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甘蓝型油菜黄籽与黑籽色素组成的差异及其相关基因的初步研究

Chromosome-specific physical localisation of expressed sequence tag loci in Corchorus olitorius L.

机构信息

Advanced Laboratory for Plant Genetic Engineering, Indian Institute of Technology, Kharagpur, India.

出版信息

Plant Biol (Stuttg). 2014 Nov;16(6):1133-9. doi: 10.1111/plb.12158. Epub 2014 Mar 13.

Abstract

Jute (Corchorus spp.), as a natural fibre-producing species, ranks next only to cotton. Inadequate understanding of its genetic architecture is a major lacuna for genetic improvement of this crop in terms of yield and quality. Establishment of a physical map provides a genomic tool that helps in positional cloning of valuable genes. In this report, an attempt was initiated to study association and localisation of single copy expressed sequence tag (EST) loci in the genome of Corchorus olitorius. The chromosome-specific association of EST was determined based on the appearance of an extra signal for a single copy cDNA probe in mitotic interphase nuclei of specific trisomic(s) for fluorescence in situ hybridisation, and validated using a cDNA fragment of the 26S rRNA gene (600 bp) as molecular probe. The probe exhibited three signals in meiotic interphase nuclei of trisomic 5, instead of two as observed in diploids and other trisomics, indicating its association with chromosome 5. Subsequent hybridisation of the same probe on the pachytene chromosomes of diploids confirmed that 26S rRNA occupies the terminal end of the short arm of chromosome 5 in C. olitorius. Subsequently, chromosome-specific association of 63 single copy EST and their physical localisation were determined on chromosomes 2, 4, 5 and 7. The study describes chromosome-specific physical localisation of genes in jute. The approach used here could be a step towards construction of genome-wide physical maps for any recalcitrant plant species like jute.

摘要

黄麻(Corchorus spp.)是一种天然纤维作物,其产量仅次于棉花。由于对其遗传结构缺乏充分了解,因此在提高该作物的产量和品质方面存在很大的局限性。建立物理图谱为基因定位提供了基因组工具,有助于克隆有价值的基因。本研究旨在探讨黄麻基因组中单拷贝表达序列标签(EST)位点的相关性和定位。基于荧光原位杂交中期分裂核中单拷贝 cDNA 探针出现额外信号,确定 EST 与染色体的特异性关联,并使用 26S rRNA 基因(600bp)的 cDNA 片段作为分子探针进行验证。该探针在 5 号三体的减数分裂中期核中显示出三个信号,而在二倍体和其他三体中则观察到两个信号,表明其与 5 号染色体相关。随后,在二倍体的粗线期染色体上对相同探针进行杂交,证实 26S rRNA 占据了 C. olitorius 5 号染色体短臂的末端。随后,确定了 63 个单拷贝 EST 在染色体 2、4、5 和 7 上的特异性关联及其物理定位。本研究描述了黄麻中基因的染色体特异性物理定位。该方法可作为构建像黄麻这样的难处理植物全基因组物理图谱的一步。

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