response of siratro (macroptilium atropurpureum urb ... papers/jtas vol...response of siratro...

PertanikaJ. Trop. Agric. Sci. 20(1): 19-29(1997) ISSN: 0126-6128© Universiti Putra Malaysia Press

Response of Siratro (Macroptilium atropurpureum Urb. Rabaceae) toVesicular-arbuscular Mycorrhizal Fungi and Rhizobium sp. in Sterilized Soil

S. GREEP, T. MUTHUKUMAR, K, UDAIYAN and V. NARMATHA BAI

Microbiology Unit, Department of BotanyBharathiar University

Coimbatore - 641 046 Tamil Nadu, India

Keywords: vesicular-arbuscular mycorrhiza, Rhizobium sp., Macroptilium atropurpureum

ABSTRAKKajian telah dijalankan terhadap kesan penginokulatan enam spesis kulat mikoriza arbuskulat vesikular iaitu,Gigaspora margarita, Glomus mossease, G. monosporum, G. versiformis, G. fascilulatum, G. deserticoladan Rhizobium sp. ke atas pertumbuhan dan kandungan NPK dalam siratro (Macroptilum atxopupuream).Spesis mikoriza telah diasingkan daripada tanah hutan rizofera Western Ghats dan digunakan dalam kulturpot bersama Sorghum bicolor sebagai tanaman perumah. Rhizobium sp. yang diasingkan daripada nodsegar Siratro telah dikultur dan dikekalkan dalam ekstrak yis bubur manitol Pengaruh kulat VAM danRhizobium sp. sama ada bersendirian atau berkombinasi ke atas pertumbuhan dan kandungan nutrien Sirartomenunjukkan pmingjmtan signifikan secara statistik selepas hanya enam hari tumbuhan ditanam. Penginokulatandual bersama kulat VAM dan Rhizobium sp. meningkatkan jumlah nod tanaman dan kandungan tisu NPKKandungan nutrien dan pertumbuhan tertinggi telah dibuktikan okh tanaman ynag diinokulat denganRhizobium bersama-sama dengan kulat VAM berganda.

ABSTRACTThe effect of inoculation of six species of vesicular-arbuscular mycorrhizal fungi viz., Gigaspora margarita,Glomus mosseae, G. monosporum, G. versiformis, G. fasciculatum, G. deserticola and Rhizobium sp. ongrowth and NPK content in siratro (Macroptilium atropurpureum) was studied. The mycorrhizal species wereisolated from the rhizosphere forest soils of the Western Ghats and multiplied in pot culture with Sorghum bicoloras the host plant. Rhizobium sp. isolated from fresh nodules of siratro was cultured and maintained in yeastextract mannitol broth. The influence of VAM fungi and Rhizobium sp. either singly or in combination on growthand nutrient contents of siratro showed statistically significant increase after only 60 days of plant growth. Dualinoculation with VAM fungi and Rhizobium increased plant nodule number and tissue NPK content. Highestgrowth and nutrient content were exhibited by plants inoculated with Rhizobium together with multiple VAM fungi.

INTRODUCTIONThe beneficial effect of vesicular-arbuscularmycorrhizal fungi (VAMF) and rhizobia onlegume growth is well documented (Barea andAzcon-Aguilar 1983; Harley and Smith 1983; El-Hassanin and Lynd 1985; Kawai and Yamamoto,1986; Ishac et al. 1987; Piccini et al. 1988).Phosphorus is often the growth-limiting nutrientsince nodulating legumes require more P andnitrogen fixation (Mosse et al 1976). ImprovedP nutrition often results in better nodulationand nitrogen fixation (Waidyanatha et al. 1979;Lynd et al. 1985). Earlier studies have shownthai dual inoculation with VAMF and rhizobia

increased plant growth, nodule size, nodule dryweight and nitrogenase activity (Smith and Daft1977: Smith et al 1979; Asimi et al 1980). Theimproved growth as a result of double symbiosiswith VAMF and rhizobial inoculation is attrib-uted to the improved phosphate uptake by VAMF(Barea and Azcon-Aquilar 1983). The effect ofVAMF and rhizobia on temperate (Abbott andRobson 1978) and tropical forage legumes in-cluding siratro (Mosse 1977; Salinas et al 1985;Ariens et al 1991) has been shown to increasegrowth of these legumes. Since Macroptiliumatropurpureum cv. Siratro is widely grown inTamil Nadu in association with pasture grasses

S, GREEP, T. MUTHUKUMAR, IC UDAIYAN AND V. NARMATHA BAI

and used as forage to supply protein to grazinganimals (Kretschmer 1972; Hodges et al 1982),the present study evaluates combinations of sixVAMF and rhizobia for successful establishmentof siratro in P-deficient soils, as a substitute forphosphorus fertilizer.

MATERIALS AND METHODSInocula of Gigaspora margarita Becker and Hall;Glomus mossease (Nicol. and Gerd.) , G.monosporum (Gerd. and Trappe); G. versiformis(Berch and Fortin); G. fasdculatum Thaxtersensu Gerd. and G. deserticola (Trappe, Blossand Menge) were isolated from the rhizosphereof forest soil in the Western Ghats. TheRhizobium strain was isolated from fresh nod-ules of siratro and maintained in yeast mannitolbroth for five days. Seeds of siratro were ob-tained from Tamil Nadu Agricultural Univer-sity, Coimbatore.

Sandy loam soil collected from the experi-mental field of Botany Department, BharathiarUniversity, Coimbatore, was collected, air driedand freed of large organic debris using a 2-mmsieve. The soil was mixed with fine sand in aratio of 4:1. The steam-sterilized soil, initiallywith pH of 7.5, N of 74 mg/g; P of 6.0 mg/g; Kof 191 mg/g was packed in 26 x 14 cm poly-thene bags. The respective bags were inocu-lated either with a single VAM species alone orin combination with Rhizobium sp.; or withmixture of all the six VAMF with and withoutRhizobium sp., or with Rhizobium sp. alone. Onehundred grams of each VAMF inocula wereplaced as a thin layer 5 cm below the soil surfacein the treatments. Steam-sterilized uninoculatedsoil served as control.

The polythene bags were arranged in aRCBD with four replications. Two surface-steri-lized (5%, H2O2) seeds of uniform size weresown in each polythene bag. Twenty ml rhizobialsuspension for yeast extract mannitol broth wheregiven to 5-day old seedlings in the respectivetreatments. The plants in the greenhouse werewatered daily to field capacity

The plants were harvested at 20-day inter-vals for a period of 80 days. Roots were washedfree of soil. Shoot length, root length, leaf area,nodule number, shoot and root dry weights(after drying at 65° C for 48 h) were recorded.VAM colonization was assessed after staining theroot samples following the method of Phillipsand Hayman (1970).

The 80-day-old plant materials were dried,ground and digested for the determination oftissue phosphorus (P) using the method appliedby Jackson (1958), nitrogen (N) using themethod of Humphries (1956) and potassium(K) using that of David (1962).

Data on plant growth, nodulation, VAMstatus and tissue nutrient content were subjectedto analysis of variance (ANOVA), and the meansseparated using Duncan's new multiple rangetest at P - 0.05 level.

RESULTSRoot lengthSiratro plants inoculated with Glomus monosporumhad longer roots at 20 and 40 days after emer-gence (DAE) but at 60 and 80 DAE, plantsixioculated with Gigaspora margarita had longerroots (Table 1).

Siratro inoculated with Rhizobium andGigaspora margarita (at 20 and 40 DAE) andRhizobium and Glomus mosseae (at 60 and 80DAE) produced longer roots than the otherendophytic inoculations. Plants inoculated witha mixture of all siz VAMF species producedlonger roots at 20 and 40 DAE than their com-bined effect with Rhizobium, whereas harvests on60 and 80 DAE gave better growth only whenassociated with Rhizobium (Table 1).

Shoot length

Shoot length at 20 and 40 DAE was greater inthe presence of Glomus deserticola. Plants inocu-lated will G. fasdculatum produced better shootlength at 60 and 80 DAE. At day 20, the longestshoot was seen in plants with the single inocula-tion of Rhizobium sp. but at subsequent stages(40, 60 and 80 DAE) G. deserticola in associationwith Rhizobium sp. produced longer shoots thanthe other five endophytes studied. Rhizobiumsp. co-inoculated with a mixture of all the sixendophytes showed better shoot length on 40and 80 DAE than with the endophytic mixturealone (Table 1).

Leaf Area

Siratro inoculated with Gigaspora margarita hadgreater leaf area than the other individual in-oculation at all stages. Rhizobium sp. in associa-tion with G. deserticola produced great leaf areathan the other five endophytes. Rhizobium sp.inoculated plants showed a more significant in-

20 PERTANIKAJ. TROP. AGRIC. SCI. VOL. 20 NO. 1, 1997

TABLE 1Effect of various inoculations on plant growth of Macroptilium atropurpureum at 20. 40. 60 and 80 days after emergence (DAE)

I

i2n

So

S

Treatment(a)

ControlVAM 1VAM 2VAM 3VAM 4VAM 5VAM 6VAM 1-6

RVAM 1+RVAM 2+RVAM 3+R

VAM 4+RVAM 5+RVAM 6+RVAM 1-6+RNPSFD

Shootlength(cm)

2.40c4.10abc5.42abc4.47abc5.97abc4.20abc7.82a5.20abc

6.92ab4.62abc5.35abc5.07abc

6.17abc5.85abc4.30abc5.57abc5.04abc2.34c

20D

Rootlength(cm)

8.31cd14.00b12.00bc14.95b12.27bc11.92bc11.37bc16.75ab

17.00ab19.25a11.32bc18.17ab

10.20cd14.10b12.82bc16.31ab13.77bc7.49d

Leaf area(sq. cm)

1.93b7.67a4.03ab5.03ab6.15ab3.12ab4.85ab5.08ab

6.35ab3.27ab4.25ab3.61ab

3.87ab3.41ab7.85a6.73ab4.31ab1.98b

Shootlength(cm)

4.90gh15.75d10.12f7.62g6.07gi

18.55bc25.75a11.60e

13.50de16.56bcd11.37ef

1.62h

13.00de13.50def19.12bc20.25b10.25ef3.53gh

40D

Rootlength(cm)

13.77b22.95ab22.00ab29.87ab17.87ab22.25ab20.00ab35.75a

18.50ab30.87ab22.37ab20.83ab

19.84ab19.84ab29.25ab30.47ab21.62ab12.19b

Leaf area(sq. cm)

2.94g9.19abcd6.26def6.83def8.92abcd6.61def6.37def7.41cdef

8.52abcd8.39abcde5.58ef5.21efg

4.22g6.28def9.80a9.46ab6.25def2.05h

Shootlength(cm)

12.17c19.42ab37.95ab23.65ab29.90ab40.25a31.70ab26.37ab

20.25b31.85ab29.25ab31.12ab

32.65ab31.37ab32.62ab29.12ab25.22ab11.53c

60D

Rootlength(cm)

36.40d59.95abc49.90abc59.50abc51.00abc59.32abc57.05abc45.87abc

25.30c51.75abc69.00a62.05ab

58.17abc49.75abc59.10abc57.20a41.30bc33.57d

Leaf area(sq. cm)

4.76g9.38abcd7.32ef7.65def9.25abcd7.83cdef7.54def8.97abcdef9.54abc9.11abed7.50def8.41abcdef8.01bcdef6.8 If9.93a9.57ab6.99f3.94g

Shootlength(cm)

24.32bc38.00abc42.40a34.25abc39.60ab43.75a39.75ab30.20abc

40.12ab33.00abc35.55abc34.33abc

37.00abc37.62abc40.35ab42.50a27.50abc22.51c

80D

Root ]length(cm)

39.13cd77.00ba59.12ab60.97ab62.87ab69.10a66.1 Sab55.02bc

34.85d58.75ab70.55ab68.57ab

67.80ab50.87bcd64.42ab67.00ab51.90bc36.25cd

Leaf area(sq. cm)

4.78c10.97a7.34b7.81b9.53ab9.41ab9.60ab8.99ab

9.69ab9.88ab8.10ab9.26ab

8.51 ab7.6 lab

lO.llab16.37ab7.84ab4.61c

(a) VAM 1 - Gigaspora VAM 2 - Glomus mosseae VAM - G. monosporum; VAM 4 - G. versiformis VAM 5 - G. fasciculatuumVAM 6 - G. deserticola R - Rhizotnum; FD - Formaldehyde; NPS Non-sterilized field soil.

Means followed by the same letter are not significantly (p <0. 05) different as determined by Duncan's new multiple range test.

I2s3

I

3I

S. GREEP, T. MUTHUKUMAR, K. UDAIYAN AND V. NARMATHA BAI

TABLE 2Effect of inoculation of various endophytes

on number of nodules per plant

Treaments(a)

20D

Days after

40D

emergence

60D 80D

ControlVAM 1VAM 2VAM 3VAM 4VAM 5VAM 1-6RVAM 1+RVAM 2+RVAM 3+RVAM 4+RVAM 5+RVAM 6+RVAM 1-6+RNPSFD

ObObObObObObOb2.75ab8.00ab3.25ab4.25ab4.50ab2.75ab6.25ab8.00a3.75ab0.0b

ObObObObObObOb3.25ab8.00a6.00ab4.50ab6.00ab7.00ab8,25a10.00a3.65ab0.0b

OeOeOeOeOeOeOe3.00de9.75ab6.50bcd6.50bcd7.20bc7.25bc9.75ab11.25ab4.00cd0.0b

Og

Og

OgOgOg3.1 Of10.50b7.25e8.25cde7.50de7.25e10.25cd14.75a6.75e0.0b

(a) see footnote to Table 1

crease in leaf area than that of the mixture of sixVAMF. However, the endophyte mixture andRhizobium sp. showed an additive effect on leafarea at all stages where the effect of endophytesis not significant when compared to Rhizobiumsp. (Table 1).

Nodule Number

Siratro in association with Rhizobium sp. and witha mixture of all the six endophytes increased thenodule number to three times that of the rhizobialinoculation alone. Gigaspora margarita on 20 DAEand G. deserticola on 40 DAE produced morenodules in the presence of Rhozobium sp, whereasat 60 and 80 DAE both inoculations had the samenodule number. At all stages of growth the nod-ules of plants with a mixture of six endophytestogether with Rhizobium sp. were four times thenumber of plants inoculated with only Rhizobiumsp. (Table 2).

Dry Weight of Roots and ShootsInoculation with Glomus mosseae significantly in-creased the root dry weight compared with allother inoculations at all stages of growth. Shootdry weight was higher in plants inoculated withG. mosseae at 20 and 40 DAE but was greater byG. monosporum at 60 and 80 DAE. Association

of Rhizobium sp. with G. deserticola producedgreater shoot dry weight than all otherendophytes with Rhizobium. The endophytemixture increased dry weight of roots at allstages and shoot dry weight at 20 and 40 DAE.The root to shoot ratio was increased by bothdual inoculation and also single VAMF inocula-tion compared to control, except at 60 DAE(Table 3).

Root Colonization

Glomus fasciculatum at 20 DAE, G. mosseae at 40DAE, G. versiformis as well as G. deserticola on 60DAE and Gigaspora margarita at 80 DAE showedhigher root colonization than the others.Rhizobium sp. in association with endophyte mix-ture increased colonization in the early stagesbut decreased later (Fig. 1).

The endophytic inoculation increased theNPK concentration in plant tissue compared touninoculated controls. Dual inoculation signifi-cantly increased the NPK concentration in planttissues. A significant increase in NPK wasobserved in plants inoculated with Rhizobium sp.and multiple VAM fungi. Nutrient accumula-tion in plants grown in sterilized field soil con-taining indigenous VAM fungi paralleled dualinoculation (Fig. 2).

DISCUSSIONThe increase in root length of siratro, inocu-lated with either G. monosporum or Gigasporamargarita compared with the remaining fourendophytes, is similar to the observations ofLopes and Olivera (1980). Gigaspora margaritaat 20 - 40 DAE and Glomus mosseae at 60 - 80DAE increased root length when co-inocu-lated with Rhizobium. Similar results have beenreported in Phaseolus vulgaris (Daniels-Hyltonand Ahmed 1994) and Vicia faba (Ishac et al1994) by dual inoculation of Rhizobium sp.with VA mycorrhiza. This supports the obser-vation that Rhizobium sp. and VA mycorrhizaincrease plant growth to a greater extent thancan be attributed to either of them whenadded singularly (Harley and Smith 1983; ElHassanin and Lynd 1985; Ishac et al. 1987;Kawai and Yamamoto 1986). The root to shootratio was increased by both dual inoculationand single inoculation of VAMF, which contra-dicts the findings that dual inoculation showedthe lowest root-to-shoot ration (Piccini et al1988).


TABLE 3Effect of various inoculations on shoot and root growth of Macroptilium atropurpureum at 20, 40, 60 and 80 days after emergence (DAE)

33

I*0

52P

$

3

s

Treatment

W

ControlVAM 1VAM 2VAM 3VAM 4VAM 5VAM 6VAM 1-6RVAM 1+RVAM 2+RVAM 3+RVAM 4+RVAM 5+RVAM 6+RVAM 1-6+RNPSFD

20D

Dry weight

Shoot

0.044a0.041a0.064a0.034a0.057a0.053a0.100a0.056a0.036a0.051a0.046a0.041a0.050a0.044a0.053a0.058a0.020a0.041a

Root

0.006b0.011b0.057b0.044a0.020b0.011b0.012b0.033b0.099b0.0044b0.064b0.064b0.006b0.081b0.014b0.122b0.606b0.007b

U /cK / o

0.144i0.245i0.867f3.33a0.336i0.223i0.115i0,559h0.275i0.856g0.372e1.586d0.116i1.82c0.255i2.35b0.290i0.165i

40D

Dry weight

Shoot

0.322cd0.204def0.620a0.612ab0.082f0.137f0.156ef0.123f0.065f0.157ef0.1240.1 OOf0.285cde0.284cde0.380c0.1460.065f0.315cd

Root

O.lOg0.030fg0.124b0.076de0.031fg0.020g0.028a0.040fO.Ollg0.059ef0.098c0.080cd0.029fg0.028g0.027g0.244a0.210aO.OlOg

0.03610.145h0.22g0.138jh0.379e0.140h0.164h0.321f0.181h0.380e0.755d0.800c0.130jh0.093k0.0751k1.660b3.277a0.0341

60D

Dry weight

Shoot

0.290c0.210bc0.668ab0.675ab0.367d0.573ab0.488ab0.498ab0.666ab0.291bc0.387b0.357b0.536ab0.468ab0.549ab0.898a0.534ab0.01 le

Root

0.057gh0.173def0.467a0.294c0.092fgh0.230cde0.237cd0.297bc0.023h0.064fgh0.137efg0.039h0.196de0.252cd0.294c0.395ab0.099fgh0.049gh

U /c

1.921c0.839d0.704e0.438hi0.259J2.017b0.487h0.476h0.036m0.256J0.353i0.11010.366i0.540g0.561fg0.442h0.183k4.350a

80D

Dry weight

Shoot

0.153i0.237hi0.685bc0.695bc0.435fg0.553de0.49bef0.667bcd0.699b0.295gh0.409f0.407fg0.628bcd0.577cde0.606bcde0.966a0.883a0.149i

Root

0.070g0.273ef0.467b0.377cd0.256ef0.264ef0.239ef0.725ab0.2120.775a0.247ef0.040g0.254ef0.281ef0.375cd0.326c0.321 de0.061 lg

U /c

0.442b1.182b0.659b0.544b0.585b0.476b0.484b1.281a0.324b2.583a0.627b0.094b0.444b0.486b0.626b0.447b0.378b0.413b

II3

(a) See footnote to Table 1


70

60

50

f 40

2 0 "

1 0 -

20 DAE

Ka)1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

Treatments

100

8 0 -

i6 0 -

4 0 -

2 0 -

40 DAE

bed bed be

• ii

II

I

•e fg

1

Jp

Iiii

• I

i i

iiiIII

1I

H 11 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

1 ( b > Treatments


RESPONSE OF SIRATRO TO VESICULAR-ARBUSCULAR MYCORRHIZAL FUNGI

100-

80H

60 H

i

20 H

60 DAE

de d e

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18Treatments

100

80 H

8)601(0

5 401

20 H

0

b

:::

:::

DAE

c

;;;;

cdd

de

cdTTT;

arr-

:\

\

: 9

be

cd cd cd

c c

a

::

1

i

1

be-7TT1

•

Kd)2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

Treatments

Fig 1. Influence of VAM fungi and rhizobia on root colonization in Macroptiliumatropurpureum

1. Control, 2. Gigaspora margarita, (VAM 1), 3. Glomus mosseae (VAM 2),4. G. monosporum (VAM 3), 5. G. versiformis (VAM 4), 6. G. Fasdculatum (VAM 5),7. G. deserticola (VAM 6) 8. VAM 1 + VAM 2 + VAM 3 + VAM 4 + VAM 5 + VAM6, 9. Rhizobium (R), 10.VAM 1 + R, 16. VAM1 + VAM2 + VAM3 + VAM4 + VAM5+ VAM6 + R, 17. Non-pasteurised soil (NPS), 18. Formaldehyde-fumigated - (FD).Bars bearing the same letters are not significantly different according to Duncan'snew multiple range test ( P < 0.05)

PERTANIKAJ. TROP. AGRIC. SCI. VOL. 20 NO. 1, 1997 25

1

o

cr o

ro

co

en

00

i:ro

«^CO

F3-^ffi

Concentration (mg/g)

01 O 01

iii;mmm

MwmwmmWrm '

Concentration (mg/g)fO 4** 0) 00 C5

o o o o o o

CO

O l

o>

H CO

55. <o

§ o

fo

CO

en -

C/5

o

I

GD


70

60-

50-

1,§,40-

"5 30-§ 20Ho

10

Kbe b c

abc

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 182(c) Treatments

Fig 2. Influence of VAM fungi and rhizobia on nutrient (NPK) status of Macroptiliumatropurpureum in 80 days

1. Control, 2. Gigaspora margarita, (VAM 1), 3. Glomus mosseae (VAM 2),4. G. monosporum (VAM 3), 5. G. versiformis (VAM 4), 6. G. Fasciculatum (VAM 5),7. G. deserticola (VAM 6) 8. VAM 1 + VAM 2 + VAM 3 + VAM 4 + VAM 5 + VAM6, 9. Rhizobium (R), 10.VAM 1 + R, 16. VAM 1 + VAM 2 + VAM 3 + VAM 4 +VAM5 + VAM6 + R, 17. Non-pasteurised soil (NPS), 18. Formaldehyde-fumigated- (FD).Bars bearing the same letters are not significandy different according to Duncan'snew multiple range test ( P < 0.05)

The presence of longer roots in endophyte-inocuiated plants rather than Rhizobium sp. at20 and 40 DAE, and the combined influenceof VAM and Rhizobium sp. at 60 and 80 DAEindicate that Rhizobium sp. might have supple-mented nutrients for root growth only at laterstages.

Unlike the roots, the shoot length increasedin G. deserticola at early stages (20 - 40 DAE) andin G. fasciculatum at later stages (60 -80 DAE).Shoot length of siratro was higher in singleinoculation with Rhizoboum sp. than that of anyendophyte studied up to 20 DAE. The G.deserticofa and Rhizobium sp. combination wasbetter than all other combinations. A plantinoculated with all the six VAMF together withRhizobium sp. showed an additive effect onshoot growth.

Gigaspora margarita among single endophyticinoculation, and G. deserticola and Rhizobiumsp. among dual inoculation, increased the leafarea whreas the endophytic mixture togetherwith Rhizobium sp. increased the leaf area morethan any of them individually. VA mycorrhizal

plants tend to have higher cytokinin activity intheir shoots (Allen et al. 1980) and cytokininspromote leaf area by cell division and cellexpansion (Bass and Kuoier 1989).

Gigaspora margarita at an early stage (20DAE) and Glomus deserticola at later stages (40DAE) and endophytic mixture (60 - 80 DAE)produced more nodules in the presence ofRhizobium sp. than other combinations. Thissupports the view that endophytes increasethe nodule number when associated withRhizobium sp. (Daniels-Hylton and Ahmed1994), In the present study, Glomus mosseaeproduced greater root dry weight than all otherendophytes at all stages. In the case of shoots,Glomus mosseae ax. early stages (20 and 40 DAE)and G. monosporum at later stages (60 and 80DAE) produce greater dry weight. Medina etal. (1988) showed that G. etunicatum and G.intraradices produce higher shoot dry weightthan other VAM inoculations. Though theendophytic mixture increased the root dryweight at all stages the shoot dry weight wasreduced at later stages (60 and 80 DAE).



The variation in colonization at differentstages of plant growth with various colonizingVAMF endophytes indicates that no specificVAMF endophyte is involved in colonizing thehost root. Increased accumulation of tissuenutrients was observed in plants inoculated witha multiple VAMF and Rhizobium sp. compared toRhizobium sp. or VAM inoculation. This sup-ports the result of Piccini et al, (1988) wheredual inoculation with Rhizobium sp. and VAMresulted in an increased accumulation of nutri-ents (N, P, K, Ca and Mg) in alfalfa than singleendophyte inoculations.

CONCLUSION

The present study clearly reveals that Gigasporamargarita and Glomus deserticola are the best forproducing quality siratro plants either singly ordually with Rhizobium. But the mixture of sixVAMF and Rhizobium was found to enrich theNPK status of the legume. Work is in progressto establish quality siratro paints under fieldconditions with an effective VAMF and Rhizobiumcombination.

ACKNOWLEDGEMENT

The first author gratefully thanks the UniversityGrants Commission (UGC), New Delhi for pro-viding financial assistance.

REFERENCES

ABBOTT, L.K. and A.D. ROBSON. 1978. Growth of

subterranean clover in relation to the forma-tion of endomycorrhizas by introduced andindigenous fungi in a field soil. New Phytologist81: 575-587.

ALLEN, M.F., T.S. MOORE and M. CHRISTENSEN. 1980.

Phytohormone changes in Boutelout gracilisinfected by vesicular-arbuscular mycorrhiza. ICytokinin increase in the host plant. CanadianJournal of Botany 58: 371.

ARIENS, R.N., T.R. THIAGARAJAN, M.H. AHMED and

W.A. MILAGHLIN. 1991. Co-selection of compat-ible rhizobia and vesicular-arbuscularmycorrhizal fungi for cowpea in sterilized andnon-sterilized soils. Biology and Fertility of Soils12: 112-116.

ASIMI, S., V. GIANINAZZI-PEARSON and S. GIANINAZZI.

1980. Influence of increasing soil phosphoruslevels on interaction between vesiculararbuscular mycorrhiza and Rhizobium insoybean. Canadian Journal of Botany 58:2200-2205.

BAREA, J.M. and C. AZCON-AGUILAR. 1983. Mycorrhizas

and their significance in nodulatiing nitrogenfixing plants. Advances in Agronomy 36: 1-54.

BASS, R. and D. KULIER. 1989. Effect of vesicular -arbuscular mycorrhizal infection and phos-phate on Plantago major Pleosperma in rela-tion to internal cytokinin concentrations.Physiologia Plantarum 76: 211-215.

DANIELS-HYLTON, K.D.M. and M.H. AHMED. 1994.

Inoculation response in kidney beans {Phaseolusvulgaris L.) to vesicular-arbuscular mycorrhizalfungi and rhizobia in non-sterilized soil. Bio-logy and Fertility of Soil 18: 95-98.

DAVID, D.J. 1962. Emission and absorpt ionspectrochemical methods. In Modern Methodsof Plant Analysis, ed. K. Peech and M.V. Tracey,vol. 5 p. 1-25. Berlin: Springer-Verlag.

EL-HASSANIM, A,S. and J.Q. LYND. 1985. Soil fertilityeffects with tripartite symbiosis for growthnodulation and nitrogenase activity of Viciafabah. Journal of Plant Nutrition 8:491-499.

HARLEY, J.C. and S.E. SMITH. 1983. Mycorrhizal Sym-

biosis, London: Academic Press.

HODGES, E.M., A.E. KRETSCHMER, P. Misu-RYjr., R.D.

ROUSH, O.C. RUELKE and G.H. SNYDER. 1982.

Production and utilization of the tropical leg-ume Aschynomene. Agricultral Experimental Sta-tion Circle S-2900.

HUMPHRIES, E.C. 1956. Mineral components andash analysis. In Modern Methods of Plant Analy-sis, ed. K. Peech and M.V. Tracey, vol. 5 p. 468-502. Berlin: Springer-Verlag.

ISHAC, Y.Z., MJ. DAFT, E.M. RAMADAN, M.E. EL-

DAMORDASH and C.N. FARES. 1987. Effect of

Rhizobium inoculation and/or endomycorrhizason peanut growth. 2nd African Assoc. Biol. Nit-rogen. Conf Cairo University, Cairo, Egyptp. 589-596.

ISHAC, Y.Z., J.S. ANGLE, M.E. EL-BOROLLOSY, M.E, EL.

DAMORDASH, M.I. MOSTAFA and C.N. FARES

1994. Growth of Vicia faba as affected by inocu-lation with vesicular-arbuscular mycorrhizaeand Rhizobium legumonisarum cv. viceae in twosoils. Biology and Fertility of Soils 17: 27-37.

JACKSON, M.L. 1958. Soil Chemical Analysis. New Delhi:Prentice-Hall.

KAWAI, Y. and Y. YAMAMOTO. 1986. Increase in the

formation and nitrogen fixation by soybeannodules by vesicular-arbuscular mycorrhiza.Plant and Cell Physiology 27: 399-405.

KRETSCHMER, A.E. Jr. 1972. Siratro (Phaseolusatropurpureus DC) A summer-growing peren-nial pasture legume for Central and South



Florida. Florida Agriculture Experimental StantionCircle.

LOPES, E.S. and E.D. OLIVERA. 1980. Efeito de spe-

cies de mycorrhizas vesicular-arbuscular emSiratro (Macroptilium atropurpureum). Bragantia39: 241-245 (English abstract).

LYND, J.Q., RJ. TYRL and A.A.C. PURCINO. 1985.

Mycorrhizal-soil fertility effects on regrowth,nodulation and nitrogenase activity of siratro(Macroptilium atropurpureum (DC Urb.) Journalof Plant Nutrition 8: 1047-1059.

MEDINA, O.A., D.M. SYLVIA and A.E. KRETSCHMER JR.

1987. Growth response of tropical forage leg-umes to inoculation with Glomus intraradices.Tropical Grasslands 21: 24-27.

MEDINA, O.A., D.M. SYLVIA and A.E. KRETSCHMER Jr.

1988. Response of siratro to vesicular-arbus-cular mycorrhizal fungi. I. Selection of effec-tive vesicular-arbuscular fungi in amended soil.Soil Science Society of American Journal52: 416419.

MOSSE, B. 1977. Plant growth responses to vesicular- arbuscular mycorrhiza. X. Responses ofStylosanthes and maize to inoculation inunsterile soils. New Phytologist 78: 277-278.

MOSSE, B., C.L. POWELL and D.S. HAYMEN. 1976.

Plant growth response to vesicular-arbuscularmycorrhiza. IX Interactions between VA my-corrhiza, rock phosphate and symbiotic nitro-gen fixation. New Phytologist 76: 331-342.

PHILLIPS, J.M. and D.S. HAYMAN. 1970. Improved

procedures for clearing roots and staining

parasitic and vesicular-arbuscular mycorrhizalfungi for rapid assessment of infection. Trans-actions of the British Mycological Society 55: 158-161.

PICCINI, D., J.A OCAMPO and EJ. BEDMAR. 1988.

Possible influence of Rhizobium on VAmycorrhiza metabolic activity in double sym-biosis of alfalfa plants (Medicago saliva L.) grownin a pot experiment. Biology and Fertility of Soils6: 65-67.

SALINAS, J.G, J.I. SANZ and E. SIEVERDING. 1985. Im-

portance of VA mycorrhizae for phosphorussupply to pasture plants in tropical oxisols.Plant and Soil 84: 347-360.

SMITH, S.E. and MJ. DAFT. 1977. Interactions be-

tween growth, phosphate content and Na fixa-tion in mycorrhizal and non-mycorrhizalMedicago saliva. Australian Journal of Plant Physi-ology 4: 403-413.

SMITH, S.E., DJ.D NICHOLAS and F.A. SMITH. 1979.

Effect of early mycorrhizal infection onnodulation and nitrogen fixation of Trifoliumsubterraneum L. Australian Journal of Plant Physi-ology 6: 305-306.

WAIDYANATHA, U.P., S.N. YOGARATNAN and W.A.

ARIYARATHE. 1979. Mycorrhizal infection ongrowth and nitrogen fixation of Pueraria andStylosanthes and uptake of phosphorus fromtwo rock phospha tes . New Phytologist82: 147-152.

(Received 29 February 1996)(Accepted 25 April 1997)


response of siratro (macroptilium atropurpureum urb ... papers/jtas vol...response of siratro...

Documents