ABSTRACT
This study was conducted to develop the protocol for artificial seed production of Stipagrostis pennata (Trin.) De Winter via somatic embryo encapsulation as well as test a temporary bioreactor system for germination and seedling growth. Embryogenic calli were encapsulated using sodium alginate and calcium chloride and then sowed in the Murashige and Skoog (MS) germination medium in in vitro cultures. The experiments were conducted as a factorial based on a completely randomized design with three replications. The treatments include three concentrations of sodium alginate (1.5%, 2.5%, and 3.5%), two ion exchange times (20 and 30 min), and two artificial seed germination media (hormone-free MS and MS supplemented with zeatin riboside and L-proline). Germination percentage and number of days needed until the beginning of germination were studied. The highest percentage of artificial seed germination was obtained when 2.5% sodium alginate was used for 30 min (ion exchange time) and when the seeds were placed on the MS germination medium supplemented with zeatin riboside and L-proline. The results of the analysis of variance in the temporary immersion bioreactor system showed that the main effects observed on the seedling growth were associated with different growth hormones in culture media and the number of feeding cycles. Experimental results also indicated that the total protein analyses of zygotic seedlings and seedlings originating from the synthetic seeds showed no statistically significant differences between these samples.
ABSTRACT
Salinity restricts seed germination and seedling growth through induction of osmotic and oxidative stresses. Therefore, this study aimed to enhance salinity tolerance in quinoa seed by pre-optimized osmo-priming treatments of CaCl2 (10 mM, 10 °C, 10 h) and KNO3 (150 mM, 5 °C, 24 h). The results showed that these treatments developed the cellular defense mechanisms in seeds as 'priming memory' that could improve the physiological and biochemical responses to salinity in post-priming stages. The germination capacity and seedling growth decreased with increasing salinity that was accompanied with a higher content of MDA and H2O2. However, the improvement of primed seed vigor against high salinity was explained by increasing the biological defense mechanisms including antioxidant enzymes (CAT, APX, SOD, GPX and PPO) and antioxidant metabolites (DPPH antioxidant activity, phenolics, flavonoids, ascorbic acid), particularly in presence of salt stress. In addition, Ca2+ and K+ priming acquired salinity tolerance in post-priming stages through a significant increase in the accumulation of proline, glycine-betaine, soluble carbohydrate. Improvement in homeostasis of K+/Na+ ratio by promoting K+ maintenance and Na+ exclusion was also found in post-priming stages. These observations may be utilized as effective methods in improving salinity tolerance of quinoa seed germination in saline agriculture by improving the antioxidant system, osmolyte accumulation and mineral nutrient homeostasis.
ABSTRACT
BACKGROUND: Stipagrostis pennata (Trin.) De Winter is an important species for fixing sand in shifting and semi-fixed sandy lands, for grazing, and potentially as a source of lignocellulose fibres for pulp and paper industry. The seeds have low viability, which limits uses for revegetation. Somatic embryogenesis offers an alternative method for obtaining large numbers of plants from limited seed sources. RESULTS: A protocol for plant regeneration from somatic embryos of S. pennata was developed. Somatic embryogenesis was induced on Murashige & Skoog (MS) medium supplemented with 3 mg·L-1 2,4-D subsequently shoots were induced on MS medium and supplemented with 5 mg·L-1 zeatin riboside. The highest shoots induction was obtained when embryogenic callus derived from mature embryos (96%) in combination with MS filter-sterilized medium was used from Khuzestan location. The genetic stability of regenerated plants was analysed using ten simple sequence repeats (SSR) markers from S. pennata which showed no somaclonal variation in regenerated plants from somatic embryos of S. pennata. The regenerated plants of S. pennata showed genetic stability without any somaclonal variation for the four pairs of primers that gave the expected amplicon sizes. This data seems very reliable as three of the PCR products belonged to the coding region of the genome. Furthermore, stable expression of GUS was obtained after Agrobacterium-mediated transformation using a super binary vector carried by a bacterial strain LBA4404. CONCLUSION: To our knowledge, the current work is the first attempt to develop an in vitro protocol for somatic embryogenesis including the SSR marker analyses of regenerated plants, and Agrobacterium-mediated transformation of S. pennata that can be used for its large-scale production for commercial purposes.
