Camacho J P M, Ruiz-Ruano F J, Martín-Blázquez R, López-León M D, Cabrero J, Lorite P, Cabral-de-Mello D C, Bakkali M
Departamento de Genética, Facultad de Ciencias, Universidad de Granada, Fuentenueva S/N, 18071, Granada, Spain.
Chromosoma. 2015 Jun;124(2):263-75. doi: 10.1007/s00412-014-0499-0. Epub 2014 Dec 5.
The desert locust (Schistocerca gregaria) has been used as material for numerous cytogenetic studies. Its genome size is estimated to be 8.55 Gb of DNA comprised in 11 autosomes and the X chromosome. Its X0/XX sex chromosome determinism therefore results in females having 24 chromosomes whereas males have 23. Surprisingly, little is known about the DNA content of this locust's huge chromosomes. Here, we use the Feulgen Image Analysis Densitometry and C-banding techniques to respectively estimate the DNA quantity and heterochromatin content of each chromosome. We also identify three satellite DNAs using both restriction endonucleases and next-generation sequencing. We then use fluorescent in situ hybridization to determine the chromosomal location of these satellite DNAs as well as that of six tandem repeat DNA gene families. The combination of the results obtained in this work allows distinguishing between the different chromosomes not only by size, but also by the kind of repetitive DNAs that they contain. The recent publication of the draft genome of the migratory locust (Locusta migratoria), the largest animal genome hitherto sequenced, invites for sequencing even larger genomes. S. gregaria is a pest that causes high economic losses. It is thus among the primary candidates for genome sequencing. But this species genome is about 50 % larger than that of L. migratoria, and although next-generation sequencing currently allows sequencing large genomes, sequencing it would mean a greater challenge. The chromosome sizes and markers provided here should not only help planning the sequencing project and guide the assembly but would also facilitate assigning assembled linkage groups to actual chromosomes.
沙漠蝗(Schistocerca gregaria)已被用作众多细胞遗传学研究的材料。其基因组大小估计为8.55Gb DNA,由11条常染色体和X染色体组成。其X0/XX性染色体决定机制导致雌性有24条染色体,而雄性有23条。令人惊讶的是,对于这种蝗虫巨大染色体的DNA含量知之甚少。在这里,我们使用福尔根图像分析密度测定法和C带技术分别估计每条染色体的DNA数量和异染色质含量。我们还使用限制性内切酶和下一代测序鉴定了三种卫星DNA。然后,我们使用荧光原位杂交来确定这些卫星DNA以及六个串联重复DNA基因家族的染色体位置。这项工作获得的结果相结合,不仅可以根据大小,还可以根据它们所含重复DNA的种类来区分不同的染色体。最近发表的飞蝗(Locusta migratoria)基因组草图是迄今为止测序的最大动物基因组,这促使人们对更大的基因组进行测序。沙漠蝗是一种造成巨大经济损失的害虫。因此,它是基因组测序的主要候选物种之一。但是这个物种的基因组比飞蝗的基因组大约大50%,虽然下一代测序目前能够对大基因组进行测序,但对其进行测序将意味着更大的挑战。这里提供的染色体大小和标记不仅有助于规划测序项目和指导组装,还将便于将组装好的连锁群分配到实际的染色体上。