| 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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| Generation of Fusarium graminearum Knockout Mutants by the Split-marker Recombination Approach
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Author:
Date:
2018-08-20
[Abstract] Fusarium graminearum is a destructive phytopathogen and shows an impressive metabolic diversity. Gene deletion is an important and useful approach for gene function study. Here we present a protocol for generating gene deletion mutant by applying “split-marker” deletion strategy (Catlett et al., 2003) with PEG-mediated protoplast transformation (Yuan et al., 2008; Martín, 2015).
[摘要] 禾谷镰刀菌是一种破坏性的植物病原体，具有令人印象深刻的代谢多样性。 基因缺失是基因功能研究的重要且有用的方法。 在这里，我们提出了一个协议，通过应用“分裂标记”删除策略（Catlett et al。，2003）与PEG介导的原生质体转化（Yuan 等。，2008;Martín，2015）。
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| Fusarium graminearum Double (Triple) Mutants Generation Using Sexual Crosses
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Author:
Date:
2018-08-05
[Abstract] Fusarium graminearum is a destructive phytopathogen that infects major cereal crops such as wheat, maize and barley. Double or triple mutants are often very useful in the phenotypic and genetic analysis of genes that function redundantly or within similar pathways. When single gene mutants are available, double or triple mutants can be generated by crossing heterothallic strains or multiple rounds of protoplast transformation. When individual mutants carry different antibiotic resistance, it is convenient to use the sexual crossing to generate desired recombinant strains. Here, we present a protocol for generating double or triple mutants by sexual crossing in one homothallic strain with further antibiotic resistance and genomic DNA PCR screening of recombinant progenies.
[摘要] 禾谷镰刀菌（Fusarium graminearum）是一种破坏性的植物病原体，可感染小麦，玉米和大麦等主要谷类作物。 双重或三重突变体通常在冗余或相似途径内起作用的基因的表型和遗传分析中非常有用。 当可获得单基因突变体时，可通过杂交异源菌株或多轮原生质体转化产生双突变体或三突变体。 当单个突变体携带不同的抗生素抗性时，使用有性杂交方便产生所需的重组菌株是方便的。 在这里，我们提出了一种方案，通过在一个同源的菌株中通过有性杂交产生双突变体或三突变体，具有进一步的抗生素抗性和重组后代的基因组DNA PCR筛选。
【背景】子囊菌真菌镰刀菌（Fusarium graminearum）是一种破坏性的植物病原体，可引起谷类食物的头部枯萎病，穗腐病，茎腐病和冠。它可以在胡萝卜琼脂体外产生perithecia 。该测定法可用于研究子囊膜发育，子囊孢子放电和性重组（Nicholson，2007）。
F。禾本科植物是同源的，具有MAT1-1和MAT1-2-1基因座;这些基因座缺失突变体中的每一个在自交中都是无菌的（Zheng et al。，2013）。为了产生双基因突变体，一个单基因突变体传统上与 MAT 缺失突变体杂交，并进一步与另一个突变体杂交（Bowden和Leslie，1999; Lee et al。 ，2011; Son et ...
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