Molecules 2010, 15
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15), 1.06 (s, 3H, H-14), 0.85~0.99 (m, 2H), 0.43 (m, 1H, H-5), 0.35 (m, 1H, H-1); HR-MS (ESI): m/z
calcd for C18H28NO4 ([M+NH4 ]+), 322.2013; found, 322.2008.
(3aR, 4aS, 5S, 5aR, 6aR)-5-(3-isopropylCarbonate-butyl)-5a-methyl-3-methylene-3a, 4, 4a, 5, 6, 6a-
18
hexahydrocyclopropa[f]benzofuran-2-one (12). Yield: 72%; a colorless oily liquid; [α]D +36.4 (C
1
0.41, CHCl3); IR (KBr) cm-1: 2974, 1757, 1722, 1660, 1148; H-NMR (400 MHz, CDCl3) δ: 6.23 (d,
J = 2.6 Hz, 1H, H-13), 5.56 (d, J = 2.4 Hz, 1H, H-13), 4.95 (m, 1H, H-8), 4.80 (m, 1H, H-4), 3.16 (m,
1H, H-7), 2.51 (m, 1H, CH(CH3)2), 2.30~2.38 (m, 2H), 1.59~1.61 (m, 2H), 1.35~1.44 (m, 2H), 1.23 (d,
J = 6.0 Hz, 3H, H-15), 1.18 (d, J = 10.8 Hz, 3H, CHCH3), 1.14 (d, J = 11.2 Hz, 3H, CHCH3), 1.07 (s,
3H, H-14), 0.87~0.98 (m, 2H), 0.43 (m, 1H, H-5), 0.34 (m, 1H, H-1); HR-MS (ESI): m/z calcd for
C19H28O4Na ([M+Na]+), 343.1880; found, 343.1875.
(3aR, 4aS, 5S, 5aR, 6aR)-5-(3-pentylCarbonate-butyl)-5a-methyl-3-methylene-3a, 4, 4a, 5, 6, 6a-
18
hexahydrocyclopropa[f]benzofuran-2-one (13). Yield: 70%; a colorless oily liquid; [α]D +49.4 (C
1
0.52, CHCl3); IR (KBr) cm-1: 2935, 1722, 1660, 1146; H-NMR (400 MHz, CDCl3) δ: 6.23 (d,
J = 2.6 Hz, 1H, H-13), 5.56 (d, J = 2.4 Hz, 1H, H-13), 4.95 (m, 1H, H-8), 4.78 (m, 1H, H-4), 3.16 (m,
1H, H-7), 3.01~3.04 (m, 1H, O=CCH2), 2.51~2.54 (m, 1H, O=CCH2), 2.30~2.38 (m, 2H), 1.59~1.61
(m, 2H), 1.27~1.51 (m, 8H), 1.23 (d, J = 6.0 Hz, 3H, H-15), 1.06 (s, 3H, H-14), 0.90~0.99 (m, 2H),
0.86~0.89 (m, 3H, CH2CH3), 0.43 (m, 1H, H-5), 0.35 (m, 1H, H-1); HR-MS (ESI): m/z calcd for
C21H36NO4 ([M+NH4]+), 366.2639; found, 366.2632.
(3aR, 4aS, 5S, 5aR, 6aR)-5-(3-phenylCarbonate-butyl)-5a-methyl-3-methylene-3a, 4, 4a, 5, 6, 6a-
18
hexahydrocyclopropa[f]benzofuran-2-one (14). Yield: 56%; a colorless oily liquid; [α]D +73.9 (C
1
0.31, CHCl3); IR (KBr) cm-1: 2942, 1757, 1711, 1485, 715; H-NMR (400 MHz, CDCl3) δ: 8.03 (m,
2H, H-3´, 5´), 7.57 (m, 1H, H-4´), 7.46 (m, 2H, H-2´, 6´), 6.24 (d, J = 2.6 Hz, 1H, H-13), 5.55 (d,
J = 2.4 Hz, 1H, H-13), 5.18 (m, 1H, H-8), 4.77 (m, 1H, H-4), 3.15 (m, 1H, H-7), 2.28~2.36 (m, 2H),
1.63~1.79 (m, 2H), 1.42~1.56 (m, 2H), 1.23 (d, J = 6.0 Hz, 3H, H-15), 1.08 (s, 3H, H-14), 0.88~0.92
(m, 2H), 0.47 (m, 1H, H-5), 0.35 (m, 1H, H-1); HR-MS (ESI): m/z calcd for C22H30NO4 ([M+NH4]+),
372.2169; found, 372.2168.
3.3. Spore Germination Assay
Microorganisms and maintenance: the strain of Colletotrichum lagenarium (36199) was provided
by Agricultural Culture Collection of China and maintained on potato dextrose agar (PDA).
Compounds 1 and 6-14 were dissolved in acetone or DMSO and added to 2% water agar medium after
sterilization to produce concentrations of 100, 75, 50, 25, 10, and 5 µg/mL or 10, 5, 2, 1, 0.5 and 0.25
µg/mL of medium. Conidial suspensions (0.2 mL) containing 1 × 105 condia/mL, derived from
cultures grown for 12 d on PDA plates, were spread on 2% water agar. Conidia were allowed to
germinate 25 ± 1 ºC for 8 h. Germination was quantified at three sites by counting 100 conidia per site.
A conidium was scored as germinated if the germ tube had reached at least half the length of the
conidium. Three plates for each concentration were used and the experiment was performed thrice,
along with 98% chlorothalonil (Syngenta Crop Protection Co., Ltd., China) as a positive control. The