<i>In Vitro</i>Conservation Tactics for a Sweet Genotype of <i>Amygdalus scoparia</i>, an Endangered Medicinally Important Tree Species by Propagation Using Synthetic Seeds

Bud stems arisen from in vitro cultures of A. scoparia were encapsulated in calcium alginate pieces for short term stockpile and germplasm interchange. The maximum frequency (88.96%) of conversion of encapsulated nodal segments into plantlets and the highest node number (7.20) was performed on a murashige and Skoog’s medium (MS medium) containing 2.2 μM banzyl adenine (BA). The highest length of re-growing shoots was achieved when MS medium was supplied with 2.2 μM BA and 0.5 μM NAA. However, the number of shoots produced was higher (5.30 and 5.10) on MS medium supplemented with 2.2 μM BA and 0.5 μM NAA and MS medium with 2.2 μM BA, respectively than on the hormone-free media. Treatment with 19.6 μM IBA resulted in the highest conversion of encapsulated nodal segments into plantlets. The frequency of conversion (89.6% - 88.6%) was retained at 25?C for up to 2 weeks without significant change. The highest frequencies (61.1%) of plantlet formation from encapsulated nodal segments were obtained by transferring synthetic seeds onto peat mass and perlite (2:1) (v/v) mixture substrate. When transplanted into the peat mass and perlite (2:1) (v/v) mixture, these plantlets showed greater plantlet high, leaf number, shoot number and root number per plantlet than those of the other substrates. The synthetic seed technology offered a promising way for short term storage without refrigerating, germplasm conservation exchange for improvement and an alternative clonal propagation method for this endangered genotype of A. scoparia.

In Vitro Synthetic Seed Production of Potato under Different Fungicide Levels and Storage Intervals

Potato propagation through tubers and seed is an inefficient way of propagation thus we propose an alternative method in this study which is in vitro propagation.Plantlets were raised from True Potato Seeds(TPS)in test tubes containing 10 mL of Murashige Skoog(MS)medium.Subculturing was done after every 21 days and after four subsequent subculturing,when there was an adequate quantity of stock material,uninodal cuttings of about 3-4 mm in length were prepared from in vitro proliferated potato plants by excising leaves of 1-2 mm on each side of the node.Prepared uninodal cuttings were cultured on 1/2 strength MS medium for root primordia development.The selected uninodal cuttings with root primordia were encapsulated with sodium alginate solution(3.5%(w/v)),mixed with 1.25%charcoal and different concentration of fungicide(thiophanate methyl,sigma)to enhance the viability percentage of synthetic seed.Furthermore,these encapsulated uninodal cuttings were subjected to different storage intervals to check maximum storage.Results showed that optimum dose for fungicide is 150 mg/L as it resulted in good viability percentage,shoot number,shoot length,rooting percentage,root number,root length and ideal for bead formation with sodium alginate.Similarly,results revealed that encapsulated uninodal cuttings can be stored at a temperature of 4℃ up to 45 days with a survival rate of 63.33%.Encapsulated uninodal cutting stored for 60 days showed a poor viability percentage of about 43.03%.It can be concluded that optimum dose of 150 mg/L with storage time of 45 days should be used to get a better outcome for synthetic potato seed production.

Embryo Rescue via Artificial Seed Technique and Long-Term Preservation of Zephyranthes

Zephyranthes is valued as a native ornamental landscaping plant and a traditional medicinal herb. Due to the low seed viability, this study was carried out to evaluate the potential of seed embryo rescue using the artificial seed technique and long-preservation in Z. atamasca and Z. grandiflora. Seed embryos were selected for encapsulation with different concentration of sodium alginate (3%, 4%, and 5%) and calcium chloride (either 25, 50, 75, and 100 mM) followed by no encapsulated embryos as a control. The greatest viability of encapsulated embryos was 95% in Z. grandiflora and 85% in Z. atamasca with the combination of 4% sodium alginate and 100 mM calcium chloride after two weeks at 5°C. The highest viability with A490nm0.12 and A490nm0.16 were achieved when embryos were cultured in pretreatment medium with 30 g/L sucrose in Z. grandiflora and Z. atamasca, respectively. The highest viability by TTC assay after cryopreservation was observed with 54% viability for Z. grandiflora and 48% viability with Z. atamasca, after 2 h of dehydration. Rain lilies embryos were successfully preserved functioned as artificial seed and cryopreservation using encapsulation-dehydration method has been established for both species that can be used for other flower species with some modifications.