Effect of exogenous quercetin on antioxidant of chestnut seedlings under drought stress
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摘要:
目的 探寻外源槲皮素（Quercetin, Q）对提高板栗苗耐旱性的生理机制和适宜喷施浓度。 方法 以燕宝（YB）板栗苗为试材，通过盆栽控水的方法，以重度干旱胁迫CK1和正常管理CK2为对照，研究在重度干旱胁迫下喷施不同浓度[600 μmol·L−1（QD1）、1200 μmol·L−1（QD2）、1800 μmol·L−1（QD3）和2000 μmol·L−1（QD4）、3000 μmol·L−1（QD5）]槲皮素对板栗幼苗叶绿素含量、抗氧化酶活性、丙二醛（MDA）和黄酮醇含量的影响。 结果 与同处于重度干旱胁迫下未经槲皮素喷施处理的CK1相比，5种不同浓度槲皮素喷施处理均能显著提高板栗叶片叶绿素含量、超氧化物歧化酶（SOD）活性、过氧化氢酶（CAT）活性、过氧化物酶（POD）活性和黄酮醇含量，同时显著降低了板栗叶片丙二醛（MDA）含量，但喷施后的板栗苗的状态仍不能恢复到正常管理的CK2。通过主成分分析及隶属函数法，得出QD1、QD2、QD3、QD4、QD5各处理的平均抗旱性度量值分别为0.735、0.641、0.738、1.389、0.828。其中，2000 μmol·L−1（QD4）槲皮素浓度处理效果更显著。 结论 外源施加槲皮素可以通过提高板栗幼苗抗氧化酶活性、叶绿素含量，减缓MDA增加速度，减缓细胞膜质过氧化的速度来减轻干旱对板栗叶片细胞的伤害，从而提高板栗幼苗耐旱性，且以2000 μmol·L−1（QD4）槲皮素浓度处理效果最佳。 Abstract:Objective To explore the physiological mechanism and suitable spraying concentration of exogenous quercetin (Q) on improving drought tolerance of chestnut seedlings. Method The chestnut seedlings of Yanbao (YB) were used as test materials. The effects of different concentrations of quercetin [600 μmol·L−1 (QD1), 1200 μmol·L−1 (QD2), 1800 μmol·L−1 (QD3) and 2000 μmol·L−1 (QD4), 3000 μmol·L−1 (QD5)] on chlorophyll content, antioxidant enzyme activity, malondialdehyde (MDA) and flavonol content of chestnut seedlings were studied by pot water control method, with severe drought stress CK1 and normal management CK2 as control. Result Compared with CK1 without quercetin spraying treatment under severe drought stress, five different concentrations of quercetin treatment could significantly increase chlorophyll content, superoxide dismutase (SOD) activity, catalase (CAT) activity, peroxidase (POD) activity and flavonol content in chestnut leaves, and significantly reduce malondialdehyde (MDA) content in chestnut leaves, but, the state of chestnut seedlings after spraying could not return to the normal management of CK2. Through principal component analysis and membership function method, the average drought resistance values of QD1, QD2, QD3, QD4 and QD5 were 0.735, 0.641, 0.738, 1.389 and 0.828, respectively. Among them, 2000 μmol·L−1 (QD4) quercetin concentration treatment effect was the most significant. Conclusion Exogenous application of quercetin could alleviate the damage of chestnut leaf cells by increasing the antioxidant enzyme activity and chlorophyll content of chestnut seedlings, slowing down the increase rate of MDA and slowing down the rate of cell membrane peroxidation, thus improving the drought tolerance of chestnut seedlings, and the effect of 2000 μmol·L−1 (QD4) quercetin concentration treatment was the best. -
Key words:
- Chestnut /
- Quercetin /
- Drought stress /
- Membership function method
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表 1 干旱胁迫下喷施槲皮素后板栗叶片叶绿素含量的变化
Table 1. Changes of chlorophyll content in chestnut leaves after spraying quercetin under drought stress (单位：mg·g−1)
处理
Handle时间
Time0 d 5 d 10 d CK1 3.53±0.01cd 3.19±0.03e 1.50±0.03f CK2 5.12±0.07a 5.13±0.04a 5.30±0.06a QD1 3.49±0.02 de 4.71±0.04 d 4.15±0.10 d QD2 3.66±0.01b 4.91±0.05c 4.15±0.01 d QD3 3.63±0.03b 5.03±0.02b 4.26±0.01c QD4 3.44±0.01e 5.05±0.01b 4.75±0.01b QD5 3.57±0.01c 4.64±0.01 d 4.05±0.01e 不同小写字母表示同列数值差异显著（P＜0.01），下同
Different lowercase letters indicate significant differences in numerical values within the same column（P＜0.01）. The same below.
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表 2 干旱胁迫下喷施槲皮素后板栗叶片MDA含量的变化
Table 2. Changes of MDA content in chestnut leaves after spraying quercetin under drought stress (单位：nmol·g−1)
处理
Handle时间
Time0 d 5 d 10 d CK1 63.12±1.23 d 131.28±2.95a 157.29±7.23a CK2 37.50±2.13e 39.90±1.04 d 41.28±0.42e QD1 68.71±6.09cd 91.07±1.72b 107.50±0.44b QD2 73.70±0.32abc 87.20±2.23bc 102.17±1.26bc QD3 76.63±3.74a 86.22±1.26c 97.65±1.79cd QD4 74.95±0.53ab 84.45±2.12c 93.22±1.49 d QD5 69.75±0.32bc 88.41±0.74bc 101.31±1.37bc
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表 3 干旱胁迫下喷施槲皮素后板栗叶片SOD活性的变化
Table 3. Changes of SOD activity in chestnut leaves after spraying quercetin under drought stress (单位：U·g−1)
处理
Handle时间
Time0 d 5 d 10 d CK1 1157.97±36.93b 1470.95±111.96 d 990.53±13.67e CK2 2034.94±61.22a 2107.45±37.58a 2054.98±42.42a QD1 1152.45±55.34b 1689.88±35.16c 1322.79±33.30 d QD2 1167.31±42.48b 1743.98±24.17c 1447.38±49.93c QD3 1080.96±55.52b 1739.51±16.11c 1431.06±21.96c QD4 1108.46±40.92b 1907.68±31.35b 1522.91±33.64b QD5 1133.02±21.24b 1753.36±44.91c 1320.27±35.56 d
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表 4 干旱胁迫下喷施槲皮素后板栗叶片POD活性的变化
Table 4. Changes of POD activity in chestnut leaves after spraying quercetin under drought stress (单位：U·g−1)
处理
Handle时间
Time0 d 5 d 10 d CK1 860.00±81.65b 893.33±57.35e 600.00±43.20f CK2 1660.00±101.98a 1646.67±24.94a 1746.67±92.86a QD1 846.67±67.99b 1066.67±9.43 de 760.00±28.28ef QD2 886.67±33.99b 1113.33±89.94 d 913.33±131.99 de QD3 893.33±77.17b 1346.67±108.73bc 1153.33±33.99bc QD4 900.00±28.28b 1506.67±133.00ab 1300.0071.18b QD5 786.67±33.99b 1206.67±73.64cd 1040.00±81.65cd
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表 5 干旱胁迫下喷施槲皮素后板栗叶片CAT活性的变化
Table 5. Changes of CAT activity in chestnut leaves after spraying quercetin under drought stress (单位：nmol·min−1·g−1)
处理
Handle时间
Time0 d 5 d 10 d CK1 164.98±3.20b 183.06±5.54 d 133.34±20.96 d CK2 316.40±17.80a 318.66±29.29a 305.10±5.54a QD1 162.72±5.54b 244.08±14.65bc 207.92±22.37bc QD2 164.98±20.96b 223.74±5.54c 176.28±19.96c QD3 155.94±5.54b 219.22±3.20c 178.54±8.46c QD4 158.20±6.39b 262.16±25.57b 212.44±13.93b QD5 166.34±2.79b 228.26±8.46bc 196.62±9.59bc
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表 6 干旱胁迫下喷施槲皮素后板栗叶片黄酮醇含量的变化
Table 6. Changes of Flavonol content in chestnut leaves after spraying quercetin under drought stress (单位：mg·g−1)
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Handle时间
Time0 d 5 d 10 d CK1 2.00±0.03a 2.74±0.25 d 2.22±0.05b CK2 1.07±0.15b 1.09±0.12e 1.10±0.02c QD1 1.95±0.20a 2.97±0.18cd 2.55±0.21ab QD2 1.96±0.08a 3.24±0.21bc 2.56±0.03ab QD3 1.99±0.08a 3.15±0.19bcd 2.36±0.17ab QD4 1.86±0.17a 3.48±0.29ab 2.71±0.07a QD5 1.79±0.12a 3.86±0.04a 2.60±0.36a
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表 7 干旱胁迫下外源喷施槲皮素5 d、10 d的主成分系数、特征值及贡献率
Table 7. The principal component coefficient, characteristic value and contribution rate of exogenous spraying quercetin for 5 days and 10 days under drought stress
时间
Time主成分
ComponentX1 X2 X3 X4 X5 X6 特征值
Eigenvalues贡献率
Contribution rate /%累积贡献率
Cumulative contribution rate /%5 d 1 0.955 0.954 −0.930 0.761 0.519 0.305 3.629 60.478 60.478 2 0.002 0.230 0.145 −0.639 0.269 0.855 1.287 21.446 81.923 10 d 1 0.498 −0.486 0.467 0.449 0.177 0.254 3.642 60.705 60.705 2 0.112 0.240 −0.220 −0.241 0.687 0.592 1.655 27.582 88.287 X1-叶绿素; X2-MDA; X3-SOD; X4-POD; X5-CAT; X6-黄酮醇。下表同。
X1- chlorophyll; X2-MDA; X3-SOD; X4-POD; X5-CAT; X6-flavonol. The same as below.
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表 8 各处理的抗旱性度量值及综合排序
Table 8. 8 The Drought resistance membership function value and comprehensive ranking of each treatment
处理
Treatment5 d抗旱性
度量值
5 days Drought-
tolerance value10 d抗旱性
度量值
10 days Drought-
tolerance value平均抗旱性
度量值
Drought-
tolerance
value排序
OrderQD1 0.927 0.544 0.735 4 QD2 0.711 0.570 0.641 5 QD3 0.855 0.621 0.738 3 QD4 0.961 1.817 1.389 1 QD5 0.983 0.672 0.828 2
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