木薯MeNF-YB转录因子的克隆及在诱导活性氧积累中的作用
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摘要
在植物的生长发育、生理活动和抗逆反应中,核因子Y的亚基NF-YB发挥着重要作用。木薯是一种具有饲用、工业利用、种植等价值的重要经济作物。为探究MeNF-YB在诱导活性氧积累中的作用,本研究克隆了华南124木薯的一个NF-YB转录因子对其进行分析,结果表明该转录因子在叶片中表达量最高,并受到H2O2诱导表达,同时MeNF-YB蛋白定位在细胞核上,而且MeNF-YB能显著提高活性氧、MDA的含量以及相对电导率。通过qRT-PCR分析表明,MeNF-YB能提高抗氧化酶基因、植物防御反应相关基因(PRs)和活性氧相关基因的表达,本研究旨在为MeNF-YB基因的功能及应用提供参考依据。
NF-YB is one of subunits of nuclear factor Y(NF-Y), which plays an important role in plant growth and development, physiological activities and stress resistance. Cassava is an important cash crop with forage,industrial use and planting value. To explore the role of MeNF-YB in inducing reactive oxygen accumulation, the NF-YB transcription factor of 124 cassava from southern China was cloned and analyzed in this research. The results showed that the expression levels of MeNF-YB was the highest in leaves, which was induced by H2 O2. At the same time, the MeNF-YB protein was located in the nucleus, and MeNF-YB could significantly increase the content of reactive oxygen and MDA as well as the relative conductivity. qRT-PCR analysis found that MeNF-YB could improve the expression of antioxidant enzymes genes, plant defense reaction related genes(PRs) and reactive oxygen related genes. The purpose of this study was to provide reference for the function and application of MeNF-YB gene.
NF-YB is one of subunits of nuclear factor Y(NF-Y), which plays an important role in plant growth and development, physiological activities and stress resistance. Cassava is an important cash crop with forage,industrial use and planting value. To explore the role of MeNF-YB in inducing reactive oxygen accumulation, the NF-YB transcription factor of 124 cassava from southern China was cloned and analyzed in this research. The results showed that the expression levels of MeNF-YB was the highest in leaves, which was induced by H2 O2. At the same time, the MeNF-YB protein was located in the nucleus, and MeNF-YB could significantly increase the content of reactive oxygen and MDA as well as the relative conductivity. qRT-PCR analysis found that MeNF-YB could improve the expression of antioxidant enzymes genes, plant defense reaction related genes(PRs) and reactive oxygen related genes. The purpose of this study was to provide reference for the function and application of MeNF-YB gene.
引文
Anke J.J.,Elena T.Y.,and Veneta M.K.,2002,A critical role for ethylene in hydrogen peroxide release during programmed cell death in tomato suspension cells,Planta,214(4):537-545
Apostol I.,Heinstein P.F.,and Low P.S.,1989,Rapid stimulation of an oxidative burst during elicitation of cultured plan cells:role in defense and signal transduction,Plant Physiol.90(1):109-116
Ballif J.,Endo S.,Kotani M.,MacAdam J.,and Wu Y.J.,2011Over-expression of HAP3b enhances primary root elonga-tion in Arabidopsis,Plant Physiol.Biochem.,49(6):579-583
Feng Z.J.,He G.H.,Zheng W.J.,Lu P.P.,Chen M.,Gong Y.M.Ma Y.Z.,and Xu Z.S.,2015,Foxtail millet NF-Y families genome-wide survey and evolution analyses identified two functional genes important in abiotic stresses,Front.Plan Sci.,6:1142
Gechev T.S.,Van B.F.,Stone J.M.,Denev I.,and Laloi C.,2006,Reactive oxygen species as signals that modulate plant stress responses and programmed cell death,Bioessays,28(11):1091-1101
Ito Y.,Thirumurugan T.,Serizawa A.,Hiratsu K.,Ohme-Takagi M.,and Kurata N.,2011,Aberrant vegetative and reproductive development by overexpression and lethality by silencing of OsHAP3E in rice,Plant Sci.,181(2):105-110
Liang M.X.,Hole D.,Wu J.X.,Blake T.,and Wu Y.J.,2012,Expression and functional analysis of nuclear factor-Y,subunit B genes in barley,Planta,235(4):779-791
Mehdy M.C.,1994,Active oxygen species in plant defense against pathogens,Plant Physiol.,105(2):467-472
Mittler R.,Vanderauwera S.,Suzuki N.,Miller G.,Tognetti V.B.,Vandepoele K.,Gollery M.,Shulaev V.,and Van B.F.,2011,ROS signaling:the new wave?Trends Plant Sci.,16(6):300-309
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Stephenson T.J.,McIntyre C.L.,Collet C.,and Xue G.P.,2011,TaNF-YB3 is involved in the regulati on of photosynthesis genes in Triticum aestivum,Funct.Integr.Genomics,11(2):327-340
Sun X.Y.,2017,Drought resistance identification of TaNF-Y1and TaNF-Y2 transgentic wheat,Thesis for M.S.,Northwest A&F University,Supervisors:Chai S.C.,and Zheng W.J.,pp.32-33(孙希媛,2017,TaNF-Y1和TaNF-Y2转基因小麦的抗旱性鉴定,硕士学位论文,西北农林科技大学,导师:柴守诚,郑炜军,pp.32-33)
Wei Y.X.,Hu W.,Wang Q.N.,Liu W.,Wu C.J.,Zeng H.Q.,Yan Y.,Li X.L.,He C.Z.,and Shi H.T.,2016,Comprehensive transcriptional and functional analyses of melatonin synthesis genes in cassava reveal their novel role in hypersensitive-like cell death,Sci.Rep.,6:35029
Yang W.J.,Wu Y.M.,and Tang Y.X.,2009,Expressing and functional analysis of GmMYBJ6 from soybean,Yichuang(Hereditas),31(6):645-653(杨文杰,吴燕民,唐益雄,2009,大豆转录因子基因GmMYBJ6的表达与功能分析,遗传,31(6):645-653)
Zhang Y.Y.,Yang C.W.,Li Y.,Zheng N.Y.,Chen H.,Zhao Q.Z.,Gao T.,Guo H.S.,and Xie Q.,2007,SDIR1 is a RING finger E3ligase that positively regulates stress-responsive abscisic acid signaling in Arabidopsis,Plant Cell,19(6):1912-1929
Apostol I.,Heinstein P.F.,and Low P.S.,1989,Rapid stimulation of an oxidative burst during elicitation of cultured plan cells:role in defense and signal transduction,Plant Physiol.90(1):109-116
Ballif J.,Endo S.,Kotani M.,MacAdam J.,and Wu Y.J.,2011Over-expression of HAP3b enhances primary root elonga-tion in Arabidopsis,Plant Physiol.Biochem.,49(6):579-583
Feng Z.J.,He G.H.,Zheng W.J.,Lu P.P.,Chen M.,Gong Y.M.Ma Y.Z.,and Xu Z.S.,2015,Foxtail millet NF-Y families genome-wide survey and evolution analyses identified two functional genes important in abiotic stresses,Front.Plan Sci.,6:1142
Gechev T.S.,Van B.F.,Stone J.M.,Denev I.,and Laloi C.,2006,Reactive oxygen species as signals that modulate plant stress responses and programmed cell death,Bioessays,28(11):1091-1101
Ito Y.,Thirumurugan T.,Serizawa A.,Hiratsu K.,Ohme-Takagi M.,and Kurata N.,2011,Aberrant vegetative and reproductive development by overexpression and lethality by silencing of OsHAP3E in rice,Plant Sci.,181(2):105-110
Liang M.X.,Hole D.,Wu J.X.,Blake T.,and Wu Y.J.,2012,Expression and functional analysis of nuclear factor-Y,subunit B genes in barley,Planta,235(4):779-791
Mehdy M.C.,1994,Active oxygen species in plant defense against pathogens,Plant Physiol.,105(2):467-472
Mittler R.,Vanderauwera S.,Suzuki N.,Miller G.,Tognetti V.B.,Vandepoele K.,Gollery M.,Shulaev V.,and Van B.F.,2011,ROS signaling:the new wave?Trends Plant Sci.,16(6):300-309
Nelson D.E.,Repetti P.P.,Adams T.R.,Creelman R.A.,Wu J.R.,Warner D.C.,Anstrom D.C.,Bensen R.J.,Castiglioni P.P.,Donnarummo M.G.,Hinchey B.S.,Kumimoto R.W.,Maszle D.R.,Canales R.D.,Krolikowsk K.A.,Dotson S.B.,Gutterson N.,Ratcliffe O.J.,and Heard J.E.,2007,Plant nuclear factor Y(NF-Y)B subunits confer drought tolerance and lead to improved corn yields on water-limited acres,Proc.Natl.Acad.Sci.USA,104(42):16450-16455
Stephenson T.J.,McIntyre C.L.,Collet C.,and Xue G.P.,2011,TaNF-YB3 is involved in the regulati on of photosynthesis genes in Triticum aestivum,Funct.Integr.Genomics,11(2):327-340
Sun X.Y.,2017,Drought resistance identification of TaNF-Y1and TaNF-Y2 transgentic wheat,Thesis for M.S.,Northwest A&F University,Supervisors:Chai S.C.,and Zheng W.J.,pp.32-33(孙希媛,2017,TaNF-Y1和TaNF-Y2转基因小麦的抗旱性鉴定,硕士学位论文,西北农林科技大学,导师:柴守诚,郑炜军,pp.32-33)
Wei Y.X.,Hu W.,Wang Q.N.,Liu W.,Wu C.J.,Zeng H.Q.,Yan Y.,Li X.L.,He C.Z.,and Shi H.T.,2016,Comprehensive transcriptional and functional analyses of melatonin synthesis genes in cassava reveal their novel role in hypersensitive-like cell death,Sci.Rep.,6:35029
Yang W.J.,Wu Y.M.,and Tang Y.X.,2009,Expressing and functional analysis of GmMYBJ6 from soybean,Yichuang(Hereditas),31(6):645-653(杨文杰,吴燕民,唐益雄,2009,大豆转录因子基因GmMYBJ6的表达与功能分析,遗传,31(6):645-653)
Zhang Y.Y.,Yang C.W.,Li Y.,Zheng N.Y.,Chen H.,Zhao Q.Z.,Gao T.,Guo H.S.,and Xie Q.,2007,SDIR1 is a RING finger E3ligase that positively regulates stress-responsive abscisic acid signaling in Arabidopsis,Plant Cell,19(6):1912-1929