McCarter James P, Mitreva Makedonka Dautova, Martin John, Dante Mike, Wylie Todd, Rao Uma, Pape Deana, Bowers Yvette, Theising Brenda, Murphy Claire V, Kloek Andrew P, Chiapelli Brandi J, Clifton Sandra W, Bird David Mck, Waterston Robert H
Genome Sequencing Center, Department of Genetics, Box 8501, Washington University School of Medicine, St, Louis, MO 63108, USA.
Genome Biol. 2003;4(4):R26. doi: 10.1186/gb-2003-4-4-r26. Epub 2003 Mar 31.
Plant parasitic nematodes are major pathogens of most crops. Molecular characterization of these species as well as the development of new techniques for control can benefit from genomic approaches. As an entrée to characterizing plant parasitic nematode genomes, we analyzed 5,700 expressed sequence tags (ESTs) from second-stage larvae (L2) of the root-knot nematode Meloidogyne incognita.
From these, 1,625 EST clusters were formed and classified by function using the Gene Ontology (GO) hierarchy and the Kyoto KEGG database. L2 larvae, which represent the infective stage of the life cycle before plant invasion, express a diverse array of ligand-binding proteins and abundant cytoskeletal proteins. L2 are structurally similar to Caenorhabditis elegans dauer larva and the presence of transcripts encoding glyoxylate pathway enzymes in the M. incognita clusters suggests that root-knot nematode larvae metabolize lipid stores while in search of a host. Homology to other species was observed in 79% of translated cluster sequences, with the C. elegans genome providing more information than any other source. In addition to identifying putative nematode-specific and Tylenchida-specific genes, sequencing revealed previously uncharacterized horizontal gene transfer candidates in Meloidogyne with high identity to rhizobacterial genes including homologs of nodL acetyltransferase and novel cellulases.
With sequencing from plant parasitic nematodes accelerating, the approaches to transcript characterization described here can be applied to more extensive datasets and also provide a foundation for more complex genome analyses.
植物寄生线虫是大多数作物的主要病原体。这些物种的分子特征以及新的防治技术的开发都可以从基因组学方法中受益。作为表征植物寄生线虫基因组的切入点,我们分析了根结线虫南方根结线虫(Meloidogyne incognita)二龄幼虫(L2)的5700个表达序列标签(EST)。
从中形成了1625个EST簇,并使用基因本体论(GO)层次结构和京都基因与基因组百科全书(KEGG)数据库按功能进行了分类。L2幼虫代表植物入侵前生命周期的感染阶段,表达多种配体结合蛋白和丰富的细胞骨架蛋白。L2在结构上类似于秀丽隐杆线虫(Caenorhabditis elegans)的 dauer 幼虫,南方根结线虫簇中编码乙醛酸途径酶的转录本的存在表明根结线虫幼虫在寻找宿主时会代谢脂质储备。在79%的翻译后的簇序列中观察到与其他物种的同源性,秀丽隐杆线虫基因组提供的信息比任何其他来源都多。除了鉴定推定的线虫特异性和垫刃目特异性基因外,测序还揭示了南方根结线虫中以前未表征的水平基因转移候选基因,这些基因与根际细菌基因具有高度同一性,包括nodL乙酰转移酶的同源物和新型纤维素酶。
随着植物寄生线虫测序的加速,本文所述的转录本表征方法可以应用于更广泛的数据集,也为更复杂的基因组分析提供基础。