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Biotechnological Importance of Piriformospora Indica Verma Et Al-A Novel Symbiotic Mycorrhiza-Like Fungus: an Overview

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Biotechnological Importance of Piriformospora Indica Verma Et Al-A Novel Symbiotic Mycorrhiza-Like Fungus: an Overview Indian Journal of Biotechnology Vol 2. January 2003. pp 65-75 Biotechnological Importance of Piriformospora indica Verma et al-A Novel Symbiotic Mycorrhiza-like Fungus: An Overview 2 3 Anjana Singh I, Archana Singh , Meera Kumari', Mahendra K Rai and Ajit Varma'", I School of Life Sciences, Jawaharlal Nehru University, ew Delhi 110067, India "Department of Biological Sciences. University of Alabama, Huntsville. AL 35899. USA 3 Department of Biotechnology, Amravati University, Amravati 444 602, India Piriforll/ospvra indica Verma et ai, a newly discovered root colonizing, AM fungi-like fungus, showed prominent positive influence on a wide range of plants of agriculture, forestry and flori-horticultural importance. Interestingly, P. indica has a wide host range of monocots and dicots including legumes, terrestrial o,~chids (Dactylorhiza lIIaculata) and members of the bryophytes (Aneura pinguis). The fungus showed potential as an agent for biological control of disease against soil-borne root pathogens. 32p experiments suggest that this fungus is important for phosphorus acquisition by the roots, especially in the arid and semi-arid regions. Mycelium could utilize a wide variety of inorganic and organic phosphate chemicals and produced acid phosphatases at the tip of the hyphae. The fungus was found to act as an excellent tool for biological hardening of tissue culture raised plants (tool for biological hardening). Fungus can be axenically grown on a wide range of synthetic simple and complex media with sucrose or glucose as carbon energy source. Mass cultivation of the fungus can be easily achieved on simplified broth culture. The growth is best obtained between 25-35°C and pH 5.8. The fungus was discovered from the rhizospheric soils of desert plants, Prosopis chilensis Stuntz and Ziziphus /lulI/lIIlllaria Burm. f. in the sandy desert of Rajasthan, North-west India. For its characteristic spore structure the isolate was named Piriforlllospora indica. Electron microscopy revealed the presence of typical doli pore septum with continuous parenthosomes, which indicated that the fungus belongs to the Hymenom)'cetes (Basidiomycota). Sequences of 188 rRNA and 28S rRNA indicate that P. indica is related to the Rhizocto/lia group and the family Sebacinaceae (Basidiomycetes). Immunofluorescence, ELISA, western blot and immuno-gold characterization indicated affinity of P. indica with the members of GlolI/eroll/ycota, namely Glolllerales, Diversisporales and Archeaosporales. Introduction (v) Ericoid mycorrhizae and (vi) The Australian lily Most terrestrial plants on earth have a symbiotic Thysanotus (Malia et aI, 2002). Here current association in their roots with soil fungi, known as hypotheses of phylogenetic relationships within mycorrhizae, which are beneficial to the growth and heterobasidiomycetous- Hymenomycetes wi th pal1icu lar health of plants and soil (Cruz et aI, 2002; Hodge et reference to Auriculariales is presented. The family aI, 2001; Jeffries & Barea, 2001; Rausch et aI, 2001). Sebacinaceae contained two genera, namely The following six different types of associations of Piriformospora and Sebacina (Weiss et aI, 2002). plants with mycorrhizae have been recognized: (i) AM fungi are the most widespread and probably Yesicular-arbuscular mycorrhizae (YAM or AM) most ancient symbionts in the world, found inmost (Smith, 1995; Walker, 1995), (ii) Ectomycorrhizae biomes and with most plant species. The co-evolution (ECM), (iii) Ectendo-, arbutoid-and monotropoid of the symbionts in this intimate relationship since mycorrhizal associations, (iv) Orchid mycorrhizae, 350 million years has involved a multitude of ecological, physiological and molecular interactions *Author for correspondence: enabling the formation of a partnership of mutual Tel: 26704511. 26107676 Ext-45 I I; Fax: 26187338, 26198234 benefit (Franken et aI, 2000; Kaldorf et aI, 1998). The E-mail: [email protected]@mail.jnu.ac.in partners in this association are members of Abbreviations: AM: arbuscular mycorrhiza; cDNA: complementary Basidiomycetes, Ascomycetes, Zygomycetes and these deoxyribonucleic acid; ECM: ectomycorrhizae; gmPGPRs: colonize most vascular plants belonging to gene'ically modified plant growth promoting rhizobacteria; Cryptogams, Gymnosperms and Angiosperms (Read. PGPRs: plant growth promoting rhizobacteria; Pitefl: plant 1999; Smith & Read, 1997). Mycorrhizal associations translation elongation factor; rRNA: rhibosomal ribonucleic acid; Ri T'-DNA: root inducing transfer DNA; YAM: vascular involve 3-way interactions between host plants. arbuscular mycorrhiza. mutualistic fungi and soil factors (Declerck et aI, 2000; Franken & Requena, 2001; Morton & Bruns, transformed as well as non-transformed roots, leading 2000; Morton & Redecker, 2001; Schuessler & to complete control of the life cycle of a few species Kluge, 2001). of AM fungi. There are also some reports of the The characteristic features of mycorrhizal enhancement of growth by in vitro culturable associations are summarized in Table 1. It is endophytes (Addy et ai, 2000; Dix & Webster, 1995; postulated that about 1.5 million fungi exist in nature, Froehlich et ai, 2000; Schulthess & Faeth, 1998). In however, only 0.7 million have been described to a nature, individual species infect plant species taxonomical status. Among them about 6000 \ belonging to different genera, families, orders and mycorrhizal species have been reported (Sutton, 1996; classes (Schuessler & Kluge, 2001). However. they Lilleskov et ai, 2002). do not establish symbiotic relationships with the AM fungi are ubiquitous, important for terrestrial species of some plant families, such as Brassicaceae, ecosystems and are obligate biotrophs (Harrison, Chenopodiaceae, Cyperaceae, ]unceaceae, Proteaceae 1999) exhibiting little host specificity (Bonfante, or with Lupinus spp (Gianinazzi-Pearson et ai, 1996; 2001). The colonization of plant roots by AM fungi Gollotte et ai, 1996). Non-mycorrhizal species and can greatly affect the pla:1t uptake of mineral genera have also been reported. in mycorrhizal nutrients. It may also protect plants from harmful families (Hirrel et ai, 1978; Trappe, 1987). Tester et elements in soil (Rufyikiri et ai, 2000). The potential al (1987) have given the details of the occurrence of of AM fungi for growth promotion of plants has been mycorrhizae in non-mycorrhizal families. well established (Azcon-Aguilar et ai, 1994; Bagyaraj Inoculum production of AM fungi presents a very & Varma, 1995; Morte et ai, 1996; Varma, 1995, difficult problem. These fungi do not grow like any 1998, 1999a; Varma & Schuepp, 1995). Mosse & other fungi, apart from with their hosts. Obligate Hepper (1975) were the first to produce a simplified symbiotic mode of growth, non-availability of pure in vitro system for the study of AM development culture and expensive means of production and their using excised roots in place of whole plants. Mugnier unreliability for the beneficial effects have greatly & Mosse (1987) modified the technique by using Ri jeopardized/undermined the mycorrhizal science. T-DNA transformed roots (hairy roots) as host tissue. Non-availability of authentic pure cultures on Becard & Piche (1992) presented an in-depth commercial scale is the greatest bottleneck in the evaluation of the root organ culture method and application of AM fungi in plant biotechnology. improved the procedures so that typical vesicular- However, mass production of several thousand viable arbuscular mycorrhiza can now be obtained on propagules of these fungi and their entrapment in Table I-Types of mycorrhizal associations AM ECM Ectendo- Arbutoid Monotropoid Ericoid Orchid Root structures Septate hyphae -(+) +- +- + + + + Hyphae in cells + -(+) + + + + + Hypha1 coils +- + + + + + Arbuscules + Fungal sheath -(+) -(+) + + Hartig net + + + + Vesicles +- Host plants Vascular plants Gymnosperms Ericales Monotropaceae Ericales Orchidaceae & Angiosperms Plant has + + + +- + +- chlorophyll Fungi Zygo-Glomales Most Basid-, but some Asco- and Zygo- Asco-(Basid- ) Basid- Note:- = absent, + = present, (+) = sqmetimes present, (-) = sometimes absent, +- = present or absent, Basid- = Basidiomycetes. Asco- = Ascomycetes, Zygo = Zygomycetes. c. f., Brundrett et aI, 1996. alginate beads has shown promise of large-scale In order to get a more precise idea about the closer application of AM fungi (Declerck et aI, 1996a,b, relati ves of P. indica, a part of 18S rRN A was 1998, 2000). amplified, sequenced and compared with corresponding data on a number of different Piriformospora indica-AM-like fungus Basidiomycota from GenBank. Sclerotinia sclerotia Verma et al (1998) have discovered a new plant (Ascomycota) and Glomus mosseae (Zygomycota) growth promoting fungus, Piriformospora indica were used as outgroups. Based on the results. a from the desert soils of North-west India. The fungus dendogram of the molecular phylogeny was grows on a wide range of synthetic and complex constructed (Fig. 3), which indicated the lowest media, e.g., minimal media, MM1, MM2, Moser B evolutionary distance of the 18S rRNA sequence of and Aspergillus (Kaefer, 1977) with 2% sucrose or the new fungus to members of the Rhizoctonia group glucose as a carbon and energy source. Young (Ceratobasidiales), namely Rhizoctonia solani Kuhn mycelia are white and almost hyaline, but conspicuous zonations (rhythmic growth) are observed in older cultures (Fig. 1a). The mycelium is mostly flat and submerged into the substratum. Hyphae are thin walled and of different
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