Oriental Lily Hybrids Engineered to Resist Aphid Attack
Oriental Lily Hybrids Engineered to Resist Aphid Attack
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DOI：
10.17660/actahortic.2009.836.36
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发表时间：
2009-08
期刊：
影响因子：
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通讯作者：
F. Krens;T. Menzel;Liu Chang;D. Dees;B. Kronenburg
中科院分区：
文献类型：
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作者：
F. Krens;T. Menzel;Liu Chang;D. Dees;B. Kronenburg
发现在我们手中建立百合品种的体外球茎规模培养物，然后在基因转移后进行愈伤组织和再生，除了‘雪之女王’之外，并不是很成功。本文所述研究的目标是确定一种更普遍适用的系统来生成具有再生能力并适合农杆菌介导的基因转移的愈伤组织。在 26 个百合品种的花柱和花丝外植体上诱导愈伤组织。这些品种选自百合属的杂交品种长花、亚洲品种和东方品种，但也有种间杂交品种。大多数品种是二倍体，但有些是三倍体水平，一个是四倍体。一般来说，愈伤组织一旦诱导，就可以相对容易地维持和繁殖以供进一步使用。在所有测试品种的两种愈伤组织类型上均观察到再生，效率范围为 40% 至 100%。在所有测试的品种中都发现了由阳性报告基因 uidA 活性证明的基因转移。在第一系列转化中可以获得转基因植物，并且无标记系统的适用性在百合中得到了证明。该方案现在可用于引入旨在赋予蚜虫抗性的基因。
Establishing in vitro bulb scale cultures of lily cultivars followed by callusing and regeneration after gene transfer was found to be not very successful in our hands, except for ‘Snow Queen’. Identifying a more generally applicable system to generate callus with the ability to regenerate and amenable to Agrobacterium-mediated gene transfer was the goal of the research described here. Callus was induced on style and filament explants of 26 cultivars of lily. The cultivars were chosen from the hybrid groups longiflorum, asiatics and orientals of the genus Lilium but also interspecific hybrids were represented. Most cultivars were diploids but some were of triploid level and one was tetraploid. In general once callus was induced, it could relatively easily be maintained and propagated for further use. Regeneration was observed on both callus types from all cultivars tested with an efficiency ranging from 40 to 100%. Gene transfer as demonstrated by positive reporter gene uidA activity was found in all cultivars tested. Transgenic plants could be obtained in the first series of transformations and the applicability of a marker-free system was proven in lily. The protocol can now be used for the introduction of genes aiming at conferring resistance to aphids.