Durable Late Blight Resistance in Potato Through Dynamic Varieties Obtained by Cisgenesis: Scientific and Societal Advances in the DuRPh Project
Durable Late Blight Resistance in Potato Through Dynamic Varieties Obtained by Cisgenesis: Scientific and Societal Advances in the DuRPh Project
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DOI：
10.1007/s11540-015-9312-6
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发表时间：
2016-03-01
期刊：
影响因子：
2.9
通讯作者：
Visser, R. G. F.
中科院分区：
文献类型：
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作者：
Haverkort, A. J.;Boonekamp, P. M.;Visser, R. G. F.
从2006年到2015年，瓦赫宁根大学和研究中心开展了一项关于马铃薯对疫霉持久抗性的研究项目(DuRPh)。它的目的是开发一种通过顺势发生持久抗晚疫病的原理证明。这个由公共资金资助的项目旨在促进基因改造的研究和公众对创新基因技术的辩论。利用根癌农杆菌介导法，在不携带非马铃薯基因的情况下，克隆和转移可杂交野生种(顺基因)的抗晚疫病基因。计划将多个R基因的堆叠插入到已建立的品种中，从而创造出一个动态变化的堆栈，其中堆叠的组成可能会随着空间和时间的变化而变化。根据所有插入的R基因的表达情况和真实性对类型进行选择。在该项目中，对野生马铃薯物种的13个R基因进行了遗传定位，其中3个基因被克隆。将1~3个R基因转化到4个品种中。这最初是利用由合成起源的可选择标记基因提供的卡那霉素抗性来完成的，以便快速测试导入的R基因和堆叠的R基因组合的性能和稳定性。一旦确认其功能，就进行无标记的转化；因此，选择了真正的顺式事件。对不同R基因的染色体定位、特异性、背景依赖性、最大堆叠大小、再生时间、体细胞无性系变异频率及其稳定性进行了研究。经过室内筛选和鉴定，在田间进行了抗晚疫病鉴定。这表明，插入的R基因能够将感病品种转变为抗病品种。通过抗药性管理研究确保了抗药性的最大寿命。研究表明，多个R基因的叠加和监测这些基因如何在空间和时间上部署，可以减少80%以上的杀菌剂使用。通过媒体和实地演示进行了多种交流，以使公众和政策制定者能够决定顺势遗传是否是使马铃薯种植更可持续的可接受工具。未来DuRPh战略的部署将在很大程度上取决于其作为转基因作物的地位或豁免。在世界范围内，接近根除晚疫病将使全球马铃薯年产量增加近8000万吨，从而大大增加所需的全球未来粮食供应。
From 2006 through 2015, a research project on Durable Resistance in potato against Phytophthora (DuRPh) was carried out at Wageningen University and Research Centre. Its objective was to develop a proof of principle for durable resistance against late blight by cisgenesis. This public-funded project aimed at stimulating research on genetic modification and public debate on innovative genetic techniques. It was decided to clone and transfer late blight resistance (R) genes of crossable wild potato species (cisgenes) by Agrobacterium tumefaciens-mediated transformation without non-potato genes. A stack of multiple R genes were planned to be inserted into established varieties, thereby creating a dynamic variety in which the composition of the stacks may vary over space and time. Cisgenic plants were selected based on the expression of all inserted R genes and trueness-to-type. Within the project, 13 R genes from wild potato species were genetically mapped and three of them were cloned. Four varieties were transformed with one to three R genes. This was initially done using kanamycin resistance provided by a selectable marker gene of synthetic origin in order to quickly test the performance and stability of the introduced R genes and stacked R gene combinations. Once the functioning thereof was confirmed, marker-free transformations were conducted; thus, true cisgenic events were selected. The results about the different R genes, their chromosomal location, their specificity, the background dependence, the maximum size of a stack, its regeneration time and associated somaclonal variation frequency and its stability were studied. After selection and characterisation in the laboratory, the best cisgenic events were assessed in field trials for late blight resistance. This showed that inserted R genes were capable of turning a susceptible variety into a resistant one. Maximising longevity of the resistance was assured through resistance management research. It was shown that stacking of multiple R genes and monitoring how to deploy these stacks spatially and temporally could reduce fungicide use by over 80%. Communications through media and field demonstrations were manifold to allow public and policymakers to decide if cisgenesis is an acceptable tool to make potato farming more sustainable. Future deployment of the DuRPh strategy will depend largely on its status as a genetically modified crop or its exemption thereof. Worldwide near eradication of late blight would increase global annual potato production by close to 80 million tons, thereby contributing considerably to the needed additional global future food supply.