Studies On Seed Biology, Isolated Culture And Germplasm Diversity Of Notopterygium Incisum By Scot Marker

Notopterygium incisum is a traditional chinese herb, which mainly grows in Sichuan, Gansu, Qinghai and Yun’nan province. Notopterygium incisum is widely used in traditional Chinese medicines.Nowadays, with the rapid growing of population, the growing environments for Notopterygium incisum is deteriorating. And with the understanding of its economy value,the demanding of this herb is climbing up.To get this herb, in present,is mainly due to digging its wild resources, however, this herb, which only grows in plateau area with high altitude and extremely bad weather has low germinating rate and long growth cycle. Driven by the economy demands, the wild resources of Notopterygium incisum is becoming less and less.The wild plants of Notopterygium incisum is facing serious challenges.Lacking of basic study such as physiology,spermology as well as genetics, the cultivations and naturalization of this species are all restrained. However, the medical use of this herb relies on its wild resources, which makes it facing the danger of extinction. To provide some science supports and genetic background knowledge for helping establishing the cultivating system of Notopterygium incisum and protecting the wild resources of this species, this research mainly studied the germplasm diversity of Notopterygium incisum, isolated culture methods for its explants and the germination of its seeds. The major results are showed as follows.1. the genomic DNA of each sample is extracted by improved2%CTAB method, and a new DNA marker, SCoT is firstly applied to Notopterygium incisum to study its genetic diversity.10primer of this DNA marker are selected in this study.each primer can amplify5to12DNA fragments,10primer of SCoT produced107DNA segments, all segments stay between750bp to2000bp. Among the107DNA segments, there are72polymorphic bands. The polymorphic amplification rate is67.29%. thus each primer can amplify7.2DNA segments.the results of cluster analysis showed that all of the samples germplasm studied can be separated by SCoT DNA marker according to its origins.the similarity coefficients ranged from0.400to0.914in SCoT analysis, which revealed that the difference of collected Notopterygium incisum had rich genetic diversity. The38applied Notopterygium incisum can be divided into5different groups on the similarity coefficient of0.67by cluster analysis. The similarity coefficient of each5groups is below0.67, and on this point,3Notopterygium incisum populatin is divided due to their origin, which revealed the rich diversity between each Notopterygium incisum population.2. In the isolated culture study of Notopterygium incisum,13medium of different hormone combination were applied in order to induce callus of different explants derived from Notopterygium incisum. The inducing rates of callus were all not high, but the most suitable hormone combination is screened:MS+6-BA2.0mg/L+NAA0.5mg/L,which had the highest inducing rate of34%. Callus can be induced from different explants of Notopterygium incisum, such as bud, stem as well as stem tips,among which,explants derived from buds can be induced more easily.3. The seeds’weight of Notopterygium incisum stayed still after24h soaking in water.the ultimate soakage is1.231g. in the first6hours, the seeds stayed in the phase of rapid imbibition, the soakage is26.27%of dried seeds’weight. After this phase, the soakage is diminishing,and slowly reached its balance of soaking. The ultimate soakage of Notopterygium incisum seeds can reach41.23%, which showed the good water absorption of seed coat. There were no obvious differences between seeds with seed coat and seeds without seed coat in the study of germination rate and speed of Notopterygium incisum seeds. Besides,the exogenous hormones like GA (Gibberellic acid) didn’t obviously affect the germination and germination process of Notopterygium incisum seeds.Only if the concentration of GA reached400mg/L, the germination rate reached12%. However, stratification did a better job breaking down the seeds dormancy, which contributed to a higher germination rate of Notopterygium incisum seeds. Combined with a400mg/L GA dispose, temperature changing stratification would get the highest germination rate of46.00%. Compared to sand stratification, this method raised16.33of percentage. In a conclusion, stratification can break down seeds dormancy of Notopterygium incisum seeds, which provide a method for raising the germination rate.