Isolation and identification of the potential allelochemicals in the aqueous extract of yellow sweet clover (Melilotus officinalis)

doi:10.11686/cyxb20140521

Abstract

Abstract:

This study aimed to isolate and identify the main allelochemicals of aqueous extract of yellow sweet clover (Melilotus officinalis). Chromatographic separation, TLC analysis and bioassay test were conducted to isolate the potential allelochemicals of the aqueous extract of yellow sweet clover. GC-MS were used to identify the potential allelochemicals of the aqueous extracts of yellow sweet clover. The results showed that the ethyl acetate phase of yellow sweet clover aqueous extracts containing a variety of allelochemicals, in which coumarin had the highest relative content, then followed by 6-(3-Hydroxy-but-1-enyl)-1,5,5-trimethyl-7-oxabicyclo-[4,1,0]heptan-2-ol, chromanone,2,6-Di-tert-butyl-4-methylphenol, 2-Bromo-1,1,3-trimethyl cyclopropane, trimethyl phosphate, and other substances. The ethyl acetate phase was further separated into three fractions, which were the upper layer, middle layer and lower layer. The chemical which had the highest relative content of the upper layer is also coumarin. In the middle layer, 2-Bromo-1,1,3-trimethyl cyclopropane, 6-(3-Hydroxy-but-1-enyl)-1,5,5-trimethy-7-oxabicyclo[4,1,0]heptan-2-ol, 1-Methylcyclopentanol, trimethyl phosphate and 2-Bromo-1,1,3-trimethyl cyclopropane had higher relative contents. In the lower layer, 2-Bromo-1,1,3-trimethyl cyclopropane, 1-Methylcyclopentanol and cyclohexyl-phosphonous dibromide had higher relative contents. The ethyl acetate phase and the fractions separated from it all showed allelopathic effect on the seed germination and seedling growth of Italian ryegrass, in which the ethyl acetate phase treatment showed the strongest inhibition on seed germination and seedling growth of Italian ryegrass, and significantly stronger than all the treatments of the fractions separated from it (P<0.05). The upper layer treatment showed the second strongest inhibition and significantly stronger than the other two layers (P<0.05). While the lower layer treatment showed significant promoting effects on seed germination and seedling growth of Italian ryegrass (P<0.05). These results suggested that many allelochemicals existing in the aqueous extract of yellow sweet clover, in which coumarin had the highest relative content, and the treatments with higher content of coumarin concentration showed stronger inhibition effect on Italian ryegrass, which also suggested that coumarin should be one of the main allelochemicals in the aqueous extract of yellow sweet clover.

CLC Number:

S551⁺.601

WU Cai-xia,LIU Su-jiao,ZHAO Guo-qi. Isolation and identification of the potential allelochemicals in the aqueous extract of yellow sweet clover (Melilotus officinalis)[J]. Acta Prataculturae Sinica, 2014, 23(5): 184-192.

References

Reference:

[1] Schlegel A J, Havlin J L. Green fallow for the central Great Plains[J]. Agronomy Journal, 1997, 89(5): 762-767.

[2] Sparrow S D, Cochran V L, Sparrow E B. Herbage yield and nitrogen accumulation by seven legume crops on acid and neutral soils in a subarctic environment[J]. Canadian Journal of Plant Science, 1993, 73(4): 1037-1045.

[3] Smith W, Gorz H. Sweetclover improvement[J]. Advances in Agronomy, 1965, 17: 163-231.

[4] Nicotra M, Meli R, Savoca F. Efficacy of Melilotus officinalis (L.) pallas in cellulitis treatment[J]. Bollettino Accademia Gioenia Scienze Naturali, 2009, 42: 20-28.

[5] Wu C X, Guo X X, Li Z H, et al. Feasibility of using the allelopathic potential of yellow sweet clover for weed control[J]. Allelopathy Journal, 2010, 25(1): 173-183.

[6] Moyer J, Blackshaw R, Huang H. Effect of sweetclover cultivars and management practices on following weed infestations and wheat yield[J]. Canadian Journal of Plant Science, 2007, 87(4): 973-983.

[7] Blackshaw R E, Moyer J R, Doram R C, et al. Yellow sweetclover, green manure, and its residues effectively suppress weeds during fallow[J]. Weed Science, 2001, 49(3): 406-413.[8] Sampietro D A, Catalan C A, Vattuone M A. Isolation, Identification and Characterization of Allelochemicals/Natural Products[M]. Boca Raton: Science Publishers, 2009.

[9] Kelton J, Price A J, Mosjidis J. Allelopathic weed suppression through the use of cover crops[J]. Weed Control, 2012, 2: 978-953.

[10] Stoker J, Bellis D. The isolation and identification of bound coumarin from Melilotus album[J]. Canadian Journal of Biochemistry and Physiology, 1962, 40(12): 1763-1768.

[11] Kang S S, Lee Y S, Lee E B. Saponins and flavonoid glycosides from yellow sweetclover[J]. Archives of Pharmacal Research, 1988, 11(3): 197-202.

[12] Macías F A, Simonet A M, Galindo J C. Bioactive steroids and triterpenes from Melilotus messanensis and their allelopathic potential[J]. Journal of Chemical Ecology, 1998, 23(7): 1781-1803.

[13] Macías F A, Simonet A M, Galindo J C, et al. Bioactive phenolics and polar compounds from Melilotus messanensis[J]. Phytochemistry, 1999, 50(1): 35-46.

[14] Wu C X, Li Z H, Shen Y X. Quantification and allelopathy potential of phenolic acids in aqueous extracts of Legumes[J]. Acta Agrestia Sinica, 2007, 15(5): 401-406.

[15] Takemura T, Kamo T, Sakuno E, et al. Discovery of coumarin as the predominant allelochemical in Gliricidia sepium[J]. Journal of Tropical Forest Science, 2013, 25(2): 268-272.[16] Bertin C, Paul R N, Duke S O, et al. Laboratory assessment of the allelopathic effects of fine leaf fescues[J]. Journal of Chemical Ecology, 2003, 29(8): 1919-1937.

[17] Chung I M, Kim K H, Ahn J K, et al. Comparison of allelopathic potential of rice leaves, straw, and hull extracts on barnyardgrass[J]. Agronomy Journal, 2003, 95(4): 1063-1070.

[18] Kato-Noguchi H, Yamamoto M, Tamura K, et al. Isolation and identification of potent allelopathic substances in rattail fescue[J]. Plant Growth Regulation, 2010, 60(2): 127-131.

[19] Bich T, Ohno O, Suenaga K, et al. Isolation and identification of an allelopathic substance from duckweed (Lemna minor L.)[J]. Allelopathy Journal, 2013, 32(2): 213-221.

[20] Salam A, Kato Noguchi H. Isolation and characterisation of two potent growth inhibitory substances from aqueous extract of Bangladeshi rice cultivar BR17[J]. Allelopathy Journal, 2011, 27(2): 207-216.

[21] Yang G Q. Main allelochemicals isolated and identified from the leachates of Ageratina adenophora Sprengel(Asteraceae) and their action mechanisms on upland rice(Oryza sativa) seedling[D]. Beijing: Chinese Academy of Agricultural Sciences, 2006.

[22] Eljarrat E, Barceló D. Sample handling and analysis of allelochemical compounds in plants[J]. TrAC Trends in Analytical Chemistry, 2001, 20(10): 584-590.

[23] El Shahawy T A, Abdelhamid M T. Potential allelopathic effect of six phaseolus vulgaris recombinant inbred lines for weed control[J]. Australian Journal of Basic&Applied Sciences, 2013, 7(1): 462-467. 

[24] Razavi S M. Plant coumarins as allelopathic agents[J]. International Journal of Biological Chemistry, 2011, 5: 86-90.

[25] Sharma R, Negi D S, Shiu W K, et al. Characterization of an insecticidal coumarin from Boenninghausenia albiflora[J]. Phytotherapy Research, 2006, 20(7): 607-609.

[26] Brooker N, Kuzimichev Y, Laas J, et al. Evaluation of coumarin derivatives as anti-fungal agents against soil borne fungal pathogens[J]. Communications in Agricultural and Applied Biological Sciences, 2006, 72(4): 785-793.

[27] Ascari J, Takahashi J A, Boaventura M A D. Phytochemical and biological investigations of Caryocar brasiliense Camb[J]. Boletín Latinoamericanoy del Caribe de Plantas Medicinalesy Aromáticas, 2010, 9(1): 20-28.

[28] Hong Y, Hu H, Sakoda A, et al. Straw preservation effects of Arundo donax L. on its allelopathic activity to toxic and bloom-forming Microcystis aeruginosa[J]. Water Science & Technology, 2011, 63(8): 1566-1573.

[29] Qiao M, Xiao J, Yin H, et al. Analysis of the phenolic compounds in root exudates produced by a subalpine coniferous species as responses to experimental warming and nitrogen fertilisation[J]. Chemistry and Ecology, 2014, 30(6): 1-11.

参考文献：

[1] Schlegel A J, Havlin J L. Green fallow for the central Great Plains[J]. Agronomy Journal, 1997, 89(5): 762-767.

[2] Sparrow S D, Cochran V L, Sparrow E B. Herbage yield and nitrogen accumulation by seven legume crops on acid and neutral soils in a subarctic environment[J]. Canadian Journal of Plant Science, 1993, 73(4): 1037-1045.

[3] Smith W, Gorz H. Sweetclover improvement[J]. Advances in Agronomy, 1965, 17: 163-231.

[4] Nicotra M, Meli R, Savoca F. Efficacy of Melilotus officinalis (L.) pallas in cellulitis treatment[J]. Bollettino Accademia Gioenia Scienze Naturali, 2009, 42: 20-28.

[5] Wu C X, Guo X X, Li Z H, et al. Feasibility of using the allelopathic potential of yellow sweet clover for weed control[J]. Allelopathy Journal, 2010, 25(1): 173-183.

[6] Moyer J, Blackshaw R, Huang H. Effect of sweetclover cultivars and management practices on following weed infestations and wheat yield[J]. Canadian Journal of Plant Science, 2007, 87(4): 973-983.

[7] Blackshaw R E, Moyer J R, Doram R C, et al. Yellow sweetclover, green manure, and its residues effectively suppress weeds during fallow[J]. Weed Science, 2001, 49(3): 406-413.

[8] Sampietro D A, Catalan C A, Vattuone M A. Isolation, Identification and Characterization of Allelochemicals/Natural Products[M]. Boca Raton: Science Publishers, 2009.

[9] Kelton J, Price A J, Mosjidis J. Allelopathic weed suppression through the use of cover crops[J]. Weed Control, 2012, 2: 978-953.

[10] Stoker J, Bellis D. The isolation and identification of bound coumarin from Melilotus album[J]. Canadian Journal of Biochemistry and Physiology, 1962, 40(12): 1763-1768.

[11] Kang S S, Lee Y S, Lee E B. Saponins and flavonoid glycosides from yellow sweetclover[J]. Archives of Pharmacal Research, 1988, 11(3): 197-202.

[12] Macías F A, Simonet A M, Galindo J C. Bioactive steroids and triterpenes from Melilotus messanensis and their allelopathic potential[J]. Journal of Chemical Ecology, 1998, 23(7): 1781-1803.

[13] Macías F A, Simonet A M, Galindo J C, et al. Bioactive phenolics and polar compounds from Melilotus messanensis[J]. Phytochemistry, 1999, 50(1): 35-46.

[14] 邬彩霞, 李志华, 沈益新. 豆科牧草水浸提液的酚酸物质含量及化感潜力[J]. 草地学报, 2007, 15(5): 401-406.

[15] Takemura T, Kamo T, Sakuno E, et al. Discovery of coumarin as the predominant allelochemical in Gliricidia sepium[J]. Journal of Tropical Forest Science, 2013, 25(2): 268-272.

[16] Bertin C, Paul R N, Duke S O, et al. Laboratory assessment of the allelopathic effects of fine leaf fescues[J]. Journal of Chemical Ecology, 2003, 29(8): 1919-1937.

[17] Chung I M, Kim K H, Ahn J K, et al. Comparison of allelopathic potential of rice leaves, straw, and hull extracts on barnyardgrass[J]. Agronomy Journal, 2003, 95(4): 1063-1070.

[18] Kato-Noguchi H, Yamamoto M, Tamura K, et al. Isolation and identification of potent allelopathic substances in rattail fescue[J]. Plant Growth Regulation, 2010, 60(2): 127-131.[19] Bich T, Ohno O, Suenaga K, et al. Isolation and identification of an allelopathic substance from duckweed (Lemna minor L.)[J]. Allelopathy Journal, 2013, 32(2): 213-221.

[20] Salam A, Kato-Noguchi H. Isolation and characterisation of two potent growth inhibitory substances from aqueous extract of Bangladeshi rice cultivar BR17[J]. Allelopathy Journal, 2011, 27(2): 207-216.

[21] 杨国庆. 紫茎泽兰淋溶主效化感物质的分离鉴定及其对旱稻幼苗的作用机理[D]. 北京: 中国农业科学院, 2006.

[22] Eljarrat E, Barceló D. Sample handling and analysis of allelochemical compounds in plants[J]. TrAC Trends in Analytical Chemistry, 2001, 20(10): 584-590.

[23] El-Shahawy T A, Abdelhamid M T. Potential allelopathic effect of six phaseolus vulgaris recombinant inbred lines for weed control[J]. Australian Journal of Basic&Applied Sciences, 2013, 7(1): 462-467. 

[24] Razavi S M. Plant coumarins as allelopathic agents[J]. International Journal of Biological Chemistry, 2011, 5: 86-90.

[25] Sharma R, Negi D S, Shiu W K, et al. Characterization of an insecticidal coumarin from Boenninghausenia albiflora[J]. Phytotherapy Research, 2006, 20(7): 607-609.

[26] Brooker N, Kuzimichev Y, Laas J, et al. Evaluation of coumarin derivatives as anti-fungal agents against soil-borne fungal pathogens[J]. Communications in Agricultural and Applied Biological Sciences, 2006, 72(4): 785-793.

[27] Ascari J, Takahashi J A, Boaventura M A D. Phytochemical and biological investigations of Caryocar brasiliense Camb[J]. Boletín Latinoamericanoy del Caribe de Plantas Medicinalesy Aromáticas, 2010, 9(1): 20-28.

[28] Hong Y, Hu H, Sakoda A, et al. Straw preservation effects of Arundo donax L. on its allelopathic activity to toxic and bloom-forming Microcystis aeruginosa[J]. Water Science & Technology, 2011, 63(8): 1566-1573.

[29] Qiao M, Xiao J, Yin H, et al. Analysis of the phenolic compounds in root exudates produced by a subalpine coniferous species as responses to experimental warming and nitrogen fertilisation[J]. Chemistry and Ecology, 2014, 30(6): 1-11.