Assessment of genetic stability on in vitro and ex vitro plants of Ficus carica var. black jack using ISSR and DAMD markers

Parab, Ankita Rajendra; Lynn, Chew Bee; Subramaniam, Sreeramanan

doi:10.1007/s11033-021-06714-1

Cited by 15 publications

(4 citation statements)

References 37 publications

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“…Our results are consistent with a recent report 31 on genetic stability evaluation of in vitro raised Ficus carica var. Black Jack plantlets using woody plant medium containing 20 μM BAP + 8 μM IAA under different light treatments (normal fluorescent white light and four different LED spectra) that showed significant (97.87%) similarity using ISSR markers.…”

Section: Resultssupporting

confidence: 94%

“…Molecular markers have been frequently utilized for capturing the genetic variations induced during in vitro propagation or conservation of plants in form of discrete banding patterns. PCR-based single primer amplification reaction (SPAR) methods including random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) have been regularly used for genetic fidelity assessment of in vitro propagates 27 – 31 . The SPAR methods need no prior sequence information, are simple, economic, rapid and require small quantities of DNA 27 .…”

Section: Resultsmentioning

confidence: 99%

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Histological and molecular insights in to in vitro regeneration pattern of Xanthosoma sagittifolium

Bansal

Sharma

Singh

et al. 2023

Sci Rep

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A study on the effect of various phytohormonal combinations on in vitro propagation of Cocoyam [Xanthosoma sagittifolium (L.) Schott] was conducted to develop an improved and efficient in vitro regeneration protocol for its mass multiplication. Histological analysis to understand the in vitro regeneration pattern and genetic fidelity assessment of regenerated plants were also carried out. Single shoots excised from in vitro established cultures of X. sagittifolium were used as explants. Among the 32 different phytohormonal combinations tested, indirect organogenesis with intervening callus phase was observed on majority of the media combinations. Meristematic clump formation was optimally achieved on all the tested media combinations with maximum 43.54 ± 0.51 shoot primordia on MS medium containing 0.2 mg/L BAP + 0.1 mg/L NAA followed by 36.44 ± 0.76 shoot primordia on MS medium having 2.5 mg/L TDZ. Micro-morphological analysis of different morphogenetic structures revealed that the regeneration of cocoyam is well executed via meristematic nodules, shoot primordia formation that may evolve in to proper shoots. Adventitious shoots (> 2 cm) were successfully (100.00 ± 0.00%) rooted on the half-strength MS medium containing IBA (0.05–1.0 mg/L) and IAA (0.05–0.5 mg/L). The number of roots ranged from 0.78 ± 0.31 on the control half-strength MS medium to 13.94 ± 0.46 on half-strength MS supplemented with 1.0 mg/L IBA. Considering somaclonal variations as a potential restriction to in vitro multiplication of plants, genetic stability was assessed using 40 ISSR primers. The PCR amplification profiles obtained from all the tested propagules (calli, meristematic clumps, regenerated plantlets) were similar to the mother plants indicating the homogeneity of the individuals raised through the regeneration protocol reported here.

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Section: Resultssupporting

confidence: 94%

Section: Resultsmentioning

confidence: 99%

Histological and molecular insights in to in vitro regeneration pattern of Xanthosoma sagittifolium

Bansal

Sharma

Singh

et al. 2023

Sci Rep

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“…The biotechnological interventions in vitro may cause somatic variation [35,36], suggesting the need for genetic stability assessment [19,37]. The genetic stability is usually estimated via morphological identification and molecular marker assessments [19,[37][38][39]. In order to confirm the genetic stability of the present shipping system, we further compared the morphology and reproduction of recovered plantlets, which germinated in vacuum-packed microplates supplemented with 100 mg/L CCC.…”

Section: Discussionmentioning

confidence: 99%

In Vitro Techniques for Shipping of Micropropagated Plant Materials

et al. 2022

Horticulturae

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Shipping of in vitro micro-cuttings in tubes or jars is a frequently used method as the plants are more likely to quickly reproduce and comply with quarantine regulations in plant germplasm distribution. However, these containers are fragile during transportation. To diminish the risk associated with the long-distance shipping of in vitro plants, a safe and widely applicable packing and conservation technique based on microplate and slow growth was developed in this study. Potato cultivar ZHB and ginger cultivar G-2 were used to optimize the system with microplates (96 wells), vacuum-sealed packaging, and slow-growth techniques. Under regular culture conditions, packing in vacuum-sealed microplates reduced the survival of ZHB and G-2 micro-cuttings to 85.8% and 20.0%, respectively, and regeneration to 61.8% and 0%, respectively. Reducing the temperature to 10 °C maintained the survival of ZHB and G-2 micro-cuttings in the range of 83.3–100% after 60 days. Exposure to darkness decreased the survival of G-2 and inhibited regrowth. Thus, conservation in darkness at 10 °C is suggested. The effects of iron concentration and plant growth retardants were further assessed. The addition of 1/4 MS medium combined with 100 mg/L chlormequat chloride (CCC) resulted in full survival and growth inhibition of plantlets, without malformation identified. Finally, incubation with 1/4 MS medium supplemented with 100 mg/L CCC in vacuum-sealed microplates at 10 °C in the dark resulted in high survival and suppressed germination. Sweet potato HXS was incubated as well to test the broad-spectrum applications of the technique; 100% survival and 6.7% germination was gained. Morphological indices of released cuttings recovered to control levels after two cycles of subculture in MS medium. A 0.1–0.2% genetic variation was detected by SSR and ISSR, suggesting genetic stability of the conserved samples. Finally, micro-cuttings were safely transported to cities located thousands of kilometers away without package and sample damage. Our results enable easy distribution of in vitro plant germplasms.

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“…SCoT markers have been reported for assessing the genetic fidelity of several plant species such as Cleome gynandra [97], Bauhinia racemosa [81], and Rauwolfia tetraphylla [38]. Similarly, the use of RAPD and ISSR markers for assessing the genetic fidelity of Ficus carica micropropagated plants was reported by several studies [34,40,87,98]. However, this is the first report using RAPD, ISSR, and SCoT primers for the assessment of the genetic fidelity of micropropagated F. palmata plants.…”

Section: Grafting Compatibility Of Micropropagated F Palmata Plantlet...mentioning

confidence: 94%

Micropropagation and Genetic Fidelity of Fegra Fig (Ficus palmata Forssk.) and Grafting Compatibility of the Regenerated Plants with Ficus carica

Al-Aizari,

Dewir,

Ghazy

et al. 2024

Plants

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Ficus palmata is an important fig species that produces edible and nutritious fruit and possesses several therapeutic uses. This study reports an effective method for the micropropagation of F. palmata using nodal explants. In vitro shoots were cultured for 7 weeks onto MS medium fortified with different concentrations of cytokinins, light intensities, sucrose concentrations, and light/dark incubation treatments. Optimal axillary shoot proliferation (10.9 shoots per explant) was obtained on a medium containing 30 g/L sucrose and supplemented with 2 mg/L 6-benzylaminopurine (BAP) under 35 μmol/m2/s light intensity. Dark incubation limited the foliage growth but favored shoot elongation and rooting compared with light incubation. Elongated shoots, under dark conditions, were rooted (100%; 6.67 roots per explant) onto MS medium containing 1 mg/L indole-3-acetic acid (IAA) and 1.5 g/L activated charcoal. The micropropagated plantlets were acclimatized with a 95% survival rate. In this study, the genetic fidelity of micropropagated F. palmata clones along with their mother plant was tested using randomly amplified polymorphic DNA (RAPD), inter-simple sequence repeats (ISSR), and start codon targeted (SCoT) molecular markers. The genetic similarity between the micropropagated plantlets and the mother plant of F. palmata was nearly 95.9%, assuring high uniformity and true-to-type regenerated plants. Using micropropagated F. palmata plantlets as a rootstock proved appropriate for the grafting F. carica ‘Brown Turkey’. These findings contribute to the commercial propagation and production of the fig crop.

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Assessment of genetic stability on in vitro and ex vitro plants of Ficus carica var. black jack using ISSR and DAMD markers

Cited by 15 publications

References 37 publications

Histological and molecular insights in to in vitro regeneration pattern of Xanthosoma sagittifolium

Histological and molecular insights in to in vitro regeneration pattern of Xanthosoma sagittifolium

In Vitro Techniques for Shipping of Micropropagated Plant Materials

Micropropagation and Genetic Fidelity of Fegra Fig (Ficus palmata Forssk.) and Grafting Compatibility of the Regenerated Plants with Ficus carica

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