生防菌sf628对梨叶片苯丙氨酸解氨酶和过氧化物酶活性的影响
|
摘要
为了从分子水平上探明生防菌抗病机制,采用化学分析方法测定生防菌sf628处理后梨叶片中苯丙氨酸解氨酶(PAL)和过氧化氢酶(CAT)的活性,并运用荧光定量PCR方法分析编码PAL和CAT基因在分子水平的表达情况。结果显示:生防菌sf628处理后,梨叶片PAL和CAT活性均显著增加,处理48 h后,PAL和CAT活性均达到最大值,比处理0 h分别提高2.44倍和4.05倍;基因PAL和CAT的表达量增加,处理24 h后,基因PAL和CAT表达量均达到最大值,分别为清水对照的2.44倍和3.51倍。说明PAL和CAT活性的变化在时间上不同步于相应基因的表达量,但两者的变化趋势一致。因此,喷施生防菌sf628可以在分子水平上调控梨叶片基因PAL和CAT的表达量,并使相应PAL和CAT活性增加,从而提高梨抗病性。
The activities of phenylalanine ammonia lyase( PAL) and catalase( CAT) in pear leaves were measured by common chemical analytic method,and expression patterns of PAL and CAT genes were analyzed by real-time PCR. The results showed that the activities of PAL and CAT were improved significantly after the antagonistic bacterium was inoculated strain sf628 and reached the maximum value after 48-h inoculation,3. 44 times and 5. 05 times greater than those after 0-h inoculation,respectively. The abundance of PAL and CAT genes was up-regulated after the strain was inoculated sf628 and reached the maximum value after 24-h inoculation,2. 44 times and 3. 51 times greater than those treated with water,respectively,indicating the expression of PAL and CAT genes did not synchronize with the change of PAL and CAT activities.It was concluded that inoculating the antagonistic bacterium strain sf628 up-regulated the expression of PAL and CAT genes in pear leaves,leading to enhanced PAL and CAT activities for disease resistance.
The activities of phenylalanine ammonia lyase( PAL) and catalase( CAT) in pear leaves were measured by common chemical analytic method,and expression patterns of PAL and CAT genes were analyzed by real-time PCR. The results showed that the activities of PAL and CAT were improved significantly after the antagonistic bacterium was inoculated strain sf628 and reached the maximum value after 48-h inoculation,3. 44 times and 5. 05 times greater than those after 0-h inoculation,respectively. The abundance of PAL and CAT genes was up-regulated after the strain was inoculated sf628 and reached the maximum value after 24-h inoculation,2. 44 times and 3. 51 times greater than those treated with water,respectively,indicating the expression of PAL and CAT genes did not synchronize with the change of PAL and CAT activities.It was concluded that inoculating the antagonistic bacterium strain sf628 up-regulated the expression of PAL and CAT genes in pear leaves,leading to enhanced PAL and CAT activities for disease resistance.
引文
[1]KOSTENYUK I A,ZON J,BURNS J K.Phenylalanine ammonia lyase gene expression during abscission in citrus[J].Physiologia Plantarum,2002,116:106-112.
[2]高俊杰,秦爱国,于贤昌.低温胁迫下嫁接对黄瓜叶片SOD和CAT基因表达与活性变化的影响[J].应用生态学报,2009,20(1):213-217.
[3]赵士诚,孙静文,马有志,等.镉对玉米幼苗活性氧代谢、超氧化物歧化酶和过氧化氢酶活性及其基因表达的影响[J].中国农业科学,2008,41(10):3025-3032.
[4]ZHENG Y H,FUNG R W M,WANG S Y,et al.Transcript levels of antioxidative genes and oxygen radical scavenging enzyme activities in chilled zucchini squash in response to superatmospheric oxygen[J].Postharvest biology and Technology,2008,47(3):151-158.
[5]ZIAEI M,SHARIFI M,BEHMANESH M,et al.Gene expression and activity of phenyl alanine amonialyase and essential oil composition of Ocimum basilicum L.at different growth stages[J].Iranian Journal of Biotechnology,2012,10(1):32-39.
[6]梁国庆,孙静文,周卫,等.钙对苹果果实超氧化物歧化酶、过氧化氢酶活性及其基因表达的影响[J].植物营养与肥料学报,2011,17(2):438-444.
[7]LUNA C M,PASTORI G M,DRISCOLL S,et al.Drought controls on H2O2accumulation,catalase(CAT)activity and CAT gene expression in wheat[J].Journal of Experimental Botany,2004,56:417-423.
[8]陈秦,薛泉宏,申光辉,等.放线菌制剂对番茄PPO活性及生物量的影响[J].西北农林科技大学学报:自然科学版,2010,38(3):184-188.
[9]唐勇军,彭丽莎,邹俊,等.几丁质酶产生茵发酵液对水稻生长及防御酶活性的影响[J].热带作物学报,2011,32(3):393-397.
[10]LI S M,HUA G G,LIU H X,et al.Analysis of defence enzymes induced by antagonistic bacterium Bacillus subtilis strain AR12 towards Ralstonia solanacearum in tomato[J].Annals of Microbiology,2008,58(4):573-578.
[11]CHAN Z L,TIAN S P.Induction of H2O2-metabolizing enzymes and total protein synthesis by antagonistic yeast and salicylic acid in harvested sweet cherry fruit[J].Postharvest Biology and Technology,2006,39:314-320.
[12]齐希猛,易润华,曾涛,等.海洋细菌TC-1对香蕉防御酶活性的影响[J].中国农学通报,2012,28(1):223-227.
[13]LIU Y Z,CHEN Z Y,LIU Y F,et al.Enhancing bioefficacy of Bacillus subtilis with sodium bicarbonate for the control of ring rot in pear during storage[J].Biological Control,2011,57(2):110-117.
[14]刘永锋,李美荣,陈志谊,等.YD4-6和NV11-4菌株抑菌活性及诱导水稻防御性相关酶活性变化[J].微生物学通报,2010,37(12):1753-1759.
[15]申宏波,胡志凤,丁俊杰,等.Harpins诱导苯丙氨酸解氨酶(PAL)活性防治大豆疫霉根腐病研究[J].大豆科学,2011,30(3):526-528.
[16]WU T M,HU Y T,LEE T M.Effects of cadmium on the regulation of antioxidant enzyme activity,gene expression,and antioxidant defenses in the marine macroalga Ulva fasciata[J].Botanical Studies,2009,50:25-34.
[17]ROMERO P M C,PALMA J M,GOMEZ M,et al.Cadmium causes the oxidative modification of proteins in pea plants[J].Plant Cell and Environment,2002,25:677-686.
[18]SAMEH S,JONATHAN N,DAVID W,et al.Stimulation of defense reactions in Medicago truncatula by antagonistic lipopeptides from Paenibacillus sp.Strain B2[J].Applied and Environmental Microbiology,2010,76(22):7420-7428.
[2]高俊杰,秦爱国,于贤昌.低温胁迫下嫁接对黄瓜叶片SOD和CAT基因表达与活性变化的影响[J].应用生态学报,2009,20(1):213-217.
[3]赵士诚,孙静文,马有志,等.镉对玉米幼苗活性氧代谢、超氧化物歧化酶和过氧化氢酶活性及其基因表达的影响[J].中国农业科学,2008,41(10):3025-3032.
[4]ZHENG Y H,FUNG R W M,WANG S Y,et al.Transcript levels of antioxidative genes and oxygen radical scavenging enzyme activities in chilled zucchini squash in response to superatmospheric oxygen[J].Postharvest biology and Technology,2008,47(3):151-158.
[5]ZIAEI M,SHARIFI M,BEHMANESH M,et al.Gene expression and activity of phenyl alanine amonialyase and essential oil composition of Ocimum basilicum L.at different growth stages[J].Iranian Journal of Biotechnology,2012,10(1):32-39.
[6]梁国庆,孙静文,周卫,等.钙对苹果果实超氧化物歧化酶、过氧化氢酶活性及其基因表达的影响[J].植物营养与肥料学报,2011,17(2):438-444.
[7]LUNA C M,PASTORI G M,DRISCOLL S,et al.Drought controls on H2O2accumulation,catalase(CAT)activity and CAT gene expression in wheat[J].Journal of Experimental Botany,2004,56:417-423.
[8]陈秦,薛泉宏,申光辉,等.放线菌制剂对番茄PPO活性及生物量的影响[J].西北农林科技大学学报:自然科学版,2010,38(3):184-188.
[9]唐勇军,彭丽莎,邹俊,等.几丁质酶产生茵发酵液对水稻生长及防御酶活性的影响[J].热带作物学报,2011,32(3):393-397.
[10]LI S M,HUA G G,LIU H X,et al.Analysis of defence enzymes induced by antagonistic bacterium Bacillus subtilis strain AR12 towards Ralstonia solanacearum in tomato[J].Annals of Microbiology,2008,58(4):573-578.
[11]CHAN Z L,TIAN S P.Induction of H2O2-metabolizing enzymes and total protein synthesis by antagonistic yeast and salicylic acid in harvested sweet cherry fruit[J].Postharvest Biology and Technology,2006,39:314-320.
[12]齐希猛,易润华,曾涛,等.海洋细菌TC-1对香蕉防御酶活性的影响[J].中国农学通报,2012,28(1):223-227.
[13]LIU Y Z,CHEN Z Y,LIU Y F,et al.Enhancing bioefficacy of Bacillus subtilis with sodium bicarbonate for the control of ring rot in pear during storage[J].Biological Control,2011,57(2):110-117.
[14]刘永锋,李美荣,陈志谊,等.YD4-6和NV11-4菌株抑菌活性及诱导水稻防御性相关酶活性变化[J].微生物学通报,2010,37(12):1753-1759.
[15]申宏波,胡志凤,丁俊杰,等.Harpins诱导苯丙氨酸解氨酶(PAL)活性防治大豆疫霉根腐病研究[J].大豆科学,2011,30(3):526-528.
[16]WU T M,HU Y T,LEE T M.Effects of cadmium on the regulation of antioxidant enzyme activity,gene expression,and antioxidant defenses in the marine macroalga Ulva fasciata[J].Botanical Studies,2009,50:25-34.
[17]ROMERO P M C,PALMA J M,GOMEZ M,et al.Cadmium causes the oxidative modification of proteins in pea plants[J].Plant Cell and Environment,2002,25:677-686.
[18]SAMEH S,JONATHAN N,DAVID W,et al.Stimulation of defense reactions in Medicago truncatula by antagonistic lipopeptides from Paenibacillus sp.Strain B2[J].Applied and Environmental Microbiology,2010,76(22):7420-7428.