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作物学报 ›› 2012, Vol. 38 ›› Issue (08): 1538-1543.doi: 10.3724/SP.J.1006.2012.01538

• 研究简报 • 上一篇    下一篇

用基因枪法获得转异天南星基因aha抗蚜虫小麦

张彦1，喻修道1，唐克轩2，夏兰琴1,*   

  1. 1中国农业科学院作物科学研究所 / 国家农作物基因资源与遗传改良重大科学工程, 北京100081; 2上海交通大学农业与生物学院植物生物技术研究中心，上海 200240
  • 收稿日期:2011-12-21 修回日期:2012-04-15 出版日期:2012-08-12 网络出版日期:2012-05-11
  • 通讯作者: 夏兰琴, E-mail: xialq@mail.caas.net.cn, Tel: 010-82105804
  • 基金资助:

    本研究由国家转基因生物新品种培育重大专项(2011ZX08002-001)资助。

Generation of Aphid Resistant Transgenic Wheat with aha From Arisaema heterophyllum by Particle Bombardment

ZHANG Yan1,YU Xiu-Dao1,TANG Ke-Xuan2,XIA Lan-Qin1,*   

  1. 1 National Key Facility for Crop Gene Resources and Genetic Improvement / Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 2 Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China
  • Received:2011-12-21 Revised:2012-04-15 Published:2012-08-12 Published online:2012-05-11
  • Contact: 夏兰琴, E-mail: xialq@mail.caas.net.cn, Tel: 010-82105804

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被引次数

13

摘要： 凝集素是一类具有特异糖结合活性的蛋白，对蚜虫等害虫有很强的抗杀作用。利用异天南星凝集素基因aha(Arisaema heterophyllum agglutinin)以及水稻Rubisco小亚基启动子，构建了aha基因植物绿色组织特异性表达载体，并采用基因枪转化方法, 与携带bar基因的pAHC20载体共转化到小麦品种科农199中。经过愈伤诱导、再生和筛选以及PCR鉴定，获得aha转基因植株42株，平均转化效率为2.41%。对转aha基因植株后代PCR鉴定表明，T1代转基因植株的分离比例基本符合孟德尔遗传规律。利用室内多目标综合判别法评定抗蚜虫特性，8个T1代转基因株系中有高抗材料1份，低抗材料3份，占参试比例44.4%。本研究为获得抗蚜虫转基因小麦新材料奠定了基础。

关键词: 小麦, 植物凝集素, Rubisco小亚基启动子, 基因枪转化, 共转化, 抗虫性

Abstract: Agglutinin is a class of mannose-binding protein, which has detrimental effect on aphids and other pests. In this study, the vector harboring aha cloned from Arisaema heterophyllum directed by rice Rubisco small subunit promoter (rbcS) was constructed. By co-transformed with pAHC20 that harbors bar selection gene, the aha gene was transferred into wheat variety Kenong 199 via bombardment. After induction, regeneration, two rounds of selection and with the conformation by PCR analysis, 42 transgenic plants with foreign aha gene were obtained, average co-transformation efficiency of 2.41%. According to PCR analysis, the segregation of the T1plants was basically consistent with Mendel’s separation. Aphid resistance bioassay was carried out using eight randomly selected transgenic lines by multiple discrimination method. One high-resistant and three low-resistant transgenic lines were identified, accounting for 44.4% of the tested materials. This study has laid a basis for application of aha gene in development of aphid resistant transgenic wheat.

Key words: Wheat, Agglutinin, rbcS promoter, Particle bombardment, Co-transformation, Insect resistance

[1] Awmack C S, Harrington R. Elevated CO2 affects the interactions between aphid pests and host plant flowering. Agric For Entomol, 2000, 2: 57–61
[2] Hilder V A, Powell K S, Gatehouse A MR, Gatehouse J A, Gatehouse L N, Shi Y, Hamliton WD O, Merryweather A, Newell C A, Timans J C, Peumans W J, Damme E V, Boulter D. Expression of snowdrop lectin in transgenic tobacco plants results in added protection against aphids. Transgenic Res, 1995, 4: 18–25
[3] Gatehouse A MR, Down R E, Powell K S, Sauvion N, Rahbe Y, Newell C A, Merryweather A, Hamiton WD O, Gatehouse J A. Transgenic potato plants with enhanced resistance to the peach-potato aphid Myzus persicae . Entomol Exp Appl, 1996, 79: 295–307
[4] Tang K X, Tinjuangjun P, Xu Y N, Sun X F, Gatehouse J A, Ronald P C, Qi H X, Lu X G, Christou P, Kohli A. Particle bombardment mediated co-transformation of elite Chinese rice cultivars with genes conferring resistance to bacterial blight and sap-sucking insect pests. Planta, 1999, 208: 552–563
[5]Xu Q-F(徐琼芳), Li L-C(李连城), Chen X(陈孝), Ma Y-Z(马有志), Ye X-G(叶兴国), Zhang Z-Y(张增艳), Xu H-J(徐惠君), Xin Z-Y(辛志勇). Study on the obtaining of transgenic wheats with GNA alien gene by biolistic particle. Sci Agric Sin (中国农业科学), 2001, 34(1): 5–8 (in Chinese with English abstract)
[6] Stoger E, Williams S, Christou P, Down R E, Gatehouse J A. Expression of the insecticidal lectin from snowdrop (Galanthus nivalis agglutinin: GNA) in transgenic wheat plants: effects on predation by the grain aphid Sitobion avenae. Mol Breed, 1999, 5: 65–73
[7] Xu Q-F(徐琼芳), Tian F(田芳), Chen X(陈孝), Hou W-S(侯文胜), Li L-C(李连城), Du L-P(杜丽璞), Xu H-J(徐惠君), Xin Z-Y(辛志勇). Inheritance of sgna gene and insect-resistant activity in transgenic wheat. Acta Agron Sin (作物学报), 2004, 30(5): 475–480 (in Chinese with English abstract)
[8] Yao J H, Pang Y Z, Qi H X, Wan B L, Zhao X Y, Kong W W, Sun X F, Tang K X. Transgenic tobacco expressing Pinellia ternate agglutinin confers enhanced resistance to aphids. Transgenic Res, 2003, 12: 715–722
[9] Yu Y, Wei Z M. Increased oriental armyworm and aphid resistance in transgenic wheat stably expressing Bacillus thuringiensis (Bt) endotoxin and Pinellia ternate agglutinin (PTA). Plant Cell Tissue Organ Cul, 2008, 94: 33–44
[10] Zhao X Y, Yao J H, Liao Z H, Zhang H Y, Chen F, Wang L, Lu Y Q, Sun X F, Yu S Q, Tang K X. Molecular cloning of a novel mannose-binding lectin gene from Arisaema heterophyllum. Plant Sci, 2003, 165: 55–60
[11] Yao J H, Zhao X Y, Qi H X, Wan B L, Chen F, Sun X F, Yu S Q, Tang K X. Transgenic tobacco expressing an Arisaema heterophyllum agglutinin gene displays enhanced resistance to aphids. Can J Plant Sci, 2004, 84: 785–790
[12] Liu Q-Q(刘巧泉), Yu H-X(于恒秀), Zhang W-J(张文娟), Wang H-M(王红梅), Gu M-H(顾铭洪). Specific expression of the foreign gene regulated by the rice rbcS promoter in transgenic rice. J Plant Physiol Mol Biol (植物生理与分子生物学学报), 2005, 31(3): 247–253 (in Chinese with English abstract)
[13] Hang H-Q(黄海群), Lin Y-J(林拥军). Cloning and functional analysis of the rice rbcS gene promoter. J Agric Biotechnol (农业生物技术学报) , 2007, 15(3): 451–458 (in Chinese with English abstract)
[14] Outchkourov N S, Peters J, Rademakers W, Jongsma M A. The promoter–terminator of chrysanthemum rbcS1 directs very high expression levels in plants. Planta, 2003, 216: 1003–1012
[15] Qin H-M(秦红敏), Guo H-N(郭洪年), Jia Y-T(贾燕涛), Li L-H(李利红), Tian Y-C(田颖川). The effect of TMV-RNA untranslation region on the expression lever of foreign gene in entire plant. Chin Sci Bull (科学通报), 2000, 45(6): 617–622 (in Chinese)
[16] Kozak M. Compilation and analysis of sequences upstream from translational site in ekaryotic mRNAs. Nucl Acid Res, 1984, 12: 857–872
[17] Mao L-Q(毛立群), Guo S-D(郭三堆). Relationship between Ω as well as the length of 3′ poly(dA) and efficiency of gene expression. Acta Bot Sin (植物学报), 1998, 40(12): 1–3 (in Chinese with English abstract)
[18] Rasco-Gaunt S, Barcelo P. Immature inflorescence culture of cereals: a highly responsive system for regeneration and transformation. In: Hall R D ed. Plant Cell Culture Protocols. Totowa NJ: Humana Press, 1999. pp 71–81
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