Production and fingerprinting of virus-free clones in a reflowering globe artichoke
Production and fingerprinting of virus-free clones in a reflowering globe artichoke
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DOI：
10.1007/s11240-009-9654-3
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发表时间：
2010-03-01
影响因子：
3
通讯作者：
Gallitelli, D.
中科院分区：
文献类型：
--
作者：
Acquadro, A.;Papanice, M. A.;Gallitelli, D.
感染地球仪朝鲜蓟的24种病毒中的大多数对作物的表现有害，并且在当前欧盟立法方面阻碍苗圃活动的发展。我们描述了一个程序，以消毒的地球仪洋蓟“Brindisino”从朝鲜蓟意大利潜伏病毒（AILV）和朝鲜蓟潜伏病毒（ArLV），而保持其宝贵的早花性状。ArLV被成功地消除了分生组织尖端培养，而AILV被删除时，两轮分生组织尖端培养间隔与体外热疗法。在体内热疗法，其次是分生组织尖端培养，也是成功的生产无病毒的材料，但效率较低的治疗后恢复植物的数量。由于目前在培养的群体的多克隆组成，选择和消毒的克隆指纹通过应用微卫星和AFLP（扩增片段长度多态性）标记。一个AFLP引物组合产生28个信息片段，用于评估研究中的克隆之间的遗传关系。我们的研究结果表明，AFLP分子指纹技术可以验证真实的克隆对应在苗圃，确保有效的对应之间的真实的和宣布的身份的克隆，以避免商业欺诈，并可能代表一个有价值的工具，评估体细胞无性系变异发生在“体外”繁殖。
Most of the 24 viruses which infect globe artichoke are detrimental to the crop's performance and hamper the development of a nursery activity in the respect of current EU legislation. We describe a procedure to sanitize globe artichoke "Brindisino" from Artichoke Italian latent virus (AILV) and Artichoke latent virus (ArLV), while preserving its valuable early flowering trait. ArLV was successfully eliminated by meristem-tip culture, while AILV was removed when two rounds of meristem-tip culture were spaced out with in vitro thermotherapy. In vivo thermotherapy, followed by meristem-tip culture, was also successful in producing virus-free material but was less efficient in terms of the number of plants recovered post treatment. Due to the multi-clonal composition of the populations at present in cultivation, the selected and sanitised clones were fingerprinted by applying microsatellite and AFLP (amplified fragment length polymorphism) markers. One AFLP primer combination produced 28 informative fragments used to evaluate genetic relatedness among the clones in study. Our results demonstrates that AFLP-based molecular fingerprinting enables to verify the true to clone correspondence in nurseries, ensure the effective correspondence between the real and the declared identity of a clone, so that to avoid commercial frauds, and might represents a valuable tool for assessing somaclonal variation occuring during 'in vitro' propagation.