Detoxed grass pea: sustainable sustenance for stressful environments

草豌豆（Lathyrus sativus）为最贫穷的人提供了可持续农业和粮食安全的绝佳机会，即使面临预测的气候变化，因为它是一种豆类，在边际土壤或恶劣的气候条件下表现良好。在许多国家，包括印度的大部分地区（其起源中心之一），欧洲和中国，草豌豆已被种植用于种子和饲料生产，因为它是一种低投入作物，廉价的蛋白质来源，特别是耐旱，耐涝和中度碱性。然而，草豌豆可以引起一种称为神经性lathyrism的毁灭性疾病，据信这是由于其β-N-草酰-l-α，β-二氨基丙酸（ODAP）的含量。草豌豆用作饲料时也会抑制动物的生长。因此，草豌豆是一种两面神的作物，因为它为那些处于生存边缘的人提供了急需的食物，但同时也有向消费者提供剧毒化合物的危险。加强育种工作，以去除抗营养毒素，改善其营养品质，提高产量，并提供对关键病原体的抗性，将使草豌豆受益。这种毒素的合成过程已被部分了解。通过常规育种开发的低毒素品系中的毒素水平是不稳定的，并且在应激环境中常常升高。我们希望利用基因组学的方法来提供安全的技术来改善这种保险作物。现代基因组学方法包括基因组测序、标记辅助育种和植物转化，已经彻底改变了作物育种。然而，许多重要的作物，如草豌豆，已经成为孤儿，因为它们不太适合改进，因为它们本质上缺乏一些所需的属性，如易处理的基因组或转化系统。传统的育种改良这些作物往往依赖于诱变，近年来，这已直接与基因组学通过反向遗传学工具TILLING。TILLING有助于在同一程序中结合正向和反向遗传学，以快速筛选表型和突变发现。TILLING特别适用于反向遗传资源有限或没有反向遗传资源的作物，因此是孤儿作物的理想选择。我们建议使用天然种质和现有的突变群体的草豌豆作为新的资源，通过非转基因途径提供改进的作物特性的突变的快速识别。用于测定ODAP的分光光度法已被调整为使用适合于筛选大量样品的微量滴定板的高通量。通过该试验，我们将筛选无毒素的品系。获得无毒品系将允许评估这些化合物在植物中（例如在抗虫性中）和在人类疾病中的真实作用。同时，诱变的植物材料将用于产生TILLING平台的DNA阵列，并在NIAB和印度用BCKV筛选胁迫反应和其他有用的农业特征（例如分枝、种子质量）。我们将研究为什么草豌豆提高了胁迫耐受性相比，其他豆类，利用这一基本的见解，以提高英国豆类，如豌豆的胁迫耐受性的目标。TILLING平台将可供研究社区revgenuk.jic.ac.uk访问。为了帮助使用TILLING平台，我们将通过RNA-seq开发转录组信息。转录组数据将有助于基因识别。我们将通过生成植物不同器官的RNA-seq数据来评估草豌豆的基因互补，包括毒素最普遍的叶子和种子。

Grass pea (Lathyrus sativus) offers an excellent opportunity for sustainable agriculture and food security for the poorest of the poor, even in the face of predicted climate change, because it is a legume and performs well in marginal soils or under harsh climatic conditions. Grass pea has been grown for seed and fodder production in many countries, including large parts of India (one of the centres of its origin), Europe and China since it is a low input crop, a cheap source of protein and is particularly tolerant to drought, water logging, and moderate alkalinity. However, grass pea can cause a devastating disease called neurolathyrism believed to be due to its content of beta-N-oxalyl-l-alpha,beta-diaminopropionic acid (ODAP) This neurological disorder is irreversible and occurs when people are dependent on the crop as a sole food source. Grass pea can also inhibit growth in animals when used as feed. Grass pea, therefore, presents as a Janus-faced crop since it provides desperately needed food for those on the edges of survival, but with the concomitant danger of delivering a highly toxic compound to its consumers. Grass pea would benefit from intensified breeding efforts to remove the anti-nutritional toxin and improve its nutritional quality, enhance yields and provide resistance to key pathogens. The synthesis of the toxin is understood in part. The toxin levels in low-toxin lines developed by conventional breeding are unstable and often elevated in stressful environments. We wish to exploit a genomics route to deliver safe technologies for improving this insurance crop.Modern genomics methods including genome sequencing, marker assisted breeding and plant transformation have revolutionised crop breeding. However, many important crops, like grass pea, have become orphaned as they are less amenable to improvement because inherently they lack some of the attributes required, like tractable genomes or transformation systems. Conventional breeding for the improvement of these crops often relies on induced mutagenesis, and in recent years, this has been linked directly to genomics via the reverse genetics tool of TILLING. TILLING facilitates the combination of forward and reverse genetics in the same programme to permit rapid screening for both phenotypes and mutation discovery. TILLING is particularly suitable for crops where there are limited or no reverse genetics resources available and hence ideal for an orphan crop. We propose to use natural germplasm and existing mutant populations of grass pea as novel resources for the rapid identification of mutations offering improved crop characteristics via a non-GM route. A spectrophotometric method for the assay of ODAP has been adapted for high-throughput using microtitre plates suitable for screening large numbers of samples. With this assay we will screen for toxin-free lines. The acquisition of toxin-free lines will allow the true roles of these compounds in plants (e.g. in insect resistance) and in human diseases to be assessed. Simultaneously the mutagenised plant material will be used to generate DNA arrays for a TILLING platform and to screen for stress responses and other useful agricultural characteristics (e.g. branching, seed quality) both at NIAB, and in India, with BCKV. We will investigate why grass pea has improved stress tolerance compared to other legumes, with the objective of using this fundamental insight to improve stress tolerance in UK legumes such as pea. The TILLING platform will be accessible to the research community revgenuk.jic.ac.uk. To aid use of the TILLING platform we will develop transcriptome information by RNA-seq. Transcriptome data will help in gene identification. We will assess the gene complement of grass pea via the generation of RNA-seq data for different organs of the plant, including leaves and seeds where the toxin is most prevalent.