Detoxed grass pea: sustainable sustenance for stressful environments

草豆（Lathyrus sativus）为可持续农业和最贫困人口的粮食安全提供了极好的机会，即使面对预测的气候变化，因为它是一种豆科植物，在边缘土壤或恶劣气候条件下表现良好。由于草豆是一种低投入作物，是廉价的蛋白质来源，并且特别耐旱、耐涝和耐碱性，许多国家（包括印度的大部分地区（草豆的起源中心之一）、欧洲和中国）种植草豆用于种子和饲料生产。然而，草豆可引起一种毁灭性的疾病，称为神经lathyrism，据信是由于其含有- n -草酰-l- α， -二氨基丙酸（ODAP）。这种神经系统疾病是不可逆转的，当人们依赖这种作物作为唯一的食物来源时，就会发生这种疾病。当草豆用作饲料时，还可以抑制动物的生长。因此，草豆是一种双面作物，因为它为那些处于生存边缘的人提供了迫切需要的食物，但同时也有向消费者提供剧毒化合物的危险。通过加强育种工作，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，草豆将从中受益。毒素的合成部分被理解。通过常规育种培育出的低毒品系的毒素水平不稳定，在紧张环境中往往会升高。我们希望利用基因组学的途径来提供安全的技术来改善这种保险作物。包括基因组测序、标记辅助育种和植物转化在内的现代基因组学方法已经彻底改变了作物育种。然而，许多重要的作物，如草豆，已经成为孤儿，因为它们天生缺乏一些必要的属性，如可处理的基因组或转化系统，因此不太适合改进。改良这些作物的传统育种通常依赖于诱导诱变，近年来，这已经通过TILLING的反向遗传学工具直接与基因组学联系起来。TILLING促进了正向和反向遗传学在同一程序中的结合，以允许快速筛选表型和突变发现。耕作法特别适用于有有限或没有反向遗传资源可用的作物，因此是孤儿作物的理想选择。我们建议利用天然种质和现有的草豆突变群体作为快速鉴定突变的新资源，通过非转基因途径改善作物特性。用于测定ODAP的分光光度法已适用于高通量，使用适合筛选大量样品的微滴板。通过这个试验，我们将筛选无毒系。获得无毒品系将能够评估这些化合物在植物（例如在抗虫方面）和在人类疾病方面的真正作用。同时，诱变的植物材料将用于NIAB和印度的BCKV耕作平台生成DNA阵列，并筛选胁迫反应和其他有用的农业特征（例如分枝、种子质量）。我们将研究为什么与其他豆科植物相比，草豆具有更好的抗逆性，目的是利用这一基本见解来提高豌豆等英国豆科植物的抗逆性。TILLING平台将对研究社区revgenuk.jic.ac.uk开放。为了帮助TILLING平台的使用，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鉴定。我们将通过生成草豆不同器官的RNA-seq数据来评估草豆的基因补体，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.