In vitro symbiotic seed germination of South Indian endemic orchid Coelogyne nervosa

Sathiyadash, Kullaiyan; Muthukumar, Thangavelu; Murugan, Shanmugaraj Bala; Ramalingam, Sathishkumar; Pandey, R. R.

doi:10.1016/j.myc.2013.08.005

Cited by 24 publications

(17 citation statements)

References 27 publications

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“…For characterization of symbiosis in orchid protocorms, many studies have employed observation of internal hyphae using a resin-embedded method [ 17 , 18 , 20 , 23 ] or a trypan blue-staining method [ 19 , 21 , 35 ]. The use of an improved ink-staining technique and crushing the protocorms allowed us to clearly and easily visualize the fungal hyphae in symbiotic protocorms and to count the total number of symbiotic cells containing pelotons by light microscopy (see Additional file 2 ).…”

Section: Resultsmentioning

confidence: 99%

“…In previous studies of symbiotic orchid protocorms, germination rate, developmental stage, size, and volume of protocorms have been used as evaluation criteria [ 7 – 16 ]. In addition, internal hyphal coils have also been observed to evaluate the symbiotic potential of orchid-mycorrhizal fungus [ 4 , 17 – 21 ]. In particular, Hadley and Williamson evaluated the number of pelotons and the volume of protocorms in the symbiosis of Dactylorhiza purpurella , revealing a positive correlation between the two parameters [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

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Quantitative evaluation of protocorm growth and fungal colonization in Bletilla striata (Orchidaceae) reveals less-productive symbiosis with a non-native symbiotic fungus

et al. 2017

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BackgroundIn nature, orchid plants depend completely on symbiotic fungi for their nutrition at the germination and the subsequent seedling (protocorm) stages. However, only limited quantitative methods for evaluating the orchid–fungus interactions at the protocorm stage are currently available, which greatly constrains our understanding of the symbiosis. Here, we aimed to improve and integrate quantitative evaluations of the growth and fungal colonization in the protocorms of a terrestrial orchid, Blettila striata, growing on a plate medium.ResultsWe achieved both symbiotic and asymbiotic germinations for the terrestrial orchid B. striata. The protocorms produced by the two germination methods grew almost synchronously for the first three weeks. At week four, however, the length was significantly lower in the symbiotic protocorms. Interestingly, the dry weight of symbiotic protocorms did not significantly change during the growth period, which implies that there was only limited transfer of carbon compounds from the fungus to the protocorms in this relationship. Next, to evaluate the orchid–fungus interactions, we developed an ink-staining method to observe the hyphal coils in protocorms without preparing thin sections. Crushing the protocorm under the coverglass enables us to observe all hyphal coils in the protocorms with high resolution. For this observation, we established a criterion to categorize the stages of hyphal coils, depending on development and degradation. By counting the symbiotic cells within each stage, it was possible to quantitatively evaluate the orchid-fungus symbiosis.ConclusionsWe describe a method for quantitative evaluation of orchid-fungus symbiosis by integrating the measurements of plant growth and fungal colonization. The current study revealed that although fungal colonization was observed in the symbiotic protocorms, the weight of the protocorm did not significantly increase, which is probably due to the incompatibility of the fungus in this symbiosis. These results suggest that fungal colonization and nutrition transfer can be differentially regulated in the symbiosis. The evaluation methods developed in this study can be used to study various quantitative aspects of the orchid-fungus symbiosis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-017-1002-x) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Quantitative evaluation of protocorm growth and fungal colonization in Bletilla striata (Orchidaceae) reveals less-productive symbiosis with a non-native symbiotic fungus

et al. 2017

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“…This might be caused by OMF differences of each orchid species, but also might be due to the small data volume of Sanger sequencing on C. viscosa , as well as the primer pairs chosen. In previous fungal studies of C. ovalis and Coelogyne nervosa , no member of Ceratobasidiaceae or Sebacinales was found ( Sathiyadash et al., 2014 ; Xing et al., 2019 ). In Tulasnellaceae, nine fungal OTUs of C. viscosa and two of C. ovalis were detected as OMF, and one was reported in a germination experiment of C. nervosa ( Sathiyadash et al., 2014 ; Xing et al., 2015 , 2019 ).…”

Section: Discussionmentioning

confidence: 84%

Similar mycorrhizal fungal communities associated with epiphytic and lithophytic orchids of Coelogyne corymbosa

Qin

Zhang

et al. 2020

Plant Diversity

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Mycorrhizal fungi are essential for the growth and development of both epiphytic (growing on trees) and lithophytic (growing on rocks) orchids. Previous studies indicate that in lowland tropical areas, orchid mycorrhizal fungal compositions are correlated with the life form (i.e., epiphytic, lithophytic, or terrestrial) of their host plants. We therefore tested if a similar correlation exists in an orchid distributed at higher elevations. Coelogyne corymbosa is an endangered ornamental orchid species that can be found as a lithophyte and epiphyte in subtropical to subalpine areas. Based on high-throughput sequencing of the fungal internal transcribed spacer 2 (ITS2)-rDNA region of mycorrhizae of C. corymbosa , we detected 73 putative mycorrhizal fungal Operational Taxonomic Units (OTUs). The OTUs of two dominant lineages (Cantharellales and Sebacinales) detected from C. corymbosa are phylogenetically different from those of other species within the genus Coelogyne , indicating that different orchid species prefer specific mycorrhizal fungi. We also found that the Non-metric multidimensional scaling (NMDS) plots of orchid mycorrhizal fungi were not clustered with life form, the variations among orchid mycorrhizal fungal communities of different life forms were not significant, and most of the OTUs detected from epiphytic individuals were shared by the lithophytic plants, suggesting that orchid mycorrhizal associations of C. corymbosa were not affected by life form. These findings provide novel insights into mycorrhizal associations with endangered ornamental orchids.

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“…This phenomenon is normal as orchid's seeds have a small embryo and lack of endosperm. In their natural habitat, orchids depend upon fungal infection for a supply of carbohydrates, nutrients and water for the seeds' development [19,20]. The supply of nutrients from the fungal colonization promotes seed germination, protocorm development and seedling growth in orchids [21].…”

Section: Discussionmentioning

confidence: 99%

Effect of Different Sound Genres on Seed Germination of and Orchids

Sulong¹,

Khalil²,

Dahari³

et al. 2016

Open Conf. Proceed. J.

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Auditory stress has been known to affect the development of plants. Recent findings have shown that the stress produced by music induced a positive effect on plants growth. However, different types of music have different kinds of stimulation on plant development. The aim of this study was to examine the in vitro seed germination of two Grammatophyllum species when exposed to five different music genres. The Grammatophyllum species used were G. hybrid and G. stapeliiflorum while the music genres were Instrumental, Rock, Hip hop, Yassin (Quranic reciting) and Ballad. Green capsules of Grammatophyllum hybrid and Grammatophyllum stapeliiflorum were obtained from Triang Botanical Valley, Pahang, Malaysia. Each group of seed cultures were exposed to different music genre for 8 hours starting from 9 am until 5 pm, each for a 6 months period. Seed cultures were kept in the dark for the first 3 months. One untreated (no music) group was kept as a control. All cultures were maintained on half-strength Murashige and Skoog (MS) media supplemented with 30 g/L sucrose, 0.5 mg/L 6-benzylaminopurine (BAP), 2 g/L peptone, 1 g/L activated charcoal and 2.5 g/L gelrite. Observations were recorded every month for the number of shoots emerged and shoot length. Data was analysed by using a one-way ANOVA. At the end of the experiment, it was found that music exposure had a positive effect on the seeds' germination as compared to the untreated control group. For G. hybrid species, the highest shoots number counted was 19.33±3.79 which was observed on seeds exposed to Yassin. The highest shoot length measured was 2.02±1.10 cm when it was exposed to Rocks music. In contrast, G. stapeliiflorum species showed the highest number of shoots of 12.00±2.64 when exposed to Ballad and the highest shoot length achieved by this species was 0.99±0.38 cm when it was exposed to Instrumental music. Analysis by using Kruskal-Wallis H and one-way ANOVA showed that p < 0.05 for all parameters. The findings showed that different species of orchids need a different type of music to influence the rate of its germination and growth.

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In vitro symbiotic seed germination of South Indian endemic orchid Coelogyne nervosa

Cited by 24 publications

References 27 publications

Quantitative evaluation of protocorm growth and fungal colonization in Bletilla striata (Orchidaceae) reveals less-productive symbiosis with a non-native symbiotic fungus

Quantitative evaluation of protocorm growth and fungal colonization in Bletilla striata (Orchidaceae) reveals less-productive symbiosis with a non-native symbiotic fungus

Similar mycorrhizal fungal communities associated with epiphytic and lithophytic orchids of Coelogyne corymbosa

Effect of Different Sound Genres on Seed Germination of and Orchids

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