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橡胶树(巴西橡胶树)微卫星标记的开发:野生橡胶树种的特征分析与交叉扩增

Microsatellite marker development for the rubber tree (Hevea brasiliensis): characterization and cross-amplification in wild Hevea species.

作者信息

Mantello Camila C, Suzuki Fernando I, Souza Livia M, Gonçalves Paulo S, Souza Anete P

机构信息

Centro de Biologia Molecular e Engenharia Genética (CBMEG) - Universidade Estadual de Campinas (UNICAMP), Cidade Universitária Zeferino Vaz, CP 6010, CEP 13083-970, Campinas, SP, Brazil.

出版信息

BMC Res Notes. 2012 Jun 25;5:329. doi: 10.1186/1756-0500-5-329.

Abstract

BACKGROUND

The rubber tree (Hevea brasiliensis) is native to the Amazon region and it is the major source of natural rubber in the world. Rubber tree breeding is time-consuming and expensive. However, molecular markers such as microsatellites can reduce the time required for these programs. This study reports new genomic microsatellite markers developed and characterized in H. brasiliensis and the evaluation of their transferability to other Hevea species.

FINDINGS

We constructed di- and trinucleotide-enriched libraries. From these two libraries, 153 primer pairs were designed and initially evaluated using 9 genotypes of H. brasiliensis. A total of 119 primer pairs had a good amplification product, 90 of which were polymorphic. We chose 46 of the polymorphic markers and characterized them in 36 genotypes of H. brasiliensis. The expected and observed heterozygosities ranged from 0.1387 to 0.8629 and 0.0909 to 0.9167, respectively. The polymorphism information content (PIC) values ranged from 0.097 to 0.8339, and the mean number of alleles was 6.4 (2-17). These 46 microsatellites were also tested in 6 other Hevea species. The percentage of transferability ranged from 82% to 87%. Locus duplication was found in H. brasiliensis and also in 5 of other species in which transferability was tested.

CONCLUSIONS

This study reports new microsatellite markers for H. brasiliensis that can be used for genetic linkage mapping, quantitative trait loci identification and marker- assisted selection. The high percentage of transferability may be useful in the evaluations of genetic variability and to monitor introgression of genetic variability from different Hevea species into breeding programs.

摘要

背景

橡胶树(巴西橡胶树)原产于亚马逊地区,是世界天然橡胶的主要来源。橡胶树育种耗时且成本高昂。然而,诸如微卫星等分子标记可以减少这些项目所需的时间。本研究报告了在巴西橡胶树中开发和表征的新的基因组微卫星标记,以及对其在其他橡胶树种中的可转移性评估。

研究结果

我们构建了富含二核苷酸和三核苷酸的文库。从这两个文库中设计了153对引物,并最初使用9种巴西橡胶树基因型进行评估。共有119对引物具有良好的扩增产物,其中90对具有多态性。我们选择了46个多态性标记,并在36种巴西橡胶树基因型中对其进行表征。预期杂合度和观察到的杂合度分别为0.1387至0.8629和0.0909至0.9167。多态性信息含量(PIC)值范围为0.097至0.8339,等位基因平均数为6.4(2至17)。这46个微卫星也在其他6种橡胶树种中进行了测试。可转移性百分比范围为82%至87%。在巴西橡胶树以及测试了可转移性的其他5个物种中发现了位点重复。

结论

本研究报告了用于巴西橡胶树的新微卫星标记,可用于遗传连锁图谱构建、数量性状位点鉴定和标记辅助选择。高可转移性百分比可能有助于评估遗传变异性,并监测不同橡胶树种的遗传变异性渗入育种项目的情况。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d62/3439345/290a53d80a8e/1756-0500-5-329-1.jpg

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