Department of Soil Science, University of Saskatchewan, 51 Campus Dr., Saskatoon SK, Canada S7N 5A8

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1 Impct of Arusculr Mycorrhizl Fungi on the Growth nd Rhizoium Symioses Development in Kuli nd Desi Chickpes Grown Under Drought Stress Conditions W. Ellouze,,, A. Nyyr,, K. Hnson, S. Bouzid nd C. Hmel, Environmentl Helth / Soil, Wter nd Air Qulity, Agriculture nd Agri-Food Cnd, Airport Rod Box, Swift Current, SK, Cnd, S9H X Deprtment of Soil Science, University of Ssktchewn, 5 Cmpus Dr., Ssktoon SK, Cnd S7N 5A8 Deprtment of Biologie, Fculté des Sciences Mthémtiques, Physiques et Nturelles, Université Tunis El Mnr, Cmpus Universitire, Tunis 6, Tunisi Key Words: chickpe, rusculr mycorrhize fungi, nodultion, symiosis, drought stress Astrct Arusculr mycorrhizl fungi (AMF) hve induced drought tolernce in severl plnts nd could increse chickpe yield under semirid climtes. Desi chickpe re more drought tolernt thn Kuli suggesting wek mycorrhizl symiosis in this chickpe type. A greenhouse experiment ws conducted to ssess the extent nd the impct of rusculr mycorrhizl symiosis on the growth nd yield in Kuli nd Desi chickpe cultivrs under well wtered nd drought stress conditions nd on their ssocition with Mesorhizoium cicer. The experiment hd split-plot design with two wter levels, % of field cpcity (deficient wter) nd 7% of field cpcity (sufficient wter) rndomized in min plots. Two inocultion tretments, M. cicer + AMF nd M. cicer only, were pplied to chickpe (Kuli, CDC Frontier nd CDC Xen; Desi, CDC Ann, nd CDC Nik). The fctoril comintions of inocultion nd cultivr were rndomized in the suplots. There were four repetitions. One set of plnts ws hrvested t the time of symioses development nd nother set ws hrvested t seed mturtion. The dt ws nlyzed with ANOVA. Results indicted tht the Kuli nd Desi chickpe mycorrhizl symioses re not different, s indicted y the sence of cultivr y inocultion interction on shoot nd root growth, nodultion nd nitrogen fixtion. AM Fungi inocultion delyed nodule development ut hd no effect on grin yield. CDC Frontier hd 4 times more nodules nd 6 times more nitrogense ctivity thn CDC Ann, Nik nd Xen. Introduction Drought stress is mjor constrint to griculture in the semi-rid tropics (Quilmo, ). It my e the single most importnt stress in chickpe (Singh, 99). Some reserchers reported stress tolernce in nodulted legumes ssocited to rusculr mycorrhizl fungi (AMF) (Sánchez-Díz, ). AMF my increse drought tolernce of plnts y mens of severl mechnisms. They cn modify plnt wter reltion nd their hyphe cn enhnce plnt wter uptke cpcity (Hrdie, 985; Fer et l., 99; Ruiz-Lozno nd Ázcón, 995). On the other hnd, the importnce of AMF in legume plnts hs een ttriuted to the high P requirements of

2 the nodultion nd N -fixtion processes (Bre nd Ázcón-Aguilr, 98). Enhnced plnt P nutrition is well known contriution of AMF to plnt productivity. There re physicl differences etween the Desi nd Kuli type of chickpe. Desi chickpe is more drought tolernt thn the Kuli (Anonymous, ). These oservtions suggest tht the level of AMF roots coloniztion nd symiosis development with Mesorhizoium cicer my differ. This work verifies the following hypotheses: () Desi nd Kuli chickpe respond differently to mycorrhizl inocultion. () Inocultion with AMF influence nodultion, nitrogense ctivity nd yield differently in Kuli nd Desi chickpe, prticulrly under drought stress conditions. The time of N -fixing symiosis development ws exmined ecuse competitive interctions etween AMF nd symiotic N-fixers were reported t this stge (Zhng et l., 995; Ctford et l., ), nd the sum of effects ws otined in finl hrvest, 7 weeks fter emergence. Mterils nd Methods The experiment ws conducted in the greenhouse. It hd split-plot design with two wter levels, % of field cpcity (deficient wter) nd 7% of field cpcity (sufficient wter) rndomized in min plots. Two inocultion tretments, M. cicer + AMF nd M. cicer only, were pplied to chickpe (Kuli, CDC Frontier nd CDC Xen; Desi, CDC Ann, nd CDC Nik). The fctoril comintions of inocultion nd cultivr were rndomized in the suplots. There were four repetitions. One set of plnts ws hrvested t the time of symioses development nd nother set ws hrvested t seed mturtion. At the first hrvest, erly morning plnt wter potentil ws determined (Scholnder et l., 965) in pressure chmer (Plnt Wter Sttus Console Model 5, Soilmoisture Equipment Corp). The youngest prt of the shoot tip cut just efore the 5th lef from the pex ws used for this determintion. Plnt shoots wter potentil ws mesured immeditely fter cutting the shoot. Mesures were tken erly morning. Nitrogense ctivity ws estimted y the cetylene reduction ssy (Crlsson nd Huss-Dnell, ). Mesures of root length nd surfce re were determined using the root nlysis system WinRhizo, fter stining the roots (Cost et l., ). Plnt shoots were dried nd weighed. Seed numer nd seed weight per plnt were determined t the second hrvest. The dt sets were nlysed y ANOVA using Network JMP v..6. A P vlue of.5 ws used s threshold to ccept the significnce of effects, except when indicted otherwise. Tretment mens were compred sed on lest significnt differences, where significnt tretment effects were found. Results After 7 weeks of growth under greenhouse conditions, the root system of Kuli chickpe ws significntly lrger thn tht of Desi. Bsed on plnt dry mss, nodule numer nd nitrogense ctivity, it ppers tht the differences etween chickpes re more relted to cultivrs thn to clsses. At symiosis development, mycorrhizl inocultion induced negtive effect on ll mesured prmeters except for nodule numer t % of field cpcity, where nodule numer ws very low nd no difference ws oserved etween mycorrhizl nd control plnts (Fig. ). As illustrted y Figure, inocultion reduced plnt wter potentil (P=.) only in the unifolite Kuli chickpe CDC Xen.

3 The negtive influence of mycorrhizl inocultion hd disppered t hrvest (Fig. ).

4 R o o t s u r f c e 4,5 Mss (g),5,5,5 5 Length (cm) Are (cm ) Numer Ethylene (ì mol min - ) 6 5 4,,5,,5 c d Ann Front ie r Nik Xe n Plnt dry,5 mss Ann Front ie r Nik Xe n Ann Front ie r Nik Xe n Ann Front ie r Nik Xe n Ann Frontier Nik Xen,5,5,5 Root length Nodule numer Nitrogense ctivity (7% FC) Root surfce ,8 c,7,6,5,4,,, %FC 7%FC c Myc + Myc - Figure. Dry weight, root length nd surfce re, nd nodule numer of chickpe, s influenced y cultivr (N = 6) nd mycorrhizl inocultion (N =). There ws no nitrogense ctivity t % FC nd this tretment ws excluded from the ANOVA. N = 8 for cultivr effect nd N = 6 for inocultion effect., mycorrhizl inoculted;, control; % FC, soil moisture mintined t % of field cpcity; 7% FC, soil moisture mintined t 7% of field cpcity. Brs with the sme letter re not significntly different ccording to LSD P =.5.

5 ( ),6 Plnt wter potentil (MP),4,,8,6,4 Mycorrhizl inocultion Control *, Ann Frontier Nik Xen Figure. Plnt wter potentil mesured erly morning 7 weeks fter emergence. A str indictes inocultion y cultivr interction ( P =.) nd significnt (P =. ccording to contrst) negtive effect of inocultion on the wter potentil of AC Xen plnts. N = 8. 9 () Yield (g plnt - ) Yield (g plnt - ) () Ann Frontier Nik Mycorrhizl inocultion Xen Control Ann Frontier Nik Xen Figure. Grin yield of the different chickpe cultivrs grown t % () nd 7% () of field cpcity. The negtive influence of mycorrhizl inocultion hd disppered t hrvest. N =4. Conclusions

6 Kuli nd Desi chickpe mycorrhizl symioses re not different, s indicted y the sence of cultivr y inocultion interction on shoot nd root growth, nodultion nd nitrogense ctivity. Arusculr mycorrhizl fungi inocultion delyed nodule development, ut hd no effect on grin yield. CDC Frontier hd 4 times more nodules nd 6 times more nitrogense ctivity thn CDC Ann, Nik nd Xen. Acknowledgements The uthors would like to thnk Agriculture nd Agri-Food Cnd for use of lortory & greenhouse fcilities. Project funding provided y Alert Pulse Growers nd Government of Tunisi grnt to Mr. Ellouze. References: Anonymous,. Development of gronomic prctices for chickpe production in Alert. On line: Bre, J.M., Azcon-Aguilr, C. 98. Mycorrhizs nd their significnce in nodulting nitrogenfixing plnts. Advnces in Agronomy 6:-54. Crlsson, G. nd Huss-Dnell, K.. Nitrogen fixtion in perennil forge legumes in the field. Plnt nd Soil 5: 5-7. Ctford, J.G., Stehelin, C., Lert, S., Piché, Y., Vierheilig, H.. Suppression of rusculr mycorrhizl coloniztion nd nodultion in split-root systems of lflf fter pre-inocultion nd tretment with Nod fctors. Journl of Experimentl Botny 54: Cost, C., L.M. Dwyer, C. Hmel, D.F. Mum, X.L. Wng, L. Nntis, nd D.L. Smith.. Root contrst enhncement for mesurement with opticl scnner-sed imge nlysis. Cndin Journl of Botny 79: -9. Fer, B.A., Zsoski, R.J., Munns, D.N., Schkel, K. 99. A method for mesuring hyphl uptke in mycorrhizl plnts. Cndin Journl of Botny 69: Hrdie, K The effect of removl of extrrdicl hyphe on wter uptke y vesiculrrusculr mycorrhizl plnts. New Phytologist : Quilmo, O.A.. The vesiculr-rusculr mycorrhizl symiosis. Africn Journl of Biotechnology : Ruiz-Lozno, J.M. nd Azcón, R Hyphl contriution to wter uptke in mycorrhizl plnts s ffected y the fungl species nd wter sttus. Physiologi Plntrum 95: Sánchez-Díz, M.. Adpttion of legumes to multiple stresses in Mediterrnen-type environments. Options Méditerrnéennes 45: Scholnder, P.F., Hmmel, H.T., Brdstreet, E.D. nd Hemingsen, E.A Sp pressure in vsculr plnts. Science 48:9-46. Singh, K.B. 99. Prolems nd prospects of stress resistnce reeding in chickpe. Pges 7-5 in: Breeding for stress tolernce in cool-seson food legumes, K.B. Singh nd M.C. Sxen (eds). ICARDA. Zhng, F., Hmel, C., Kinmehr, H., nd Smith, D.L Root-zone temperture nd soyen (Glycine mx. (L.) Merr.) vesiculr-rusculr mycorrhize: Development nd interctions

7 with the nitrogen fixing symiosis. Environmentl nd Experimentl Botny 5:

Received for publication: September 25, 2012; Accepted: October 25, 2012.

Received for publication: September 25, 2012; Accepted: October 25, 2012. ISSN: 2287-688X Annls Of Plnt Sciences Issue 1; Volume 1(Novemer); 6-11 Growth, Wter Use Efficiency nd Antioxidnt Defense Responses of Mycorrhizl nd Non Mycorrhizl Allium stivum L. Under Drought Stress