天麻特异DNA序列的克隆及其在天麻鉴定中的应用

应用改进的RAPD方法测定了名贵中药材天麻基因组DNA指纹图谱;通过选择和回收各种天麻种群共有和优良种群特有的DNA片段,加以克隆、测序和生物信息学分析,证明其中5个DNA序列是未报道的,已被美国基因数据库收录,并运用高效液相色谱技术测定了天麻样本的有效成分天麻素含量。运用PCR技术研究了这些DNA序列在9个天麻种群中的分布及其与天麻素含量的关系。结果表明这5个DNA序列在这些天麻种群中的分布各不相同,其中DNA序列1是所研究的全部天麻种群共有、而其伪品没有的特异DNA分子标记;DNA序列2可能与天麻的天麻素含量高有关。这些DNA标记序列可用于天麻的真伪鉴别、品种鉴定和优选优育等。

Cloning of Distinguishing DNA Sequences of Gastrodia elata Blume and Application of Them in Identifying Gastrodia Tuber

Gastrodia elata Bl. is a famous and costful traditional Chinese medicine. Their genomic DNA fingerprints were investigated using a modified Randomly Amplified Polymorphic DNA method. DNA fragments common to all or to fine populations were identified and recovered. Five DNA fragments were proven not to be reported through DNA cloning, PCR identifying, nucleotide sequencing and bioinformatics analyses and were received in and recorded by NCBI GenBank. Gastrodine contents of the Gastrodia tuber samples were determined using high performance liquid chromatography technique. The distribution of the five DNA fragments in 9 Gastrodia elata Blume populations and the correlation with gastrodine content were studied. The results show the distribution of these DNA sequences varied greatly among the populations whereby DNA Sequence 1 was the common and distinguishing molecular marker for all the populations studied and DNA Sequence 2 may relate to higher gastrodine content. In conclusion, these DNA marker sequences can be employed to identify genuine gastrodia tubers, better varieties and optimize their selection and cultivating.