Using of DNA-Barcoding, SCoT and SDS-PAGE Protein to Assess Soma-Clonal Variation in Micro-Propagated Fig (Ficus carica L.) Plant.

<b>Background and Objective:</b> <i>In vitro</i> propagation of fig (<i>Ficus carica</i> L.) is one of the possible approaches that may be used to maximize the diversity of plant species. The current work was carried out to evaluate genetic stability of micropropagated fig plantlets and to determine the effect of <i>in vitro </i>propagation on genomic content of Saudi fig. <b>Materials and Methods:</b> The start codon-targeted (SCoT), DNA-barcoding chloroplast gene RNA polymerase1 (<i>rpoC1</i> sequencing) and total protein profiling assays (SDS-PAGE) techniques were used to detect genetic stability in micropropagated fig plantlets. <b>Results:</b> The Scorable PCR bands were produced with 10 SCoT primers used, where the total number of bands was 135 bands. Twenty polymorphic bands were generated with 18.4% of a polymorphism percentage. According to the result, no visual unique bands were generated which confirmed the genetic homogeneity of micropropagated plantlets samples compared to the control sample (mother plant). Sequence analysis and phylogenetic tree generated using fig <i>rpoC1</i> sequence showed high similarity between control and plantlets samples of fig plant. The protein profiling results revealed no remarkable changes between micropropagated plantlets and the mother plant. <b>Conclusion:</b> The results indicate that using SCoT, DNA barcoding and protein profiling have demonstrated their utility to detect genetic homogeneity in micropropagated fig plantlets, which suggests using of micropropagation protocol of plants applied on the plantlets in the current study as a reliable protocol for <i>in vitro</i> culture and conservation of fig plant.

Production of Virus-Free Rose Plants using Meristem-Tip Culture and in vitro Thermotherapy.

<b>Background and Objective:</b> Tissue culture and thermotherapy were proved to be suitable in eliminating viruses of many plants. This study was designed in an attempt to produce virus-free Al-Taif rose plants (<i>Rosa damascena</i> Trigintipetala Dieck) through the practical application of the tissue culture approach and thermotherapy. <b>Materials and Methods:</b> Double Antibody Sandwich-Enzyme-Linked Immunosorbent Assay ( DAS-ELISA) and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) techniques were used to detect the presence of <i>Apple mosaic virus</i> (ApMV) and <i>Strawberry latent ringspot virus</i> (SLRV) in rose plant materials collected from Taif, KSA. RT-PCR was more sensitive than DAS-ELISA in detecting the 2 viruses. <b>Results:</b> Three different meristem-tip sterilization methods were compared and results revealed that treatment 3 (T₃: 70% Ethanol for 1.0 min and 15% Clorox (Sodium hypochlorite 5.25%) for 10 min) was the most suitable as 97.78% of cleaned meristem tips survived. Meristem tips with different lengths were thermotherapy-treated for different durations. It was indicated that meristem tips of 0.5 or 1.0 cm and heat-treated at 37^oC for four weeks gave the highest percentage of meristems that were able to differentiate into micro-shoots. <b>Conclusion:</b> RT-PCR detection of ApMV and SLRV revealed that using thermotherapy-treatment, for 4 weeks, of 0.5 cm long meristem tips was successfully applied to eliminate the 2 viruses in 92 and 96% of regenerated plantlets, respectively.

Molecular Assessment of Genetic Stability Using CDDP and DNA-barcoding Assays in Long-term Micropropagated Rose Plant.

BACKGROUND AND OBJECTIVE: Roses are the world's best-known garden plants, established as ornamental plants cultivated for their blooms. Taif rose (Rosa damascena trigintipetala) refers to the Damascus Rose species and is regarded one of Taif Governorate's most significant financial goods, which produces an extremely fragrant commercially precious essential oil. The objective of current study was to assess the genetic stability of micropropagated Taif rose and to assess the usefulness of Conserved DNA Derived Polymorphism (CDDP) and DNA-barcoding genes such as; rpoC1 (chloroplast gene RNA polymerase1) in the detection of somaclonal variation. MATERIALS AND METHODS: Ten combinations of CDDP PCR primers were employed and the rpoC1 gene region was sequenced for mother plant (control) and micropropagated plantlets of Taif rose plant. RESULTS: Based on CDDP data, phylogenetic divergence indicated that the distinct specimens of Taif rose micro-propagated plantlets and control were genetically differentiated by a difference of 1% of genetic dissimilarity. Phylogenetic tree which developed using rpoC1 DNA showed that rpoC1 DNA sequencing discovered a genetic difference between the control and micro-propagated plantlets of Taif rose. CONCLUSION: Furthermore, CDDP and DNA barcoding using rpoC1 gene have demonstrated their usefulness in investigating the genetic history of Rosa species and their ability to explore genetic mutation.