Optimizing micropropagation of Indonesian conserved orchid Vanda celebica using organic compounds and a temporary immersion system

Vanda celebica is an orchid species endemic and protected in North Sulawesi, Indonesia. To overcome such difficulties, micropropagation methods have been widely used, incorporating modified media and culture systems to improve success rates. Therefore, this study aimed to evaluate the viability of...

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Main Authors: Ainna N. LAILY, Awang MAHARIJAYA, Dyah R. WULANDARI, Dwi M. PUSPITANINGTYAS, Siti R. ARIATI, Andri F. MARTIN, Betalini W. HAPSARI
Format: Article
Language:English
Published: AcademicPres 2024-12-01
Series:Notulae Botanicae Horti Agrobotanici Cluj-Napoca
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Online Access:https://www.notulaebotanicae.ro/index.php/nbha/article/view/14065
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author Ainna N. LAILY
Awang MAHARIJAYA
Dyah R. WULANDARI
Dwi M. PUSPITANINGTYAS
Siti R. ARIATI
Andri F. MARTIN
Betalini W. HAPSARI
author_facet Ainna N. LAILY
Awang MAHARIJAYA
Dyah R. WULANDARI
Dwi M. PUSPITANINGTYAS
Siti R. ARIATI
Andri F. MARTIN
Betalini W. HAPSARI
author_sort Ainna N. LAILY
collection DOAJ
description Vanda celebica is an orchid species endemic and protected in North Sulawesi, Indonesia. To overcome such difficulties, micropropagation methods have been widely used, incorporating modified media and culture systems to improve success rates. Therefore, this study aimed to evaluate the viability of dry seeds stored at -20 °C for five years and the micropropagation of their protocorms using organic compounds, enhanced by a temporary immersion system (TIS) bioreactor. The genetic similarity between regenerants and the parental plant was also assessed using inter-sequence simple repeats (ISSR) markers to support conservation efforts. The results showed that V. celebica seeds stored for five years retained viability, with germination rates ranging from 56.16% to 73.45%. The addition of 40 g L-1 potato puree to the germination media significantly enhanced protocorm development. For further proliferation, ½-MS media supplemented with 150 ml L-1 coconut water and 50 ml L-1 carrot juice was found to be the most effective, resulting in a protocorm-like body (PLB) induction rate of 2.22 per explant, with an initiation time of 14.43 weeks, a 96.54% survival rate, and 30.56% of explants successfully induced. Dividing stage-4 protocorms yielded the best response, producing an average of 4.54 induced explants with an initiation time of 8.18 weeks. The TIS bioreactor system showed optimal results for shoot regeneration, achieving 64.84% and 37.07% regeneration rates. Genetic analysis indicated a similarity coefficient of 0.74-0.89 between regenerants and the parental plant, suggesting a genetic diversity comparable to Vanda species in their natural habitat.
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publishDate 2024-12-01
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series Notulae Botanicae Horti Agrobotanici Cluj-Napoca
spelling doaj-art-1c712f5586d94a4abd09f9130f2d22d22024-12-20T18:14:18ZengAcademicPresNotulae Botanicae Horti Agrobotanici Cluj-Napoca0255-965X1842-43092024-12-0152410.15835/nbha52414065Optimizing micropropagation of Indonesian conserved orchid Vanda celebica using organic compounds and a temporary immersion systemAinna N. LAILY0Awang MAHARIJAYA1Dyah R. WULANDARI2Dwi M. PUSPITANINGTYAS3Siti R. ARIATI4Andri F. MARTIN5Betalini W. HAPSARI6Bogor Agricultural University, Faculty of Agriculture, Department of Agronomy and Horticulture, BogorBogor Agricultural University, Faculty of Agriculture, Department of Agronomy and Horticulture, Bogor; Bogor Agricultural University, Center for Tropical Horticulture Studies, BogorResearch Center for Applied Botany, National Research and Innovation Agency, BogorResearch Center for Applied Botany, National Research and Innovation Agency, BogorResearch Center for Biosystematics and Evolution-National Research and Innovation Agency, BogorResearch Center of Genetic Engineering, National Research and Innovation Agency, BogorResearch Center of Genetic Engineering, National Research and Innovation Agency, Bogor Vanda celebica is an orchid species endemic and protected in North Sulawesi, Indonesia. To overcome such difficulties, micropropagation methods have been widely used, incorporating modified media and culture systems to improve success rates. Therefore, this study aimed to evaluate the viability of dry seeds stored at -20 °C for five years and the micropropagation of their protocorms using organic compounds, enhanced by a temporary immersion system (TIS) bioreactor. The genetic similarity between regenerants and the parental plant was also assessed using inter-sequence simple repeats (ISSR) markers to support conservation efforts. The results showed that V. celebica seeds stored for five years retained viability, with germination rates ranging from 56.16% to 73.45%. The addition of 40 g L-1 potato puree to the germination media significantly enhanced protocorm development. For further proliferation, ½-MS media supplemented with 150 ml L-1 coconut water and 50 ml L-1 carrot juice was found to be the most effective, resulting in a protocorm-like body (PLB) induction rate of 2.22 per explant, with an initiation time of 14.43 weeks, a 96.54% survival rate, and 30.56% of explants successfully induced. Dividing stage-4 protocorms yielded the best response, producing an average of 4.54 induced explants with an initiation time of 8.18 weeks. The TIS bioreactor system showed optimal results for shoot regeneration, achieving 64.84% and 37.07% regeneration rates. Genetic analysis indicated a similarity coefficient of 0.74-0.89 between regenerants and the parental plant, suggesting a genetic diversity comparable to Vanda species in their natural habitat. https://www.notulaebotanicae.ro/index.php/nbha/article/view/14065conservation (SDG 15)germination; in vitro cultureISSRprotocorm-like bodyseed storage
spellingShingle Ainna N. LAILY
Awang MAHARIJAYA
Dyah R. WULANDARI
Dwi M. PUSPITANINGTYAS
Siti R. ARIATI
Andri F. MARTIN
Betalini W. HAPSARI
Optimizing micropropagation of Indonesian conserved orchid Vanda celebica using organic compounds and a temporary immersion system
Notulae Botanicae Horti Agrobotanici Cluj-Napoca
conservation (SDG 15)
germination;
in vitro culture
ISSR
protocorm-like body
seed storage
title Optimizing micropropagation of Indonesian conserved orchid Vanda celebica using organic compounds and a temporary immersion system
title_full Optimizing micropropagation of Indonesian conserved orchid Vanda celebica using organic compounds and a temporary immersion system
title_fullStr Optimizing micropropagation of Indonesian conserved orchid Vanda celebica using organic compounds and a temporary immersion system
title_full_unstemmed Optimizing micropropagation of Indonesian conserved orchid Vanda celebica using organic compounds and a temporary immersion system
title_short Optimizing micropropagation of Indonesian conserved orchid Vanda celebica using organic compounds and a temporary immersion system
title_sort optimizing micropropagation of indonesian conserved orchid vanda celebica using organic compounds and a temporary immersion system
topic conservation (SDG 15)
germination;
in vitro culture
ISSR
protocorm-like body
seed storage
url https://www.notulaebotanicae.ro/index.php/nbha/article/view/14065
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