High frequency plant regeneration in Indian Siris via cyclic somatic embryogenesis with biochemical, histological and SEM investigations

Saeed, Taiba; Shahzad, Anwar

doi:10.1016/j.indcrop.2015.07.060

Cited by 30 publications

(11 citation statements)

References 63 publications

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“…The embryo germination was enhanced and root initiation and conversion of embryos to complete plantlets was increased by the addition of GA 3 . The requirement of GA 3 to trigger germination of SE has also been reported in Magnolia obovata Thunberg (Kim et al 2007), Albizia lebbeck (Saeed and Shahzad 2015). In the present study, germination with enhanced root formation observed may be due to the endogenous level of plant growth regulators.…”

Section: Discussionsupporting

confidence: 83%

Assessment of genetic stability in somatic embryo derived plantlets of Pterocarpus marsupium Roxb. using inter-simple sequence repeat analysis

Tippani

Nanna

Mamidala

et al. 2018

Physiol Mol Biol Plants

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An efficient plantlet regeneration protocol using immature zygotic embryos (IZEs) via somatic embryogenesis has been developed in Pterocarpus marsupium Roxb. The regenerated plantlets were evaluated for their genetic stability. IZEs were incubated on Murashige and Skoog (MS) media augmented with 1.07-16.11 lM naphthalene acetic acid (NAA) or 0.90-13.97 lM 2,4-dichlorophenoxyacetic acid. The optimum callus induction (96.6%) was observed on MS medium augmented with 5.37 lM NAA. Induction of somatic embryos (SEs) was observed after subculture of calli on medium with decreased concentrations of NAA (0.54-5.37 lM), either alone or 2.69 lM NAA in combination with 2.22-8.90 lM benzyladenine (BA) or 2.32-9.30 lM Kinetin. Maximum number (33.4 ± 0.85) of SEs occurred on MS medium augmented with 2.69 lM NAA ? 4.40 lM BA ? 3% sucrose. Highest percentage (67.3 ± 0.37) of SEs matured and developed into cotyledonary stage by subsequent subculture on the same medium. SE formation and maturation decreased when sucrose concentrations were higher than 3%. Seventy percent of mature somatic embryos developed into plantlets on half strength MS medium augmented with 5.80 lM gibberellic acid. The various stages of development during somatic embryogenesis include globular, heart, torpedo and mature stages as revealed by the stereomicroscopic and histological studies of explants. Plantlets derived from SEs were successfully acclimatized in the greenhouse with a survival rate of 78%. Among the survived plantlets, 9 plantlets were randomly selected for inter-simple sequence repeat (ISSR) analysis. Of the 13 primers used, 8 produced reproducible and monomorphic bands. ISSR analysis revealed a homogenous amplification profile for all regenerated plantlets analyzed validating the genetic stability of somatic embryo derived plantlets.

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Section: Discussionsupporting

confidence: 83%

Assessment of genetic stability in somatic embryo derived plantlets of Pterocarpus marsupium Roxb. using inter-simple sequence repeat analysis

Tippani

Nanna

Mamidala

et al. 2018

Physiol Mol Biol Plants

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“…Moreover, sprouting probability was higher (∼0.25) in the somatic embryos of this work cultured with 10 μM Mtop compared to the rest of the concentrations evaluated. This aromatic cytokinin has been used to promote maturation of Albizia lebbeck somatic embryos at 7.5 μM and it has shown a reduction in the number of abnormal shoots of Barleria greenii compared to equimolar concentrations of BA (Amoo et al, 2010; Saeed and Shahzad, 2015). This is the first work known to the authors about the use of Mtop in papaya.…”

Section: Discussionmentioning

confidence: 99%

Explant Type, Culture System, 6-Benzyladenine, Meta-Topolin and Encapsulation Affect Indirect Somatic Embryogenesis and Regeneration in Carica papaya L.

2018

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A protocol to propagate papaya hybrid plants through indirect somatic embryogenesis was developed considering the effect of explant type, culture system, particular cytokinins and encapsulation, in different stages of the process. Optimal 2,4-dichlorophenoxyacetic acid (2,4-D) concentrations for non-embryogenic callus formation ranged between 9.0 and 27.1 μM in half-cut seeds, while higher concentrations were harmful. Non-embryogenic callus was also obtained with 22–158 μM 2,4-D from hypocotyl segments. Callus with embryogenic structures was only obtained in half-cut seeds cultured in the darkness on half-strength Murashige and Skoog culture medium supplemented with 2,4-D, while hypocotyl segments and isolated zygotic embryos failed to produce this type of callus regardless of the 2,4-D and sucrose (30 and 70 g l-1) concentrations tested in this study. Both, embryogenic callus development and quantity of somatic embryos formed per embryogenic callus, which ranged between 11 and 31 units after 14 months, required 2,4-D, but without any effect of the concentration. Histological studies confirmed the multicellular origin of the somatic embryos. In further steps, liquid medium induced over four times more somatic embryos than agar-gelled medium and showed significantly higher production of globular somatic embryos (85 vs. 57%). Both, 6-benzyladenine (BA) and meta-topolin (Mtop) stimulated sprouting (40–45%) of the somatic embryos (development of shoots only) in concentrations of up to 2.7 and 10 μM, respectively. Sprouting probability showed a 2nd order polynomial trend despite the range of concentration used for each cytokinin. This is the first report about the positive effect of Mtop on the apical shoot development of Carica papaya somatic embryos known to the authors. Radicle growth was observed in 5% or less of the cultivated embryos, regardless of the BA concentration. Finally, all encapsulation conditions tested (2.5, 3.5, and 4.5% sodium alginate, combined with 50 and 100 mM CaCl2) reduced sprouting of somatic embryos when compared to the non-encapsulated ones, whereas capsule hardness showed low correlation with embryo sprouting. Embryos were further cultivated until they became plantlets approximately 5 cm long. They were acclimatized and afterward planted in the field, where they flowered and produced fruit.

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“…Numerous studies have shown that the occurrence of plant embryoids is affected by factors such as genotypes, explants, hormone types and concentrations, metal ions, and other nutrients [9, 20, 21]. Plant callus can be induced to induce effective embryonic development only by determining the suitable culture conditions for plant embryoids.…”

Section: Discussionmentioning

confidence: 99%

Ecological occurrence and plant regeneration of embryoid of the endangered and endemic plantDysosma versipellisin China

Tan

Zhou

et al. 2018

Preprint

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In this study, the effective callus culture, somatic embryogenesis, and plant 9 regeneration system of Dysosma versipellis, which is an endangered and endemic plant in China, 10 were established under specific culture conditions. Using the D. versipellis leaves, petioles, and 11 roots as explants, DPS software orthogonal design method and SPSS Duncan's multiple range test 12 were used to investigate their effects of D. versipellis on callus formation, embryoid induction,13 and plant regeneration by adding different phytohormones. Results showed that leaves and 14 petioles were the most suitable materials in inducing callus. The effect of phytohormone on callus 15 formation followed the order of 2,4-dichlorophenoxyacetic acid (2,4-D)>thidiazuron (TDZ)> 16 kinetin>naphthylacetic acid (NAA)>2-ip. The best medium for callus formation was MS+2,4-D 1 17 mg/L+NAA 0.05 mg/L+TDZ 0.5 mg/L+2-ip 1 mg/L. The optimal medium to induce the 18 formation of granular callus embryoid was MS+0.5 mg/L 6-BA+0.1 mg/L NAA, and the 19 induction rate was 71.33%. The embryoid rooting and plant regeneration medium was MS+0.5 20 mg/L IBA+0.5 mg/L GA3. The optimal medium formula obtained in this study was suitable for 21 the rapid induction of callus, embryoid, and plant regeneration of D. versipellis under in vitro 22culture conditions. Further study on the action mechanism, signal regulation mechanism, and 23 artificial seed production of fungal elicitors affecting the accumulation of podophyllotoxin is 24 important. 25

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High frequency plant regeneration in Indian Siris via cyclic somatic embryogenesis with biochemical, histological and SEM investigations

Cited by 30 publications

References 63 publications

Assessment of genetic stability in somatic embryo derived plantlets of Pterocarpus marsupium Roxb. using inter-simple sequence repeat analysis

Assessment of genetic stability in somatic embryo derived plantlets of Pterocarpus marsupium Roxb. using inter-simple sequence repeat analysis

Explant Type, Culture System, 6-Benzyladenine, Meta-Topolin and Encapsulation Affect Indirect Somatic Embryogenesis and Regeneration in Carica papaya L.

Ecological occurrence and plant regeneration of embryoid of the endangered and endemic plantDysosma versipellisin China

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