Transforma tion of C ry I Ac and AP I Two Insect-resistantGenes to Poplar( Popu lus alba ×P. glandu losa )

LI Ke-you1; FAN Jun-feng; ZHAO Zhong; LI Ling; ZHU Hai-lan

Volume 20 Issue 5

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Article Navigation > Forest Research > 2007 > 20(5): 699-704

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Transforma tion of C ry I Ac and AP I Two Insect-resistantGenes to Poplar( Popu lus alba ×P. glandu losa )

1.
College of Forestry, Northwest Sci-Tech of Agricultural and Forestry University, Yangling　712100, Shaanxi, China
2.
Research Institute of Forestry, CAF,Beijing　100091, China

Received Date: 2007-03-16

Abstract

Based on the established leaf exp lant regeneration system of pop lar ( Popu lus alba ×P. glandu losa cv.84K) , Cry I Ac and API fused genes were introduced into the genome of pop lar 84K by Agrobacterium tum efaciensmediated transformation. Through successive selection in the period of shoot and root induction under high level kanamycin-p ressure, 400 kanamycin-resistant regenerated p lantswere obtained, 70 of them were positive resulted byPCR detection. Southern hybridization and EL ISA detection were further showed that Cry I Ac and API gene hadbeen integrated into the genome of 84Kwith one or two cop ies and exp ressed. The transformation p lantswere giveninsect feeding testwith larvae of Clostera anachoreta. The results showed the mortality of insect larvae was accountedfor 60. 0%～80. 0% , and the growth of the survival larvae were seriously inhibited. These p lants could p rovide newresources for pop lar insect-resistant breeding.
- poplar ( P. a lba ×P. glandulosa cv. 84K),
- Cry I Ac and API gene,
- transformation,
- resistance to insect

References

[1]	王永芳, 高宝嘉,郑均宝,等. 杨树抗虫基因工程进展[J]. 河北林果研究, 2001, 16 (1) : 83～90
[2]	贺成林, 李云. 杨树抗虫基因工程进展[J]. 河北林果研究,2004, 16 (1) : 82～89
[3]	王忠华,舒庆尧,崔海瑞, 等. B t杀虫基因与B t转基因抗虫植物研究进展[J]. 植物学通报, 1999 , 16 (1) : 51～58
[4]	伍宁丰,范云六. 含苏云金芽孢杆菌杀虫蛋白基因的杨树工程植株的建立[J]. 科学通报, 1991, 9: 705～708
[5]	田颖川,李太元, 韩一凡, 等. 抗虫转基因欧洲黑杨的培育[J].生物工程学报, 1993, 9 (4) : 291～297
[6]	Wang G J, Stefano C, Chen Y. et al. pop lar ( Populus nigra L. )p lants transformation with a B acillus thuringensis toxin gene: insecticidal activity and genome analysis[J]. Transgenic Res. , 1996, 5:289～301
[7]	McCown B H, McCabe D E, Russel D R, et al. Transformation ofPopulus and incorporation of pest resistance by electric discharge particle acceleration[J]. Plant Cell Rep, 1991, 9: 590～595
[8]	饶红宇,黄敏仁. 杨树NL-80106转B t基因植株的获得及抗虫性[J]. 植物资源与环境学报, 2000, 9 (2) : 1～5
[9]	Howe G T, Goldfarb B, Strauss S H. Agrobacterum -mediated transformation of hybrid pop lar suspension cultures and regeneration oftransformed p lants[J]. Plant cell, Tissue and Organ Culture, 1994,36: 59～71
[10]	郑均宝,张玉满,杨文芝,等. 741杨离体叶片再生及抗虫基因转化[J]. 河北农业大学学报, 1995, 18 (3) : 20～25
[11]	陈颖, 韩一凡, 田颖川, 等. 苏云金杆菌杀虫晶体蛋白基因转化美洲黑杨的研究[J]. 林业科学, 1995, 21 (2) : 97～103
[12]	王学聘,韩一凡, 戴莲韵, 等. 抗虫转基因欧美杨的培育[J]. 林业科学, 1997, 33 (1) : 69～74
[13]	McNabb H S. Genetic transformation and its app lication to fultureforests[J]. TrendsBiotechnol, 1987, 5 (4) : 155～159
[14]	Klopfenstein N B. Transformation of Populus hybrid exp resses awound2inducible potato p roteinase inhibiritor Ⅱ-CAT gene fusion[J]. Can J For Res, 1991, 21: 1 321～1 328
[15]	Klopfenstein N B, Mcnabb J H, Hart E R. Transformation of Populus hybrids to study and imp rove pest resistance[J]. Silvae Genetica, 1993, 42: 86～90
[16]	Lep le J C. Toxicity to Chrysom ela trem ulae (Coleop tera: Chrysomelidae) of trandgenic pop lars exp ressing a cysteine p roteinase inhibitor[J]. Molecular Breeding, 1995, 1 (4) : 319～328
[17]	Delledonne M. Transformation of white pop lar ( Populus alba L. )with a novel A rabidopsis thaliana cysteine p roteinase inhibitor andanalysis of insect pest resistance[J]. Molecular Breeding, 2001, 7(1) : 35～42
[18]	郝贵霞,朱祯,朱之悌. 豇豆蛋白酶抑制剂基因转化毛白杨的研究[J]. 植物学报, 1999, 41 (12) : 1 276～1 282
[19]	Barton K A. Insecticidal toxins in p lant-exp ress an insect-specifictoxin from a scorp ion[P]. C12N015 /82, European patent. patentNo, EP0431829A I, 1990
[20]	郑均宝,梁海永,田颖川, 等. 转双价抗虫741毛白杨的选择及抗虫性[J]. 林业科学, 2000, 36 (2) : 13～19
[21]	李明亮,张辉,胡建军,等. 转B t基因和蛋白酶抑制剂基因杨树抗虫性的研究[J]. 林业科学, 2000, 36 (2) : 93～97
[22]	杨敏生,李志兰,王颖,等. 双价抗虫基因对三倍体毛白杨的转化和抗虫性表达[J]. 林业科学, 2006, 42 (9) : 61～68
[23]	田颖川,郑均宝,虞红梅,等. 转双价抗虫基因杂种741毛白杨的研究[J]. 植物学报, 2000, 42 (3) : 263～268
[24]	王关林,方宏筠. 植物基因工程(第二版) [M]. ,北京:科学出版社, 2002: 744
[25]	Sambrook J, Russel D W. 分子克隆实验指南(第三版) [M]. 黄培堂,王嘉宝,朱厚础,等译. 北京:科学出版社, 2005: 487～509

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Transforma tion of C ry I Ac and AP I Two Insect-resistantGenes to Poplar( Popu lus alba ×P. glandu losa )

1. College of Forestry, Northwest Sci-Tech of Agricultural and Forestry University, Yangling　712100, Shaanxi, China

2. Research Institute of Forestry, CAF,Beijing　100091, China

Received Date: 2007-03-16

Abstract: Based on the established leaf exp lant regeneration system of pop lar ( Popu lus alba ×P. glandu losa cv.84K) , Cry I Ac and API fused genes were introduced into the genome of pop lar 84K by Agrobacterium tum efaciensmediated transformation. Through successive selection in the period of shoot and root induction under high level kanamycin-p ressure, 400 kanamycin-resistant regenerated p lantswere obtained, 70 of them were positive resulted byPCR detection. Southern hybridization and EL ISA detection were further showed that Cry I Ac and API gene hadbeen integrated into the genome of 84Kwith one or two cop ies and exp ressed. The transformation p lantswere giveninsect feeding testwith larvae of Clostera anachoreta. The results showed the mortality of insect larvae was accountedfor 60. 0%～80. 0% , and the growth of the survival larvae were seriously inhibited. These p lants could p rovide newresources for pop lar insect-resistant breeding.

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Reference (25)

[1]	王永芳, 高宝嘉,郑均宝,等. 杨树抗虫基因工程进展[J]. 河北林果研究, 2001, 16 (1) : 83～90
[2]	贺成林, 李云. 杨树抗虫基因工程进展[J]. 河北林果研究,2004, 16 (1) : 82～89
[3]	王忠华,舒庆尧,崔海瑞, 等. B t杀虫基因与B t转基因抗虫植物研究进展[J]. 植物学通报, 1999 , 16 (1) : 51～58
[4]	伍宁丰,范云六. 含苏云金芽孢杆菌杀虫蛋白基因的杨树工程植株的建立[J]. 科学通报, 1991, 9: 705～708
[5]	田颖川,李太元, 韩一凡, 等. 抗虫转基因欧洲黑杨的培育[J].生物工程学报, 1993, 9 (4) : 291～297
[6]	Wang G J, Stefano C, Chen Y. et al. pop lar ( Populus nigra L. )p lants transformation with a B acillus thuringensis toxin gene: insecticidal activity and genome analysis[J]. Transgenic Res. , 1996, 5:289～301
[7]	McCown B H, McCabe D E, Russel D R, et al. Transformation ofPopulus and incorporation of pest resistance by electric discharge particle acceleration[J]. Plant Cell Rep, 1991, 9: 590～595
[8]	饶红宇,黄敏仁. 杨树NL-80106转B t基因植株的获得及抗虫性[J]. 植物资源与环境学报, 2000, 9 (2) : 1～5
[9]	Howe G T, Goldfarb B, Strauss S H. Agrobacterum -mediated transformation of hybrid pop lar suspension cultures and regeneration oftransformed p lants[J]. Plant cell, Tissue and Organ Culture, 1994,36: 59～71
[10]	郑均宝,张玉满,杨文芝,等. 741杨离体叶片再生及抗虫基因转化[J]. 河北农业大学学报, 1995, 18 (3) : 20～25
[11]	陈颖, 韩一凡, 田颖川, 等. 苏云金杆菌杀虫晶体蛋白基因转化美洲黑杨的研究[J]. 林业科学, 1995, 21 (2) : 97～103
[12]	王学聘,韩一凡, 戴莲韵, 等. 抗虫转基因欧美杨的培育[J]. 林业科学, 1997, 33 (1) : 69～74
[13]	McNabb H S. Genetic transformation and its app lication to fultureforests[J]. TrendsBiotechnol, 1987, 5 (4) : 155～159
[14]	Klopfenstein N B. Transformation of Populus hybrid exp resses awound2inducible potato p roteinase inhibiritor Ⅱ-CAT gene fusion[J]. Can J For Res, 1991, 21: 1 321～1 328
[15]	Klopfenstein N B, Mcnabb J H, Hart E R. Transformation of Populus hybrids to study and imp rove pest resistance[J]. Silvae Genetica, 1993, 42: 86～90
[16]	Lep le J C. Toxicity to Chrysom ela trem ulae (Coleop tera: Chrysomelidae) of trandgenic pop lars exp ressing a cysteine p roteinase inhibitor[J]. Molecular Breeding, 1995, 1 (4) : 319～328
[17]	Delledonne M. Transformation of white pop lar ( Populus alba L. )with a novel A rabidopsis thaliana cysteine p roteinase inhibitor andanalysis of insect pest resistance[J]. Molecular Breeding, 2001, 7(1) : 35～42
[18]	郝贵霞,朱祯,朱之悌. 豇豆蛋白酶抑制剂基因转化毛白杨的研究[J]. 植物学报, 1999, 41 (12) : 1 276～1 282
[19]	Barton K A. Insecticidal toxins in p lant-exp ress an insect-specifictoxin from a scorp ion[P]. C12N015 /82, European patent. patentNo, EP0431829A I, 1990
[20]	郑均宝,梁海永,田颖川, 等. 转双价抗虫741毛白杨的选择及抗虫性[J]. 林业科学, 2000, 36 (2) : 13～19
[21]	李明亮,张辉,胡建军,等. 转B t基因和蛋白酶抑制剂基因杨树抗虫性的研究[J]. 林业科学, 2000, 36 (2) : 93～97
[22]	杨敏生,李志兰,王颖,等. 双价抗虫基因对三倍体毛白杨的转化和抗虫性表达[J]. 林业科学, 2006, 42 (9) : 61～68
[23]	田颖川,郑均宝,虞红梅,等. 转双价抗虫基因杂种741毛白杨的研究[J]. 植物学报, 2000, 42 (3) : 263～268
[24]	王关林,方宏筠. 植物基因工程(第二版) [M]. ,北京:科学出版社, 2002: 744
[25]	Sambrook J, Russel D W. 分子克隆实验指南(第三版) [M]. 黄培堂,王嘉宝,朱厚础,等译. 北京:科学出版社, 2005: 487～509

Transforma tion of C ry I Ac and AP I Two Insect-resistantGenes to Poplar( Popu lus alba ×P. glandu losa )

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通讯作者: 陈斌, bchen63@163.com

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Transforma tion of C ry I Ac and AP I Two Insect-resistantGenes to Poplar( Popu lus alba ×P. glandu losa )

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