Molecular Phylogeny Of Ziziphus Mill In China Based On ITS And PsbA-trnH Sequence Data

Ziziphus Mill, is the most economic value among Rhamnaceae. Ziziphus Mill. Classification research can trace back to early in the eighteenth century, but always since then the Ziziphus Mill. Classification problems last more confusion, once having 40 species, 60 species, and 100 species etc. different ideas, and do not know if it is natural. ITS sequence (including ITS1, 5.8S rDNA and ITS2) of nuclear ribosomal DNA and psbA-trnH sequence of cpDNA from 12 species of Ziziphus in China and a outgroup genus P. spina-christi Mill, were determined by directly sequencing of PCR products, and molecular phylogeny of Ziziphus in China based on ITS and psbA-trnH sequences was studied. The extracting methods of genomic DNA from dry leaf of Ziziphus stored in silica were compared.1. Improved CTAB, high salt precipitation, and high salt and low pH method were employed to extract genomic DNA from the leaves of 3 species of Ziziphus Mill, stored in silica for 15 days and half a year. The extracted DNA was tested using the value of A₂₆₀/A₂₈₀, agarose gel electrophoresis and PCR amplification. The DNA from the leaves stored in silica for half a year is good for PCR amplification and the method of high salt precipitation is best.2. The size of ITS of Ziziphus Mill, is from 601 to 611bp, and the length variation is 10bp.The paiwise distances are 0.331% to 10.000% between species, and 14.919% to 17.944% between genus Ziziphus Mill, and outgroup. Phylogenetic analysis using Fitch parsimony showed that: Z. pubinervis Rehd. , Z. apetaia Hook. And Z. fungii Merr. were resolved as sisters to Z.jujuba Mill. And Z.acidojujuba C. Y. Cheng et M. J. Liu; Zxiangchengensis Y.L.Chen et P.K.Chou, Zmontana W.W.Smith and Z.mairei Dode were resolved as sisters to ZAncurva Roxb. and Z.rugosa Lam.; both Z.mauritiana Lam. and...