Pollinator Visit, Characteristics of Secreted Nectar and Analysis of Nectar Sugar and Amino Acid Contents in Flower of Dendropanax morbifera Lev. Kim Moon-Sup, Kim Hyeusoo, Kim So-Dam 1, Park Sang-Jin 1, Song Jeong-Ho and Kim Sea-Hyun* Division of Special Purpose Trees, National Institute of Forest Science, Suwon 16631, Korea 1 Department of Forest Resources, Sunchon National University, Suncheon 57922, Korea (Received 3 November 2015; Revised 12 November 2015; Accepted 12 November 2015) Abstract This study was conducted to provide a basic data such as honeybee visit and secreted nectar for possibility of development as a honey plant, Dendropanax morbifera Lev. The surveyed tree s flowers bloom during 23 days in U-san park, Gwang-yang city in 2014. It is showed that more Apis mellifera visited flowers in priority than other pollinators, so the percentage of A. mellifera which visited flowers was 69%. And number of visited honeybees per flowering lateral bunch can be assumed 47.3 honeybees for a day. Nectar volume secreted by nectary was 4.5µl and dry nectar volume was 0.48µl from one flower. Nectar concentration showed 20.9% and sugar contents of a flower is 79.4µg, averagely. So, the minimum estimates of honey harvest for a flower was 93.4µg. Analysis of amino acid showed that Phenylalanine, Glutamine and Proline were more abundant. Especially, Phenylalanine ratio shows 67.2%. Finally, the surveyed D. morbifera is considered as possible sub-honey plant because of its visited honeybee data and nectar characteristics such as nectar sugar and amino acid contents. Key words: Nectar secretion, Sugar contents, Honey potential, Amino acid contents, Phenylalanine (Honey), (Nectary) (Nectar).. *Corresponding author. E-mail: goldtree@korea.kr 307
308 (Parenchyma cell),,. 80%,,,,,, (Pacini et al., 2003). 1950 Beutler(1953), (1984) Microcapillary tube., (, 2009; Han et al., 2010; Kim et al., 2011;, 2013;, 2014a;, 2014b; Kim et al., 2015), (Kang et al., 2009)., (Nicolon et al., 2007), Sucrose 6 Fructose Glucose. (Petanidou et al., 1996)., (Dynamic) (Wolff et al., 2006)., (Nicolon et al., 2007).,. (Dendropanax morbifera Lev.),. 8~9 3~5cm, 5~10mm. 5 5. 5, 5 (, 2006)., (, 1998).,. 4,176 555 (, 2014), 250 (, 1987),.. 15 Fig. 1. Pictures of D. morbifera shape and flowers.
, 4.5m, 14.5cm 3. 4. 2 24. 08 16 1, 6 10 (Apis mellifefa). 1,, 8 15 8 17 3. 20 10 4 3µl capillary tube(drummond, USA) 4. 4. 2 (15~16, 16~17 ) 3µl capillary tube (FO- 600M, JEIO TECH, Korea) 80 C 24,..,, 500µl microliter syringe(hamilton, USA) (GMK-703T, Korea). HPLC 0.45µm membrane filter 80% Eppendorf vial ( 70 C). 0.45µm syringe filter(millipore, Billerica, MA, USA) HPLC(Dionex ultimate 3000, Dionex, USA). 3, 0.5ml/min, 80 C. Shodex Ri-101 detector(japan), Aminex 87P column(300mm 7.8mm/Bio-rad, USA)., Sucrose, Glucose, Fructose(Sigma, USA). O-phthalaldehyd (OPA)-Fluorenylmethyl chloroformate(fmoc). Borate buffer, OPA/Mercaptopropionic acid(mpa), FMOC, 0.45µm syringe filter (Millipore, Billerica, MA, USA) HPLC(1200 series, Aglient, USA). 10mM Na 2 HPO 4, 10mM Na 2 B 4 O 7 10H 2 O A (ph 8.2) Water: Acetonitrile:Me thanol=10:45:45 B A :B 100:0(v/v, %) 26~28 55:45, 28~30.5 0:100, 30.5 100:0. 1.5ml/min 1ml 40 C Inno C18 column (Innopia, Korea, 4.6mm 150mm, 5µm)., 338nm, OPA 450nm, 340nm, FMOC 305nm, 309
310 266nm. 2014 8 8 8 31 23. 8 8 8 22 15 8 15 17 16 (Fig. 2).,. (Apis mellifera), (Bombus ignitus), (Vespidae), (Syrphidae). Flowering rate (%) 100 90 80 70 60 50 40 30 20 10 0 8th 9th 10th 11th 12th 13th 14th 15th 16th 17th 18th 19th 20th 21st 22nd 23rd August Date Fig. 2. Flowering rate of D. morbifera. Number of pollinator / Flowering lateral bunch (ea) 18 16 14 12 10 8 6 4 2 0 8:00 A. mellifera Other pollinators 9:00 10:00 11:00 12:00 13:00 14:00 15:00 16:00 Time Fig. 3. Visited number of pollinators on flowering lateral bunch of D. morbifera. A : A.mellifera B : Vespa nalis Fig. 4. Picture of major pollinators and ratio of visited pollinators on flowering lateral bunch of D. morbifera. (Fig. 3). 8 16 47.3, 21.8, 13 14. 372,,, 458, 488, 290, 220, 160 (, 2009; Han et al., 2010;, 2013;, 2014b; Kim et al., 2015),. 69%, (Vespidae), (Syrphidae), (B. ignitus), (Fig. 4).,.,,. 15% 4% 8% 4% 3 2 69% A. mellifera B.ignitus Vespidae Syrphidae Etc.
Table 1. Pattern of secreted nectar volume of total and dry nectar per flower during the 2 days Characteristics Total nectar 1.58 2.49* 0.49 0.90 0.47 0.70 1.49 1.51 4.03 Dry nectar 0.09 0.13 0.05 0.08 0.05 0.06 0.22 0.14 0.40 Total nectar 3.01 2.51 1.72 2.10 0.03 0.08 0.23 0.56 4.99 Dry nectar 0.27 0.18 0.24 0.27 0.00 0.01 0.04 0.09 0.55 *expressed Mean S.D. 8/15 8/16 8/17 10h 16h 10h 16h 10h 16h Total (unit: ul) Table 1. (Kim et al., 2011; Kim et al., 2012;, 2013;, 2014b),. 8 15 4.03µl, 8 16 4.99µl 4.5µl. 15 12µl 13.8µl 10 18.4µl (Han et al., 2010; Kim et al., 2011;, 2013).,. (Pacini et al., 2003)., 88.1%, 0.40µl 0.55µl 0.48µl. 2.0µl, 1.0µl. Nepi and Stpiczynsk(2007),. (Kim et al., 2012). 8 16 17 Microliter syringe, HPLC Table 2. 16 17 60 240µl 4.0µl Table 2. Nectar composition and estimated sugar contents and honey production based on nectar characteristics in the floral nectar of D. morbifera Characteristics Sample size (flower) 60 21 61 Collected nectar volume (µl/flower) 4.0 2.4 2.1 2.8 Nectar concentration (w/w%) 23.6 16.0 23.2 20.9 Amount of free sugars (µg/µl) 29.1 25.6 28.6 27.8 Sugar contents of flower (µg/flower)* 116.3 61.0 61.0 79.4 Honey potential (µg/flower) 136.8 71.7 71.7 93.4 *Collected nectar volume (µl/flower) Amount of free sugars (µg/µl) Date / (Time) 8.16 / (17:00) 8.17 / (10:00) 8.17 / (17:00) Average 311
312 Table 3. Amino acids on nectar in flower of D. morbifera Amino acid Date / (Time) 8.16 / (17:00) 8.17 / (10:00) 8.16 / (17:00) Average of percentage on total amino acid Percentage of amino acid (%) Aspartate 0.9 0.7 0.9 0.8 0.1 Glutamate 1.1 1.2 2.2 1.5 0.6 Asparagine 2.4 1.5 3.5 2.5 1.0 Serine 3.4 2.0 3.1 2.9 0.7 Glutamine 10.8 8.8 13.5 11.0 2.3 Histidine 1.5 1.6 2.2 1.8 0.4 Glycine 1.0 0.3 0.5 0.6 0.3 Threonine 1.0 0.7 1.1 0.9 0.2 Arginine 0.2 0.1 0 0.1 0.1 Alanine 1.3 1.1 1.3 1.2 0.1 GABA 0.5 0.2 1.0 0.6 0.4 Tyrosine 1.9 1.0 1.6 1.5 0.4 Valine 1.0 0.5 1.0 0.8 0.3 Methionine 0 0 0 0 Tryptophan 0 0 0 0 Phenylalanine 66.8 76.4 58.3 67.2.9.1 Isoleucine 1.0 0.5 0.9 0.8 0.3 Leucine 1.2 0.7 1.1 1.0 0.3 Lysine 0 0 0 0 Hydroproline 0 0 0 0 Proline 4.2 2.5 7.8 4.8 2.7 Sum 100 100 100 100 23.6%. 17 10 17 2.4µl 2.1µl 16.0%, 23.2%. 3, 27.8µg/µl. Microliter syringe HPLC 8 16 17 116.3µg, 17 10 17 61.0µg 79.4µg. 48.0µg, 37.8µg, 71.0µg (, 2014a; Kim et al., 2015).,. Petanidou(2003) (85:100), 85%. 93.4µg. 1..,.
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