Combining late blight resistance and better tuber quality with resistance to potato virus Y (PVY) to improve Maris Piper potato

将晚疫病抗性和更好的块茎品质与马铃薯 Y 病毒 (PVY) 抗性相结合，以改良 Maris Piper 马铃薯

• 批准号: BB/W017903/1
• 负责人: Jonathan Jones
• 金额: $ 52.66万
• 依托单位: University of East Anglia
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW017903%2F1
• 关键词: Combining; late; blight; resistance; better

The potato crop is susceptible to many diseases and pests, notably late blight (LB) caused by Phytophthora infestans. This disease causes at least $6B in losses worldwide every year, costs £50M per year in the UK and led to the Irish potato famine in the 1840s.Potato tubers are also prone to problems. Storage in cool conditions elevates levels of reducing sugars, and after high temperature cooking such as frying or roasting, these sugars can be converted into toxic acrylamide. Potato tubers are damaged by bruising if dropped or crushed, resulting in discolouration by an enzyme reaction, leading to significant food wastage. Tubers can also be lost to tuber blight in storage.Finally, potato virus Y can result in severe yield losses or rejection of seed potato tubers, imposing an additional burden and expense on the potato industry.We cloned three very effective LB Resistance genes called Rpi-vnt1, Rpi-amr3 and Rpi-amr1 and have shown that when deployed together in GM potato, they confer complete resistance in the field to all currently circulating LB races. This should provide a durable solution to the late blight problem in the UK.We also obtained commercially deployed and validated genes from the American potato company Simplot Inc that reduce levels of the enzymes that elevate reducing sugars and that mediate the discolouration upon bruising, thus alleviating the problem. In a currently running grant, we introduced the 3 Rpi genes and the tuber quality traits, into UK-favoured variety Maris Piper (MP), using Agrobacterium, creating PiperPlus 1.0. However, all lines that we generated are susceptible to PVY, greatly impairing our capacity to bulk them up for more extensive field trials. PVY is aphid transmitted, and difficult to control, especially as insecticides used to control PVY-transmitting aphids are being increasingly restricted, and global warming means that aphids and PVY are an increasing problem for seed potato production in Scotland. Also, Scotland is still opposed to planting GM crops, meaning that seed potato production outside of Scotland will be needed.With colleagues, we cloned an extremely effective PVY resistance gene, Ry-sto. We aim in this project to combine PVY resistance with the LB resistance and tuber quality traits of PiperPlus 1.0, to create PiperPlus 2.0. We have made a DNA construct to introduce all traits together on one T-DNA via Agrobacterium and are producing transgenic Maris Piper lines using the GM method. We expect to be able to field-trial ~20 PiperPlus 2.0 candidate lines in Norwich in summer 2022, and another ~20 in summer 2023. We can easily identify lines in which Ry and all 3 Rpi genes are expressed and we will also select those plant lines with the improved tuber quality traits. The best lines from batch 1 in the Norwich trial in 2022 will be multiplied up for field trials in summer 2023 at NIAB in Cambridge, and the best lines from batch 2 in Norwich 2023 will be multiplied up for NIAB trials summer 2024, to select the best performing PiperPlus 2.0 lines that will be indistinguishable from normal Maris Piper, except that they will be LB resistant, PVY resistant and have the improved tuber quality traits. These lines will also be evaluated to verify comparable yield to that of normal Maris Piper, and to ensure that use of the GM method has not resulted in acquisition of any deleterious traits, such as elevated production of steroidal glycoalkaloids. We will also define exactly where in the potato genome the DNA that we added has inserted, and that the final selected PiperPlus 2.0 line carries a simple, single copy insertion event.The project will thus enable us to make by 2024 a commercially viable line with these added traits, and we anticipate that by 2024 changes in the regulatory framework will enable this improved line to be approved for commercial production and public consumption.

马铃薯作物易受多种病虫害的影响，特别是由致病疫霉引起的晚疫病 (LB)。这种疾病每年在全球造成至少 60 亿美元的损失，在英国每年造成 5000 万英镑的损失，并导致 1840 年代的爱尔兰马铃薯饥荒。马铃薯块茎也容易出现问题。在阴凉条件下储存会增加还原糖的含量，而在油炸或烘烤等高温烹饪后，这些糖会转化为有毒的丙烯酰胺。如果掉落或压碎，马铃薯块茎会因碰伤而受损，导致酶反应变色，从而导致大量食物浪费。块茎也可能因储存过程中的块茎枯萎病而损失。最后，马铃薯Y病毒可导致严重的产量损失或种薯块茎的排斥，给马铃薯产业带来额外的负担和费用。我们克隆了三种非常有效的LB抗性基因，称为Rpi-vnt1、Rpi-amr3和Rpi-amr1，并表明当在转基因马铃薯中一起部署时，它们在田间赋予对当前流行的所有抗性的完全抗性。 LB 比赛。这应该为英国的晚疫病问题提供持久的解决方案。我们还从美国马铃薯公司 Simplot Inc 获得了商业部署和验证的基因，这些基因降低了提高还原糖并介导瘀伤变色的酶的水平，从而缓解了这个问题。在当前正在运行的拨款中，我们使用农杆菌将 3 个 Rpi 基因和块茎品质性状引入英国喜爱的品种 Maris Piper (MP)，创建了 PiperPlus 1.0。然而，我们产生的所有品系都容易受到 PVY 的影响，这极大地削弱了我们将它们扩大以进行更广泛的现场试验的能力。 PVY 是通过蚜虫传播的，难以控制，特别是用于控制传播 PVY 的蚜虫的杀虫剂越来越受到限制，而且全球变暖意味着蚜虫和 PVY 成为苏格兰种薯生产中日益严重的问题。此外，苏格兰仍然反对种植转基因作物，这意味着需要在苏格兰以外的地方生产马铃薯种薯。我们与同事一起克隆了一种极其有效的 PVY 抗性基因 Ry-sto。我们在该项目中的目标是将 PiperPlus 1.0 的 PVY 抗性与 LB 抗性和块茎品质性状结合起来，创建 PiperPlus 2.0。我们已经构建了一种 DNA 构建体，通过农杆菌将所有性状一起引入到一个 T-DNA 上，并使用 GM 方法生产转基因 Maris Piper 品系。我们预计能够于 2022 年夏季在诺维奇对约 20 个 PiperPlus 2.0 候选品系进行田间试验，并于 2023 年夏季对约 20 个候选品系进行田间试验。我们可以轻松识别表达 Ry 和所有 3 个 Rpi 基因的品系，我们还将选择那些具有改善的块茎品质性状的植物品系。 2022 年诺维奇试验中第 1 批的最佳品系将在 2023 年夏季在剑桥的 NIAB 进行现场试验，2023 年诺维奇第 2 批的最佳品系将在 2024 年夏季 NIAB 试验中进行倍增，以选择性能最佳的 PiperPlus 2.0 品系，这些品系与正常的 Maris Piper 没有区别，只是它们具有 LB 抗性、PVY 抗性和有改进的 块茎品质性状。还将对这些品系进行评估，以验证其与正常 Maris Piper 的产量相当，并确保使用转基因方法不会导致获得任何有害性状，例如甾体配糖生物碱产量增加。我们还将准确定义我们添加的 DNA 在马铃薯基因组中的插入位置，以及最终选定的 PiperPlus 2.0 品系携带简单的单拷贝插入事件。因此，该项目将使我们能够在 2024 年之前制造出具有这些附加性状的商业上可行的品系，并且我们预计到 2024 年，监管框架的变化将使该改进品系能够获得商业生产和公共消费的批准。