Construction Of Molecular Linkage Map And Analysis Of The Genetic Diversity Of The Populations In P.Taiwanensis Hayata.

Random amplified polymorphic DNA (RAPD) markers were used to construct molecular genetic linkage map of a single tree (P.Taiwanensis Hayata.) from a cross 18 X 28 (each from the superior tree from Xianju county and Pan ' an county in Zhejiang province). 236 random oligonucleotide primers were screened and 33 primers were selected to generate RAPD markers within a sample of 60 megagametophyte DNAs. Specially, amplification of the mix primers of SI427 and the other primer appeared to be effective. A total of 52 segregating loci were identified, including 10 loci which appeared to be experencing segregation distortion. Using mapmaker version 3.0, the 39 markers formed 9 main linkage groups and 13 markers were not assigneded to any linkage group. The resulting linkage map of P. Tarwanensis Hayata (including 39 markers) spanned 770.7 cM with an average distance of 2fl.ZcM between markers, linkage groups in size from 1.7 cM (2 markers included )to 306.3 cM(13 markers included),adjacent markers map disance 1.7 cM to 40.0 cM. This map should be the first framework and provide a basis for analysis of genome in P. Tarwanensis Hayata.The analysis of genetic diversity of 10 families in P.Taiwanensis Hayata. was carried out by RAPD molecular markers, which may provide the theoretical base for breeding. 17 primers were screened from 206 random oligonucleotide primers and 38 polymorphic loci were obtained from PCR of 17 primers. The mean Shannon Wevieis index was 4.5251,and Shannon Weveis value of phenotypic diversity varied from 0.0102 to 0.0504. The level of genetic diversity of the family (yuyao X wenshi) was highest,and its Shannon Wevieis index was 7.9658.The genetic distance was caculated according to the 38 polymorphic loci and the cluster map was constructed. The cluster analysis indicated that 10 families could be divided into three groups at the point of genetic distance 0.42.Meanwhile we have proposed the use of an integrated microfluidic chip-based system as a new tool in the analysis of polymorphic RAPD fragments in P.Taiwanensis Hayata.. The chip-based method was found to be very sensitive, requiring much less sample and only quarter the time compared to the agarose gel method. The automated data analysis sizes and quantitates the DNA fragments, thus yielding a more thorough, reproducible, sensitive, and rapid analysis.