| Phytophthora infestans (Late blight) Infection Assay in a Detached Leaf of Potato
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Author:
Date:
2021-02-20
[Abstract] Phytophthora infestans is a hemibiotroph oomycete that primarily infects potato and tomato. It infects stems, leaves, and tubers and fruits of potato and tomato. High throughput and reproducible infection assays are prerequisites to find sources of resistance in any crop. In this protocol, we describe a detached leaf assay (DLA) for conducting the virulence assay of P. infestans in potato leaves. A late blight infection assay using a potato detached leaf is a semi-high throughput assay in which hundreds of plants can be screened in a laboratory setting.
[摘要] [摘要]疫霉菌是一种主要感染马铃薯和番茄的半生营养菌卵菌。它感染马铃薯和番茄的茎，叶，块茎和果实。高通量和可重复的感染测定是寻找任何农作物抗药性来源的先决条件。在此协议中，我们描述了分离叶分析（DLA），用于进行马铃薯叶中致病疫霉的毒力测定。使用马铃薯离体叶的晚疫病感染测定是一种半高通量测定，其中可以在实验室环境中筛选数百种植物。
[背景]就食品和营养价值而言，马铃薯（Solanum tuberosum L.）是最重要的非谷物粮食作物之一（Zhang等人，2017）。晚疫病引起卵菌病原体马铃薯的晚疫病菌是世界上最具破坏性的马铃薯的疾病之一，是在马铃薯生产中最重要的产量限制因素（Haverkort等人，2008和2016 ;费舍尔等人， 2012 ）。后期抗白叶枯病的育种被认为是抗击这种疾病的重要因素。为此，在可用种质中鉴定新的抗性来源是至关重要的一步。已经开发了几种测试方法，例如田间测试，整株植物测定和离体叶片测定（DLA）。DLA提供的感染增加，并且马铃薯叶表现出对疫病菌的敏感性高于田间和整个植物的检测方法（Stewart ，1990 ；Vleeshouwers等，1999 ...
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| Optimize Highly Efficient Genetic Transformation Method of Spring Wheat (Triticum aestivum L.) Cultivars Using Mature Embryo
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Author:
Date:
2020-11-20
[Abstract] Genetic engineering is a powerful tool to develop desired traits in crop plants to make the crops suitable for future demand and changing environments. For the genetic engineering of wheat cultivars, the development of efficient transformation and regeneration systems has always been a primary requirement. Immature embryos have been used as explants for callus generation and genetic transformation of wheat cultivars, but the availability of healthy immature embryos as an explant throughout the year is difficult. In contrast, mature embryos are relatively easy to use throughout the year. The following protocol describes the genetic transformation of wheat cultivars using a mature embryo as explants by the Agrobacterium-mediated transformation method.
[摘要] [摘要]基因工程是开发农作物所需性状，使农作物适合未来需求和环境变化的有力工具。对于小麦品种的基因工程，开发高效的转化和再生系统一直是首要要求。未成熟的胚已被用作小麦品种的愈伤组织生成和遗传转化的外植体，但是全年都很难获得健康的未成熟胚作为外植体。相反，成熟的胚胎一年四季都相对容易使用。以下方案描述了使用小麦品种的遗传转化一个成熟胚由外植体的土壤杆菌介导的转化方法。
[背景]遗传转化是通过基因工程在小麦分子遗传学和改善使用的最重要的技术。微粒轰击和农杆菌介导的转化是小麦中遗传转化最广泛使用的两种方法（Vasil等，1992 ；Ding等，2009 ...
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| Quantification of Methylglyoxal Levels in Cowpea Leaves in Response to Cowpea Aphid Infestation
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Author:
Date:
2020-10-20
[Abstract] Aphids are a serious pest of crops across the world. Aphids feed by inserting their flexible hypodermal needlelike mouthparts, or stylets, into their host plant tissues. They navigate their way to the phloem where they feed on its sap causing little mechanical damage to the plant. Additionally, while feeding, aphids secrete proteinaceous effectors in their saliva to alter plant metabolism and disrupt plant defenses to gain an advantage over the plant. Even with these arsenals to overcome plant responses, plants have evolved ways to detect and counter with defense responses to curtail aphid infestation. One of such response of cowpea to cowpea aphid infestation, is accumulation of the metabolite methylglyoxal. Methylglyoxal is an α,β-dicarbonyl ketoaldehyde that is toxic at high ...
[摘要] [摘要] 蚜虫是世界各地农作物的严重害虫。蚜虫通过将它们柔软的针状口器插入宿主植物组织中取食。它们会进入韧皮部，在那里吃韧皮部的汁液，对植物的机械损伤很小。此外，蚜虫在进食时，会在唾液中分泌蛋白质效应器来改变植物的新陈代谢，破坏植物的防御，从而获得对植物的优势。即使有了这些武器来克服植物的反应，植物也进化出了探测和反击的方式，以减少蚜虫的侵扰。豇豆蚜虫感染的这种反应之一是代谢物甲基乙二醛的积累。甲基乙二醛是一种对、对二羰基酮醛，在高浓度下有毒。在暴露于许多不同的非生物和生物胁迫后，甲基乙二醛水平略有上升，并已被证明在低水平上作为一种新兴的防御信号分子。在此，我们描述了一种在豇豆蚜虫感染后，利用高氯酸提取工艺在豇豆叶片中测量甲基乙二醛的方案。将提取的上清液用碳酸钾中和，丙酮醛与N-乙酰- l-半胱氨酸反应量化，生成N-乙酰- s -(1-羟基-2-oxo-prop-1-yl)半胱氨酸，并进行分光光度定量。
[背景] ...
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