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| alfAFP和spCEMA融合基因表达载体的构建及其对小麦的遗传转化 |
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张立1,2, 王建锋1,3, 王晓杰1,3, 康振生1,3, 韩德俊1,2
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| (1.西北农林科技大学, 旱区作物逆境生物学国家重点实验室, 陕西 杨凌 712100;2.西北农林科技大学, 农学院, 陕西 杨凌 712100;3.西北农林科技大学, 植物保护学院, 陕西 杨凌 712100) |
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| 摘要: |
| 以小麦品种西农1376为受体,利用Overlapping PCR的方式构建了防御素基因alfAFP和抗菌肽基因spCEMA融合基因的表达载体,与pAHC20共转化幼胚愈伤组织,获得转基因再生T0代植株269株,春化后移栽成活188株。对叶片基因组DNA进行目的基因和bar基因PCR扩增表明其中7株是共整合植株,共整合频率为0.1%,初步证明融合基因已成功通过共转化方式导入受体小麦品种中。影响共整合率的因素分析表明,在使用除草剂PPT对bar基因进行筛选的过程中,筛选时间的缩短并不会明显降低筛选的效率。 |
| 关键词: 小麦 转基因 融合基因 共转化 |
| DOI：10.11841/j.issn.1007-4333.2012.05.003 |
| 投稿时间：2012-04-17 |
| 基金项目:国家重大专项(2011ZX08002-001);高等学校学科创新引智计划资助项目(B07049) |
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| Co-transformation of wheat with fusion genes alfAFP and spCEMA using biolistics |
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ZHANG Li1,2, WANG Jian-feng1,3, WANG Xiao-jie1,3, KANG Zhen-shen1,3, HAN De-jun1,2
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| (1.The National Key Laboratory for Arid Regions Crop Adversity Biology, Northwest Agriculture and Forestry University, Yangling 712100, China;2.College of Agronomy, Northwest Agriculture and Forestry University, Yangling 712100, China;3.College of Plant Protection, Northwest Agriculture and Forestry University, Yangling 712100, China) |
| Abstract: |
| Transgenic wheat variety XiNong 1376 containing the plant defensin gene alfAFP and antimicrobial peptide gene spCEMA fusion gene were obtained using cotransformation with bar as a selective marker via biolistics.In total,269 plantlets were generated through screening,and 188 of which were alive after vernalization.Leaf genome DNA PCR array showed that 7 co-transformed plants were obtained,and the frequency of co-integration was 0.1%.Studies also illustrated that the shortened time of the bar gene screening process did not significantly reduce the efficiency of PPT screening. |
| Key words: wheat transgenic gene fusion co-transformation |
