| Identification the relationship of important traits in allopolyploids and their parents and proving the allopolyploids genetic variation of a tri-species hybrid MTF-1 (tetraploid maize ×Tripsacum dactyloides L × zea perennis) (2n=76) and tri-species hybrid with zea perennis backcross progeng MTZZ-1 (2n=96) based on morphological, anatomical, molecular biology and molecular genetics analysis. The main results are as follows:1. Three parents and two allopolyploids morphological traits and some organ anatomical traits indicated that, MTZ-1 most of the traits, such as stem diameter, root number, root volume and so on, ranging between three parents, a few traits such as filigree cross area has Hpb, there are some traits such as flowery vascular area produces degradation; MTZZ-1 most of the traits, such as leaf area, stem diameter, root cross-section conduits and so on was not significant with MTZ-1, a few traits such as stem ear numbers was significantly higher than MTZ-1, show the " polyploidized " phenomenon.2. Three parents and two Allopolyploid all observations traits cluster analysis showed that, the genetic distance between MTZ-1 and tripsacum is closest, maize secondly,9475 farthest; the genetic distance between MTZZ-1 and 9475 is closest, maize secondly, tripsacum farthest. Description MTZ-1 gene influenced by friction Wo, while MTZZ-1 gene influenced by 9475. Demonstrating that the gene of MTZ-1 is most influenced by tripsacum and MTZZ-1 is most influenced by 9475.3. Three parents and two allopolyploid TrnL-F and ITS direct sequence analysis showed that, MTZ-1 and MTZZ-l’s TrnL-F sequence identical with maize, indicating that maize is the female of hybrids, changes by backcross in the nucleus is little effect on TrnL-F; two allopolyploids ITS sequence length between the three parents; G+C content decreased, higher incidence of GC mutation; Three parents and two allopolyploid ITS information locus combined with the peaks analysis showed that the peak which corresponds the informative sites appeares set peak, and bear with three parental informative locus. Integrated ITS direct sequencing results showed that three parental ITS sequences in the same cell occur interaction, and through this analysis method can identify Allopolyploid parental components.4. ITS monoclonal sequence analysis showed that, there are only 2.4% tripsacum type sequence in MTZ-1, and MTZZ-1 has not tripsacum type sequence, suggesting that the tripsacum type ITS sequences in two Allopolyploid disappear faster; in MTZ-1, the maize has a larger role on the 9475 ITS sequences, but in MTZZ-19475 ITS sequence has a larger role on maize, and the tripsacum sequence in two allopolyploid relatively small role, indicating that two allopolyploid ITS occurs interaction between maize and 9475 mainly, and the interaction of the size base on number of chromosomes; two hybrids of the information loci analysis on the bit showed that comparing the same loci with MTZ-1,9475 informative loci increased in MTZZ-1, indicating that the 9475 DNA adding increased the detection rate of 9475 loci; all monoclonal sequences of MTZ-1, MTZZ-1, Zea (maize and teosinte) and tripsacum constructe phylogenetic tree, MTZ-1 monoclonal sequences in the phylogenetic tree is divided into three categories, corresponing with Section Luxuriantes, Section Zea and tripsacum respectively, but corresponing little with tripsacum; while MTZZ-I is mainly divided into two categories, only corresponing with Section Luxuriantes and Section Zea respectively. Indicating that three parental ITS sequences occurs coevolution mainly between maize and 9475.5. MTZ-1 and MTZZ-1ITS direct sequencing as well as Zea (maize and teosinte) and tripsacum monoclonal ITS Sequence Clustering analysis showed that there is different clustering results of two allopolyploid morphological and anatomical traits cluster, combined ITS sequence site analysis, indicating that the newly synthesized allopolyploid ITS sequence have more variation sites, ITS sequence can not be used on the new synthetic allopolyploid cluster analysis.6. MTZ-1 FISH-GISH in situ hybridization during mitosis and meiosis showed that chromosomes during mitosis occurs incompatibility, appearing partial chromosome fragments missing,45sDNA which including ITS sequences occurred aggregation and separation, indicating that in some MTZ-1 mitosis cells the three parental chromosomal instability and interaction; in Meiosis, the 45sDNA signal loci aggregation and the chromosome fragments missing also occurred, combined with ITS sequence analysis, in the same cell the three parents occur coevolution, which provides the basis of rich character variation in triples hybrid. |
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