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利用重复 DNA 探针在 Beta L. 后代家系中选择单体添加植物

Selection of monosomic addition plants in offspring families using repetitive DNA probes in Beta L.

机构信息

DLO-Centre for Plant Breeding and Reproduction Research (CPRO-DLO), PO Box 16, NL-6700, AA Wageningen, The Netherlands.

出版信息

Theor Appl Genet. 1996 May;92(7):891-7. doi: 10.1007/BF00221903.

Abstract

The distribution of two repetitive DNA probes Sat-121 and PB6-4, specific for the section Procumbentes of the genus Beta, was tested in 16 B. patellaris monosomic addition families using a dot-blot hybridization procedure. All monosomic additions were accurately distinguished from diploid sib plants with both DNA probes. The probe PB6-4, with the strongest signal after hybridization, was selected for rapid screening of an extensive number of putative monosomic additions in B. patellaris or B. procumbens addition families using a squash-blot hybridization procedure. The probe PB6-4 detected 118 monosomic additions in 640 plants (18.4%) in eight different B. procumbens addition families. The addition family with chromosome 4 of B. procumbens was semi-lethal and could not be tested. The distribution of PB6-4 in B. patellaris addition families was confirmed in 63 addition families using the squash-blot procedure. In 4580 plants of these addition families, 628 individual monosomic additions (13.7%) were found. The relationship of the morphological characteristics of monosomic addition plants to the results of the squash-blot hybridization (plants with signal) using probe PB6-4 is quite rigorous but not complete. The correlation between plants with a signal and chromosome number (2n=19) is complete. These results indicate that sequences present on PB6-4 are probably present on all chromosomes of B. patellaris and B. procumbens. The possibility of utilizing the sequence information of Sat-121 for a PCR-based assay to screen for putative monosomic addition plants was also investigated as an alternative to chromosome counting. The DNA-amplification profiles using the primers REP and REP.INV clearly distinguished monosomic addition plants from their diploid sibs.

摘要

使用斑点杂交程序,我们测试了两种特定于贝塔属匍匐组的重复 DNA 探针 Sat-121 和 PB6-4 在 16 个 B. patellaris 单体添加家系中的分布。所有单体添加都可以通过这两种 DNA 探针从二倍体同窝植物中准确区分出来。杂交后信号最强的探针 PB6-4 被选中,用于使用斑点杂交程序快速筛选大量 B. patellaris 或 B. procumbens 单体添加家系中的假定单体添加。探针 PB6-4 在 8 个不同的 B. procumbens 单体添加家系的 640 株植物(18.4%)中检测到 118 个单体添加。含有 B. procumbens 第 4 号染色体的单体添加家系是半致死的,无法进行测试。通过斑点杂交程序,我们在 63 个单体添加家系中证实了 PB6-4 在 B. patellaris 单体添加家系中的分布。在这些添加家系的 4580 株植物中,发现了 628 个单体添加个体(13.7%)。使用探针 PB6-4 进行的单体添加植物形态特征与斑点杂交(有信号的植物)结果之间的关系非常严格但不完整。具有信号的植物与染色体数(2n=19)之间的相关性是完整的。这些结果表明,PB6-4 上存在的序列可能存在于 B. patellaris 和 B. procumbens 的所有染色体上。我们还研究了利用 Sat-121 的序列信息进行基于 PCR 的测定以筛选假定的单体添加植物的可能性,作为染色体计数的替代方法。使用引物 REP 和 REP.INV 的 DNA 扩增谱可以清楚地区分单体添加植物与其二倍体同窝植物。

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