Centre for Mycorrhizal Culture Collection

Development of green cover at solid waste dumping site of a soda lime industry by mycorrhizal technology
A n oop S in g h, R een a S in g h, an d A lok A d holeya C e n tr e fo r M y c o r r h iz a l R e s e a r c h , T E R I , D a r b a r i S e th B lo c k , I n d ia H a b ita t C e n tr e , L o d h i R o a d , N e w D e lh i – 1 1 0 0 0 3 , I n d ia

I ntroduction
S o il m ic ro o rg a n is m s a re im p o r ta n t in th e re c o v e r y o f d is tu rb e d a n d p o te n tia lly to x ic e n v iro n m e n ts b e c a u s e th e y p ro d u c e p la n t g ro w th s tim u la tin g s u b s ta n c e s s u c h a s h o r m o n e s a n d v ita m in s , im m o b iliz e h e a v y m e ta ls in th e s o il, b in d s o il p a r tic le s in to s ta b le a g g re g a te s w h ic h im p ro v e s o il s tr u c tu re , re d u c e e ro s io n p o te n tia l, a n d c a n c o n tr ib u te to n u tr ie n t a v a ila b ility to p la n ts ( G a d d 1 9 9 3 ; S h e tty , H e tr ic k , F ig g e et al. 1 9 9 4) . T h e A M F ( a rb u s c u la r m y c o r rh iz a l fu n g i) a re a n im p o r ta n t c o m p o n e n t o f th e s o il m ic ro b ia l b io m a s s. T h e s y m b io s is is m u tu a lis tic b a s e d o n b i-d ire c tio n a l n u tr ie n t tra n s fe r b e tw e e n th e s y m b io n ts. T h e p la n t b e n e fits p a r tic u la rly th ro u g h e n h a n c e d p h o s p h o r u s , a n d w a te r a n d m in e ra l n u tr ie n t u p ta k e ( S m ith a n d R e a d 1 9 8 1 ) , w h ic h o fte n re s u lts in b e tte r g ro w th . T h e A M F c a n p ro te c t p la n ts a g a in s t th e to x ic e ffe c ts o f e x c e s s iv e c o n c e n tra tio n s o f h e a v y m e ta ls ( H e g g o , A n g le , a n d C h a n e y et al. 1 9 9 0 ; M a rsc h n e r 1 9 9 5 ). W a s te la n d s a re th e d ra s tic a lly d is tu rb e d la n d s w h e re n a tiv e v e g e ta tio n a n d a n im a l c o m m u n itie s h a v e b e e n re m o v e d a n d th e to p s o il h a s b e e n lo s t, a lte re d , o r b u r ie d . E x a m p le s o f w a s te la n d s a re s u rfa c e m in e d la n d s fo r m in e ra ls ( c o a l, p a u lin e , p h o s p h a te , b a u x ite , g ra v e l, s a n d , e tc .) , m in in g w a s te s , o rg a n ic /in o rg a n ic s o lid w a s te d u m p in g s ite s o f v a r io u s in d u s tr ie s , to p s o il re m o v e d fo r u s e e ls e w h e re c re a tin g ‘b u r ro w p its ’, s a lin e s o ils , d e g ra d e d m a te r ia l, a n d w in d a n d w a te r e ro d e d s ite s. N a tu ra l re h a b ilita tio n o f s u c h la n d s is a g ra d u a l p ro c e s s . A M ( a rb u s c u la r m y c o r rh iz a ) is a p o te n tia l to o l fo r re c la m a tio n o f w a s te la n d s o w in g to its b e n e fic ia l e ffe c ts. S u c h a s s o c ia tio n in te r m s o f th e ir g ro w th ra te , to le ra n c e a g a in s t b io tic a n d a b io tic s tre s s e s , a n d s e e d lin g s u r v iv a l b e n e fit th e p la n t s p e c ie s. T h e s e m ic ro o rg a n is m s p ro v id e n u tr itio n to th e p la n t b y s e q u e s te r in g th e n u tr ie n ts fro m th e s o il a n d tra n s lo c a tin g th e m to th e p la n t, a n d , in re tu r n , g e t c a rb o n fro m th e p la n ts. T h is m a k e s th e u tiliz a tio n o f th e n u tr ie n ts h ig h ly e ffic ie n t a n d re d u c e s th e d e p e n d e n c e o n e x te r n a l c h e m ic a l in p u ts. T h e re fo re ,

th e m y c o r rh iz a l te c h n o lo g y o ffe r s a b io lo g ic a l m e a n s o f a s s u r in g p la n t p ro d u c tio n a t a lo w c o s t w ith o u t c h e m ic a l fe r tiliz e r s. P la n ts c a n b e u s e d in th e re m e d ia tio n o f c o n ta m in a te d s o ils. In fa c t, p la n ts h a v e m e c h a n is m s fo r a c c u m u la tio n , to le ra n c e , o r a lle v ia tio n o f h ig h le v e ls o f h e a v y m e ta ls in c o n ta m in a te d s o il ( K h a n , K u e k , C h a u d h r y et al. 20 0 0 ) . U n d e r fie ld c o n d itio n s , d iffe re n t p la n t s p e c ie s liv e to g e th e r a n d h y p h a e o f A M F in te rc o n n e c t th e ro o t s y s te m s o f a d ja c e n t p la n ts , c h a n g in g th e le v e l o f A M c o lo n iz a tio n . A M h y p h a e c a n m e d ia te n u tr ie n t tra n s fe r b e tw e e n p la n ts ( B e th le n fa lv a y , S c h re in e r, M ih a ra et al. 1 9 9 6 ; O c a m p o 1 9 8 6 ). T h e s tu d y d e s c r ib e d in th is p a p e r fo c u s e d o n th e c o n tr ib u tio n o f A M s y m b io s is to re c la im th e s o lid w a s te g e n e ra te d b y s o d a a s h in d u s tr y a n d d e v e lo p a g re e n c o v e r b y g ro w in g d iffe re n t p la n t s p e c ie s.

M aterials and methods
T h e p re s e n t s tu d y w a s c a r r ie d o u t in th e p re m is e s o f T a ta C h e m ic a l L td , M ith a p u r, w h ic h is lo c a te d in th e D w a rk a s u b -d iv is io n o f G u ja ra t s ta te o n th e w e s t c o a s t o f In d ia . T a ta C h e m ic a ls p ro d u c e s 240 0 T P D ( to n n e s p e r d a y ) s o d a a s h , 1 5 0 0 T P D o f v a c u u m -e v a p o ra te d s a lt, a n d 3 3 o th e r p ro d u c ts , a n d g e n e ra te a h u g e a m o u n t o f s o lid w a s te . T h e s e s o lid w a s te s a re h ig h ly a lk a lin e in n a tu re , w ith a la rg e q u a n tity o f s a lts , a n d v e r y lo w in n itro g e n , p h o s p h o r u s , a n d p o ta s s iu m (T a b le 1 ) . P la n t s p e c ie s v iz . d e s i b a b o o l, ra m b a b o o l, c a s u r in a , a n d p a ra s p a p a l w e re s e le c te d fo r p la n ta tio n a t a s o lid w a s te d u m p in g s ite o n to p o f th e r id g e s ( 1 .0 m b a s e , 0 .7 5 m h e ig h t, 0 .7 5 m to p ) . P la n ts w e re p la n te d in 0 .45 m , 0 .45 m , 0 .45 m s iz e p its , u s in g 3 k g a e ro b ic c o m p o s t a lo n g w ith 20 0 p ro p a g u le s / p la n t. T h e d r ip s y s te m w a s in s ta lle d fo r ir r ig a tio n a n d 2 lite r s o f w a te r w a s p ro v id e d d a ily to e a c h p la n t. S o il s a m p le s w e re ta k e n b e fo re p la n ta tio n a n d a fte r o n e y e a r o f p la n ta tio n , a n d w e re a n a ly s e d fo r p H , E C , O C ( o rg a n ic c a rb o n ) , n itro g e n , p h o s p h o r u s , a n d

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Tabl 1 Changes in physico-chemical characteristics of e solid waste before and after plantation.
Parameters Initial Desibabool After one year Rambabool Casurina Paras pipal 7.26 62.40 2.62 0.21 29.12 161.54 7.15 7.35 23.65 65.45 2.75 2.61 0.24 0.37 38.65 37.15 175.65 165.45

pH 11.45 EC (m s/cm ) 74.10 OC (% ) 1.02 N (% ) 0.01 P (m g/kg) 7.64 K (m g/kg) 68.00

7.35 53.25 2.56 0.75 39.62 318.65

N – nir t ogen;P – phosphor K – pot um us; assi

potassium by following standard methods. The height and girth of plants at 37 cm height was observed after one year.

Results and discussion
The Mycorrhiza biofertilizer is necessary for augmenting the nutritional capabilities of the plant roots. Bio-inputs at various stages of greening operation of wasteland overburdens can provide additional benefits. For example, decomposition of green vegetation serving as green manure can be quickened by specific bio-inputs. Mycorrhiza has benefited both the plants and the soil. Plant benefits include augmentation of the supply of phosphorus and trace elements (iron, boron, zinc, copper, etc.), protection of plant roots from root diseases, high soil temperatures, and high salt concentrations, amongst others. Fungal elements in mycorrhiza biofertilizer bind soil particles, improve their aggregating capabilities, stabilize soil aggregates, and check leaching of important elements. Fungal elements also free the solids of the heavy metals by absorbing them and binding them in their cell walls. As symbionts of plant roots, AMF (arbuscular mycorrhizal fungi) are critical components of soil microbial communities which influence above-ground productivity and plant community development (Sanders, Clapp, and Wiemken 1996; van der Heijden, Klironomos, Ursic et al. 1998), confer improved water relations (Allen and Allen 1986; Neumann and George 2004), increase nutrient uptake and stress tolerance (Lapointe and Molard 1997), and assist in stable aggregate formation and enhanced carbon and phosphorus dynamics in the rhizosphere (Coleman and Crossley 1996; Sanders, Clapp, and Wiemken 1996; Jeffries, Gianinazzi, Perotto et al. 2003). In the present study, after one year of plantation, the pH of solid waste is reduced up to 7.15, and salts are also reduced in a great quantity (Table 1). Increase in organic carbon, nitrogen, phosphorus, and potassium content was also observed with each plant species used

in the experiment. It might be due to improve in aggregating capabilities and reduction in leaching of important elements by mycorrhiza. Medina, V assileva, Barea et al. (2006) reported that mycorrhizal treatment resulted in an increase of essential nutrients and a decrease of the contaminant metal. The mycorrhizal colonization in amended soils had an enhancing effect for specific absorption rates of nutrients (nitrogen, phosphorus, and potassium) and a non-significant effect for the pollutant mineral studied (zinc). This selective and contrasting AM effect for nutrients and pollutant acquisition indicates the benefit of this symbiosis under contaminated conditions (Medina, V assileva, Barea et al. 2006). In fact, mycorrhizal plants in amended soil have an increased amount of nutrients (nitrogen, phosphorus, and potassium) absorbed per unit of root mass (Koide 1993). Figure 1 clearly showed the green cover on the solid dumping site after one year of plantation of four different plant species with mycorrhizal technology. All plant species got their average height and stem girth (Table 2). In an experiment, Bi, Li, Christie et al. (2003) reported that root colonization by AMF gave higher maize yields, higher plant uptake of most of the

A

B

Figure 1 (A) Site at the time of plantation and (B) after completion of one year

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Table 2 Average height and girth of plants grown at solid waste dumping site.
Height Number of plants 14 5 172 438 Average height (feet) 5.3 5.2 5.8 5.4 Girth at 37 cm

Plant species Desibabool Rambabool Casurina Paras pipal

M aximum M inimum Average M aximum M inimum height height girth girth girth (feet) (feet) (cm) (cm) (cm) 7.4 9.0 11.2 10.2 3.10 2.11 1.00 1.10 6.5 8.2 12.7 12.0 14 16 20 20 2 4 2 3

nutrients studied, and may have protected the plants from excessive accumulation of sodium in the shoots when grown in soil overlying fly ash. Mycorrhizal colonization represents an energetic cost to the host plant in the form of carbon supplied to the mycosimbiont (Douds, Pfeffer, and Shachar-Hill 2000). This cost could be compensated by the functionality of this symbiosis since AMF absorbs photosynthates, but nutrients are supplied in return (Medina, Vassileva, Barea et al. 2006). The establishment of mycorrhiza in disposed solid waste by the chemical industry may require the addition of AMF strains more suited for such an environment. selection, culture, and inoculation of soil organisms highly adapted to the types of plants, and soil conditions of a severely disturbed landscape may be an effective means for ensuring success of the re-vegetation effort. This study indicates that successful growth of different plant species is possible in solid waste generated from chemical industries and can be improved by colonization of the plant roots by mycorrhizal fungi. Thus, remediation of areas infilled with solid waste may be possible using either undisturbed soil containing viable propagules of indigenous AMF or disturbed soil inoculated with effective strains of AMF. The fungi can assist plants in the exploitation of soil and solid waste’s nutrients and may help them to resist excessive salt (sodium) accumulation. AMF may also contribute to the re-establishment of a general soil microflora, and of a sustainable agricultural system combined with the appropriate use of fertilizers.

Bi Y L, Li, X L, Christie P, Hu Z Q , and Wong M H. 2003 G r o w th a n d n u tr ie n t u p ta k e o f a r b u s c u la r m y c o r r h iz a l m a iz e in d iffe r e n t d e p th s o f s o il o v e r ly in g c o a l fly a s h Chemosphere 5 0 : 863–869 Coleman D C and Crossley Jr. D A. 1996 F u n d a m e n ta ls o f s o il e c o lo g y Academic Press, Amsterdam. Douds D D, Pfeffer P E, and Shachar-Hill Y . 2000 A p p lic a tio n o f in v itr o m e th o d s to s tu d y c a r b o n u p ta k e a n d tr a n s p o r t b y A M fu n g i Plant Soil 2 2 6 : 255–261 Gadd G M. 1993 I n te r a c tio n s o f fu n g i w ith to x ic m e ta ls New Phytologist 12 4 : 25–60 Heggo A, Angle J S, and Chaney R L. 1990 E ffe c ts o f v e s ic u la r a r b u s c u la r m y c o r r h iz a l fu n g i o n h e a v y - m e ta l u p ta k e b y s o y b e a n s Soil Biology and Biochemistry 2 2 : 865–869 Jeffries P, Gianinazzi S, Perotto S, Turnau K, and Barea J M. 2003 T h e c o n tr ib u tio n o f a r b u s c u la r m y c o r r h iz a l fu n g i in s u s ta in a b le m a in te n a n c e o f p la n t h e a lth a n d s o il fe r tility Biol. Fertil. Soils 37 : 1–16 Khan A G, Kuek C, Chaudhry T M, Khoo C S, and Hayes W J. 2000 R o le o f p la n ts , m y c o r r h iz a e a n d p h y to c h e la to r s in h e a v y m e ta l c o n ta m in a te d la n d r e m e d ia tio n Chemosphere 4 1: 197–207 Koide R T. 1993 P h y s io lo g y o f th e m y c o r r h iz a l p la n t Adv. Plant Pathol. 9: 33–54 Lapointe L and Molard J. 1997 C o s ts a n d b e n e fits o f m y c o r r h iz a l in fe c tio n in a s p r in g e p h e m e r a l, E r y th r o n iu m a m e r ic a n u m New Phytologist 135 : 491–500 Marschner H. 1995 M in e r a l n u tr itio n o f h ig h e r p la n ts Academic Press, London Medina A, Vassileva M, Barea J M, and Azcon R. 2006 T h e g r o w th - e n h a n c e m e n t o f c lo v e r b y A s p e r g illu s tr e a te d s u g a r b e e t w a s te a n d G lo m u s m o s s e a e in o c u la tio n in Z n c o n ta m in a te d s o il Applied Soil Ecology 33: 87–98

References:
Allen E B and Allen M F. 1986 W a te r r e la tio n s o f x e r ic g r a s s e s in th e fie ld : in te r a c tio n s o f m y c o r r h iz a s a n d c o m p e titio n New Phytologist 10 4 : 559–571 Bethlenfalvay G J, Schreiner R P, Mihara K L, and McDaniel H. 1996 M y c o r r h iz a e , b io c id e s , a n d b io c o n tr o l 2 . M y c o r r h iz a l fu n g i e n h a n c e w e e d c o n tr o l a n d c r o p g r o w th in a s o y b e a n – c o c k le b u r a s s o c ia tio n tr e a te d w ith th e h e r b ic id e b e n ta z o n Applied Soil Ecology 3: 205–214

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Neumann E and George E. 2004 Colonisation with the arbuscular mycorrhizal fungus Glomus mosseae ( N icol. and Gerd.) enhanced phosphorus uptake from dry soil in Sorghum bicolor ( L .) Plant and Soil 261, 245–255 Ocampo J A. 1986 V esicular arbuscular mycorrhizal infection of host and nonhost plants— effect on the growth-responses of the plants and competition between them Soil Biology and Biochemistry 18: 607–610 Sanders I R, Clapp J P, and Wiemken A. 1996 The genetic diversity of arbuscular mycorrhizal fungi in natural ecosystems— a key to understanding the ecology and functioning of the mycorrhizal symbiosis New Phytologist 33: 123–134

Shetty K G, Hetrick B A D, Figge D A H, and Schwab A P. 1994 Effects of mycorrhizae and other soil microbes on revegetation of heavy-metal contaminated mine spoil Environmental Pollution 86: 181–188 Smith S E and Read D J. 1981 Mycorrhizal symbiosis Academic Press, San Diego van der Heijden M G A, Klironomos J N, Ursic M., Moutoglis P, Streitwolf-Engel R, Boller T, Wiemken A, and Sanders I R. 1998 Mycorrhizal fungal diversity determines plant productivity, plant biodiversity, ecosystem variability and productivity

Recent references
The latest additions to the network’s database on mycorrhiza are published here for members’ information. The Mycorrhiza Network will be pleased to supply any of the available documents to bonafide researchers at a nominal charge. This list consists of papers from the following journals. J ou rnal of H azardou s Materials J ou rnal of P lant Nu trition and Soil ScienceZ eitschrift F u r P flanzenernahru ng U nd B odenku nde P J ou rnal of Su stainab le Agricu ltu re P Molecu lar P lant- Microb e I nteractions P Mycological P rogress P P Mycological Research P P Mycorrhiza P P New P hytologist P P Nu clear I nstru m ents and Methods in P hysics P Research Section B - B eam I nteractions with P Materials and Atom s P P P hytochem istry Copies of papers published by mycorrhizologists during this quarter may please be sent to the Network for inclusion in the next issue.
Name of the author(s) and year of publication Title of the article, name of the journal, volume no., issue no., page nos (address of the first author or of the corresponding author, marked with an asterisk)

P P P P P

Agronom y for Su stainab le D ev elopm ent Annals of F orest Science Aq u atic B otany B iocontrol Science and T echnology C om m u nications in Soil Science and P lant Analysis E nv ironm ental P ollu tion E x perim ental Agricu ltu re F em s Microb iology E cology H ereditas J ou rnal of B asic Microb iology J ou rnal of B iotechnology J ou rnal of F orest Research

P P

Alexander I J. 2006

Ectomycorrhizas - O ut of Africa? New P hytologist 172(4): 589–591 [ Alexander I J, University Aberdeen, School of Biological Science, Aberdeen AB9 1FX, Scotland] Mycorrhization helper bacteria: A case of specificity for altering ectomycorrhiza architecture but not ectomycorrhiza formation Mycorrhiza 16(8): 533–541 [ * Aspray, T J, Warwick HRI, Warwick CV35 9EF, England]

Aspray T J* , Frey-Klett P, Jones J E, Whipps J M, Garbaye J, Bending G D. 2006

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