Green-tissue-specific, C4-PEPC-promoter-driven expression of Cry1Ab makes transgenic potato plants resistant to tuber moth (Phthorimaea operculella, Zeller)
Green-tissue-specific, C4-PEPC-promoter-driven expression of Cry1Ab makes transgenic potato plants resistant to tuber moth (Phthorimaea operculella, Zeller)
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DOI：
10.1007/s00299-009-0790-3
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发表时间：
2009-12-01
影响因子：
6.2
通讯作者：
Mahna, Naser
中科院分区：
文献类型：
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作者：
Hagh, Ziba Ghasimi;Rahnama, Hassan;Mahna, Naser
获得更安全的转基因植物的一个重要策略可能是使用空间或组织特异性启动子，而不是组成性启动子。在本研究中，我们利用光诱导玉米PEPC启动子调控cry1Ab基因，旨在生产抗马铃薯块茎蛾（Phthorimaea operculella, Zeller）的转基因马铃薯。在60个表型正常的再生品系中，55个品系的cry1Ab和nptII基因均为pcr阳性。Southern对三个假定的转基因品系的分析显示，它们只有一个完整的cry1Ab基因拷贝。对Cry1Ab蛋白在转基因株系叶片和光暴露（LE）块茎中的表达进行了研究，发现该蛋白存在于叶片和绿色块茎中，而不存在于光暴露（LNE）块茎中。对随机选择的9个品系的切下叶片进行生物测定分析，8个品系的PTM幼虫死亡率为100%。采用温室全株生物测定法对6个选育品系进行了验证。LE转基因块茎也表现出100%的幼虫死亡率；然而，对转基因LNE块茎的损害程度很高，在统计上与非转基因块茎相同。基于这些结果，我们认为利用光诱导PEPC启动子空间表达Cry1Ab可以控制PTM在田间的侵染，并显著减少污染对储存马铃薯的传播。
An important strategy for obtaining a safer transgenic plant may be the use of a spatial- or tissue-specific promoter, instead of a constitutive one. In this study, we have used a light-inducible maize PEPC promoter to regulate the cry1Ab gene, aiming to produce transgenic potatoes that are resistant to potato tuber moth (PTM) (Phthorimaea operculella, Zeller). Out of 60 regenerated lines having normal phenotypes, 55 lines were PCR-positive for both the cry1Ab and nptII genes. Southern analysis on three selected putative transgenic lines revealed that they have only a single intact copy of the cry1Ab gene. An investigation of the Cry1Ab protein in the leaves and light-exposed (LE) tubers of the transgenic lines demonstrated the presence of the protein in the foliage and green tubers but not in the light-not exposed (LNE) tubers. A bioassay analysis of excised leaves of nine randomly selected lines showed that eight lines had 100% PTM larval mortality. Confirming results were obtained in six selected lines using the whole plant bioassay in the greenhouse. LE transgenic tubers also exhibited 100% larval mortality; however, the levels of damage to the LNE transgenic tubers were high and statistically the same as those incurred by the non-transgenic ones. Based on the results, we believe that this spatial expression of Cry1Ab using the light-inducible PEPC promoter can control PTM infestation in the field and significantly reduce pollution transmission to storage potatoes.