Preparation and Bioassay of Harpin Nanoparticles

LI Lin; WAN San-lian; LIU Wen-bo; ZHANG Yu; MOU Wei-guo; ZHENG Fu-cong

doi:10.15886/j.cnki.rdswxb.2012.02.011

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Volume 3 Issue 2

Jun. 2012

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Article Navigation > Journal of Tropical Biology > 2012 > 3(2): 147-154

LI Lin, WAN San-lian, LIU Wen-bo, ZHANG Yu, MOU Wei-guo, ZHENG Fu-cong. Preparation and Bioassay of Harpin Nanoparticles[J]. Journal of Tropical Biology, 2012, 3(2): 147-154. doi: 10.15886/j.cnki.rdswxb.2012.02.011

Citation:

LI Lin, WAN San-lian, LIU Wen-bo, ZHANG Yu, MOU Wei-guo, ZHENG Fu-cong. Preparation and Bioassay of Harpin Nanoparticles[J]. Journal of Tropical Biology, 2012, 3(2): 147-154. doi: 10.15886/j.cnki.rdswxb.2012.02.011

Preparation and Bioassay of Harpin Nanoparticles

doi: 10.15886/j.cnki.rdswxb.2012.02.011

College of Environment and Plant Protection, Hainan University, Haikou 570228, China

Received Date: 2012-05-07

Abstract

Harpin_Xoo and Harpin_Xm are derived from Xanthomonas oryzae pv.oryzae and Xanthomonas citri subsp.malvacearum,respectively,and have the characteristics of Harpin.Harpin_Xoo and Harpin_Xm were prepared as nanoparticle agents.Both the two proteins and their protein nanoparticles induced hypersensitive response and oxidative burst in tobacco leaves,playing a beneficial role in the process of tobacco seed growing,especially Harpin_Xm nanoparticles which promoted oxidative burst of tobacco by 104.5%.Harpin_Xoo and Harpin_Xm nanoparticles showed a burst release at the initial stage,and their release rates increased to 83.9% and 84.5%,respectively,within 48 h.These two nanoparticles played a role in promoting protein activity continuously.
- Harpin_Xoo,
- Harpin_Xm,
- nanoparticles,
- hypersensitive response,
- oxidative burst

References

[1]	缪卫国,宋从凤,乔子辰等.转hpa1_Xoo基因棉花活性氧产生及防卫相关基因的表达[J].南京农业大学学报,2011,34(2):61-66.
[2]	李平,陆徐忠,邵敏,等.水稻黄单胞细菌Harpin蛋白的遗传多样性及其诱导烟草过敏反应和抗病性功能[J].中国科学,2004,34(2):136-143.
[3]	MIAO W G,SONG C F,ZHENG F C,et al.The defense function and secondary structure analysis of hpa Xm from Xanthomonas citri subsp.malvacearum[J].Amino Acids,2011(41):69-70.
[4]	李明.HarpinX蛋白的纯化、抗体制备及转hrfX基因棉花的研究[D].南京:南京农业大学,2005.
[5]	龚晓崇.生长素对Harpin蛋白激发HR的调控机制初步研究[D].南京:南京农业大学,2011.
[6]	ZHANG S,KLESSIG D F.MAPK cascades in plant defense signaling[J].Trends Plant Sci,2001(6):520-527.
[7]	SAMUEL M A,MILES G P,ELLIS B E.Ozone treatment rapidly activates MAP kinase signaling in plants[J].Plant J,2000,22(4):367-376.
[8]	李俊芳,闫妍,卢晓静,等.纳米粒子的生物安全性研究进展[J].材料导报,2011,1(25):49-52.
[9]	陈红丽,吕洁丽,晏杰,等.壳聚糖修饰的PLGA纳米粒作为蛋白多肽类药物载体的研究[J].功能材料,2011(2):202-205.
[10]	冯博华,张子勇.鱼藤酮-壳聚糖接枝物农药纳米粒子水分散制剂研究[J].农药学学报,2011,13(1):64-70.
[11]	湛江,杨扬,宁辉,等.载Harpin_Xooc蛋白纳米粒的制备及生物学效应测定[J].南京农业大学学报,2010,33(3):57-61.
[12]	缪卫国.转hpa1_Xoo基因棉花抗病虫防卫反应与全基因组转录谱分析及棉花角斑病菌hpa Xm基因的功能[D].南京:南京农业大学,2009.
[13]	JANES K A,CALVO P,ALONSO M J.Polysaccharide colloidal particles as delivery systems for macromolecules[J].Ad-vanced Drug Delivery Reviews,2001,47:83-97.
[14]	TANG Z X,SHI L E,QIAN J Q.Neutral lipase from aqueous solutions on chitosan nano-particles[J].Biochemical Engi-neering Journal,2007,34:217-223.
[15]	GAN Q,WANG T.Chitosan nanoparticle as protein delivery carrier-systematic examination of fabrication conditions for effi-cient loading and release[J].Colloids and Surfaces B:Biointerfaces,2007,59(1):24-34.
[16]	JIANG M,ZHANG J.Effect of abscisic acid on active oxygen species,antioxidative defense system and oxidase damage in leaves of maize seedlings[J].Plant Cell Physiol,2001,42:1265-1273.
[17]	TRUJILLO M,KOGEL K H,HüCKELHOVEN R.Superoxide and hydrogen peroxide play different roles in the nonhost inter-action of barley and wheat with inappropriate formae speciales of Blumeria graminis[J].Mol Plant-Microbe Interact,2004,17:304-312.
[18]	刘爱新,李多川,董汉松,等.Harpin_Xoo诱发烟草过敏性反应早期H₂O₂变化及有关基因表达[J].植物病理学报,2006,36(2):163-168.
[19]	胡重怡,郑少清,陈兴江,等.烟草无菌苗培养前的种子消毒技术研究[J].中国烟草科学,2007,28(2):45-46.
[20]	桂耀林.培养基的配置与简化-植物组织培养技术(Ⅱ)[J].生命世界,1985(5):39-41.
[21]	WEI Z M,LABY R J,ZUMOFF C H,et al.Harpin,elictor of the hypersensitive response produced by the plant pathogen Erwinia amylovora[J].Science,1992,257:85-88.
[22]	LEVINE A,TENHAKEN R,DIXON R,et al.H₂O₂ from the oxidative burst orchestrayes the plant hypersensitive disease re-sistance response[J].Cell,1994,79:583-593.
[23]	MIAO W G,WANG X B,LI M,et al.Genetic transformation of cotton with a harpin-encoding gene hpaxoo confers an en-hanced defense response against different pathogens through a priming mechanism[J].BMC Plant Biology,2010,10:67.
[24]	MIAO W G,WANG J S.Hpa Xm from Xanthomonas smithii pv.smithii is a novel harpin protein with signal peptide[J].Amino acids,2008(37):S120-S121.
[25]	BAKER C J,ORLANDI E W,MOCK N M.Harpin,an elicitor of the hypersensitive response in tobacco caused by Erwinia amylovora,elicits active oxygen production in suspension cells[J].Plant Physiology,1993,102:1341-1344.
[26]	NЁOL L,THIEME F,NENNSTIEL D,et al.Two novel typeⅢ-secreted proteins of Xanthomonas campestris pv.vesicatoria are encoded within the hrp pathogenicity island[J].J Bacteriol,2002,184:1340-1348.
[27]	ZHU W,MAGBANUA M M,WHITE F F.Identification of two novel hrp-associated genes in the hrp gene cluster of Xan-thomonas oryzae pv.oryzae[J].J Bacteriol,2000,182:1844-1853.
[28]	PENG J L,BAO Z L,REN H Y,et al.Expression of harpin in transgenic tobacco induces pathogen defense in the absence of hypersensitive cell death[J].Phytopathology,2004,94:1048-1055.
[29]	WANG X Y,SONG C F,MIAO W G,et al.Mutations in the N-terminal coding region of the harpin protein hpa1from Xan-thomonas oryzae cause loss of hypersensitive reaction induction in tobacco[J].Appl Microbiol Biotechnol,2008,81:359-369.
[30]	陈惠萍,徐朗莱.壳聚糖调节植物生长发育及诱发植物抗病性研究进展[J].云南植物研究,2005,27(6):613-619.
[31]	CHIRKOV S N.The antiviral activity of chitosan[J].Applied Biochemistry and Microbiology,2002,38(1):1-8.
[32]	惠娜娜.壳寡糖诱导烟草抗烟草花叶病的若干特点研究[D].杨凌:西北农林科技大学,2007.

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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Preparation and Bioassay of Harpin Nanoparticles

College of Environment and Plant Protection, Hainan University, Haikou 570228, China

Received Date: 2012-05-07

Key words:

Abstract: Harpin_Xoo and Harpin_Xm are derived from Xanthomonas oryzae pv.oryzae and Xanthomonas citri subsp.malvacearum,respectively,and have the characteristics of Harpin.Harpin_Xoo and Harpin_Xm were prepared as nanoparticle agents.Both the two proteins and their protein nanoparticles induced hypersensitive response and oxidative burst in tobacco leaves,playing a beneficial role in the process of tobacco seed growing,especially Harpin_Xm nanoparticles which promoted oxidative burst of tobacco by 104.5%.Harpin_Xoo and Harpin_Xm nanoparticles showed a burst release at the initial stage,and their release rates increased to 83.9% and 84.5%,respectively,within 48 h.These two nanoparticles played a role in promoting protein activity continuously.

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Reference (32)

[1]	缪卫国,宋从凤,乔子辰等.转hpa1_Xoo基因棉花活性氧产生及防卫相关基因的表达[J].南京农业大学学报,2011,34(2):61-66.
[2]	李平,陆徐忠,邵敏,等.水稻黄单胞细菌Harpin蛋白的遗传多样性及其诱导烟草过敏反应和抗病性功能[J].中国科学,2004,34(2):136-143.
[3]	MIAO W G,SONG C F,ZHENG F C,et al.The defense function and secondary structure analysis of hpa Xm from Xanthomonas citri subsp.malvacearum[J].Amino Acids,2011(41):69-70.
[4]	李明.HarpinX蛋白的纯化、抗体制备及转hrfX基因棉花的研究[D].南京:南京农业大学,2005.
[5]	龚晓崇.生长素对Harpin蛋白激发HR的调控机制初步研究[D].南京:南京农业大学,2011.
[6]	ZHANG S,KLESSIG D F.MAPK cascades in plant defense signaling[J].Trends Plant Sci,2001(6):520-527.
[7]	SAMUEL M A,MILES G P,ELLIS B E.Ozone treatment rapidly activates MAP kinase signaling in plants[J].Plant J,2000,22(4):367-376.
[8]	李俊芳,闫妍,卢晓静,等.纳米粒子的生物安全性研究进展[J].材料导报,2011,1(25):49-52.
[9]	陈红丽,吕洁丽,晏杰,等.壳聚糖修饰的PLGA纳米粒作为蛋白多肽类药物载体的研究[J].功能材料,2011(2):202-205.
[10]	冯博华,张子勇.鱼藤酮-壳聚糖接枝物农药纳米粒子水分散制剂研究[J].农药学学报,2011,13(1):64-70.
[11]	湛江,杨扬,宁辉,等.载Harpin_Xooc蛋白纳米粒的制备及生物学效应测定[J].南京农业大学学报,2010,33(3):57-61.
[12]	缪卫国.转hpa1_Xoo基因棉花抗病虫防卫反应与全基因组转录谱分析及棉花角斑病菌hpa Xm基因的功能[D].南京:南京农业大学,2009.
[13]	JANES K A,CALVO P,ALONSO M J.Polysaccharide colloidal particles as delivery systems for macromolecules[J].Ad-vanced Drug Delivery Reviews,2001,47:83-97.
[14]	TANG Z X,SHI L E,QIAN J Q.Neutral lipase from aqueous solutions on chitosan nano-particles[J].Biochemical Engi-neering Journal,2007,34:217-223.
[15]	GAN Q,WANG T.Chitosan nanoparticle as protein delivery carrier-systematic examination of fabrication conditions for effi-cient loading and release[J].Colloids and Surfaces B:Biointerfaces,2007,59(1):24-34.
[16]	JIANG M,ZHANG J.Effect of abscisic acid on active oxygen species,antioxidative defense system and oxidase damage in leaves of maize seedlings[J].Plant Cell Physiol,2001,42:1265-1273.
[17]	TRUJILLO M,KOGEL K H,HüCKELHOVEN R.Superoxide and hydrogen peroxide play different roles in the nonhost inter-action of barley and wheat with inappropriate formae speciales of Blumeria graminis[J].Mol Plant-Microbe Interact,2004,17:304-312.
[18]	刘爱新,李多川,董汉松,等.Harpin_Xoo诱发烟草过敏性反应早期H₂O₂变化及有关基因表达[J].植物病理学报,2006,36(2):163-168.
[19]	胡重怡,郑少清,陈兴江,等.烟草无菌苗培养前的种子消毒技术研究[J].中国烟草科学,2007,28(2):45-46.
[20]	桂耀林.培养基的配置与简化-植物组织培养技术(Ⅱ)[J].生命世界,1985(5):39-41.
[21]	WEI Z M,LABY R J,ZUMOFF C H,et al.Harpin,elictor of the hypersensitive response produced by the plant pathogen Erwinia amylovora[J].Science,1992,257:85-88.
[22]	LEVINE A,TENHAKEN R,DIXON R,et al.H₂O₂ from the oxidative burst orchestrayes the plant hypersensitive disease re-sistance response[J].Cell,1994,79:583-593.
[23]	MIAO W G,WANG X B,LI M,et al.Genetic transformation of cotton with a harpin-encoding gene hpaxoo confers an en-hanced defense response against different pathogens through a priming mechanism[J].BMC Plant Biology,2010,10:67.
[24]	MIAO W G,WANG J S.Hpa Xm from Xanthomonas smithii pv.smithii is a novel harpin protein with signal peptide[J].Amino acids,2008(37):S120-S121.
[25]	BAKER C J,ORLANDI E W,MOCK N M.Harpin,an elicitor of the hypersensitive response in tobacco caused by Erwinia amylovora,elicits active oxygen production in suspension cells[J].Plant Physiology,1993,102:1341-1344.
[26]	NЁOL L,THIEME F,NENNSTIEL D,et al.Two novel typeⅢ-secreted proteins of Xanthomonas campestris pv.vesicatoria are encoded within the hrp pathogenicity island[J].J Bacteriol,2002,184:1340-1348.
[27]	ZHU W,MAGBANUA M M,WHITE F F.Identification of two novel hrp-associated genes in the hrp gene cluster of Xan-thomonas oryzae pv.oryzae[J].J Bacteriol,2000,182:1844-1853.
[28]	PENG J L,BAO Z L,REN H Y,et al.Expression of harpin in transgenic tobacco induces pathogen defense in the absence of hypersensitive cell death[J].Phytopathology,2004,94:1048-1055.
[29]	WANG X Y,SONG C F,MIAO W G,et al.Mutations in the N-terminal coding region of the harpin protein hpa1from Xan-thomonas oryzae cause loss of hypersensitive reaction induction in tobacco[J].Appl Microbiol Biotechnol,2008,81:359-369.
[30]	陈惠萍,徐朗莱.壳聚糖调节植物生长发育及诱发植物抗病性研究进展[J].云南植物研究,2005,27(6):613-619.
[31]	CHIRKOV S N.The antiviral activity of chitosan[J].Applied Biochemistry and Microbiology,2002,38(1):1-8.
[32]	惠娜娜.壳寡糖诱导烟草抗烟草花叶病的若干特点研究[D].杨凌:西北农林科技大学,2007.

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Preparation and Bioassay of Harpin Nanoparticles

doi: 10.15886/j.cnki.rdswxb.2012.02.011

Abstract

References

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通讯作者: 陈斌, bchen63@163.com

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