Abstract
To explore the potential of in vitro rapid regeneration, three varieties (Golpaygan-181, Orumieh-1763, and Gorgan-1601) of sainfoin (Onobrychis viciifolia Scop. syn. Onobrychis sativa L.) were evaluated. For the first time, an encapsulation protocol was established from somatic embryogenic callus in torpedo and cotyledonary stages to create artificial seeds. Callus derived from different concentrations of Kinetin (0–2.0 mg L−1) and Indole-3-acetic acid (0–2.0 mg L−1) was coated with sodium alginate and subsequently cultured either in Murashige and Skoog (MS) medium or in soil substrate. Adventitious shoots from synthetic beads developed into rooting in full and half strength MS medium supplemented with various concentrations of auxin and cytokinin. Prolonged water conservation of black and red soils (1 : 1) had the highest rate of survival plantlets in the acclimatization process. Diverse resistance techniques in Onobrychis viciifolia were evaluated when the plants were subjected to water deficiency. Higher frequency of epicuticular waxes was observed in in vivo leaves compared to in vitro leaves. Jagged trichomes nonsecreting glands covered by spines were only observed in the lower leaf side. Ultimately, stomata indices were 0.127 (abaxial), 0.188 (adaxial) in in vivo and 0.121 (abaxial), 0.201 (adaxial) in in vitro leaves.
Highlights
Despite the fact that sainfoin (Onobrychis viciifolia) is an important forage species, it has received little attention and assessment for in vitro studies
Leaf and stem explants of O. viciifolia were cultured in Murashige and Skoog (MS) media supplemented with different concentrations of Kinetin and IAA
Callus percentage was low in the control culture, MS medium supplemented with 1.5 mg L−1 Kinetin and 2 mg L−1 IAA had the highest percentage in both stem and leaf explants
Summary
Despite the fact that sainfoin (Onobrychis viciifolia) is an important forage species, it has received little attention and assessment for in vitro studies. Among the attributes of sainfoin, it improves soil fertility, where the environmental conditions limit the cultivation of alfalfa, and produces safe bloat forage. The progress of this species by genetic engineering techniques will contribute significant advantages for plant breeding objectives. A basic prerequisite of genetic engineering is advance of an efficient adventitious shoot regeneration system for the desired species. Rapid multiplication of shoot tips is notable to reduce the cost and genetic purity of micropropagated plants. Different auxins and cytokinins concentrations in MS medium play an important role in achieving a desired rate of multiple shoot formation. Ratio of regeneration depends on culture type, composition of the medium, and the variety used [1]
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