for priming of seeds. The 20% solution of castor oil was prepared by adding 10 ml oil into 40 ml of distilled water. The seeds were kept overnight into solution for soaking. On the next day, the seeds were dried into shade and sown into respective plots. Sowing The seeds were sown done on 15 November, 2018 by manually with 2 seeds at a location, at a depth of 3-4 cm in the rows which were prepared before sowing. Irrigation Land preparation Through ploughing, the ground was carried to a good tilth, and two crosses were harrowing. The beds were ready and channels for adequate irrigation were supplied. The crop was raised in winter season, so much irrigation was not needed. The light irrigation was given after sowing followed by after ten days of interval. Thereafter, irrigations were given as per the requirement. 3486 Int.J.Curr.Microbiol.App.Sci (2020) 9(7): 3484-3499 recorded at 60 and 90 DAS and at maturity under irrigated and rainfed conditions. Thinning The thinning was done at 35 days after sowing of seeds to obtain optimum plant population in each experimental plot. Number of branches Intercultural operations Five plants from each replication were selected and number of branches was counted and the mean was calculated. Hoeing and weeding operations were done whenever required. Necessary plant protection measures were being taken up to protect the crop from pest and diseases. Harvesting Harvesting was done after the seeds attained physical maturity. The harvesting of the crops was done on 9 March, 2019. Observations recorded Five plants were randomly selected from each plot. The following observations were recorded on the different growth and yield parameters. The characters studied under the present experiment are listed below. Growth parameters Plant Fresh weight Five plants from each replication were uprooted at 60 DAS and their weight was measured with the help of balance machine and expressed in gram. Total Dry matter Five plants from each replication were uprooted at 60 DAS and oven dried at 65°C till constant weight was obtained. Plant dry weight was expressed as g plant-1. Days to 50% flowering The time taken to produce flower by 50% of plants in each treatment of each replication was recorded and days counted by date of sowing. Germination percentage (%) Stem dry weight Number of germinated seeds were recorded every 48 h after initiation of germination until 10 days. Germination percentage was calculated (Association of Official Seed Analysis 1983) as follows: Germination percentage = (No. of seed germinated/ Total No. of seed sown) × 100 Five plants from each plot were uprooted at the time of 50% flowering and their main stem was separated. The stem than oven dried at 65 oC for 24 h and their weight was measured at balance machine and expressed in gram. Leaf dry weight Plant height at 60 and 90 DAS Height of the selected plants was measured from the base of the plant (soil surface) up to the growing tip of the main stem with meter scale and expressed in centimetres. It was Five plants from each plot were uprooted at the time of 50% flowering and their leaves were separated. The leaves than oven dried at 65 oC for 24 h and their weight was measured at balance machine and expressed in gram. 3487 Int.J.Curr.Microbiol.App.Sci (2020) 9(7): 3484-3499 level of significance for each character was worked out. Yield Parameters Number of pods per plant Results and Discussion Pods of 5 plants were selected randomly at maturity from each treatment and counted and average number of pods plant-1was calculated. Number of seeds per pod After threshing the bunch of five plants, number of seeds were counted and divided with total number of pods recorded from these five plants. Yield per plant (g) After threshing, the total yield of five tagged plants per plot was weighed and the resultant was divided by the total tagged plants and the seed yield per plant was obtained. Yield per plot (g) Seed yield of each net plot was weighed and recorded after threshing and winnowing. Yield per hactere (kg) Straw yield was calculated in kg by subtracting the grains yield from the biological yield. Test weight (100) seeds (g) One hundred seeds were randomly taken from the harvest (8% seed moisture) and weighed on a precision balance to record their mass. Statistical analysis The statistical analysis for all the characters studied was done by method recommended by Panse and Sukhatme, (1961) for Randomized Block Design and for the statistical analysis of data. The CD (Critical Difference) at 5% In further studies garlic was not use after germination because 50 Percent of garlic reduces germination to 0%. In this study results of the experiment entitled Effect of Seed Priming with Botanical Extracts on Plant Growth and Yield of Lentil (Lens culinaris M.) obtained during the course of investigation are summarized with the help of suitable table and graphs. The botanical seed treatments were found significant for different morphological fetures of lentil. Growth parameters Germination percentage It is evident from Table 4.1 that the seed priming with neem leaf extracts resulted as maximum germination percentage (90.80 %) followed by T5 (89.23 %) and T4 (88.44 %) when compared to control (86.01%). No seeds were germinated when the seeds were primed with 50% garlic extracts (Fig.2). Plant height The data pertaining to the plant height given in the Table 4.2indicated thatthere was significant difference in plant height at90 DAS (Fig.3). Maximum plant height (34.27 cm) was observed in treatment T4, followed by T5 (33.80 cm) and T3 (31.93 cm). Whereas, least plant height (28.80) was observed in control. There was no significant difference in plant height at 60 days after sowing. Number of branches per plant It is revealed from observed data presented in Table 4.2 that there was significant difference 3488 Int.J.Curr.Microbiol.App.Sci (2020) 9(7): 3484-3499 between treatments in relation to number of branches per plant (Fig.4). Higher number of branches (5.20) were recorded in T2 followed by T4 (4.53) and T3 (4.33). While the lowest number of branches (3.60) were observed in which was at par with T5 (4.40). Plant freshweight(g) Various seed priming treatments shown significant difference in fresh weight of plant at 60 DAS (Fig.5). Higher plant weight was observed in T3 (25.77 g) when compared to control (19.12 g). Treatments T5, T2 and T4 were recorded plant fresh weight 24.13 g, 20.45 g and 20.20 g, respectively (Table 4.3). Total dry matter (g) Seed priming with various botanical extracts revealed significant difference in total dry matter of plant at 60 DAS (Fig. 4.5). Maximum total dry matter (6.50 g) was observed in T3, while the least total dry matter (4.75 g) was recorded in control which was at par with T5, T2 and T4, respectively in ascending order (Table 4.3). Days to 50% flowering It is evident from the Table 4.4 that there was significant difference between the treatments in relation to days to 50% flowering (Fig. 5). Maximum days to produce 50% of flowers (63.67 days) were taken by control. Seed priming with onion extracts was resulted as minimum days (56.33 days) for producing 50% of flowers. Stem dry weight at 50% flowering The data in Table 4.4 indicating that maximum stem dry weight (641.67 mg) at the time of 50 % flowering was observed in T4 followed by T5 (511 mg), T2 (500.33 mg) and T3 (492.67 mg). Minimum dry weight (438.67 mg) was observed in control. Leaf dry weight at 50% flowering The data in relation to leaf dry weight at the time 50% flowering was revealed significant difference between treatments (Fig.7). The maximum leaf dry weight was observed in T5 (3.57 g) followed by T4 (3.54 g), T2(3.32 g) and T3(2.50 g). Whereas, the minimum leaf dry weight (2.46 g) was observed in control (Table 4.4). Yield parameters Pods per plants The data on number of pods per plant, presented in Table 4.5 indicated significant differences between various treatments (Fig. 8).Among the treatments, T5 recorded significantly higher number of pods per plant (174.73), followed by T2 (157.47), T4(149.40) and T3 (135.80). Whereas, minimum number of pods per plants was found in T1 (118.40). Seeds per pods It is evident from Table 4.5 that result indicated significant differences between various treatments (Fig.9). Among the treatments, T2 and T4 recorded similar and higher number of seeds per pods (1.87), which were followed by T5 (1.86), T3 (1.81). Whereas, minimum number of seeds per pods (1.76) was found in T1. Yield per plant (g) Plant (g) The data on yield per plant, presented in Table 4.6 indicated significant differences between various treatments (Fig.10).Among the treatments, T5 recorded significantly higher yield per plant (3.90 g), followed by T2 (3.51 g), T3 (3.46 g) and T4 (135.80 g). Whereas, minimum yields per plant (3.27 g) was found in T1. 3489 Int.J.Curr.Microbiol.App.Sci (2020) 9(7): 3484-3499 Yield per plot (g) Yield per hectare (q) The data with respect to yield per plot, given in Table 4.6has shown significant differences between various treatments (Fig.11). Among the treatments, T2 recorded significantly higher yield per plot (167 g), followed by T5 (160 g), T3 (148 g) and T4(144 g). Whereas, minimum yield per plot (139 g) was found in T1. In relation to yield per hectare, T2 recorded significantly higher yield per hectare (9.94 q), followed by T5 (9.56 q), T3 (8.83 q) and T2 (8.57 q). Whereas, minimum yield per hectarewas found in T1(8.29 q). Table.1 Soil analysis of experimental plot S.No. 1 2 3 4 5 6 Properties Texture Soil Ph Organic carbon (%) Available nitrogen (kg/ha) Available phosphorus (kg/ha) Available potassium (kg/ha) Value Sandy clay 6.3 0.85 94.3 3.55 131 Experimental details 1 2 3 4 5 6 7 8 9 Experimental design Number of replications Number of treatments Total number of plots Spacing Plot size Variety used Date of sowing Date of Harvesting Randomized Block Design 3 6 18 30 × 10 cm 1.2 × 1.4 metres PL-8 15-November-2018 9 March, 2019 Details of treatments Sr. No. 1. 2. 3. 4. 5. 6. Treatment No. T1 T2 T3 T4 T5 T6 3490 Treatments Control Neem leaf extract Castor oil Ginger extract Onion extract Garlic extract Int.J.Curr.Microbiol.App.Sci (2020) 9(7): 3484-3499 Table.4.1 Effect of seed priming with botanicals on germination percentage of lentil Treatments T1 T2 T3 T4 T5 T6 S.EM. CD at 5% Germination percentage (%) 86.01 90.80 81.25 88.44 89.23 0.00 1.04 3.28 Control Neem leaf extract Castor oil Ginger extract Onion extract Garlic extract Table.4.2 Effect of seed priming with botanical extracts on Plant Height at 60 DAS and 90 DAS and number of branches Treatments T1 T2 T3 T4 T5 S.EM. CD at 5% Control Neem leaf extract Castor oil Ginger Onion Plant height at 60 DAS 12.67 13.00 13.20 14.73 12.80 0.90 NS Plant height at 90 DAS 28.80 30.07 31.93 34.27 33.80 0.71 2.33 Number of branches per plant 3.60 5.20 4.33 4.53 4.40 0.13 0.44 Table.4.3 Effect of seed priming with botanicals on Plant fresh weight and dry weight at 60 DAS Treatments Control Neem leaf extract Castor oil Ginger Onion S.EM. CD at 5% Plant fresh weight at 60 DAS 19.12 20.45 25.77 20.20 24.13 0.49 1.60 Total dry matter at 60 DAS 4.75 5.40 6.50 4.90 5.87 0.35 1.16 Table 4.4 Effect of seed priming with botanicals on Days to 50% flowering, Stem dry weight at 50% flowering and Leaf dry weight at 50% flowering Treatments Control T1 Neem leaf extract T2 Castor oil T3 Ginger T4 Onion T5 S.EM. CD at 5% Days to 50% flowering 63.67 60.67 58.33 59.67 56.33 0.94 3.07 3491 Stem dry weight at 50% flowering (mg) 438.67 500.33 492.67 641.67 511.00 6.88 22.43 Leaf dry weight at 50% flowering (g) 2.46 3.32 2.50 3.54 3.57 0.10 0.32 Int.J.Curr.Microbiol.App.Sci (2020) 9(7): 3484-3499 Table.4.5 Effect of seed priming with botanicals on Pods per plant and Seeds per pod Treatments Control T1 Neem leaf extract T2 Castor oil T3 Ginger T4 Onion T5 S.EM. CD at 5% Pods per plant 118.40 157.47 135.80 149.40 174.73 4.33 3.84 Seeds per pod 1.76 1.87 1.81 1.87 1.86 0.02 0.08 Table.4.6 Effect of seed priming with botanicals on yield per plant,yield per plot and yield per hector Treatments Control T1 Neem leaf extract T2 Castor oil T3 Ginger T4 Onion T5 S.EM. CD at 5% yield per plant (g) 3.27 3.51 3.46 3.42 3.90 0.12 0.38 Y yield per plot(g) 139.33 167.00 148.33 144.00 160.67 5.81 18.95 yield per hector(qt) 8.29 9.94 8.83 8.57 9.56 0.35 1.13 Table.4.7 Effect of seed priming with botanicals on 1000 seeds weight(g) Treatments T1 T2 T3 T4 T5 1000 seeds weight(g) 17.83 20.24 19.07 18.59 18.73 0.45 1.46 Control Neem leaf extract Castor oil Ginger Onion S.EM. CD at 5% Fig.1 100 90 80 70 60 50 40 30 20 10 0 November December January February March Max. Temp. Mini. Temp. RH Max. 3492 RH Mini. Rainfall (mm) Int.J.Curr.Microbiol.App.Sci (2020) 9(7): 3484-3499 Fig.2 Effect of seed priming with botanical extracts on Plant Height at 60 DAS and 90 DAS 40 35 30 25 20 15 10 5 0 T1 T2 T3 Plant height at 60 DAS T4 T5 Plant height at 90 DAS Fig.3 Effect of seed priming with botanicals on fresh weight of plant at 60 DAS 30 20 10 0 T1 Plant fresh weight at 60 DAS T2 T3 T4 T5 Fig.4 Effect of seed priming with botanicals on total dry matter of plant at 60 DAS 8 6 4 2 0 T2 Total dry matter T3 at 60 DAS T1 T4 T5 Fig.5 Effect of seed priming with botanicals on Days to 50% flowering 66 64 62 60 58 56 54 52 T1 T2 Days to 50%T3flowering 3493 T4 T5