During seed germination and plantlet growth, an important aspect of Lupinus β-conglutin proteolysis is the accumulation of blad in the cotyledons. Blad, a 20.4 kDa, 173 residue polypeptide which inhibits fungal growth, is the main subunit of BCO (Blad-containing oligomer), a Lupinus bioactive polypeptide oligomer, which underwent a successful translational research during the last thirty years. The development of this recent broad-spectrum biological fungicide for plant disease control made evident the advantage of being non-toxic to the environment, plants, humans and other animals, an interesting characteristic given the increasing consumer’s concern about food safety. The industrial-scale production of the edible fungicide BCO for agricultural purposes involves germinating and growing for ca. 8 days massive amounts of Lupinus plantlets. Therefore, it becomes economically relevant to maximize/increment the amount of extractable BCO. Although BCO represents an alternative pathway for lupin production in Europe, there is no previous evidence on lupin cultivars concerning BCO activity or its cotyledonary concentration, and therefore on the most promising cultivars for BCO extraction and fungicide production. In this work, the amount of BCO and its level of fungicide activity was evaluated in seven sweet cultivars of L. albus (cvs. Amiga, Energy, Estoril, Ludic, Misak, Multitalia and Rumbo), along 20 days after the onset of germination. In addition, four distinct lots of cv. Energy and six lots of cv Misak, with different harvest years (1997, 2010, 2013 and 2014) and/or different sites of production in Portugal (Alto Alentejo, Baixo Alentejo and Beira Litoral) were also analysed. Quantitative assays demonstrated maximal accumulation of BCO in the cotyledons of 4-days-old L. albus plantlets, apart from significant differences between seeds’ harvest year or production site. The assays also showed that cv. Energy had significant higher accumulation of BCO (31.67 μg mg⁻¹ wet weight) than Rumbo (25.67 μg mg⁻¹) and Misak (22.12 μg mg⁻¹), respectively second and third cultivars with highest accumulation of the oligomer. Additionally, very significant differences were also observed among seeds’ harvest year or production site. These observations reduced dramatically the original purpose of this work on the study of BCO variations at the level of sweet L. albus cultivars, since the ‘noise’ detected in BCO levels caused by the year and location of their production was far greater than its variation among cultivars. The observation that storing for several years the seeds at -20 °C did not seem to decrease BCO activity and suggests that the variation in cotyledonary BCO accumulation is essentially controlled by the prevailing edaphoclimatic conditions during seed formation. Furthermore, the activity tests exhibited significant antifungal activity between 3 and 5 days-old plantlets, with absence of antifungal activity on 8 days-old plantlets or older. Overall this study is of considerable importance to maximize BCO extraction from the cotyledons of sweet L. albus cultivars.

在种子发芽和幼苗生长过程中，羽扇豆β-凝集素蛋白水解的一个重要方面是子叶中叶片的积累。Blad是20.4 kDa的173个残基的多肽，可抑制真菌的生长，是Lupinu BCO（含Blad的低聚物）的主要亚基。的生物活性多肽寡聚物，在过去的30年中进行了成功的翻译研究。这种最近用于植物病害防治的广谱生物杀真菌剂的开发证明了对环境，植物，人类和其他动物无毒的优势，鉴于消费者对食品安全的日益关注，这是一个有趣的特性。用于农业目的的食用杀菌剂BCO的工业规模生产涉及发芽和生长约。8天大量的羽扇豆苗。因此，最大化/增加可提取的BCO的量在经济上是重要的。尽管在欧洲，BCO代表了羽扇豆生产的替代途径，但以前没有关于羽扇豆品种有关BCO活性或其子叶浓度的证据，因此没有关于BCO提取和杀真菌剂生产的最有前途的品种的证据。在这项工作中，评估了L. albus的七个甜品种中BCO的含量及其杀真菌活性的水平。（与Amiga，Energy，Estoril，Ludic，Misak，Multitalia和Rumbo相对），在发芽开始后的20天之内。此外，还有四个不同的简历。还分析了能源和六批具有不同收获年限（1997年，2010年，2013年和2014年）和/或葡萄牙不同生产地点（Alto Alentejo，Baixo Alentejo和Beira Litoral）的简历Misak。定量分析表明，除种子收获年或生产地点之间的显着差异外，在4天大的白。幼苗的子叶中，BCO的积累量最大。该测定还显示出cv。能量有BCO的显著更高积累（31.67微克毫克^-1湿重）比RUMBO（25.67微克毫克^-1）和Misak（22.12微克毫克^-1），分别是寡聚体含量最高的第二和第三品种。另外，在种子的收获年或生产地点之间也观察到非常显着的差异。这些观察结果大大降低了这项研究在甜L水平上BCO变异研究的初衷。阿不思品种，因为由年份和生产地点引起的BCO水平检测到的“噪声”远大于其在品种之间的变化。将种子在-20°C下保存数年的观察似乎并未降低BCO的活性，并表明子叶BCO积累的变化基本上受种子形成过程中普遍存在的气候条件控制。此外，活性测试在3至5天大的幼苗之间显示出显着的抗真菌活性，而对8天或更老的幼苗没有抗真菌活性。总体而言，这项研究对于最大限度地从甜L的子叶中提取BCO具有重要意义。白bus品种。