水杨酸和茉莉酸甲酯对远志愈伤组织生长和相关酶活性及化学成分的影响

Effects of salicylic acid and methyl jasmonate on growth, activity of related enzymes and chemical components in Polygala tenuifolia Willd.

目的研究水杨酸(salicylic acid,SA)和茉莉酸甲酯(methyl jasmonate,MeJA)对远志愈伤组织生长和相关酶活性及化学成分的影响。方法以远志继代愈伤组织为材料,加入SA(0、4、8、12、16、20、24、28、32 mg/L)和MeJA(0、100、200、400、600、800、1 000 μmol/L)不同浓度下暗培养30 d后,测定远志愈伤组织生长量、丙二醛(MDA)含量、抗氧化酶活性、总酚、总黄酮含量及远志[口山]酮Ⅲ、3,6'-二芥子酰基蔗糖含量。结果 MeJA对远志愈伤组织的生长具有抑制作用,12 mg/L水杨酸(SA)对远志愈伤组织的生长具有促进作用。SA和MeJA对远志愈伤组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)活性及丙二醛(MDA)含量均具有促进作用,且随着SA和MeJA浓度的增大,SOD、CAT、POD活性先上升后下降,MDA含量持续上升。SA质量浓度为20 mg/L时,CAT、SOD活性达到最大值,分别为248.45、4 451.06 U/mg,SA质量浓度为16 mg/L时,POD活性达到最大值,为7.22 U/mg;SA质量浓度为32 mg/L时,MDA含量达到最大值,为25.09 nmol/mg;MeJA浓度为600μmol/L时,CAT、SOD、POD活性达到最大值,分别为273.30、1 451.06、15.27 U/mg;MeJA浓度为1 000 μmol/L时,MDA含量达到最大值,为27.10 nmol/mg。SA对远志愈伤组织中总黄酮的积累具有抑制作用,对总酚无显著影响;MeJA对远志愈伤组织中总酚、总黄酮的积累具有促进作用,MeJA浓度为600 μmol/L时,总黄酮含量最高;MeJA浓度为400 μmol/L时,总酚含量最高。SA和MeJA均对远志愈伤组织中远志[口山]酮Ⅲ、3,6'-二芥子酰基蔗糖的积累具有促进作用,SA浓度为32 mg/L时,远志[口山]酮Ⅲ、3,6'-二芥子酰基蔗糖含量最高;MeJA浓度为1 000 μmol/L时远志[口山]酮Ⅲ、3,6'-二芥子酰基蔗糖含量最高。结论 MeJA对远志愈伤组织生长具有抑制作用,12 mg/L的SA对其具有促进作用。SA和MeJA对SOD、POD、CAT活性,MDA含量及远志[口山]酮Ⅲ、3,6'-二芥子酰基蔗糖的积累均具有促进作用。SA对远志愈伤组织中总黄酮的积累具有抑制作用,对总酚无显著影响,MeJA对远志愈伤组织中总酚、总黄酮的积累具有促进作用。

Objective To study the effects of salicylic acid (SA) and methyl jasmonate (MeJA) on growth, activity of related enzymes and chemical components in Polygala callus. Methods The callus of Polygala was used as material. After 30 d of dark culture at different concentrations of SA (0, 4, 8, 12, 16, 20, 24, 28, 32 mg/L) and MeJA (0, 100, 200, 400, 600, 800, 1 000 μmol/L), the growth of callus, anti-oxidant enzyme activity, the content of MDA, total phenolic, total flavonoid, polygalaxanthone Ⅲ, and 3,6'-disinapoyl sucrose were determined. Results MeJA inhibited the growth of Polygala callus, and 12 mg/L SA promoted the growth of Polygala callus. SA and MeJA promoted the activity of SOD, CAT, POD, and MDA in the callus of Polygala. With the increase of SA and MeJA concentration, the activities of SOD, CAT and POD increased first and then decreased, and the content of MDA continued to rise. When the concentration of SA was 20 mg/L, the activities of CAT and SOD reached the maximum, which were 248.45 U/mg and 4451.06 U/mg, respectively. When the concentration of SA was 16 mg/L, the activity of POD reached the maximum, which was 7.22 U/mg. When the concentration of SA was 32 mg/L, the MDA content reached the maximum value of 25.09 nmol/mg. When the concentration of MeJA was 600 μmol/L, the activities of CAT, SOD, and POD reached the maximum, which were 273.30, 1451.06 and 15.27 U/mg, respectively. When the concentration of MeJA was 1000 μmol/L, the MDA content reached the maximum value of 27.10 nmol/mg. SA inhibited the accumulation of total flavonoids in Polygala callus, and had no significant effect on total phenols. MeJA promoted the accumulation of total phenols and total flavonoids in Polygala callus. When MeJA concentration was 600 μmol/L, the content of total flavonoids was the highest. When the concentration of MeJA was 400 μmol/L, the total phenolic content was the highest. Both SA and MeJA promoted the accumulation of polygalaxanthone Ⅲ and 3,6'-disinapoyl sucrose in Polygala callus. When the concentration of SA was 32 mg/L, the concentration of polygalaxanthone Ⅲ, 3,6'-disinapoyl sucrose was the highest. When the concentration of MeJA was 1 000 μmol/L, the content of polygalaxanthone Ⅲ, 3,6'-disinapoyl sucrose was the highest. Conclusion MeJA had an inhibitory effect on the growth of Polygala callus, and 12 mg/L SA can promote this effect. SA and MeJA promoted the activity of SOD, POD, CAT, the content of MDA and the accumulation of polygalaxanthone Ⅲ and 3,6'-disinapoyl sucrose in Polygala callus. SA inhibited the accumulation of total flavonoids in Polygala callus, and had no significant effect on total phenolics. MeJA promoted the accumulation of total phenolics and total flavonoids in Polygala callus.