马铃薯耐疫霉转基因品系RB基因的遗传稳定性分析

Q4 Agricultural and Biological Sciences Indonesian Journal of Agricultural Science Pub Date : 2015-10-01 DOI:10.21082/ijas.v16n2.2015.pp.51-58
E. Listanto, E. I. Riyanti, T. Santoso, T. Hadiarto, A. D. Ambarwati
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引用次数: 5

摘要

培育广谱抗性高的马铃薯品种是马铃薯防治晚疫病的关键长期治理策略。根据农艺性状和对晚疫病的抗性,选择了转基因马铃薯Katahdin SP951与非转基因格兰诺拉和大西洋杂交的6个子代。本研究旨在分析转基因马铃薯品系中RB基因的插入数量及其稳定性。通过植物DNA提取、southern blot分析和聚合酶链反应(PCR)进行研究。采用Southern blot法检测整合到马铃薯基因组中的插入物数量,采用PCR法检测RB基因世代间的稳定性。结果表明,经nptII检测,Atlantic与转基因Katahdin SP951(20号和27号品系)、Granola与转基因Katahdin SP951(69号品系)杂交获得的后代含有1个拷贝数的RB基因。结果与亲本转基因Katahdin SP951中插入RB基因的结果相似。RB基因在4代(G0、G1、G2和G3)中均存在,表明该基因已稳定整合到植物基因组中。RB基因的单拷贝数会抑制沉默基因表达的发生。通过对RB基因的稳定性分析,可以确定该基因在几代后仍然存在于植物基因组中。
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GENETIC STABILITY ANALYSIS OF RB GENE IN GENETICALLY MODIFIED POTATO LINES TOLERANT TO Phytophthora infestans
Development of potato cultivars with high levels of broad spectrum resistance is a key long-term management strategy against late blight disease caused by Phytophthora infestans. Six progeny lines of hybridization between transgenic potato Katahdin SP951 with non-transgenic Granola and Atlantic were selected based on agronomical characteristics and resistance to late blight disease. The study aimed to analyze the number of insertions and stability of inserted RB gene in the transgenic potato lines. The research was carried out through plant DNA extraction, southern blot analysis and polymerase chain reaction (PCR). Southern blot analysis was used to detect the number of inserts integrated into potato genome, while PCR analysis was used to detect stability of RB gene from generation to generation. The results showed that the progenies obtained from hybridization between Atlantic and transgenic Katahdin SP951 (lines No. 20 and 27) and between Granola and transgenic Katahdin SP951 (line No. 69) contained one copy number of RB gene, according to the probing of nptII. The result is similar to that of inserted RB gene found in the parental transgenic Katahdin SP951. The presence of RB gene in four different generations (G0, G1, G2 and G3) showed stable integration of the gene into the plant genome. The single copy number of RB gene will repress the occurrence of silencing gene expression. The stability analysis of RB gene can determine that the gene is still present in plant genome after several generations.
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来源期刊
Indonesian Journal of Agricultural Science
Indonesian Journal of Agricultural Science Agricultural and Biological Sciences-Soil Science
CiteScore
1.00
自引率
0.00%
发文量
5
审稿时长
12 weeks
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