Articles
Influencing in vitro clonal propagation of aquatic plant, Cryptocoryne walkeri, by plant growth regulators
Article number
1455_23
Pages
189 – 200
Language
English
Abstract
Currently aquatic plants are gaining more popularity in traditional aquariums and water gardens.
Due to its economic value, this study aimed to establish a new and reproducible protocol for the in vitro propagation of aquatic plant, Cryptocoryne walkeri. We developed an efficient and cost-effective method for rapid and high-quality shoot multiplication and in vitro rooting of Cryptocoryne walkeri using shoot tip explants derived from mature plants.
A four-step method was developed to induce successful plant regeneration on MS medium augmented with suitable plant growth regulators (PGRs). Step 1 (initiation of aseptic cultures): the shoot tips were surface sterilized using 8% Clorox® (5.25% sodium hypochlorite, NaOCl) for 15 min, followed by rinsing three times with sterile distilled water.
They were again surface sterilized for another 4% Clorox® (5.25% sodium hypochlorite, NaOCl) for 5 min.
Step 2 (shoot multiplication): shoot tip explants (0.5 cm in length) of 7 days old in vitro were cultured on MS medium fortified with 3.0 mg L‑1 kinetin (KIN) with a maximum of 6.70±0.49 shoots per explant within 60 d of culture period.
Step 3 (in vitro rooting): shoots were then separated and transferred individually to MS medium supplemented with 1.0 mg L‑1 α-naphthalene acetic acid (NAA) for 30 days.
This medium yielded maximum number of roots (6.10±0.14 roots shoot‑1) with 9.95±0.97 mm average length.
Step 4 (hardening and field transfer of tissue culture raised plants): the 30-day rooted plantlets were hardened and established in pots containing a mixture of organic soil and sand (1:1) overlaid with tap water under greenhouse conditions for acclimatization for 30 days.
The acclimatization of in vitro raised plantlets was successfully undertaken with 100% survival rate in greenhouse conditions.
This complete study has successfully outlined a rapid, high frequency direct shoot organogenesis induction of an ornamental aquatic plant, Cryptocoryne walkeri from shoot tip explants inclusive of shoot proliferation, rooting, and acclimatization.
Due to its economic value, this study aimed to establish a new and reproducible protocol for the in vitro propagation of aquatic plant, Cryptocoryne walkeri. We developed an efficient and cost-effective method for rapid and high-quality shoot multiplication and in vitro rooting of Cryptocoryne walkeri using shoot tip explants derived from mature plants.
A four-step method was developed to induce successful plant regeneration on MS medium augmented with suitable plant growth regulators (PGRs). Step 1 (initiation of aseptic cultures): the shoot tips were surface sterilized using 8% Clorox® (5.25% sodium hypochlorite, NaOCl) for 15 min, followed by rinsing three times with sterile distilled water.
They were again surface sterilized for another 4% Clorox® (5.25% sodium hypochlorite, NaOCl) for 5 min.
Step 2 (shoot multiplication): shoot tip explants (0.5 cm in length) of 7 days old in vitro were cultured on MS medium fortified with 3.0 mg L‑1 kinetin (KIN) with a maximum of 6.70±0.49 shoots per explant within 60 d of culture period.
Step 3 (in vitro rooting): shoots were then separated and transferred individually to MS medium supplemented with 1.0 mg L‑1 α-naphthalene acetic acid (NAA) for 30 days.
This medium yielded maximum number of roots (6.10±0.14 roots shoot‑1) with 9.95±0.97 mm average length.
Step 4 (hardening and field transfer of tissue culture raised plants): the 30-day rooted plantlets were hardened and established in pots containing a mixture of organic soil and sand (1:1) overlaid with tap water under greenhouse conditions for acclimatization for 30 days.
The acclimatization of in vitro raised plantlets was successfully undertaken with 100% survival rate in greenhouse conditions.
This complete study has successfully outlined a rapid, high frequency direct shoot organogenesis induction of an ornamental aquatic plant, Cryptocoryne walkeri from shoot tip explants inclusive of shoot proliferation, rooting, and acclimatization.
Authors
S. Rittirat, S. Klaocheed, S. Kalawong, K. Thammasiri
Keywords
aquatic plant, micropropagation, plant growth regulators, plant tissue culture
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