Improved protoplast fusion technologies using metabolic inhibitors and microfluidics

使用代谢抑制剂和微流体改进原生质体融合技术

• 批准号: RGPIN-2016-06252
• 负责人: Jones, Andrew
• 金额: $ 2.04万
• 依托单位: University of Guelph
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=710234
• 关键词: Improved; protoplast; fusion; technologies; using

Plant breeding has dramatically increased agricultural productivity and will play an important role in adapting to a changing climate and feeding the growing population. In many cases the desired trait, be it disease/insect resistance, or adaptation to cold, heat, or drought stress, is not present in the crop but can be incorporated by hybridization with closely related wild relatives. However, this requires that the species are sexually compatible with each other, which is often not the case. Protoplast fusion technologies, in which cells from two species can be fused together, can overcome sexual barriers and allow crops to be crossed with more distantly related species. This technology is particularly attractive for tree breeding, where traditional approaches can take decades and are not feasible. However, trees present a number of technical challenges that have not been fully addressed and limit the application of this technology. One example of this is American elm, where the native population was nearly eradicated by the invasive Dutch elm disease (DED). Early researchers found that the American elm population does not contain genes for DED resistance and despite over 50 years of traditional breeding there remains a need for true DED resistant trees. While some Asiatic elms are resistant, early attempts at hybridization failed due to sexual incompatibility and somatic hybridization was proposed as early as 1980 to overcome this. However, a number of problems related to isolation and regeneration of protoplasts have prevented success. In recent work, we have identified the primary source of the problem and developed a novel approach that resolved some of these issues. This has led to the first protoplast to plant regeneration system for American elm and provides a framework for protoplast fusion. The key breakthrough that facilitated this was through manipulating one of the plant's metabolic pathways to alter the chemical composition of the cell wall, making it more amenable to the process. However, an in depth investigation of this system has not been conducted and a number of issues require further study to understand the underlying mechanism in American elm, evaluate if this approach will work in other tree species, and develop new approaches to improve fusion efficiency. This proposal will establish a research program to refine this technique and develop more controlled protoplast fusion technologies using a microfluidic approach for economically and ecological important plants, with a focus on woody species. These technologies could play an important role in the genetic improvement of plants to adapt to new climatic conditions and help feed the growing population.

植物育种极大地提高了农业生产力，并将在适应气候变化和养活不断增长的人口方面发挥重要作用。 在许多情况下，所需的性状，无论是抗病性/抗虫性，还是对冷、热或干旱胁迫的适应性，都不存在于作物中，但可以通过与密切相关的野生近缘种杂交来引入。 然而，这需要物种之间的性相容，而事实往往并非如此。 原生质体融合技术，即两个物种的细胞可以融合在一起，可以克服性别障碍，使作物与更远的相关物种杂交。 这项技术对树木育种特别有吸引力，传统方法可能需要数十年时间，并且不可行。 然而，树木提出了一些尚未完全解决的技术挑战，并限制了这项技术的应用。 其中一个例子是美国榆树，当地人口几乎被入侵的荷兰榆树病（DED）消灭。 早期的研究人员发现，美国榆树种群不包含抗DED的基因，尽管经过50多年的传统育种，仍然需要真正的抗DED树木。 虽然一些亚洲榆树是抗性的，但早期的杂交尝试由于性不相容而失败，早在1980年就提出了体细胞杂交来克服这一点。 然而，与原生质体的分离和再生相关的许多问题阻碍了成功。 在最近的工作中，我们已经确定了问题的主要来源，并开发了一种新的方法来解决其中的一些问题。 这导致了第一个美国榆树原生质体到植物再生系统，并为原生质体融合提供了框架。 促成这一点的关键突破是通过操纵植物的一种代谢途径来改变细胞壁的化学成分，使其更容易适应这一过程。 然而，这个系统还没有进行深入的调查，一些问题需要进一步研究，以了解美国榆树的潜在机制，评估这种方法是否适用于其他树种，并开发新的方法来提高融合效率。 该提案将建立一个研究计划，以完善这一技术，并使用微流体方法开发更可控的原生质体融合技术，用于经济和生态上重要的植物，重点是木本植物。 这些技术可以在植物遗传改良方面发挥重要作用，以适应新的气候条件，并帮助养活不断增长的人口。