Acta Phytotaxonomica Sinica 33(4), 350—356 (1995)
Preliminary Report of RAPD Analysis in Paeonia suffruticosa subsp. spontanea and Paeonia rockii
Pei Yan-long, Zou Yu-ping, Yin Zhen, Wang Xiao-quan, Zhang Zhi-xian, Hong De-yuan
(Laboratory of Systematic & Evolutionary Botany, Institute of Botany, Chinese Academy of Sciences, Beijing 100093)
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Abstract Ten different oligonucleotide primers of arbitrary sequence were used in Capillary and Air Thermocycler to amplify genomic total DNA of Paeoiaa suffruticosa subsp. spontanea and P. rockii isolated from several local populations in Shanxi, Shaanxi and Gansu provinces. Under the strictly standardized amplification condition for all the primers these primers yielded clear and reproducible bands corresponding to amplified products and separable by agarose gel electrophoresis. Among a total of 71 bands amplified, 16(22. 5%) were polymorphic in a single individual of P. suffruticosa subsp. spontanea, while among a total of 76 bands 21 (27. 6%) were polymophic in a single individual of P. rockii. On an average, the pairwise marker difference between band profiles of conspecific individuals (different populations) was 7. 9 for P. suffruticosa subsp. spontanea and 8. 7 for P. rockii respectively. The average marker difference between P. suffruticosa subsp. spontanea and P. rockii was 10. 3. Obviously, greater number of plants and primers will be required to detect satisfactorily level of genetic diversity. These preliminary results showed that intraspecific genetic diversity was low for the two endangered species. RAPD as a molecular marker was useful and feasible for detecting the genetic variation within species of wild moutans. And it was also potential for studying evolution and relationships between species.
Key words P. suffruticosa subsp. spontanea; P. rockii; genetic variation; RAPD
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Explanation of Plate 1
Plate 1
A. Effect of different concentrations of template on amplification profiles of a single individual P. suffruticosa subsp. spontanea.
M: pGEM DNA Marker (36-2645bp) ;
2,3,4,8,9,10: Amplification with 10ng/micro-l primer 3 in different concentrations of template:
2,8: 15ng/micro-l ;
3,9: 30ng/micro-l;
4,10: 60ng/micro-l;
5,6,7,11,12,13: Amplification with 10ng/micro-l primer 5 in different concentrations of template: 5,11: 15ng/micro-l;
6,12: 10ng/micro-l;
7,13: 60ng/micro-l;
B. Effect of different concentrations of primers on amplification profiles of a single individual of P. suffruticosa subsp. spontanea.
M: pGEM DNA Marker (36 2645bp);
1: 160ng/micro-l primer 5,
2: 80ng/micro-l primer 5,
3: 40ng/micro-l primer 5,
4: 20ng/micro-l primer 5,
5: 10ng/micro-l primer 5.
templete concentration was 30ng/micro-l for each one.
C. Amplification in P. suffruticosa subsp. spontanea and P. rockii with Primer 2.
M: pGEM DNA Marker (36 - 2645bp)
0: PCR Control without template
1 - 7: P. suffruticosa subsp. spontanea
8 – 14: P. rockii.
D. Amplification in P. suffruticosa subsp. spontanea and P. rockii genomic DNA with primer 3. M: pGEM DNA Marker (36 – 2645bp);
0: PCR Control without template;
1 - 7: P. suffruticosa subsp. spontanea
8 – 14: P. rockii.
E. Amplification in P. suffruticosa subsp. spontanea and P. rockii genomic DNA with primer 5. M: pGEM DNA Marker (36-2645bp);
1 - 7: P. suffruticosa subsp. spontanea
8 – 14: P. rockii.
