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山茶品种花色变异与花青苷的关系

花匠小妙招
2024-12-16 00:23

摘要:

应用高效液相色谱-光电二极管阵列检测（HPLC-DAD）和超高效液相色谱-四极杆-飞行时间质谱（UPLC-Q-TOF-MS）联用技术，分析山茶（Camellia japonica）白色、粉色、红色和黑色4个色系22个品种花瓣中花青苷成分与含量；按照CIE L*a*b*表色系法测量其花色变异，运用多元线性回归方法研究其花色变异与花青苷之间的关系，以期为山茶花色育种提供理论依据。结果表明，山茶品种花瓣中共检测到7种花青苷，分别为矢车菊素-3-O-β-半乳糖苷（Cy3Ga）、矢车菊素-3-O-β-葡萄糖苷（Cy3G）、矢车菊素-3-O-[6-O-（E）-咖啡酰]-β-半乳糖苷（Cy3GaECaf）、矢车菊素-3-O-[6-O-（E）-咖啡酰]-β-葡萄糖苷（Cy3GECaf）、矢车菊素-3-O-[6-O-（Z）-p-香豆酰]-β-葡萄糖苷（Cy3GZpC）、矢车菊素-3-O-[6-O-（E）-p-香豆酰]-β-半乳糖苷（Cy3GaEpC）和矢车菊素-3-O-[6-O-（E）-p-香豆酰]-β-葡萄糖苷（Cy3GEpC）。山茶品种白色花瓣中均未检测到花青苷，粉色、红色和黑色花瓣中主要花青苷均为Cy3G、Cy3GEpC和Cy3Ga；红色花瓣中花青苷总量及Cy3G、Cy3GEpC和Cy3Ga含量远高于粉色，黑色花瓣中花青苷总量及Cy3G、Cy3GEpC和Cy3Ga含量远高于红色和粉色花瓣。从粉色、红色到黑色，花瓣中主要花青苷含量及花青苷总量明显增加，Cy3Ga和Cy3G比例升高，Cy3GEpC比例降低。Cy3G和Cy3GEpC是决定山茶品种花色的主要花青苷，其含量的积累可导致花瓣红色程度增加。

Abstract:

The flower colors, anthocyanin components and contents in 22 cultivars of C. japonica, which were categorized into 4 color series including white, pink, red and dark red, were determined. Flower colors were measured by CIE L*a*b* scale, and anthocyanin components and contents were measured by high-performance liquid chromatography coupled with diode array detection (HPLC-DAD) and ultra-performance liquid chromatography quadrupole-time-of-flight massspectrometry (UPLC-Q-TOF-MS).The relationship between the variation of flower colors and anthocyanin components was explored by multiple liner regression analyses, which provided a theoretical basis for flower color breeding of C.japonica.The results show that seven anthocyanins were detected in cultivars of C. japonica, including cyanidin-3-O-β-galactoside(Cy3Ga), cyanidin-3-O-β-glucoside(Cy3G), cyanidin-3-O-[6-O-(E)-caffeoyl]-β-galactoside(Cy3GaECaf), cyanidin-3-O-[6-O-(E)-caffeoyl]-β-glucoside(Cy3GECaf), cyanidin-3-O-[6-O-(Z)-p-coumaroyl]-β-glucoside(Cy3GZpC), cyanidin-3-O-[6-O-(E)-p-coumaroyl]-β-galactoside(Cy3GaEpC) and cyanidin-3-O-[6-O-(E)-p-coumaroyl]-β-glucoside(Cy3GEpC). No anthocyanins were detected from white petals in cultivars of C. japonica, and the main anthocyanins from pink, red and dark red petals were Cy3G, Cy3GEpC and Cy3Ga. Total anthocyanin and the contents of Cy3G, Cy3GEpC and Cy3Ga from red petals were far higher than that from pink petals. Total anthocyanin and the contents of Cy3G, Cy3GEpC and Cy3Ga from dark red petals were far higher than that from red and pink petals. From pink, red to dark red, total anthocyanins and the main anthocyanins from petals in cultivars of C. japonica increased obviously, and the proportion of Cy3G and Cy3Ga also increased but that of Cy3GEpC decreased. It is concluded that Cy3G and Cy3GEpC were the main anthocyanins which determined the colors of petals in cultivars of C. japonica, and the accumulation of their contents enhanced the red color of petals.

图 1 山茶品种花色分布

Figure 1. Distribution of flower colors in cultivars of Camellia japonica

图 2 山茶品种花青苷的HPLC图谱

Figure 2. HPLC chromatogram of anthocyaninsin cultivars of Camellia japonica

表 1 山茶品种花色数据

Table 1 The data of flower colors in cultivars of Camellia japonica

山茶品种花色 L* a* b* C* h/(°) ‘雪塔’ 白色 89.14 -0.31 1.97 1.99 102.55 ‘白桃’ 白色 83.63 -1.07 6.21 8.95 93.52 ‘白牡丹’ 白色 86.39 -3.05 7.04 7.64 103.07 ‘水晶白’ 白色 90.62 -1.64 5.07 5.33 107.99 ‘绿珠球’ 白色 90.02 -0.43 2.57 2.62 101.08 ‘白鱼尾椿’ 白色 89.59 -1.08 6.76 6.86 99.38 ‘白孔雀椿’ 白色 89.10 -0.51 3.21 3.27 101.21 平均值 88.36 -1.15 4.69 5.24 101.26 ‘粉西施’ 粉色 64.65 37.51 2.85 39.00 3.61 ‘粉玲珑’ 粉色 65.34 42.15 6.25 42.16 5.14 ‘粉明天’ 粉色 69.70 34.41 8.86 37.10 5.56 ‘粉三学士’ 粉色 70.53 29.30 3.33 29.49 6.66 ‘粉鱼尾椿’ 粉色 65.84 35.71 2.70 34.48 4.50 平均值 67.21 35.82 4.80 36.45 5.09 ‘双喜’ 红色 45.21 52.96 12.90 54.52 13.71 ‘孔雀椿’ 红色 40.41 51.65 15.69 53.99 16.89 ‘红绣球’ 红色 45.04 50.08 12.87 51.71 14.40 ‘大朱砂’ 红色 41.87 52.35 12.65 53.87 13.59 ‘紫酱金花’ 红色 41.03 52.71 19.44 56.19 30.27 平均值 42.44 51.36 13.74 53.19 14.96 ‘霍伯’ 黑色 31.58 40.53 15.98 43.56 21.68 ‘皇家天鹅绒’ 黑色 31.90 38.21 15.97 41.41 22.68 ‘黑樱桃’ 黑色 32.25 34.16 12.45 36.43 21.65 ‘午夜魔幻’ 黑色 26.64 31.92 9.89 33.43 17.25 ‘黑魔法’ 黑色 24.95 30.02 12.82 32.79 23.18 平均值 29.46 34.97 13.42 37.52 21.29 L*为明度; a*为红绿色相值; b*为黄蓝色相值; C*为彩度; h为色调角。

表 2 山茶品种花青苷成分的紫外-可见吸收光谱与质谱数据

Table 2 Chromatographic and spectral data of anthocyanins in cultivars of Camellia japonica

色谱峰保留时间/min 吸收波长λmax/nm A440/Avis，max/% 分子离子m/z 碎片离子m/z 推定结果 P1 8.37 281, 514 32 449 287 Cy3Ga P2 9.46 283, 516 32 449 287 Cy3G P3 21.34 282, 316, 515 34 611 449, 287 Cy3GaECaf P4 24.83 283, 316, 517 34 611 449, 287 Cy3GECaf P5 25.71 284, 312, 516 35 595 449, 287 Gy3GZpC P6 26.14 285, 313, 518 36 595 449, 287 Cy3GaEpC P7 29.96 284, 313, 518 35 595 449, 287 Cy3GEpCA440/Avis，max为花青苷花440 nm与可见光区最大吸收波长(λvis, max)处的吸收值之比。Cy3Ga为矢车菊素-3-O-β-半乳糖苷; Cy3G为矢车菊素-3-O-β-葡萄糖苷; Cy3GaECaf为矢车菊素-3-O-[6-O-(E)-咖啡酰]-β-半乳糖苷; Cy3GECa为矢车菊素-3-O-[6-O-(E)-咖啡酰]-β-葡萄糖苷; Cy3GZpC为矢车菊素-3-O-[6-O-(Z)-p-香豆酰]-β-葡萄糖苷; Cy3GaEpC为矢车菊素-3-O-[6-O-(E)-p-香豆酰]-β-半乳糖苷; Cy3GEpC为矢车菊素-3-O-[6-O-(E)-p-香豆酰]-β-葡萄糖苷。

表 3 山茶品种花青苷含量

Table 3 The contents of anthocyanins in cultivars of Camellia japonica

μg·(100 mg)-1 山茶品种 Cy3Ga Cy3G Cy3GaECaf Cy3GECaf Gy3GZpC Cy3GaEpC Cy3GEpC 合计 ‘粉西施’ 1.46±0.03 9.36±0.46 0.31±0.00 0.24±0.00 0.56±0.12 0.91±0.27 4.91±0.36 17.75 ‘粉玲珑’ 1.25±0.05 11.62±0.38 0.23±0.00 0.18±0.00 0.84±0.08 0.65±0.02 5.19±0.12 19.96 ‘粉明天’ 1.85±0.12 15.34±0.54 — 0.25±0.00 2.69±0.11 0.78±0.04 6.56±0.03 27.47 ‘粉三学士’ 1.89±0.11 7.44±0.26 — — 1.78±0.00 1.39±0.00 6.95±0.25 19.45 ‘粉鱼尾椿’ 2.50±0.21 8.16±0.31 — — — 2.21±0.17 5.37±0.22 18.32 平均值 1.81 10.38 0.11 0.13 1.17 1.19 5.80 20.59 ‘双喜’ 9.28±0.28 42.12±0.62 — — 6.22±0.11 6.58±0.16 25.36±0.43 89.56 ‘孔雀椿’ 11.32±0.28 56.48±0.65 — — 3.12±0.06 4.96±0.15 38.56±0.58 114.44 ‘红绣球’ 12.57±0.26 110.56±1.32 3.67±0.09 2.96±0.00 3.87±0.01 4.32±0.06 19.23±0.18 157.18 ‘大朱砂’ 31.22±0.27 121.09±1.25 2.51±0.03 — 2.76±0.00 11.24±0.43 25.31±0.58 194.13 ‘紫酱金花’ 25.15±0.55 142.35±1.16 2.81±0.01 2.34±0.00 3.62±0.11 4.80±0.09 21.15±0.63 202.22 平均值 17.91 94.52 1.80 1.06 3.92 6.38 25.92 151.51 ‘霍伯’ 45.24±0.65 312.16±1.26 — — 4.42±0.06 6.52±0.00 15.32±0.13 383.66 ‘皇家天鹅绒’ 91.91±0.63 271.45±1.45 — — — — 4.25±0.00 367.61 ‘黑樱桃’ 123.24±0.97 492.66±1.63 23.17±0.26 26.32±0.18 9.24±0.11 23.75±0.31 53.56±0.69 751.93 ‘午夜魔幻’ 152.42±0.75 682.33±2.18 14.78±0.16 — 6.65±0.09 28.17±0.28 52.09±0.72 936.44 ‘黑魔法’ 262.21±1.12 871.22±2.27 11.26±0.32 — 7.21±0.00 23.24±0.35 51.54±0.54 1 226.68 平均值 135.00 525.96 9.84 5.26 5.50 16.34 35.35 733.27“—”表示未鉴定出。Cy3Ga为矢车菊素-3-O-β-半乳糖苷; Cy3G为矢车菊素-3-O-β-葡萄糖苷; Cy3GaECaf为矢车菊素-3-O-[6-O-(E)-咖啡酰]-β-半乳糖苷; Cy3GECa为矢车菊素-3-O-[6-O-(E)-咖啡酰]-β-葡萄糖苷; Cy3GZpC为矢车菊素-3-O-[6-O-(Z)-p-香豆酰]-β-葡萄糖苷; Cy3GaEpC为矢车菊素-3-O-[6-O-(E)-p-香豆酰]-β-半乳糖苷; Cy3GEpC为矢车菊素-3-O-[6-O-(E)-p-香豆酰]-β-葡萄糖苷。 [1]

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