ISSR-PCR technique: a useful method for characterizing new allotetraploid somatic hybrids of mandarin
ISSR-PCR technique: a useful method for characterizing new allotetraploid somatic hybrids of mandarin
复制标题
DOI：
10.1007/s00299-002-0450-3
复制
发表时间：
2002-06
影响因子：
6.2
通讯作者：
M. Scarano;L. Abbate;S. Ferrante;S. Lucretti;N. Tusa
中科院分区：
文献类型：
--
作者：
M. Scarano;L. Abbate;S. Ferrante;S. Lucretti;N. Tusa
以红背柚和邓肯葡萄柚（CitrusparadisiMacfadyen）为母本，分别与普通柑橘（C.c.）deliciosaTenore）和Fortune mandarin（C. reticulataBlanco）进行原生质体对称融合。采用同工酶带型分析和流式细胞术进行早期筛选。然后提取杂交后代的DNA，并使用锚定在5′和3′末端的不同引物进行简单重复序列间聚合酶链反应（ISSR-PCR）。PCR产物在1.5%琼脂糖凝胶上解析并用溴化乙锭染色。体细胞杂种中亲本的所有带型都出现了。本文还讨论了ISSR-PCR方法作为柑桔体细胞杂种筛选技术的实用性和有效性。
Somatic hybrids between Redblush and Duncan grapefruit (CitrusparadisiMacfadyen) with Avana, Tardivo di Ciaculli mandarin (C. deliciosaTenore) and Fortune mandarin (C. reticulataBlanco) were obtained by symmetric protoplast fusion. Isozyme banding pattern analysis and flow cytometry were used for early screening. DNA of the hybrids was then extracted and subjected to inter-simple sequence repeat polymerase chain reaction (ISSR-PCR) using different primers anchored at the 5′ and 3′ ends. PCR products were resolved on 1.5% agarose gels and stained with ethidium bromide. All the bands from the parental cultivars were present in the somatic hybrids. The usefulness and efficiency of the ISSR-PCR method as a screening technique forCitrussomatic hybrids are discussed.