Tang Mingyong, Chen Zhiwen, Grover Corrinne E, Wang Yumei, Li Shuangshuang, Liu Guozheng, Ma Zhiying, Wendel Jonathan F, Hua Jinping
Department of Plant Genetics and Breeding /Key Laboratory of Crop Heterosis and Utilization of Ministry of Education /Beijing Key Laboratory of Crop Genetic Improvement, China Agricultural University, Beijing, 100193, China.
Department of Ecology, Evolution and Organismal Biology, Iowa State University, Ames, IA50011, USA.
BMC Genomics. 2015 Oct 12;16:770. doi: 10.1186/s12864-015-1988-0.
The mitochondrial genome from upland cotton, G. hirsutum, was previously sequenced. To elucidate the evolution of mitochondrial genomic diversity within a single genus, we sequenced the mitochondrial genome from Sea Island cotton (Gossypium barbadense L.).
Mitochondrial DNA from week-old etiolated seedlings was extracted from isolated organelles using discontinuous sucrose density gradient method. Mitochondrial genome was sequenced with Solexa using paired-end, 90 bp read. The clean reads were assembled into contigs using ABySS and finished via additional fosmid and BAC sequencing. Finally, the genome was annotated and analyzed using different softwares.
The G. barbadense (Sea Island cotton) mitochondrial genome was fully sequenced (677,434-bp) and compared to the mitogenome of upland cotton. The G. barbadense mitochondrial DNA contains seven more genes than that of upland cotton, with a total of 40 protein coding genes (excluding possible pseudogenes), 6 rRNA genes, and 29 tRNA genes. Of these 75 genes, atp1, mttB, nad4, nad9, rrn5, rrn18, and trnD(GTC)-cp were each represented by two identical copies. A single 64 kb repeat was largely responsible for the 9 % difference in genome size between the two mtDNAs. Comparison of genome structures between the two mitochondrial genomes revealed 8 rearranged syntenic regions and several large repeats. The largest repeat was missing from the master chromosome in G. hirsutum. Both mitochondrial genomes contain a duplicated copy of rps3 (rps3-2) in conjunction with a duplication of repeated sequences. Phylogenetic and divergence considerations suggest that a 544-bp fragment of rps3 was transferred to the nuclear genome shortly after divergence of the A- and D- genome diploid cottons.
These results highlight the insights to the evolution of structural variation between Sea Island and upland cotton mitochondrial genomes.
陆地棉(G. hirsutum)的线粒体基因组先前已完成测序。为阐明单个属内线粒体基因组多样性的进化,我们对海岛棉(Gossypium barbadense L.)的线粒体基因组进行了测序。
使用不连续蔗糖密度梯度法从分离的细胞器中提取一周龄黄化幼苗的线粒体DNA。线粒体基因组采用Solexa技术进行双末端90 bp读长测序。利用ABySS软件将干净的读段组装成重叠群,并通过额外的fosmid和BAC测序完成组装。最后,使用不同软件对基因组进行注释和分析。
海岛棉线粒体基因组已完成全序列测定(677,434 bp),并与陆地棉的线粒体基因组进行了比较。海岛棉线粒体DNA比陆地棉多7个基因,共有40个蛋白质编码基因(不包括可能的假基因)、6个rRNA基因和29个tRNA基因。在这75个基因中,atp1、mttB、nad4、nad9、rrn5、rrn18和trnD(GTC)-cp各有两个相同的拷贝。一个64 kb的单一重复序列在很大程度上导致了两个线粒体DNA基因组大小9%的差异。两个线粒体基因组的结构比较揭示了8个重排的同线性区域和几个大的重复序列。陆地棉主染色体上缺失了最大的重复序列。两个线粒体基因组都包含rps3的一个重复拷贝(rps3-2)以及重复序列的重复。系统发育和分歧分析表明,在A基因组和D基因组二倍体棉花分化后不久,rps3的一个544 bp片段转移到了核基因组中。
这些结果突出了对海岛棉和陆地棉线粒体基因组结构变异进化的认识。