4.7 Article

M-brigde- and elicitor-assisted enhanced post-storage germination of Rauvolfia serpentina synthetic seeds, their genetic fidelity assessment and reserpine estimation

Journal

INDUSTRIAL CROPS AND PRODUCTS
Volume 180, Issue -, Pages -

Publisher

ELSEVIER
DOI: 10.1016/j.indcrop.2022.114732

Keywords

Alginate-encapsulaion; HPTLC; RAPD; Secondary metabolite; Silver nitrate; Storage

Funding

  1. Plant Tissue Culture laboratory at Regional Nuclear Agricultural Research Center

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The present study demonstrates that the germination efficiency of synthetic seeds of Rauvolfia serpentina can be improved by using filter paper soaked in (1/2)Murashige and Skoog liquid medium. The involvement of M-bridge resulted in the fastest germination and maximum proliferation. Storage at 8°C showed the highest germination efficiency and clonal fidelity.
The present study reports an amelioration of synthetic seed germination efficiency with a simple intervention of filter paper (M-bridge) drenched in (1/2)Murashige and Skoog liquid ((1/2)AMSL) medium in Rauvolfia serpentina. Synthetic seeds were developed from in vitro shoot tips using 3% (w/v) sodium alginate and 75 mM calcium chloride. Media for synthetic seed gemination were tested in cotton bed drenched with sterile water (SW), cotton bed drenched with (MSL)-M-1/2 medium, M-bridge drenched with (MSL)-M-1/2 medium, and (MS)-M-1/2 semisolid ((MSS)-M-1/2) medium alone. Synthetic seeds inoculated on (MSL)-M-1/2 or SW or (MSS)-M-1/2 media, exhibited slower germination and inferior performance; however, with the involvement of M-bridge, fastest germination (5.6 +/- 0.4 days) and maximum proliferation (4.8 +/- 0.7 shoots with 42.8 +/- 2.4 mm length, and 19.8 +/- 2.7 leaves/synthetic seed after 42 days of inoculation) were recorded. Among the tested elicitors (silver nitrate, salicylic acid and sodium nitroprusside) at their different concentrations supplemented in (MSL)-M-1/2 medium (with M-bridge), 50 mu M silver nitrate resulted in 100% germination frequency and resulted in germination within 5 days. Further, the synthetic seeds were stored for 42 days at -20 degrees C, 8 degrees C, and 25 degrees C. Maximum germination efficiency was obtained in case of seeds stored at 8 degrees C with respect to earliness in germination (6.9 +/- 0.3 days), germination percentage (71.8 +/- 3.4%) and subsequent proliferation (4.9 +/-; 0.6 shoots with 18.9 +/- 0.9 mm length, and 8.9 +/- 0.7 leaves/synthetic seed). Clonal fidelity analysis of the in vitro regenerants performed using RAPD markers exhibited monomorphic banding pattern. The reserpine content of the regenerants was found to be maximum following 42 days-storage at 8 degrees C(126.9 mu g/g), followed by a substantial decline in the synthetic seed-derived regenerants following 42-days-storage at 25 degrees C(120.7 mu g/g) or 20 degrees C(121.8 mu g/g).

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