Clonal Diversity Study Of Tupistra Chinensis Baker

Tupistra chinensis is a perennial herb within the family Liliaceae, alias Zhu Genqi [Shaanxi], Yan Qi. The plant distributes in South China, such as Hubei, Hunan, Fujian, Jiangxi, Zhejiang, Henan, Anhui, Shaanxi(Qinling south), Sichuan and other places, grows in the moist valleys with altitude from 1000 to 2000 meter. Within Tupistra alcohol extract containing a large amount of steroidal saponins and sapogenin and other active chemical components, and minimal side effects, it has great medicinal value. With a wide range of medicinal resources Tupistra get attention, and the impact of excessive dredging and environmental damage, drastically reduced the number of wild Tupistra will lead to depletion of the germplasm. Therefore, it should take urgent measures to protect and restore this precious natural resource.(1)Development of SSR Primers;This study was based on the second-generation sequencing to establish a gene library Tupistra screened out microsatellite zone and 31 pairs of primers were designed, and these primers for amplification and detection, and ultimately selected 13 pairs of bands clear, highly polymorphic primers. Meanwhile, Tupistra grandistigma, Tupistra delavay, Tupistra wattii, Tupistra ensifolia, Reineckia carnea, Rohdea japonica Collected from Hubei, Shaanxi, Yunnan, altogether 6 natural populations of 27 individuals as test materials, test batch of primers among their relatives versatility. The results showed that: Tupistra delavay, Tupistra wattii, Tupistra ensifolia population, 13 pairs of primers able to detect allele, primers general rate of 100%. In the curved core, flag leaf, tubular flowers Tupistra population 12 pairs of primers can be detected allele primer was 92.3%. There are 9 pairs of primers can be detected allele primer was 69.2% in Reineckia carnea and Rohdea japonica population.(2)Analysis of genetic diversity; The study in five populations of Tupistra chinensis dry leaves as experimental material, extraction of genomic DNA amplified, electrophoresis and software analysis draws the following conclusion, Tupistra chinensis at the species level genetic diversity,(NA) for 4.5385,(HO) for 0.4228.(I) is 0.5922. Significant genetic differentiation(GST) was 0.3152.(3)Clonal diversity analysis; In this paper,five populations from 280 Tupistra dried leaves of the experimental material, the use of microsatellite DNA markers(SSR) technology Tupistra of clonal diversity, clonal structure, spatial genetic structure and genetic diversity were studied. Studies have shown that: On the population level, DDS(NC = 2.000, PD = 0.500, H = 2.300, D = 0.970, E = 0.836), TXL(NC = 1.372, PD = 0.729, H = 3.359, D = 0.957, E = 0.920),XGH(NC = 1.125, PD = 0.889, H = 3.860, D = 0.950, E = 0.955) and GHZ(NC = 1.181, PD = 0.847, H = 3.831, D = 0.956, E = 0.954). Four populations mean clone size NC average 1.420, genotype ratio of PD average 0.730, Shannon Wiener index h average 3.338, Simpson index D average 0.958, gene cloning of homogeneous index e average 0.916. At the species level, size NC clone is 2.639, genotype PD ratio is 0.379, Shannon index H is 5.416, Simpson index D 0.987 cloning genotype homogeneity index E 0.982. shannon information index(I) range 0.3599-0.5595, with an average of 0.4616, 0.5922 at the species level. Coefficient of genetic differentiation(GST) ranged from 0.2734-0.3608, with an average of 0.3152.Spatial autocorrelation analysis shows that DDS individuals do not exist significant spatial genetic structure in all spatial distances grades, TXL individuals had significant positive correlation at 2.7m, XGH individuals had significant positive correlation at 1.8m, there is a significant genetic structure in the negative space between 8.2m-10.3m, GHZ individual significant positive correlation at 2.1m, at 3.1m- there is a significant genetic structure of the negative space between 5.4m. Space four populations of autocorrelation coefficients(r) were 0.049,0.059,0.060 and 0.052.This study also revealed that the spatial distribution pattern of the population of Tupistra chinensis is related to the ecological factors(altitude, light, humidity, and grading) of the habitat. The Tupistra chinensis has abundant genetic diversity in the species level, the genetic differentiation is more remarkable and has high clonal diversity, and it is suggested that in situ conservation, artificial breeding and so on.