Int.J.Curr.Microbiol.App.Sci (2019) 8(2): 2118-2123

International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 8 Number 02 (2019) Journal homepage: http://www.ijcmas.com

Original Research Article https://doi.org/10.20546/ijcmas.2019.802.245

Sphagnum Moss as Growing Medium in Phalaenopsis Orchid

M.M. Kaveriamma1*, P.K. Rajeevan2, D. Girija2 and K. Nandini2

1DST Young Scientist University of Agricultural and Horticultural Sciences College of Forestry, Kodagu, India 2Kerala Agricultural University College of Horticulture, Kerala, India

*Corresponding author

ABSTRACT

Phalaenopsis orchids occupy a top position in the international market as pot plant. Commonly used media for growing Phalaenopsis orchid are coconut husk chips, tree bark, fir bark, cocopeat and sphagnum moss. Growing medium is important as it provides

K e yw or ds anchorage, retains moisture, nutrients and aerates the roots. In addition to these functions,

a study was carried out to assess the possible growth promoting effects of sphagnum moss Phalaenopsis on orchids when used as growing medium. Eight month old hardened plants of Orchid, Klebsiella Phalaenopsis ‘Magic Kiss’ were used for the trial. They were planted in translucent plastic pneumoniae, Wilt, pots with three different growing media viz., coarse coconut husk chips, cocopeat (fine Moss coconut husk bits) and sphagnum moss in combination with charcoal and tile bits. Plants Article Info grown in sphagnum moss were significantly superior to those grown in coconut husk chips and coconut husk bits as media, both in terms of vegetative and floral attributes. Root cross Accepted: section of sphagnum moss grown plants revealed healthy velamen and their root 15 January 2019 Available Online: parenchyma tissues were rich in chlorophyll. From the moss surface, bacterial isolates of 10 February 2019 Bacillus thuringensis and B. aryabhattai were identified. Endophytic nitrogen fixing bacteria Klebsiella pneumoniae was isolated from leaves and roots grown in moss. Plants grown in moss had no incidence of wilt caused by Fusarium oxysporum. The isolates tested positive for IAA production.

Introduction Growing medium is important as it provides Phalaenopsis, commonly called the moth anchorage, retains moisture, nutrients and orchids is the most important commercial aerates the roots. In addition to these genus of orchids. Phalaenopsis are short functions, a study was carried out to assess stemmed monopodial orchids. Plants are slow the possible growth promoting effects of growing and mature plants attain an average sphagnum moss on orchids when used as height of twelve to fifteen centimeters, growing medium. In the Western Ghats of although a few individuals may grow taller India, epiphytic orchids are found growing in (Sahavacharin, 1981). abundance on trees covered with sphagnum moss. Sphagnum leaves are highly 2118

Int.J.Curr.Microbiol.App.Sci (2019) 8(2): 2118-2123 specialised; they form a special tissue of The plants were grown in the respective living, chlorophyll-containing chlorocytes and media along with charcoal and tile bits. dead cell content-free hyalocytes, which are Transluscent plastic pots were used as responsible for their huge potential to store container. Sphagnum moss growing as water. Galacturonic acid is rich in carboxylic parasites on coffee plants in Kodagu district acid groups that give Sphagnum its high nestled in Western Ghats were used. Coconut cation exchange capacity (Spearing, 1972). husk was collected locally, chipped into 2 Acidity generally retards microbial inches blocks, soaked in water for 48 hours to metabolism. Sphagnum species also produce remove tannins and later used. Commercial bioactive secondary metabolites influencing grade cocopeat blocks were used after microbial colonisation. Living Sphagnum soaking for 48 hours and draining out excess mosses are colonised in high abundances with moisture. Microbial load was analyzed in specific microorganisms, which fulfil media, in phyllosphere and rhizosphere for important functions like nutrient supply and plants grown in sphagnum moss and without pathogen defence for moss growth and health moss. It was done using serial dilution (Opelt et al., 2007). Tens of thousands of technique. Bacterial identification was done microbial species associate with plants, and using 16S rDNA sequencing at the plant–microbe interactions are crucial to plant Department of Microbiology, Kerala health (Lundberg et al., 2012). Microbes have Agricultural University. the potential to benefit plants through nutrient acquisition, disease suppression, and Results and Discussion modulation of host immunity (Mendes et al., 2011). Orchids exhibit a wide range of habitats, terrestrial and epiphytic plants occupying the Materials and Methods major share of plant genera. In the epiphytic orchids, a variety of media are used, the main The present study was carried out at the top functions of such media being, providing ventilated rain-shelter greenhouse of the All good aeration, holding moisture and allowing India Co-ordinated Floriculture Improvement good drainage. Media like tree bark, coconut Project at the Department of Pomology and husk, charcoal, brick pieces, tile bits etc are Floriculture, College of Horticulture, popular with epiphytic orchids. The media is Vellanikkara. The site is situated at a latitude also reported to be genus specific. Wang of 10°31' N and longitude of 76°13' E. The (2005) reported that, while growing area lies 22.25m above MSL. The phalaenopsis in containers filled with artificial experimental site enjoyed a humid tropical medium, important considerations are climate with maximum and minimum average aeration, capillary action, water and nutrient- temperatures of 36.59°C and 21.30°C during holding capacities, stability and weight of the the period of investigation. The mean relative medium components, as well as cost and humidity varied from 46.86 per cent to 78.70 consistency. In the present study, three per cent. The light intensities varied between different growing media were used namely 6000 and 11000 Lux. Eighteen months old coconut chips, cocopeat and sphagnum moss. plants of Phalaenopsis ‘Magic Kiss’ were Coconut husk chips and cocopeat are used to assess the influence of three different commonly used for growing orchids. There types of growing media, namely coconut husk have been reports on sphagnum moss being a chips, cocopeat and sphagnum moss. Fifteen good media for orchids in terms of good plants were included under each treatment. moisture holding capacity but not in terms of

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Int.J.Curr.Microbiol.App.Sci (2019) 8(2): 2118-2123 nutrient supply and microbial load. Although sphagnum moss could probably be the single moss from New Zealand is of higher quality, best material for growing young phalaenopsis many growers use Chilean moss because it is in warm (tropical and subtropical) conditions. less expensive. For the trial, high quality long threaded moss growing on trees and shrubs Flowering and floral characters especially from coffee plantations (where it is considered a parasite) from Kodagu in the A strong base of vegetative characters Western Ghats were used. They were superior resulted in better flowering characters of and very well suited for phalaenopsis plants grown in moss, compared to those growing. Vegetative and floral characters using coconut chip bits and coconut husk bits. were studied and also plant tissue and media Inflorescence emergence occurred at the were analysed for nutrients, chlorophyll earliest (124.90 days) in the treatment plants content and bacterial colonies (Table 1 and 2). where moss was used as the medium. Time taken for emergence of inflorescences was on Vegetative characters par when coconut husk chips and cocopeat were used with values 153.30 days and Good vegetative growth is an indication of the 155.90 days, respectively. Significant photosynthetic ability of plants. Among the differences were observed among treatments treatment plants, plants grown using in the number of inflorescences produced. sphagnum moss as media was found to have a Inflorescence count of 1.80 per year was significantly better vegetative growth. Plants recorded in plants grown in moss. This was were significantly taller, with an increase of significantly superior compared to coconut 1.64 cm when sphagnum moss was used as chips bits and cocopeat which recorded 0.90 medium. Plants grown in cocopeat and and 0.80, respectively and were on par. The coconut husk chips were on par with an inflorescence produced was significantly increase of 0.21 cm and 0.40 cm respectively. longer. Significant differences were observed Significant differences were observed with with regard to flowering duration among regard to leaf characters among treatments. treatments. It was maximum (112.70 days) in Leaf length, leaf breadth and leaf area were plants where moss was used and the values maximum with 16.72 cm, 9.87cm and 87.78 were on par when coconut husk chips and cm2, respectively, in plants where sphagnum cocopeat were used (73.40 and 70.10 days, moss was used. It was minimum in plants respectively). Inflorescence length recorded grown with cocopeat, recording 8.94 cm, 5.32 highest value (28.17 cm) in plants where cm and 42.00 cm2 which were on par with moss was used as media. It was significantly plants grown in coconut husk chips as superior to coconut husk chips and cocopeat growing medium. Remarkable differences as media wherein the length of inflorescence were noted with regard to the interval of was 14.80 cm and 14.03 cm, respectively. production of leaves in plants where Number of flowers per inflorescence was the sphagnum moss was used as the growing maximum (7.30) in sphagnum moss as medium, compared to the other two (Table 1). medium. The values were significantly New leaves were produced at an interval of superior to coconut husk chips and cocopeat 91.40 days in treatment with sphagnum moss, (3.30 and 2.10, respectively) which were on which was significantly superior to coconut par. Difference in flower size among chips (221.90 days) and cocopeat (260.50 treatments was not significant though flower days). This is in agreement with the studies of size was better in moss grown plants. Wang (2005) who reported that pure

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Table.1

Growing medium Plant Leaf Character Inflorescence Character

Height Length leaf Interval of Days for Infloresce- Flower area leaf emergence nce length count (cm) (cm2) production (cm) (number) (Days)

Sphagnum moss 1.64a 16.72a 87.78a 91.40a 124.90a 28.17a 7.30a

Coconut husk chips 0.40b 9.87b 39.30b 221.90b 153.30b 14.80b 3.30b

Cocopeat 0.21b 8.94b 42.00b 260.50b 155.90b 14.03b 2.10b

Table.2 Details of bacteria in Sphagnum moss and Phalaenopsis orchid grown without moss

NO. Name of Source Colony morphology Cell shape IAA production isolate (μg/ml) 1. M1-MM1 Moss -surface Circular, entire, G+ve 44 smooth, glistening rods 2. MD1 Moss -surface Rough G+ve 40 undulate chain 3. MY Orchid in presence of Yellow fluidal G-ve 72 moss-Surface of leaf & Convex, rods root ; endophytic in leaf Entire, circular 4. MW Orchid leaf surface Circular Entire, G+ve 30 (grown with moss) Smooth, glistening rods

5. LWMY Orchid leaf surface Yellow circular G-ve 47 (grown in coconut husk)

6. LWMW Orchid leaf surface White Circular, G-ve 64 (grown in coconut husk) transparent

7. Moss-red Moss- surface Red, Undulate G+ve NA (imprinting)

8 Moss- Surface of moss white undulate G+ve NA (imprinting)

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The endogenous growth regulators could grown in the other two media. In vitro studies possibly be responsible for the high growth with cowpea seeds indicated enhancement of rate in these plants. Hiller et al., (1979) germination percentage and seedling vigour. reported that the changes in endogenous Sachdev et al., (2009) report that six IAA gibberellin like activity were related with producing strains of Klebsiella significantly stem elongation, but not with floral initiation. increased root length and shoot height of Flower count was significantly superior in inoculated wheat seedlings over control. plants grown in moss. Stoessl and Arditti Bacillus thuringensis and B. aryabhatti were (1984) reported that orchids are known to recorded as well in plants grown in moss as produce various phenolic compounds and media. The plants grown using sphagnum phytoalexins, which were shown to suppress a moss had no incidence of disease. Plants number of different microorganisms. Plant grown in coconut based media recorded exudates also supply the rhizosphere with higher incidence of collar rot caused by tryptophan that is the main precursor in Fusarium oxysporum. Besides the provision microbial IAA biosynthesis (Kravchenko et of nurients, plant-associated bacteria are al., 2004). IAA producing bacteria transform important for supporting growth, health and it into auxin, increasing its exogenous level. stress resistance of plants. Nitrogen-fixing bacteria belonging to PGPB (Plant Growth Promoting Bacteria) can fix Acknowledgement atmospheric nitrogen and supply it to plants. Here we use the term PGPB as bacteria Authors are grateful to All India Co-ordinated including diazotrophic bacteria or plant Research Project in Floriculture for the growth-promoting rhizobacteria (PGPR). financial assistance received for the research PGPB can competitively colonize plant root, project. promote plant growth, and reduce plant diseases. In order to further understand the References superiority shown by sphagnum moss, the population and types of microflora (surface as Hiller, L., Kelly, W.C. and Powell, L.E. 1979. well as endophytic) associated with the three Temperature interactions with growth substrates were assessed. A Gram negative regulators and endogenous rod shaped bacterium that formed highly gibberellin-like activity during seed fluidal yellow colonies on nutrient agar was stalk elongation in carrots. Plant found to be the most predominant in moss. Physiol. 63: 1055-1061. This isolate fixes nitrogen, solubilizes Kravchenko, L.V., Azarova, T.S., Makarova, insoluble P and produces HCN. It also N.M. and Tikhonovich, I.A. 2004. The produces IAA from tryptophan, to the extent effect of tryptophan present in plant root of 72 µg ml-1 of the medium, in a period of exudates on the phytostimulating two weeks, in pure culture. This isolate was activity of rhizobacteria. Microbiol. 73: identified as Klebsiella pneumoniae by 16S 156–158. rDNA sequencing. The isolate was associated Lindow, S.E., and Brandle, M.T., 2003. with phalaenopsis, both in leaves and roots as Microbiology of the phyllosphere. lAppl endophytes and also on the surface of leaves, Environ Microbio. 2003 Apr; when the orchids were grown on moss. 69(4):1875-83. Population was more in leaf (203x 103cfu/ g) Lundberg DS, Lebeis SL, Paredes SH, than in roots (11x103cfu). The bacterium Yourstone S, Gehring J, Malfatti S, could not be detected when the plants were Tremblay J, Engelbrektson A, Kunin V,

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del Rio TG et al., 2012. Defining the growth. Ind. J Exp Biol. 47(12): 993- core Arabidopsis thaliana root 1000. microbiome. Nature 488: 86–90. Sahavachrin, O.1981. Induction of plantlets Mendes, R., Kruijt, M., De Bruijn, I., on inflorescence of Phalaenopsis by Dekkers, E., Van der Voort, M., application of N-6-benzyl adenine. Schneider, J. H. M., Piceno, Y. M., Kasetsart Journal of Natural science DeSantis, T. Z., Andersen, G. L., 15(2):54-64 Bakker, P. A. H. M., and Raaijmakers, Spearing, A.M., 1972. Cation-Exchange J. M. 2011. Deciphering the rhizosphere Capacity and Galacturonic Acid microbiome for disease-suppressive Content of Several Species of bacteria. Science 332:1097-1100. Sphagnum in Sandy Ridge Bog, Central Opelt, 2007 Investigations of the structure and New York State. The Bryologist 75(2): function of bacterial communities 154-158 associated with Sphagnum mosses. Stoessl, A. and Arditti, J. 1984. Orchid Environ Microbiol. 2007 Nov; 9(11): phytoalexins. In: Arditti J (ed) Orchid 2795-809. biology: reviews and perspectives. Sachdev, D.P., Chaudhari, H.G., Kasture, Cornell university press, Ithaca, NY. V.M., Dhavale, D.D. and Chopade, 151-175. BA.2009. Isolation and characterization Wang, Y.T. 2005. Phalaenopsis mineral of indole acetic acid (IAA) producing nutrition. ISHS Acta Horticulturae 878: Klebsiella pneumoniae strains from 7 rhizosphere of wheat (Triticum aestivum) and their effect on plant

How to cite this article:

Kaveriamma, M.M., P.K. Rajeevan, D. Girija and Nandini, K. 2019. Sphagnum Moss as Growing Medium in Phalaenopsis Orchid. Int.J.Curr.Microbiol.App.Sci. 8(02): 2118-2123. doi: https://doi.org/10.20546/ijcmas.2019.802.245

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