The use of DNA sequencing (ITS and trnL-F), AFLP, and fluorescent in situ hybridization to study allopolyploid Miscanthus (Poaceae)
The use of DNA sequencing (ITS and trnL-F), AFLP, and fluorescent in situ hybridization to study allopolyploid Miscanthus (Poaceae)
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DOI：
10.3732/ajb.89.2.279
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发表时间：
2002-02-01
影响因子：
3
通讯作者：
Renvoize, SA
中科院分区：
文献类型：
--
作者：
Hodkinson, TR;Chase, MW;Renvoize, SA
芒草的两个克隆，分别以 M. Xgiganteus 和 M. sacchariflorus 的名称种植，已在欧洲用于生物量试验，但这些克隆的身份及其起源尚未确定。DNA 测序、扩增片段长度多态性 (AFLP) 和染色体研究均未证实芒草。是一种异源三倍体 (2n = 3x = 57)，结合了来自 M. sinensis (2n = 2x = 38) 和 M. sacchariflorus (2n = 38 或 76) 的基因组。在 M. Xgiganteus 的聚合酶链反应产物中发现了 18S-25S 核糖体 DNA (ITS) 内转录间隔区的两个等位基因。对它们的克隆显示，其中一个与 M. sinensis 相匹配，另一个与 M. sacchariflorus 相匹配。质体trnL、内含子和trnL-F间隔序列表明M. Xgiganteus的母系谱系是M. sacchariflorus。荧光原位杂交 (FISH) 用于研究芒草中的基因组组织，但无法区分 M. Xgiganteus 中存在的不同亲本基因组，这表明两个亲本基因组在重复 DNA 水平上仍然极其相似。这项研究是一个例子，其中 rDNA 序列和 AFLP 指纹允许识别杂种中的亲本基因组，但 FISH 方法在重复 DNA 水平上（包括基因组原位杂交、 GISH），但无法这样做，因为它们的序列仍然太相似。
Two clones of Miscanthus, grown under the names M. Xgiganteus and M. sacchariflorus, have been used in biomass trials in Europe, but neither the identity of these clones nor their origin has been established DNA sequencing, amplified fragment length polymorphism (AFLP), and chromosome studies confirm that M. Xgiganteus. is an allotriploid (2n = 3x = 57) combining genomes from M sinensis (2n = 2x = 38) and M. sacchariflorus (2n = 38 or 76). Two alleles of the internal transcribed spacer of 18S-25S nuclear ribosomal DNA (ITS) were discovered in polymerase chain reaction products of M. Xgiganteus. Cloning of these revealed that one matched M. sinensis and the other M. sacchariflorus. Plastid trnL, intron and trnL-F spacer sequences showed that the maternal lineage of M. Xgiganteus was M. sacchariflorus. Fluorescent in situ hybridization, FISH, was used to investigate genome organization in Miscanthus but was unable to differentiate between the different parental genomes present in M. Xgiganteus, indicating that two parental genomes are still extremely similar at the repetitive DNA level This study is an example in which rDNA sequences and AFLP fingerprints permit identification of the parental genomes in a hybrid, but FISH methods, at the repetitive DNA level (including genomic in situ hybridization, GISH), were unable to do so because their sequences remain too similar.