Common Protocols in Orchid Micropropagation

Park, So‐Young; Huh, Yoon-Sun; Paek, Kee-Yoeup

doi:10.1007/978-1-4939-7771-0_8

Cited by 7 publications

(3 citation statements)

References 16 publications

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“…Organogenesis and somatic embryogenesis are both morphogenetic processes leading to plantlet regeneration. PLBs are generally considered “somatic embryos” of the orchids because of their similarities in morphology and cell wall composition [ 8 , 9 , 57 , 58 ]. To investigate whether PLB development of Paphiopedilum follows the somatic embryogenesis program, the expression patterns of the classic embryo markers including SERK , BBM , CLV , and WUS were examined.…”

Section: Discussionmentioning

confidence: 99%

Characterization of phytohormone and transcriptome profiles during protocorm-like bodies development of Paphiopedilum

et al. 2021

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Background Paphiopedilum, commonly known as slipper orchid, is an important genus of orchid family with prominent horticultural value. Compared with conventional methods such as tillers and in vitro shoots multiplication, induction and regeneration of protocorm-like bodies (PLBs) is an effective micropropagation method in Paphiopedilum. The PLB initiation efficiency varies among species, hybrids and varieties, which leads to only a few Paphiopedilum species can be large-scale propagated through PLBs. So far, little is known about the mechanisms behind the initiation and maintenance of PLB in Paphiopedilum. Results A protocol to induce PLB development from seed-derived protocorms of Paphiopedilum SCBG Huihuang90 (P. SCBG Prince × P. SCBG Miracle) was established. The morphological characterization of four key PLB developmental stages showed that significant polarity and cell size gradients were observed within each PLB. The endogenous hormone level was evaluated. The increase in the levels of indoleacetic acid (IAA) and jasmonic acid (JA) accompanying the PLBs differentiation, suggesting auxin and JA levels were correlated with PLB development. Gibberellic acid (GA) decreased to a very low level, indicated that GA inactivation may be necessary for shoot apical meristem (SAM) development. Comparative transcriptomic profiles of four different developmental stages of P. SCBG Huihuang90 PLBs explore key genes involved in PLB development. The numbers of differentially expressed genes (DEGs) in three pairwise comparisons (A vs B, B vs C, C vs D) were 1455, 349, and 3529, respectively. KEGG enrichment analysis revealed that DEGs were implicated in secondary metabolite metabolism and photosynthesis. DEGs related to hormone metabolism and signaling, somatic embryogenesis, shoot development and photosynthesis were discussed in detail. Conclusion This study is the first report on PLB development in Paphiopedilum using transcriptome sequencing, which provides useful information to understand the mechanisms of PLB development.

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Section: Discussionmentioning

confidence: 99%

Characterization of phytohormone and transcriptome profiles during protocorm-like bodies development of Paphiopedilum

et al. 2021

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“…In vitro micropropagation is very common in orchid propagation because of the complexity in vegetative and sexual propagations. Several in vitro techniques are followed for orchid micropropagation, and propagation by PLB (protocorm-like-body) is one of them [3][4][5]. In vitro regeneration of PLB was studied in different Dendrobium sp.…”

Section: Introductionmentioning

confidence: 99%

Low Concentration of Anti-Auxin and Anti-Fungal Agent Accelerates the PLB Regeneration of Dendrobium okinawense under Green LED

Mumivand

Shimasaki

Mehraj

2022

Plants

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Dendrobium okinawense is an endangered epiphytic orchid, and there has been no scientific report so far on its propagation. Protocorm is a mass of cells, and protocorm-like bodies (PLBs) are lookalike protocorms produced by vegetative explants in vitro. Regeneration of PLBs is a widely used technique for orchid micropropagation. We used different light-emitting diodes (LEDs) for the PLB regeneration of D. okinawense. The number of PLBs and fresh weight were increased by 81.1% and 80.8%, respectively, under green LED over the white fluorescent (WF) light. We added different concentrations of PCIB (p-Chlorophenoxyisobutyric acid, an anti-auxin) and HMI (3-Hydroxy-5-methyl isoxazole, an anti-fungal agent) in culture media. The number of PLBs was increased in media having 0.01 mg/L of PCIB (35.9%) compared to control (no PCIB), whereas 19.3% increased in media having 0.01 mL/L of HMI compared to control (no HMI). Green LED in combination with 0.01 mg/L of PCIB significantly increased the number of PLBs (69.0%) compared to the WF–without PCIB combination. In LEDs-PCIB and LED-HMI combinations, HMI did not show better PLBs regeneration compared with PCIB. The results suggested that a combination of low concentrations of PCIB and green LED have the potential to accelerate PLB regeneration of D. okinawense.

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“…In vitro micro-propagation is very common in orchid regenerations because of its complexity in vegetative and sexual propagations. Several in vitro techniques are followed for orchid micro-propagation, and propagation by PLB (protocorm-like-body) is one of them [1][2][3]. In vitro organogenesis of PLB was studied in different Dendrobium sp.…”

Section: Introductionmentioning

confidence: 99%

Low Concentration of Putative Anti-Auxin and Anti-Fungal Agent Accelerates the PLB Organogenesis of <em>Dendrobium okinawense</em> Under Green LED

Mehbub¹,

Shimasaki²,

Mehraj³

2021

Preprint

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Dendrobium okinawense is an endangered epiphytic orchid, and there has been no scientific report so far on its propagation. Protocorm is mass of cell, and protocorm-like bodies (PLBs) look alike protocorm produced by vegetative explants in vitro. Regeneration of PLBs is the most efficient technique for the orchids micro-propagation. We used different light emitting diodes (LEDs) for the efficient PLB organogenesis of D. okinawense. PLBs regenartion under green and red LED surpassed respectively 81.1% and 71.6% in numbers, and respectively 80.8% and 57.8% in fresh weight over white fluorescent light. We manipulated the culture media by different concentrations of PCIB and HMI. PLBs organogenesis promoted by low concentration, it increased respectively 35.9% and 19.3% over control by 0.01 mg/L PCIB and 0.01 ml/L HMI in numbers. Green LED and PCIB independently produced mostly similar numbers of new PLBs. Interestingly, culture media with 0.01 mg/L of PCIB further increased 8.5% of the numbers of PLBs under green LED, whereas the culture media with 0.01 mg/L HMI reduced the number of PLBs under green LED. PLBs culture under green LED with very low concentrations of PCIB manipulated culture media can significantly increase their organogenesis of D. okinawense.

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Common Protocols in Orchid Micropropagation

Cited by 7 publications

References 16 publications

Characterization of phytohormone and transcriptome profiles during protocorm-like bodies development of Paphiopedilum

Characterization of phytohormone and transcriptome profiles during protocorm-like bodies development of Paphiopedilum

Low Concentration of Anti-Auxin and Anti-Fungal Agent Accelerates the PLB Regeneration of Dendrobium okinawense under Green LED

Low Concentration of Putative Anti-Auxin and Anti-Fungal Agent Accelerates the PLB Organogenesis of <em>Dendrobium okinawense</em> Under Green LED

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