Design and testing of a plant-specific PCR primer for ecological and evolutionary studies
Design and testing of a plant-specific PCR primer for ecological and evolutionary studies
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DOI：
10.1111/j.1365-294x.1992.tb00182.x
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发表时间：
1992-12-01
影响因子：
4.9
通讯作者：
Cullings, K. W.
中科院分区：
文献类型：
--
作者：
Cullings, K. W.
利用聚合酶链反应（PCR）引物28KJ（5′-GGCGGTAAATTCCGTCC-3′）特异性扩增植物DNA。该引物位于28S核糖体RNA基因5'端下游约250个碱基处，与通用引物28C结合使用。该引物组合对31种被子植物、9种针叶树、1种藻类和30种真菌（21种担子菌和9种子囊菌）进行了特异性检测。对植物标本馆标本和新鲜标本进行了测试。28KJ/28C引物组合成功扩增了所有植物和针叶树的dna，但没有扩增真菌和藻类的dna。从植物/真菌共生体（针叶树的外生菌根和菊科植物的ericoid菌根）中扩增植物DNA，并通过将菌根DNA序列或限制性内切酶酶切模式与已知植物样品的DNA序列或酶切模式进行比较，鉴定参与这些共生体的植物。这些方法可以在根系不明确的情况下快速准确地鉴定复杂植物/真菌系统中的植物伴生物。
A polymerase chain reaction (PCR) primer, 28KJ (5'-GGCGGTAAATTCCGTCC-3'), was developed to specifically amplify plant DNA. This primer is located approximately 250 bases downstream of the 5' end of the 28S ribosomal RNA gene, and it was used in combination with the universal primer 28C. The specificity of this primer combination was tested against 31 angiosperms, 9 conifers, 1 alga and 30 fungi (21 basidiomycetes and 9 ascomycetes). Both herbarium specimens and fresh samples were tested. The 28KJ/28C primer combination successfully amplified all angiosperm and conifer DNAs, but no fungal or algal DNAs. Plant DNA was amplified from plant/fungal symbioses (ectomycorrhizae of conifers and ericoid mycorrhizae of Ericaceous plants), and the plants involved in these symbioses were identified by comparing DNA sequences or restriction enzyme digest patterns of the mycorrhizal DNAs to those of known plant samples. These methods allow rapid and accurate identification of plant associates in complex plant/fungal systems when the identity of the roots is unclear.