Ahmad Rafiq, Tomaszewska Paulina, Shah M Kausar Nawaz, Khan Nadeem, Ibrar Danish, Akhtar Naeem, Heslop-Harrison J S Pat
Department of Genetics, Genomics and Cancer Sciences, University of Leicester, Leicester, LE1 7RH, UK.
Department of Plant Breeding and Genetics, PMAS-Arid Agriculture University, Rawalpindi, Pakistan.
Mol Biol Rep. 2024 Dec 26;52(1):84. doi: 10.1007/s11033-024-10184-6.
Molecular cytogenetics, utilizing DNA probes, serves as a critical tool for mapping genes to the physical structures of chromosomes.
In this study, we examined three Allium species: A. cepa L., A. sativum L., and A. fistulosum L., using in situ hybridization to localize 45S rDNA and 5S rDNA genes.
We observed variation in both the chromosomal localization and signal intensity of the 45S and 5S rDNA probes across the species. Notably, in A. sativum, additional 5S rDNA signals were detected on chromosome 8, in a heterozygous condition. Additionally, we aimed to explore the feasibility of localizing genes associated with pigment biosynthesis in A. cepa, specifically the PAL and FLS genes. For this, we employed TSA-FISH on both meiotic and mitotic chromosomes. Preliminary results suggested that the PAL gene was localized to meiotic metaphase chromosomes, while the single-copy FLS gene was detected on mitotic chromosomes.
The TSA-FISH technique proved neither routine nor robust for consistent localization of these specific probes in plant chromosomes. The findings based on rDNA analysis also offer insights into potential evolutionary implications among the different Allium species studied.
利用DNA探针的分子细胞遗传学是将基因定位到染色体物理结构的关键工具。
在本研究中,我们检测了三种葱属植物:洋葱、大蒜和大葱,采用原位杂交技术定位45S rDNA和5S rDNA基因。
我们观察到45S和5S rDNA探针在不同物种间的染色体定位和信号强度存在差异。值得注意的是,在大蒜中,在8号染色体上检测到额外的5S rDNA信号,呈杂合状态。此外,我们旨在探索在洋葱中定位与色素生物合成相关基因的可行性,特别是苯丙氨酸解氨酶(PAL)和黄酮醇合酶(FLS)基因。为此,我们在减数分裂和有丝分裂染色体上采用了酪胺信号放大荧光原位杂交(TSA-FISH)技术。初步结果表明,PAL基因定位于减数分裂中期染色体,而单拷贝的FLS基因在有丝分裂染色体上被检测到。
TSA-FISH技术对于在植物染色体中一致定位这些特定探针既不常规也不稳定。基于rDNA分析的结果也为所研究的不同葱属物种之间潜在的进化意义提供了见解。