Highlights
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Paphiopedilum villosum (Lindl.) Stein is an endangered epiphytic slipper orchid in Thailand.
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In vitro asymbiotic and symbiotic seed germination of P. villosum were investigated.
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The fungal isolate PVCP01 isolated from roots of P. villosum significantly increased the GRI and DRI of every protocorm developmental stage within 16 weeks.
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The fungal isolates PVCP01 were identified as Tulasnella sp. based on morphological characters and ITS sequence.
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This study firstly reported that Tulasnella sp. isolate PVCP01promoted seed germination and development of P. villosum in vitro.
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- Paphiopedilum villosum (Lindl.) Stein is an endangered epiphytic slipper orchid in Thailand.
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- In vitro asymbiotic and symbiotic seed germination of P. villosum were investigated.
- •
- The fungal isolate PVCP01 isolated from roots of P. villosum significantly increased the GRI and DRI of every protocorm developmental stage within 16 weeks.
- •
- The fungal isolates PVCP01 were identified as Tulasnella sp. based on morphological characters and ITS sequence.
- •
- This study firstly reported that Tulasnella sp. isolate PVCP01promoted seed germination and development of P. villosum in vitro.
Abstract
Paphiopedilum villosum
(Lindl.) Stein is a native epiphytic slipper orchid in Thailand. This
species is now being threatened and endangered. Propagation of this
species is essential for conservation and reintroduction purposes. In
this study, the propagation of P. villosum was achieved through the in vitro asymbiotic and symbiotic seed germination. Seeds of P. villosum
sown on asymbiotic media, Murashige and Skoog (MS), Vacin and Went (VW)
and Thomale GD (TH), did not germinate within 16 weeks. Seven different
fungal strains were isolated from roots of this orchid species. The
germination rate index (GRI) and the development rate index (DRI) of P. villosum
seeds in treatments inoculated with fungal isolates PVCP01, PVCP05, and
PVCP06 was significantly higher than uninoculated control treatments.
Fungal isolate PVCP01 significantly increased the GRI and DRI of every
stage of protocorm development, whereas fungal isolates PVCP05 and
PVCP06 were only able to promote seed germination and protocorm
development to stage 2. As for the wild orchid species, P. villosum,
a compatible fungus is therefore required for promoting seed
germination and protocorm development. Based on analysis of
morphological characters and sequences of the nuclear ribosomal
transcribed spacer (ITS), fungal isolates PVCP01, PVCP 05, and PVCP06
were identified as Tulasnella sp., Ceratobasidium sp., and Flavodon
sp., respectively. The information obtained from this study will be
used to propagate other threatened Thai orchids for conservation and
reintroduction programs.
Paphiopedilum villosum
(Lindl.) Stein is a native epiphytic slipper orchid in Thailand. This
species is now being threatened and endangered. Propagation of this
species is essential for conservation and reintroduction purposes. In
this study, the propagation of P. villosum was achieved through the in vitro asymbiotic and symbiotic seed germination. Seeds of P. villosum
sown on asymbiotic media, Murashige and Skoog (MS), Vacin and Went (VW)
and Thomale GD (TH), did not germinate within 16 weeks. Seven different
fungal strains were isolated from roots of this orchid species. The
germination rate index (GRI) and the development rate index (DRI) of P. villosum
seeds in treatments inoculated with fungal isolates PVCP01, PVCP05, and
PVCP06 was significantly higher than uninoculated control treatments.
Fungal isolate PVCP01 significantly increased the GRI and DRI of every
stage of protocorm development, whereas fungal isolates PVCP05 and
PVCP06 were only able to promote seed germination and protocorm
development to stage 2. As for the wild orchid species, P. villosum,
a compatible fungus is therefore required for promoting seed
germination and protocorm development. Based on analysis of
morphological characters and sequences of the nuclear ribosomal
transcribed spacer (ITS), fungal isolates PVCP01, PVCP 05, and PVCP06
were identified as Tulasnella sp., Ceratobasidium sp., and Flavodon
sp., respectively. The information obtained from this study will be
used to propagate other threatened Thai orchids for conservation and
reintroduction programs.
Keywords
- Orchid mycorrhiza;
- Mycorrhizal compatibility;
- Epiphytic orchid;
- Symbiotic germination;
- Endophyte
- Orchid mycorrhiza;
- Mycorrhizal compatibility;
- Epiphytic orchid;
- Symbiotic germination;
- Endophyte
