磷脂酶参与二萜类化合物冬凌草甲素(oridonin)对拟南芥的化感作用
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摘要
为探究二萜类化合物冬凌草甲素(oridonin)对拟南芥的化感作用机制,本研究以冬凌草甲素为供体,以哥伦比亚野生型拟南芥WT和3个拟南芥磷脂酶A_1(PLA_1)、磷脂酶A_2(PLA_2)、磷脂酶C(PLC)的突变体株系pla_1、pla_2和plc_1为受体植物,通过施加不同浓度的冬凌草甲素(10、60μmol·L~(-1)),对4种拟南芥株系的种子萌发率、幼苗生长状况、相对电导率(EL)、丙二醛(MDA)含量以及3种磷脂酶活性进行了检测。结果表明:外施不同浓度的冬凌草甲素后,拟南芥种子萌发受到抑制,PLA_1和PLA_2参与此抑制过程;根长和下胚轴生长呈现典型的"低促高抑"趋势,PLA_1参与其对根的调控,PLC参与其对下胚轴的调控; 3种磷脂酶PLA_1、PLA_2和PLC酶活性均有不同程度的变化; EL和MDA增大且与浓度和时间呈正相关,PLA_2正调控10μmol·L~(-1)冬凌草甲素增大EL的过程,PLA_1和PLA_2共同响应60μmol·L~(-1)冬凌草甲素增大EL的过程; PLA_1和PLA_2共同作用正调控MDA积累过程;冬凌草甲素对植物的化感作用与磷脂酶有关,植物通过调控磷脂酶响应了冬凌草甲素的部分化感作用,且不同的磷脂酶在调控植物响应冬凌草甲素的生长过程中具有不同的作用。
We investigated the allelopathy mechanism of the diterpenoid oridonin to Arabidopsis thaliana.The germination rate,seedling growth,electrolyte leakage(EL),malondialdehyde(MDA),and three phospholipase activities(PLA_1,PLA_2 and PLC) of four Arabidopsis thaliana lines(the Colombia ecotype WT,and three phospholipase deletion mutants pla_1,pla_2 and plc_1)were measured under different oridonin concentrations(10 and 60 μmol · L~(-1)).The results showed that germination was inhibited by oridonin,and PLA_1 and PLA_2 participated in this process.The growth of root and hypocotyl showed a "low-promotion and high-inhibition"effect.PLA_1 was involved in the regulation of roots.PLC was involved in the regulation of hypocotyl.The activities of different phospholipases changed greatly under different oridonin concentrations.The EL and MDA increased and were positively correlated with treatment concentration and duration.The stimulation of EL by 10 μmol·L~(-1) of oridonin was regulated by PLA_2.The stimulation of EL by 60 μmol·L~(-1) of oridonin was co-regulated by PLA_1 and PLA_2.PLA_1 and PLA_2 regulated the MDA accumulation under oridonin treatment.Our results showed that phospholipase involves in the allelopathy of diterpenoid oridonin to Arabidopsis thaliana.Different phospholipases have different roles in regulating the response of plant to oridonin during its growth.
We investigated the allelopathy mechanism of the diterpenoid oridonin to Arabidopsis thaliana.The germination rate,seedling growth,electrolyte leakage(EL),malondialdehyde(MDA),and three phospholipase activities(PLA_1,PLA_2 and PLC) of four Arabidopsis thaliana lines(the Colombia ecotype WT,and three phospholipase deletion mutants pla_1,pla_2 and plc_1)were measured under different oridonin concentrations(10 and 60 μmol · L~(-1)).The results showed that germination was inhibited by oridonin,and PLA_1 and PLA_2 participated in this process.The growth of root and hypocotyl showed a "low-promotion and high-inhibition"effect.PLA_1 was involved in the regulation of roots.PLC was involved in the regulation of hypocotyl.The activities of different phospholipases changed greatly under different oridonin concentrations.The EL and MDA increased and were positively correlated with treatment concentration and duration.The stimulation of EL by 10 μmol·L~(-1) of oridonin was regulated by PLA_2.The stimulation of EL by 60 μmol·L~(-1) of oridonin was co-regulated by PLA_1 and PLA_2.PLA_1 and PLA_2 regulated the MDA accumulation under oridonin treatment.Our results showed that phospholipase involves in the allelopathy of diterpenoid oridonin to Arabidopsis thaliana.Different phospholipases have different roles in regulating the response of plant to oridonin during its growth.
引文
陈锡莲.2013.二萜化合物Leukamenin E对拟南芥愈伤组织PLD的调控作用研究(硕士学位论文).兰州:西北师范大学.
邓治,校现周.2004.植物体中磷脂酶A2的研究进展.华南热带农业大学学报,3(10):16-19.
邓治.2004.伤害对巴西橡胶树胶乳中磷脂酶A2的影响(硕士学位论文).海南:华南热带农业大学.
丁兰,景宏伟,王涛,等.2010.甘肃产毛叶香茶菜二萜成分及其化感潜能评估.西北师范大学学报:自然科学版,46(2):88-93.
丁兰,祁林林,景宏伟,等.2008.四种对映-贝壳杉烷二萜化合物化感作用潜能的初步评价.西北师范大学学报:自然科学版,44(6):74-78.
高福元,张吉立,刘振平,等.2010.持续低温胁迫对园林树木电导率和丙二醛含量的影响.山东农业科学,(2):47-49.
何文平,杨鹏军,张旭强,等.2017.低温胁迫下内源性ROS对磷脂酶D活性影响的研究.草地学报,25(3):567-573.
孔垂华,胡飞,王朋.2016.植物化感(相生相克)作用.北京:高等教育出版.
李忠,江立庚,唐荣华,等.2015.花生根系分泌物的鉴定和化感作用研究.华南农业大学学报,36(5):48-53.
梁丽,常明,刘睿杰,等.2013.磷脂酶研究进展.食品工业科技,34(4):393-396.
马丹炜,王亚男,王煜,等.2015.化感胁迫诱导植物细胞损伤研究进展.生态学报,35(5):1640-1645.
彭少麟,南蓬,钟扬.2002.高等植物中的萜类化合物及其在生态系统中的作用.生态学杂志,21(3):33-38.
宋颖琦,杨谦.2006.磷脂酶在植物逆境胁迫信号传导中的作用.哈尔滨工业大学学报,38(2):243-246.
王明道,陈红歌,刘新育,等.2009.地黄对芝麻的化感作用及其化感物质的分离鉴定.植物生态学报,33(6):1191-1198.
王庆玲,董涛,张子龙.2015.三七对小麦的化感作用.生态学杂志,34(2):431-437.
王新霞,安炎黄,陈璐,等.2017.冬凌草甲素对拟南芥种子萌发和幼苗生长的化感作用.西北植物学报,37(12):2467-2473.
乌凤章,刘桂丰,姜静.2008.种子萌发调控的分子机理研究进展.北方园艺,(2):54-58.
谢敬宇,谭世强,郭帅,等.2013.人参根系腐解物对十字花科植物的化感作用.中国植保导刊,33(2):18-21.
杨玲.2013.三种对映-贝壳杉烷型二萜化合物对拟南芥幼苗根系生长及根中生长素极性运输的影响的初步研究(硕士学位论文).兰州:西北师范大学.
杨宁,丁芳霞,陈锡莲,等.2013.对映-贝壳杉烷型二萜化合物Leukamenin E对拟南芥愈伤组织磷脂酶D及抗氧化物酶活性的影响.兰州大学学报:自然科学版,49(6):822-827.
杨宁,王程亮,陈霞,等.2014.高山离子芥磷脂酶活性测定及酶动力学分析.草地学报,22(2):375-379.
杨期和,叶万辉,廖富林,等.2005.植物化感作用对种子萌发的影响.生态学杂志,24(12):1459-1465.
张秋菊,张爱华,孙晶波,等.2012.植物体中萜类物质化感作用的研究进展.生态环境学报,21(1):187-193.
Ding L,Qi LL,Jing HW,et al.2008.Phytotoxic effects of leukamenin E(an Ent-kaurene diterpenoid)on root growth and root hair development in Lactuca sativa L.seedlings.Journal of Chemical Ecology,34:1492-1500.
Distéfano AM,Scuffi D,Garcíamata C,et al.2012.Phospholipase Dδis involved in nitric oxide-induced stomatal closure.Planta,236:1899-1907.
Jincy M,Djanaguiraman M,Jeyakumar P,et al.2017.Inhibition of phospholipase D enzyme activity through hexanal leads to delayed mango(Mangifera indica L.)fruit ripening through changes in oxidants and antioxidant enzymes activity.Scientia Horticulturae,218:316-325.
Kaya Y,Aksakal O,Sunar S,et al.2015.Phytotoxical effect of Lepidium draba L.extracts on the germination and growth of monocot(Zea mays L.)and dicot(Amaranthus retroflexus L.)seeds.Toxicology&Industrial Health,31:247-254.
Kunrath-Lima M,de Miranda MC,da Fonseca Ferreira A,et al.2018.Phospholipase C delta 4(PLCδ4)is a nuclear protein involved in cell proliferation and senescence in mesenchymal stromal stem cells.Cellular Signalling,49:59-67.
Liu GA,Ding L,Yang Y,et al.2006.Antioxidative action of ent-kaurene diterpenoids.Research on Chemical Intermediates.32:787-794.
Martin VL,McCoy EL,Dick WA.1990.Allelopathy of crop residues influences com seed germination and early growth.Agronomy Journal,82:555-560.
Molisch H.1937.Der Einfluss einer Pflanze auf die andere,Allelopathie.Jena,Germany:Gustav Fischer.
Piel MS,Peters GHJ,Brask J.2017.Chemoenzymatic synthesis of fluorogenic phospholipids and evaluation in assays of phospholipases A,C and D.Chemistry and Physics of Lipids,202:49-54.
Rice EL.1984.Allelopathy.New York:Academic Press.
Tsai HC,Chung KR.2014.Calcineurin phosphatase and phospholipase C are required for developmental and pathological functions in the citrus fungal pathogen Alternaria alternata.Microbiology,160:1453-1465.
Welti R,Li W,Li M,et al.2002.Profiling membrane lipids in plant stress responses.Role of phospholipase D alpha in freezing-induced lipid changes in Arabidopsis.Journal of Biological Chemistry,277:31994-312002.
Williamson GB,Richardson D.1988.Bioassays for allelopathy,measuring treatment response with independent controls.Journal of Chemical Ecology,14:181-187.
Yakle GA,Cruse RM.1984.Effects of fresh and decomposing corn plant residues extracts on corn seedling development.Soil Science Society of America Journal,48:1143-1146.
Yang N,Chen XL,Wu GF,et al.2013.Characterization of phospholipase D from Arabidopsis thaliana callus in response to Ent-kaurene diterpenoid leukamenin E.Journal of Plant Growth Regulation,32:628-635.
邓治,校现周.2004.植物体中磷脂酶A2的研究进展.华南热带农业大学学报,3(10):16-19.
邓治.2004.伤害对巴西橡胶树胶乳中磷脂酶A2的影响(硕士学位论文).海南:华南热带农业大学.
丁兰,景宏伟,王涛,等.2010.甘肃产毛叶香茶菜二萜成分及其化感潜能评估.西北师范大学学报:自然科学版,46(2):88-93.
丁兰,祁林林,景宏伟,等.2008.四种对映-贝壳杉烷二萜化合物化感作用潜能的初步评价.西北师范大学学报:自然科学版,44(6):74-78.
高福元,张吉立,刘振平,等.2010.持续低温胁迫对园林树木电导率和丙二醛含量的影响.山东农业科学,(2):47-49.
何文平,杨鹏军,张旭强,等.2017.低温胁迫下内源性ROS对磷脂酶D活性影响的研究.草地学报,25(3):567-573.
孔垂华,胡飞,王朋.2016.植物化感(相生相克)作用.北京:高等教育出版.
李忠,江立庚,唐荣华,等.2015.花生根系分泌物的鉴定和化感作用研究.华南农业大学学报,36(5):48-53.
梁丽,常明,刘睿杰,等.2013.磷脂酶研究进展.食品工业科技,34(4):393-396.
马丹炜,王亚男,王煜,等.2015.化感胁迫诱导植物细胞损伤研究进展.生态学报,35(5):1640-1645.
彭少麟,南蓬,钟扬.2002.高等植物中的萜类化合物及其在生态系统中的作用.生态学杂志,21(3):33-38.
宋颖琦,杨谦.2006.磷脂酶在植物逆境胁迫信号传导中的作用.哈尔滨工业大学学报,38(2):243-246.
王明道,陈红歌,刘新育,等.2009.地黄对芝麻的化感作用及其化感物质的分离鉴定.植物生态学报,33(6):1191-1198.
王庆玲,董涛,张子龙.2015.三七对小麦的化感作用.生态学杂志,34(2):431-437.
王新霞,安炎黄,陈璐,等.2017.冬凌草甲素对拟南芥种子萌发和幼苗生长的化感作用.西北植物学报,37(12):2467-2473.
乌凤章,刘桂丰,姜静.2008.种子萌发调控的分子机理研究进展.北方园艺,(2):54-58.
谢敬宇,谭世强,郭帅,等.2013.人参根系腐解物对十字花科植物的化感作用.中国植保导刊,33(2):18-21.
杨玲.2013.三种对映-贝壳杉烷型二萜化合物对拟南芥幼苗根系生长及根中生长素极性运输的影响的初步研究(硕士学位论文).兰州:西北师范大学.
杨宁,丁芳霞,陈锡莲,等.2013.对映-贝壳杉烷型二萜化合物Leukamenin E对拟南芥愈伤组织磷脂酶D及抗氧化物酶活性的影响.兰州大学学报:自然科学版,49(6):822-827.
杨宁,王程亮,陈霞,等.2014.高山离子芥磷脂酶活性测定及酶动力学分析.草地学报,22(2):375-379.
杨期和,叶万辉,廖富林,等.2005.植物化感作用对种子萌发的影响.生态学杂志,24(12):1459-1465.
张秋菊,张爱华,孙晶波,等.2012.植物体中萜类物质化感作用的研究进展.生态环境学报,21(1):187-193.
Ding L,Qi LL,Jing HW,et al.2008.Phytotoxic effects of leukamenin E(an Ent-kaurene diterpenoid)on root growth and root hair development in Lactuca sativa L.seedlings.Journal of Chemical Ecology,34:1492-1500.
Distéfano AM,Scuffi D,Garcíamata C,et al.2012.Phospholipase Dδis involved in nitric oxide-induced stomatal closure.Planta,236:1899-1907.
Jincy M,Djanaguiraman M,Jeyakumar P,et al.2017.Inhibition of phospholipase D enzyme activity through hexanal leads to delayed mango(Mangifera indica L.)fruit ripening through changes in oxidants and antioxidant enzymes activity.Scientia Horticulturae,218:316-325.
Kaya Y,Aksakal O,Sunar S,et al.2015.Phytotoxical effect of Lepidium draba L.extracts on the germination and growth of monocot(Zea mays L.)and dicot(Amaranthus retroflexus L.)seeds.Toxicology&Industrial Health,31:247-254.
Kunrath-Lima M,de Miranda MC,da Fonseca Ferreira A,et al.2018.Phospholipase C delta 4(PLCδ4)is a nuclear protein involved in cell proliferation and senescence in mesenchymal stromal stem cells.Cellular Signalling,49:59-67.
Liu GA,Ding L,Yang Y,et al.2006.Antioxidative action of ent-kaurene diterpenoids.Research on Chemical Intermediates.32:787-794.
Martin VL,McCoy EL,Dick WA.1990.Allelopathy of crop residues influences com seed germination and early growth.Agronomy Journal,82:555-560.
Molisch H.1937.Der Einfluss einer Pflanze auf die andere,Allelopathie.Jena,Germany:Gustav Fischer.
Piel MS,Peters GHJ,Brask J.2017.Chemoenzymatic synthesis of fluorogenic phospholipids and evaluation in assays of phospholipases A,C and D.Chemistry and Physics of Lipids,202:49-54.
Rice EL.1984.Allelopathy.New York:Academic Press.
Tsai HC,Chung KR.2014.Calcineurin phosphatase and phospholipase C are required for developmental and pathological functions in the citrus fungal pathogen Alternaria alternata.Microbiology,160:1453-1465.
Welti R,Li W,Li M,et al.2002.Profiling membrane lipids in plant stress responses.Role of phospholipase D alpha in freezing-induced lipid changes in Arabidopsis.Journal of Biological Chemistry,277:31994-312002.
Williamson GB,Richardson D.1988.Bioassays for allelopathy,measuring treatment response with independent controls.Journal of Chemical Ecology,14:181-187.
Yakle GA,Cruse RM.1984.Effects of fresh and decomposing corn plant residues extracts on corn seedling development.Soil Science Society of America Journal,48:1143-1146.
Yang N,Chen XL,Wu GF,et al.2013.Characterization of phospholipase D from Arabidopsis thaliana callus in response to Ent-kaurene diterpenoid leukamenin E.Journal of Plant Growth Regulation,32:628-635.