EVALUATION OF SILICA NANOPARTICLES (SiO2NP) AND SOMACLONAL VARIATION EFFECTS ON GENOME TEMPLATE STABILITY IN RICE USING RAPD AND SSR MARKERS
Abstract
Assessment of DNA changes and mutations at molecular level are important in plant breeding. In this study DNA changes in four rice genotypes (Sakha-107, Giza-179, Sakha-106 and Sakha-101) induced by silica nanoparticles (0, 150, 300 and 450 ppm) and somaclonal variations were determined using RAPD and SSR analyses. The potential effects of SiO2NP (< 100 nm) on rice plant growth were studied and the results showed positive and negative effects. Application of SiO2NP enhanced the fresh weight, shoot length and root length of the drought-sensitive rice varieties, but number of roots/seedling was increased with high concentration (450 ppm) in all studied genotypes. Mature embryos of four rice genotypes were used as explant source for callus induction and plant regeneration system. The obtained results showed significant effect of genotype on callus induction and plant regeneration in rice. For assessment of genome template stability percentage (GTS%), five RAPD primers were used and produced a total number of bands ranged from 25 to 51 in the studied genotypes. Results indicated that the GTS% was lower in somaclonal variations for all studied genotypes. Also, the changes occurred in DNA included gain or loss of bands compared with the control plants. On the other hand, among different concentrations of silica nanoparticles, the drought tolerant genotypes (Sakha-107 and Giza-179) gave the highest percentage of GTS in 150 ppm, while the drought sensitive genotypes (Sakha-106 and Sakha-101) gave the highest percentage on 300 and 450 ppm for Sakha-106 and sakha-101, respectively. These results confirmed the effects of SiO2NP and somaclonal variations on mutation and DNA instability and suggested that genomic template stability (GTS) reflecting changes in RAPD profiles was the most sensitive endpoint compared with the traditional indices such as root and shoot growth. On the other hand, two SSR markers (RM215 and RM518) were applied and generated 93.75 and 75% polymorphism, respectively. These variations among rice varieties and their treatments could help in rice plant breeding for drought tolerance by the selection of the suitable genotypes which are able to tolerate high drought stress conditions.References
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