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寄生杂草独脚金全长 cDNA 文库和表达序列标签分析。

A full-length enriched cDNA library and expressed sequence tag analysis of the parasitic weed, Striga hermonthica.

机构信息

Plant Science Center, RIKEN, Tsurumi-ku, Yokohama, Japan.

出版信息

BMC Plant Biol. 2010 Mar 30;10:55. doi: 10.1186/1471-2229-10-55.

Abstract

BACKGROUND

The obligate parasitic plant witchweed (Striga hermonthica) infects major cereal crops such as sorghum, maize, and millet, and is the most devastating weed pest in Africa. An understanding of the nature of its parasitism would contribute to the development of more sophisticated management methods. However, the molecular and genomic resources currently available for the study of S. hermonthica are limited.

RESULTS

We constructed a full-length enriched cDNA library of S. hermonthica, sequenced 37,710 clones from the library, and obtained 67,814 expressed sequence tag (EST) sequences. The ESTs were assembled into 17,317 unigenes that included 10,319 contigs and 6,818 singletons. The S. hermonthica unigene dataset was subjected to a comparative analysis with other plant genomes or ESTs. Approximately 80% of the unigenes have homologs in other dicotyledonous plants including Arabidopsis, poplar, and grape. We found that 589 unigenes are conserved in the hemiparasitic Triphysaria species but not in other plant species. These are good candidates for genes specifically involved in plant parasitism. Furthermore, we found 1,445 putative simple sequence repeats (SSRs) in the S. hermonthica unigene dataset. We tested 64 pairs of PCR primers flanking the SSRs to develop genetic markers for the detection of polymorphisms. Most primer sets amplified polymorphicbands from individual plants collected at a single location, indicating high genetic diversity in S. hermonthica. We selected 10 primer pairs to analyze S. hermonthica harvested in the field from different host species and geographic locations. A clustering analysis suggests that genetic distances are not correlated with host specificity.

CONCLUSIONS

Our data provide the first extensive set of molecular resources for studying S. hermonthica, and include EST sequences, a comparative analysis with other plant genomes, and useful genetic markers. All the data are stored in a web-based database and freely available. These resources will be useful for genome annotation, gene discovery, functional analysis, molecular breeding, epidemiological studies, and studies of plant evolution.

摘要

背景

专性寄生植物独脚金(Striga hermonthica)感染高粱、玉米和小米等主要谷类作物,是非洲最具破坏性的杂草害虫。了解其寄生特性将有助于开发更复杂的管理方法。然而,目前用于研究独脚金的分子和基因组资源有限。

结果

我们构建了独脚金全长富集 cDNA 文库,从文库中测序了 37710 个克隆,获得了 67814 条表达序列标签(EST)序列。EST 被组装成 17317 个基因,其中包括 10319 个连续序列和 6818 个单序列。独脚金基因数据集与其他植物基因组或 EST 进行了比较分析。大约 80%的基因在其他双子叶植物中都有同源物,包括拟南芥、杨树和葡萄。我们发现 589 个基因在半寄生三蕊草属物种中保守,但在其他植物物种中不存在。这些是专门参与植物寄生的基因的良好候选者。此外,我们在独脚金基因中发现了 1445 个推定简单序列重复(SSR)。我们测试了 64 对侧翼 SSR 的 PCR 引物,以开发用于检测多态性的遗传标记。大多数引物对都能从单个地点采集的单个植物中扩增出多态性条带,表明独脚金的遗传多样性很高。我们选择了 10 对引物来分析从不同宿主物种和地理位置采集的田间独脚金。聚类分析表明,遗传距离与宿主特异性无关。

结论

我们的数据为研究独脚金提供了第一个广泛的分子资源集,包括 EST 序列、与其他植物基因组的比较分析以及有用的遗传标记。所有数据都存储在基于网络的数据库中,并免费提供。这些资源将有助于基因组注释、基因发现、功能分析、分子育种、流行病学研究和植物进化研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8949/2923529/5d3452dbfb23/1471-2229-10-55-1.jpg

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