Recent advances in genome sequencing technologies, especially next generation sequencing, have facilitated rapid sequencing of a complete genome, transcriptome or large number of cDNA sequences from non-model organism, providing a massive amount of information for phylogenetic analyses have been widely used to reconstruct the three of life, including to resolve the phylogenetic position of Gnetales. It is interesting that most studies based on concatenated protein-coding nuclear genes (ESTs) support the gnetales-other gymnosperms hypothesis whereas the studies based on chloroplast genes support the Gnepine hypothesis after data treatment. This incongruence could have two explanations. First, for gymnosperms, the nuclear genome sequence is currently unavailable, and available EST sequence cannot well represent the nuclear genome except in some species Pinaceae. Therefore, it is difficult to obtain enough real orthologous nuclear genes from all main lineages of gymnosperms for phylogenetic reconstruction. For example, analyzed 12,469,970 amino acid site from 150 species across land plants. However , according to our reanalysis of the data, none of the site was left after removing the poorly aligned positions in the alignment using Gblocks. Moreover, only 3688 and 1864 sites can be well aligned between Gnetophytes and angiosperms, and between Gnetophytes and Pinaceae, respectively, and in particular the two alignmeny have only about 130 overlapped sites .