Filatov Dmitry A
School of Biosciences, University of Birmingham, Birmingham B12 2TT, United Kingdom.
Methods Enzymol. 2005;395:418-42. doi: 10.1016/S0076-6879(05)95023-4.
Few plant species are dioecious and only a small fraction of these species are known to have sex chromosomes. Considerable efforts to isolate sex-linked genes from dioecious Silene latifolia (Caryophillaceae) have resulted in the isolation of surprisingly few sex-linked genes, suggesting that the methods used previously were not efficient in plants. This chapter analyzes the methods that have been and can be used for isolation of genes from plant sex chromosomes. The most successful method used for the isolation of Y-linked genes included the screening of a male complementary DNA (cDNA) library with the probe obtained by degenerate oligonucleotide-primed polymerase chain reaction (PCR) of the microdissected Y chromosomes. However, chromosome microdissection requires sophisticated equipment and is difficult to apply to species with cytologically indistinguishable sex chromosomes. Genome and cDNA library subtraction methods were surprisingly unsuccessful, probably because of low divergence between the homologous X- and Y-linked genes in plants. Segregation testing and genomics-based methods are increasingly popular and are the most promising approaches for isolation of multiple genes from plant sex chromosomes.
很少有植物物种是雌雄异株的，而且已知这些物种中只有一小部分具有性染色体。人们付出了巨大努力从雌雄异株的石竹科植物白麦瓶草中分离性连锁基因，但令人惊讶的是，分离出的性连锁基因数量极少，这表明之前使用的方法在植物中效率不高。本章分析了已有的以及可用于从植物性染色体中分离基因的方法。用于分离Y连锁基因的最成功方法包括用通过显微切割的Y染色体的简并寡核苷酸引物聚合酶链反应（PCR）获得的探针筛选雄性互补DNA（cDNA）文库。然而，染色体显微切割需要精密的设备，并且难以应用于性染色体在细胞学上难以区分的物种。基因组和cDNA文库消减方法出人意料地不成功，可能是因为植物中同源的X连锁和Y连锁基因之间的差异较小。分离测试和基于基因组学的方法越来越受欢迎，并且是从植物性染色体中分离多个基因最有前景的方法。
