Objective To establish a tissue culture system of Pueraria thomsonii and to cultivate a new breed autotetraploid.Methods Using the MS medium as the basic medium,the explants shoot apexes and stem segments were cultured to differentiate the regenerations.The autotetraloid with genetic material doubled was achieved by treating shoot tips with colchicine of different concentrations for different long times.Results The proper media MS+6-BA 1.0 mg/L + 2,4-D 1.0 mg/L+NAA 0.2 mg/L is for pro-(tocorn) inducing,MS+6-BA 1.0 mg/L+2,4-D 0.2 mg/L for diffrentiation and propagation,and 1/2(MS+) IBA 0.2 mg/L for rooting.The best effect of the autotetraploid induction can be achieved by treating the 0.3-0.5 cm shoot tips cut from the 1 cm plantlets with 0.4%-0.5% colchicine for 48 h,and it is unlikely to succeed by treating shoot tips with colchicine of very low or high concentration or for very long or short time.Conclusion The autotetraploid gives a chance to improve the content of medical materials in the root of P.thomsonii.

Plants with alien genomic components (alien chromosomes / chromosomal fragments / genes) are important materials for genomic research and crop improvement. To date, four strategies based on trait observation, chromosome analysis, specific proteins, and DNA sequences have been developed for the identification of alien genomic components. Among them, DNA sequence-based molecular markers are mainly used to identify alien genomic components. This review summarized several molecular markers for identification of alien genomic components in wheat, cabbage and other important crops. We also compared the characteristics of nine common molecular markers, such as simple sequence repeat (SSR), insertion-deletion (InDel) and single nucleotide polymorphism (SNP). In general, the accuracy of using a combination of different identification methods is higher than using a single identification method. We analyzed the application of different combination of identification methods, and provided the best combination for wheat, brassica and other crops. High-throughput detection can be easily achieved by using the new generation molecular markers such as InDel and SNP, which can be used to determine the precise localization of alien introgression genes. To increase the identification efficiency, other new identification methods, such as microarray comparative genomic hybridization (array-CGH) and suppression subtractive hybridization (SSH), may also be included.
Chromosomes, Plant ; Comparative Genomic Hybridization ; Genome, Plant/genetics* ; Genomics ; Triticum/genetics*