The Effects of Different Plant Activators on Protein, Lipid and Fatty Acids in Snack-Seed Pumpkin

. In this study, Nusem and Beppo snack seed pumpkin cultivars were used to determine the effects of different plant activators on seed protein, lipid and fatty acids contents. In the context of study, plant activators consist of Crop-set (CR), EM1, ERS, Vitormone-Plus Drip (VIT), Bacillus subtilis (OSU 142), Bacillus megatorium (M3), Azospirillum sp . (SP 245), Spirulina platensis (SIP), Ecocompost (EKO), Camli Botanica liquid organic fertilizer (BOT) and Zincon (ZIN) were used as organic fertilizer. In the experiment, the plant activators were applied to the plants alone or in combination with each other and organic fertilizer. Two separate control groups which were organic and conventional (CONV.) fertilizer have been identified. As a result of the use of different plant activators, the highest protein content was obtained from CONV. application (35.50%), M3+SP 245 (33.09%) and M3 (33.04%); the highest lipid content was observed from SP 245+OG (45.90%), CR (44.48%) and SIP+OG (44.26%) applications. The use of different plant activators effected the fatty acid contents of seeds. Total 11 fatty acids were identified. Among the fatty acids, C16:0 (Palmitic acid), C18:0 (Stearic


Introduction
Snack pumpkin product is the seed of plant from Cucurbita pepo and Cucurbita moschata. Seeds not only contain snack food but are also sources for food raw materials. Turkey produces about 42 181 tons Snack pumpkin in 628 441 area [1]. Pumpkin seed (Cucurbita pepo L.) is high in oil, protein, and total unsaturated fatty acids and provides an important source of nutrition [2]. Snack pumpkin showed rice produce both area and quantity. But, despite the increase in production quantities, the amount of product received from the unit area did not reach adequate levels. For this purpose the increase yield and quality used microbial fertilizer in addition plant activators [3]. Plant activators contains alone such as Plant Growth Promoting Rhizobacteria= PGPR; Azotobacter, Azospirillum and Bacillus subtilis, Bacillus firmus, Pseudomonas striata etc.), mycorrhiza (Glomus fasciculatum, Glomus intraradices, Glomus aggregatum etc.), microalgae (Spirulina platensis) microorganism or combination these microorganism with mineral element, vitamin, plant extract etc. The present study was carried out to examine the effect of different plant activators on total proteins, total lipid, fatty acids of snack-seed pumpkin.

Materials and Methods
This study was conducted at the experimental field in Cukurova University Pozanti Agricultural Research and Application Center. "Nusem" snack seed pumpkin and "Beppo" naked seed pumpkin were used as plant material. These snack pumpkins were cut and seeds extracted in 07 July 2015 and 21 July 2016 respectively, the seeds dried about ten days for analysis. 5-10 g powdered seed sample was taken for determination of lipid and 1 g for protein analysis.
As organic fertilizers (OG); Ecocompost, Camlı Botanica Liquid organic fertilizer and Zincon have been used. In the experiment, the plant activators were applied to the plants alone or in combination with each other and organic fertilizer. Two separate control groups which were organic and conventional (CONV.) fertilizer have been identified. The following methods were used: Kjeldahl method [4] for total protein, and Bligh and Dyer method [5] was used for total oil; powdered seed samples were measured in quantities ranging from 5-10 g and placed in to the cellulose cartridges. Over the cartridge was pressed well with cotton on the purpose of that the samples should not diffuse. The cellulose cartridge was placed on a glass ballon and extractioned with diethyl ether solution for 3 hours. Diethyl ether was evaporated from the diethyl ether-lipid fraction which have been remained in the glass balloons using the method of a rotary evaporator in a water bath at 65 ° C. After that, the balloons were waited at stand at 60 ° C for 20 minutes in the stove to evaporate of the diethyl ether, they cooled until the room temperature in a desiccator and weighed on a 0.1 mg sensitive precision scale. The following formula has been used in the calculation of the lipid ratio Kenar [6].
Lipid ratio (%) = [Glass balloon weight (g)+ Lipid (g)]-[Glass balloon (g)] x100/ Sample weight (g) Fatty acid profiles of fat extracted from the seed snack pumpkin samples were determined according to the method described by Ichibara et al. [7] with minor modification by using by gas chromatography (GC).
Statistical analyses were performed by the use of the JMP 5.1 program and Tukey test was used to compare the averages.

Results and Discussion
At in the percent of protein, lipid and fatty acid, significant different between applications average were recorded (Table 1, 2 and 3). The protein content ranged from 28.32 % to 35.50 %. The highest value was obtained from application of CONV. whereas applications ERS+OG and VIT+OG had the lowest protein content (28.32%) ( Table 1).  Table 2 shows the lipid contents of seeds. The highest value was obtained from SP 245+OG (45.90%), CR (44.48%) and SIP+OG (44.26%) whereas the lowest value was observed from applications CONV. (37.24%) and SP 245 (39.41%).  Table 3 shows dominant fatty acids found in oils of seeds. Total 11 fatty acids were identified. Among them, the highest proportions in oil of snack seed pumpkin were C16:0 (Palmitic Acid Methyl Ester), C18:0 (Stearic Acid Methyl Ester), C18:1 (Oleic Acid Methyl Ester) and C18:2 (Linoleic Acid Methyl Ester).

International Letters of Natural Sciences Vol. 72
Ermis [8] reported that the pumpkin seeds contained high lipid (35%-48%), and also oleic acid (40%-58%) and linoleic acid (30%-40%) were dominant fatty acids. He also reported that it contained 35%-40% of protein. These results were similar to the results of our study. However, the values were found to be different for oleic acid, linoleic acid and protein. Stevenson et al. [9] reported that the oil content of pumpkin cultivars ranged from 10.9 to 30.9%. Researchers also reported the predominant fatty acids in pumpkin seed oil from all 12 cultivars studied as linoleic acid, oleic acid, palmitic acid, and stearic acid. These results were similar to findings of our study but values for oil content were found to be different. Lazoz [10] investigated the nutritional, fatty acid, and oil characteristics of pumpkin and melon seeds. As a result, they found the defatted flour of the pumpkin and melon seeds contained, respectively: crude protein, 55.4 and 39.4%; and the major fatty acid of the oils was linoleic (18:2) at concentrations of 43.1 and 64.6%, followed by oleic (18:1), 37.8 and 20.1%, respectively. Habib et al [11] reported that the seeds of pumpkin (Curcurbita maxima) contained 36.70% total lipid, 34.56% total protein and pumpkin seed oil contained the highest amount of oleic acid (40.58%) while stearic acid, palmitic acid and linoleic acid content were found to be 27.06, 17.39 and 14.97%. These result were similar to the results of our study but values for protein, lipid, palmitic and linoleic acid were found to be different.

Conclusions
In our study eight plant activators and three organic fertilizer were used in snack seed pumpkin with or without organic fertilizers. Total protein ranged from 28.32% to 35.50%, and total lipid ranged from 37.24% to 45.90%. 11 fatty acid were identified. C16:0, C18:0, C18:1n9 and C18:2 were found dominant compared to other fatty acids. As a result, applied plant activators on Nusem and Beppo varieties had no effect total protein content, total lipid and fatty acid in snack seed pumpkin.

Conflict of Interest
The authors declare that there is no conflict of interest.