Metabolite profiles of Chinese native Citrus species and their
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Metabolite profiles of Chinese native Citrus species and their taxonomic implications Li Jing 1,2,3, Zhentian Lei 2, Guiwei Zhang 1, 3, Alan Cesar Pilon 2, 4, David V Huhman 2, Rangjin Xie 5, Wanpeng Xi 1, 3, Zhiqin Zhou1, 3*, Lloyd W Sumner 2* 1 College of Horticulture and Landscape Architecture, Southwest University, Chongqing 400716, China 2 Plant Biology Division, the Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK 73401 3 Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education, Chongqing 400715, China 4 Nucleus of Bioassays, Biosynthesis and Ecophysiology of Natural Products -NuBBE, São Paulo State University - UNESP, Rua Prof. Francisco Degni 55, Araraquara, 14800-900, São Paulo, Brazil 5 Citrus research institute, Chinese Academy of Agricultural Sciences/Southwest University, Chongqing 400712, China *Corresponding authors: Plant biology division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, OK 73401. Tel: 1-580-224-6710; Fax: 1-580-224-6692; Email: lwsumner@noble.org or College of Horticulture and Landscape Architecture, Southwest University, Chongqing 400716, China. Tel: 86-23-68250229; Fax: 86-23-68251274; Email: zzqswu@yahoo.com. Supplemental Table 1 Phytochemical components of citrus essential oils # RT Ri-RI(lib) Score Library (min) (min) 1 15.0376 -7.8 83 Adam 2 15.4750 -5.1 98 3 17.3848 4.4 4 18.0693 5 Name Citrus essential oil Mi Ta Su Dao Go We Ba Dai Chi Zh α-Thujene 0.01±0.00 0.01±0.00 0.01±0.01 0.88±0.01 1.23±0.01 0.45±0.04 tr 0.01±0.00 0.01±0.00 1.51±0.14 Adam α-Pinene 0.98±0.07 1.19±0.04 1.23±0.15 3.11±0.10 4.07±0.08 2.04±0.13 1.23±0.01 0.85±0.08 1.22±0.08 4.93±0.31 97 Adam Sabinene 1.12±0.23 0.74±0.07 0.59±0.28 0.21±0.04 2.54±0.10 0.19±0.03 0.15±0.03 0.11±0.02 0.36±0.03 0.17±0.06 2.1 95 Adam Myrcene 5.70±0.44 5.85±0.21 5.83±0.35 4.95±0.12 4.67±0.10 4.81±0.13 5.63±0.02 11.05±0.13 5.50±0.11 3.87±0.13 18.8887 2.3 79 Noble α-Phellandrene 0.02±0.01 0.01±0.00 0.08±0.01 tr tr tr tr tr tr tr 6 19.3785 2.3 79 Noble α-Terpinene 0.05±0.01 0.03±0.00 0.04±0.00 0.19±0.03 0.29±0.01 0.10±0.01 0.01±0.00 0.01±0.00 0.02±0.00 0.41±0.02 7 19.9738 3.6 95 Noble R-(+)-Limonene 90.21±0.98 90.87±0.00 91.06±1.05 68.93±0.86 62.72±0.32 78.87±0.35 90.31±1.02 85.09±0.18 90.03±0.12 57.99±0.71 8 20.2929 6.2 78 Noble Eucalyptol tr tr tr tr 0.01±0.00 tr tr 0.01±0.00 tr tr 9 21.1755 -2.2 67 Adam para-Mentha-3,8-diene 0.19±0.01 0.11±0.01 0.19±0.03 18.45±0.20 21.83±0.10 11.39±0.20 0.04±0.01 0.11±0.03 0.10±0.02 28.04±0.43 10 22.1857 0 95 Noble Terpinolene 0.09±0.01 0.06±0.00 0.12±0.00 0.72±0.01 1.05±0.04 0.40±0.01 0.04±0.01 0.04±0.00 0.09±0.01 1.44±0.09 11 22.5936 Unknown 0.61±0.10 0.42±0.04 0.03±0.02 1.49±0.20 0.79±0.12 1.20±0.20 1.42±0.18 0.11±0.05 0.65±0.11 0.30±0.06 12 22.8511 Unknown 0.09±0.00 0.10±0.00 0.04±0.01 0.64±0.97 0.11±0.01 0.10±0.01 0.61±0.88 0.11±0.00 0.12±0.01 0.04±0.00 13 23.1121 Unknown — — tr — — — — tr tr tr 14 23.7966 Unknown tr tr tr 0.01±0.00 0.01±0.00 tr 0.01±0.00 tr 0.01±0.00 0.01±0.00 15 24.0737 Unknown 0.01±0.00 tr 0.01±0.00 0.01±0.00 0.02±0.00 0.01±0.00 0.01±0.00 0.01±0.00 0.01±0.00 0.02±0.00 16 24.3212 Unknown tr 0.01±0.00 0.01±0.00 0.01±0.01 0.02±0.00 0.01±0.00 0.01±0.00 tr 0.01±0.00 0.02±0.02 17 24.5631 5.2 82 Adam Isopulegol tr tr tr tr tr tr tr — — tr 18 26.3636 3.7 64 Adam Octanol acetate 0.36±0.04 0.34±0.01 0.24±0.06 0.13±0.02 0.30±0.03 0.24±0.01 0.03±0.01 0.76±0.07 0.47±0.01 0.22±0.03 19 27.9811 Unknown 0.01±0.00 0.01±0.00 0.01±0.00 0.01±0.00 0.01±0.00 tr 0.01±0.00 0.02±0.00 0.02±0.00 0.01±0.00 20 28.1059 Unknown 0.01±0.00 0.01±0.00 0.01±0.00 0.01±0.00 0.02±0.00 0.01±0.00 tr 0.03±0.00 0.02±0.00 0.01±0.00 21 28.3492 Unknown 0.21±0.10 0.06±0.02 0.08±0.06 0.01±0.00 0.01±0.00 0.01±0.00 0.01±0.01 0.11±0.01 0.14±0.00 0.01±0.00 22 29.4755 Unknown 0.01±0.00 0.01±0.00 0.01±0.00 0.01±0.00 0.02±0.00 0.01±0.00 0.01±0.00 0.03±0.00 0.02±0.00 0.01±0.00 23 30.3954 Unknown — — — tr — tr tr — tr — 24 30.5727 Citronellyl acetate 0.01±0.00 0.01±0.00 0.01±0.01 0.03±0.02 0.04±0.00 0.03±0.01 0.06±0.01 0.07±0.01 0.04±0.00 0.05±0.00 25 30.7500 Unknown tr — tr tr tr tr tr tr tr tr 26 30.8450 Unknown 0.01±0.00 0.01±0.00 0.01±0.00 0.01±0.00 0.30±0.00 0.02±0.01 0.15±0.03 0.29±0.03 0.28±0.01 0.28±0.02 27 31.3843 Unknown tr tr tr 0.01±0.01 0.20±0.00 tr 0.06±0.01 0.79±0.14 0.60±0.02 0.01±0.00 28 31.5269 Unknown tr tr tr tr tr tr tr tr tr tr 29 31.6301 7-epi-Sesquithujene tr — tr tr tr tr tr tr tr tr 30 31.9340 Unknown 0.02±0.00 0.01±0.00 0.01±0.00 0.07±0.02 0.60±0.00 tr 0.01±0.00 tr tr tr 31 32.3138 Unknown tr tr tr tr 0.01±0.01 tr tr 0.06±0.01 0.04±0.00 0.02±0.01 32 32.5449 Unknown 0.01±0.00 tr 0.01±0.01 0.01±0.00 0.01±0.00 0.01±0.00 0.01±0.00 0.03±0.01 0.01±0.00 0.01±0.00 33 32.8427 Unknown 0.02±0.00 0.01±0.00 0.03±0.01 0.01±0.00 0.03±0.00 0.01±0.00 0.02±0.00 0.09±0.01 0.04±0.00 0.01±0.00 34 32.9142 -2.6 82 Noble (-)-trans-Caryophyllene tr tr tr tr tr tr tr tr tr tr 35 33.0699 -1.5 79 Adam cis-Prenyl limonene tr — tr tr tr tr — tr tr tr 36 33.1566 Unknown 0.03±0.00 0.01±0.00 0.01±0.00 tr tr tr 0.02±0.01 tr tr 0.01±0.00 37 33.1999 Unknown 0.02±0.00 0.01±0.00 0.01±0.00 tr tr 0.01±0.00 0.02±0.00 0.02±0.01 0.01±0.00 0.01±0.00 38 33.4176 Unknown 0.04±0.04 tr 0.01±0.01 tr tr tr 0.02±0.01 0.04±0.01 0.07±0.00 0.26±0.05 39 33.8017 Unknown tr tr tr 0.01±0.00 0.01±0.00 tr tr tr tr tr 40 33.8684 Unknown tr tr tr tr tr tr tr tr tr tr 41 34.4533 Unknown 0.01±0.00 0.01±0.00 0.01±0.01 0.02±0.01 0.01±0.00 0.03±0.01 0.02±.001 0.14±0.01 0.05±0.01 0.15±0.03 42 34.5122 trans-Muurola-4(14),5-diene 0.01±0.00 0.01±0.00 0.01±0.00 0.01±0.01 tr 0.03±0.00 0.02±0.01 0.01±0.00 0.05±0.00 0.15±0.03 43 34.5742 Unknown — — tr — — — — — — — -0.3 -2.7 3.6 61 63 90 Adam Adam Adam 44 34.7472 1 89 45 34.8604 46 34.9696 1.1 80 47 35.3234 1.6 64 α-Farnesene 0.07±0.00 0.06±0.01 0.17±0.06 tr tr tr 0.01±0.00 tr 0.01±0.00 0.01±0.00 Unknown 0.03±0.01 0.02±0.01 0.07±0.03 tr tr tr 0.01±0.00 tr tr tr Adam β-Curcumene tr tr 0.01±0.00 tr tr tr tr tr tr tr Adam Zonarene 0.02±0.01 0.01±0.00 0.02±0.01 0.02±0.01 0.02±0.00 0.01±0.00 0.02±0.01 tr 0.01±0.00 0.01±0.00 Adam Metabolite derivatives were tentatively identified by matching retention time as well as mass spectra with those in NF-GOLM-ADAM-2011 Mass Spectral library. Metabolites were considered tentatively identified with a spectral match score higher than 60 and RI-deviation (Ri-RI(lib)) lower than 8. Data were normalized to the total area for each chromatogram. Values are expressed as mean ± SD of three determinations. tr, trace amount (<0.01%); -: not detected. Supplemental Table 1 Phytochemical components of citrus essential oils (continued) # RT Ri-RI(lib) Score Library (min) (min) 1 15.0376 -7.8 83 Adam 2 15.4750 -5.1 98 3 17.3848 4.4 4 18.0693 5 Name Citrus essential oils Gu Ji Vo Me Al Ra Bu St Cht Chp α-Thujene 1.11±0.02 0.08±0.01 0.15±0.02 1.05±0.13 0.05±0.02 0.94±0.11 0.67±0.01 tr 0.30±0.02 0.01±0.00 Adam α-Pinene 5.26±0.15 1.35±0.04 1.82±0.14 3.24±0.50 1.21±0.11 4.30±0.56 3.67±0.34 0.49±0.28 1.74±0.03 0.82±0.04 97 Adam Sabinene 3.19±0.44 7.54±1.98 13.64±2.06 0.28±0.01 4.48±1.71 2.39±0.17 0.06±0.01 0.23±0.19 0.07±0.03 0.20±0.04 2.1 95 Adam Myrcene 3.94±0.03 5.65±0.12 5.86±0.25 4.40±0.06 7.07±1.50 3.62±0.17 7.82±0.24 9.31±2.72 9.02±0.15 11.32±0.19 18.8887 2.3 79 Noble α-Phellandrene tr tr 0.01±0.00 tr tr tr tr tr tr tr 6 19.3785 2.3 79 Noble α-Terpinene 0.41±0.08 0.18±0.05 0.51±0.14 0.22±0.02 0.19±0.07 0.31±0.03 0.02±0.01 0.03±0.01 0.07±0.02 tr 7 19.9738 3.6 95 Noble R-(+)-Limonene 61.24±1.28 83.08±1.95 75.51±1.23 70.37±0.27 84.38±0.78 64.09±0.40 55.05±2.26 84.28±0.75 64.28±0.74 85.67±0.36 8 20.2929 6.2 78 Noble Eucalyptol 0.36±0.12 tr 0.01±0.00 0.12±0.02 tr 0.20±0.05 tr 0.01±0.00 tr 0.01±0.00 9 21.1755 -2.2 67 Adam para-Mentha-3,8-diene 21.42±0.73 0.70±0.12 1.53±0.42 18.00±0.22 1.03±0.38 21.26±0.50 19.18±1.33 0.40±0.10 16.09±0.11 0.16±0.08 10 22.1857 0 95 Noble Terpinolene 0.95±0.08 0.16±0.03 0.31±0.06 0.90±0.02 0.18±0.03 0.81±0.03 0.47±0.09 0.10±0.04 0.42±0.02 0.03±0.00 11 22.5936 Unknown 0.01±0.00 tr 0.03±0.01 0.06±0.01 tr 0.01±0.00 0.03±0.00 0.09±0.08 0.01±0.00 0.03±0.00 12 22.8511 Unknown 0.06±0.00 0.03±0.01 0.05±0.02 0.04±0.00 0.02±0.00 0.05±0.00 0.04±0.00 0.17±0.08 0.05±0.01 0.02±0.00 13 23.1121 Unknown tr — tr tr — tr tr tr tr tr 14 23.7966 Unknown 0.03±0.01 tr tr tr tr 0.02±0.00 0.01±0.00 tr tr 0.01±0.00 15 24.0737 Unknown 0.01±0.00 0.01±0.00 0.01±0.00 0.02±0.00 0.01±0.00 0.01±0.00 0.05±0.01 0.01±0.00 0.01±0.00 0.01±0.00 16 24.3212 Unknown 0.04±0.04 tr tr 0.01±0.00 0.01±0.00 0.02±0.01 0.07±0.023 tr 0.01±0.00 — 17 24.5631 5.2 82 Adam Isopulegol 0.02±0.01 tr 0.02±0.01 tr tr 0.02±0.00 tr tr tr 18 26.3636 3.7 64 Adam Octanol acetate 0.23±0.01 0.10±0.02 0.10±0.01 0.03±0.01 0.10±0.04 0.18±0.00 0.11±0.05 1.91±1.26 0.46±0.03 0.02±0.01 19 27.9811 Unknown 0.02±0.00 0.01±0.00 0.01±0.00 0.01±0.00 0.02±0.01 0.01±0.00 0.09±0.02 0.03±0.02 0.01±0.01 0.01±0.00 20 28.1059 Unknown 0.04±0.00 0.01±0.00 0.02±0.00 0.01±0.00 0.01±0.01 0.07±0.01 0.07±0.00 0.03±0.01 0.01±0.00 0.01±0.00 21 28.3492 Unknown 0.04±0.01 0.01±0.00 0.01±0.00 0.10±0.02 0.01±0.01 0.07±0.01 5.24±1.55 0.20±0.18 0.02±0.00 0.05±0.02 22 29.4755 Unknown 0.04±0.01 0.02±0.00 0.02±0.01 0.03±0.00 0.02±0.01 0.03±0.01 0.07±0.01 0.06±0.04 0.06±0.01 0.01±0.00 23 30.3954 Unknown tr tr — tr tr tr tr tr 0.01±0.00 tr 24 30.5727 Citronellyl acetate 0.13±0.00 0.50±0.03 0.15±0.05 0.02±0.00 0.59±0.28 0.14±0.01 0.19±0.01 0.19±0.13 0.20±0.03 0.15±0.02 25 30.7500 Unknown tr — tr 0.01±0.00 — tr tr tr tr tr 26 30.8450 Unknown 0.02±0.00 0.30±0.02 0.01±0.00 0.06±0.00 0.29±0.06 0.02±0.00 2.52±0.21 0.08±0.06 0.27±0.01 0.29±0.02 27 31.3843 Unknown 0.02±0.00 0.08±0.01 0.01±0.00 0.01±0.00 0.09±0.05 0.02±0.00 4.10±0.89 0.56±0.47 0.15±0.02 0.69±0.04 28 31.5269 Unknown tr tr tr tr tr tr tr tr tr tr 29 31.6301 7-epi-Sesquithujene tr tr tr 0.02±0.00 tr tr tr tr tr — 30 31.9340 Unknown 0.05±0.01 tr tr 0.09±0.00 tr 0.04±0.01 tr 0.04±0.05 0.05±0.03 tr 31 32.3138 Unknown 0.02±0.00 tr 0.01±0.01 0.01±0.00 0.01±0.01 0.01±0.00 0.01±0.00 0.09±0.10 0.08±0.01 0.01±0.00 32 32.5449 Unknown 0.05±0.00 0.03±0.00 0.02±0.00 0.02±0.00 0.03±0.01 0.05±0.00 0.07±0.00 0.02±0.01 0.01±0.00 0.01±0.00 33 32.8427 Unknown 0.25±0.04 0.05±0.00 0.07±0.02 0.01±0.00 0.05±0.01 0.20±0.01 0.05±0.01 1.02±0.79 0.10±0.00 0.25±0.01 34 32.9142 -2.6 82 Noble (-)-trans-Caryophyllene 0.01±0.00 tr tr tr tr 0.01±0.00 tr 0.03±0.04 tr tr 35 33.0699 -1.5 79 Adam cis-Prenyl limonene tr tr tr tr tr tr tr 0.01±0.01 tr tr 36 33.1566 Unknown 0.03±0.00 0.02±0.00 0.01±0.01 0.01±0.00 0.01±0.00 0.03±0.01 0.01±0.00 0.01±0.01 0.03±0.01 tr 37 33.1999 Unknown 0.04±0.01 0.02±0.00 0.01±0.00 0.02±0.01 0.03±0.02 0.04±0.01 0.05±0.00 0.02±0.01 0.06±0.01 0.01±0.00 38 33.4176 Unknown 0.06±0.00 0.01±0.00 0.01±0.00 0.24±0.00 0.01±0.01 0.06±0.00 0.13±0.00 0.17±0.16 0.73±0.09 0.10±0.01 39 33.8017 Unknown 0.04±0.00 0.02±0.00 0.01±0.00 0.02±0.00 0.02±0.00 0.04±0.00 0.02±0.00 0.02±0.02 0.02±0.00 tr 40 33.8684 Unknown tr tr tr tr tr tr tr tr tr tr 41 34.4533 Unknown 0.29±0.12 0.03±0.00 0.01±0.00 0.19±0.03 0.03±0.01 0.31±0.04 0.06±0.01 0.18±0.10 3.84±0.73 0.05±0.00 42 34.5122 trans-Muurola-4(14),5-diene 0.29±0.12 0.02±0.00 tr 0.18±0.01 0.01±0.01 0.29±0.03 tr 0.06±0.07 1.63±0.59 0.01±0.00 43 34.5742 Unknown tr — — — — tr tr — — — 44 34.7472 α-Farnesene 0.03±0.00 tr tr 0.01±0.00 tr 0.20±0.00 tr 0.02±0.03 0.04±0.03 0.02±0.00 45 34.8604 Unknown 0.05±0.06 tr 0.02±0.02 0.03±0.02 tr 0.13±0.01 0.02±0.00 0.01±0.01 0.01±0.01 0.01±0.00 -0.3 -2.7 3.6 1 61 63 90 89 Adam Adam Adam Adam 46 34.9696 1.1 80 Adam β-Curcumene 0.12±0.02 0.03±0.00 0.02±0.01 0.03±0.00 0.02±0.01 0.12±0.00 0.02±0.00 tr 0.01±0.00 tr 47 35.3234 1.6 64 Adam Zonarene 0.08±0.01 tr tr 0.03±0.00 tr 0.08±0.00 tr 0.13±0.13 0.15±0.05 0.02±0.00 Metabolite derivatives were tentatively identified by matching retention time as well as mass spectra with those in NF-GOLM-ADAM-2011 Mass Spectral library. Metabolites were considered tentatively identified with a spectral match score higher than 60 and RI-deviation (Ri-RI(lib)) lower than 8. Data were normalized to the total area for each chromatogram. Values are expressed as mean ± SD of three determinations. tr, trace amount (<0.01%); -: not detected.
