代红艳;闫玉娇;李晓明;李贺;张志宏;

• 1:沈阳农业大学园艺学院
• 2:沈阳农业大学生物科学技术学院

摘要(Abstract)：

【目的】从山楂(Crataegus pinnatifida)中克隆过氧化物酶(POD)编码基因,通过转基因验证其在木质素合成中的作用,为揭示山楂内果皮形成分子机制奠定基础。【方法】利用RT-PCR技术从山楂克隆基因,以p RI 101-AN为构建基因过量表达载体的基础载体,通过农杆菌介导的遗传转化方法将目的基因导入烟草。【结果】从山楂中克隆出编码序列长度为996 bp的POD72基因,命名为Cp POD72。山楂POD72与木本和草本植物POD72的氨基酸序列一致性均较高。构建了Cp POD72基因的过量表达载体,通过农杆菌介导的转化获得了66个烟草转化植株。RT-PCR检测结果显示Cp POD72基因在烟草转基因植物中异位表达,组织化学染色分析表明转Cp POD72基因烟草植株中木质素含量增加。【结论】Cp POD72基因可能参与木质素合成。

关键词(KeyWords)： 山楂;过氧化物酶基因;木质素;转基因烟草;异位表达

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31170635)

作者(Author): 代红艳;闫玉娇;李晓明;李贺;张志宏;

Email:

DOI: 10.13925/j.cnki.gsxb.20150324

参考文献(References)：

• [1]PHIPPS J B,OKENNON R J,LANCE R W.Hawthorns and medlars[M].Royal Horticultural Society,Cambridge,UK,2003.
• [2]RIGELSKY J M,SWEET B V.Hawthorn:Pharmacology and therapeutic uses[J].American Journal of Health-System Pharmacy,2002,59:417-422.
• [3]ZHAO Huanchun,FENG Baotian.China Fruit-plant Monograph·Hawthorn(Crataegus)Flora[M].Beijing:China Forest Press,1996:94-152.赵焕谆,丰宝田.中国果树志·山楂卷[M].北京:中国林业出版社,1996:94-152.
• [4]GUITIáN P.Latitudinal variation in the fruiting phenology of a bird-dispersed plant(Crataegus monogyna)in Western Europe[J].Plant Ecology,1998,137:139-142.
• [5]QIAO Yanchun,LüDeguo,SUN Wenwen,LIU Miao.Comparison of accumulation status of lignin between soft core and hard core of hawthorn[J].Subtropical Plant Science,2009,38(4):9-11.乔燕春,吕德国,孙文文,刘淼.软核与硬核山楂种核木质素积累状况比较[J].亚热带植物科学,2009,38(4):9-11.
• [6]DAI H,HAN G,YAN Y,ZHANG F,LIU Z,LI X,LI W,MA Y,LI H,LIU Y,ZHANG Z.Transcript assembly and quantification by RNA-Seq reveals differentially expressed genes between soft-endocarp and hard-endocarp hawthorns[J].PLo S ONE,2013,8(9):e72910.
• [7]TAKAHAMA U,ONIKI T,SHIMOKAWA H.A possible mechanism for the oxidation of sinapyl alcohol by peroxidase-dependent reaction in the apoplast:Enhancement of the oxidation by hydroxycinnamic acid and component of the apoplast[J].Plant and Cell Physiology,1996,37:499-504.
• [8]XIAO Min,ZHANG Zhihong,DAI Hongyan,YANG Hongyi,LI He.Enhancing the stability of detection of Strawberry vein banding virus by PCR[J].Journal of Fruit Science,2005,22(5):483-487.肖敏,张志宏,代红艳,杨洪一,李贺.PCR检测草莓镶脉病毒的稳定性研究[J].果树学报,2005,22(5):483-487.
• [9]CHANG L,ZHANG Z,YANG H,LI H,DAI H.Detection of strawberry RNA and DNA viruses by RT-PCR using total nucleic acid as a template[J].Journal of Phytopathology,2007,155:431-436.
• [10]FRANCOZ E,RANOCHA P,NGUYEN-KIM H,JAMET E,BURLAT V,DUNAND C.Roles of cell wall peroxidases in plant development[J].Phytochemistry,2015,112:15-21.
• [11]CHRISTENSEN J H,BAUW G,WELINDER K G,Van MONTAGU M,BOERJAN W.Purification and characterization of peroxidases correlated with lignification in poplar xylem[J].Plant Physiology,1998,118(1):125-135.
• [12]ABELES F B,BILES C L.Characterization of peroxidases in lignifying peach fruit endocarp[J].Plant Physiology,1991,95(1):269-273.
• [13]CHENG Hua,LI Linling,WANG Yan,CHENG Shuiyuan.Molecular cloning,characterization and expression of POD1 gene from Ginkgo biloba L.[J].Acta Agriculturae Boreali-Sinica,2010,25(6):44-51.程华,李琳玲,王燕,程水源.银杏过氧化物酶基因POD1的克隆及表达分析[J].华北农学报,2010,25(6):44-51.
• [14]MANSOURI I E,MERCADO J A,SANTIAGO-DOMENECH N,PLIEGO-ALFARO F,VALPUESTA V,QUESADA M A.Biochemical and phenotypical characterization of transgenic tomato plants overexpressing a basic peroxidase[J].Physiologia Plantarum,1999,106(4):355-362.
• [15]HERRERO J,FERNáNDEZ-PéREZ F,YEBRA T,NOVOUZAL E,POMAR F,PEDRE?O Má,CUELLO J,GUéRA A,ESTEBAN-CARRASCO A,ZAPATA JM.Bioinformatic and functional characterization of the basic peroxidase 72 from Arabidopsis thaliana involved in lignin biosynthesis[J].Planta,2013,237(6):1599-1612.
• [16]RALPH J,LUNDQUIST K,BRUNOW G,LU F,KIM H,SCHATZ P F,MARITA J M,HATFIELD R D,RALPH SA,CHRISTENSEN J H.Lignins:Natural polymers from oxidative coupling of 4-hydroxyphenylpropanoids[J].Phytochemistry Reviews,2004,3(1-2):29-60.
• [17]VANHOLME R,DEMEDTS B,MORREEL K,RALPH J,BOERJAN W.Lignin biosynthesis and structure[J].Plant Physiology,2010,153(3):895-905.