5 . IR e S3 xj. 1 t s St r'l cii D ± s c; xj. s s :i- c> ra 5.1 Collection of leguminous samples A total number of 87 nodulated leguminous species, collected during the course of investigation were identified to species level and a lis t was prepared. When the li s t was checked against the global listin g of nodulated and non-nodulated leguminous species (Allen and Allen 1981 and personal communication) 19 species were found to be new reports to the global list and thus were selected for detailed studies. The details regarding their taxonomic position, habit and place of their collection are given in Table 2. It could be seen from the table that the species comprised one from Caesalpinioideae, seventeen from Papilionoideae and one from Mimosoideae. They included 13 shrubs, 5 arboreal trees and 1 climber. It may be mentioned here that of the 19 species studied, 11 can be considered as rare plants because of the enormous d iffic u ltie s encountered during their collection. The 19 species were studied in detail for their nodule morphology. A special mention may be made here of the nodulation of 5 tree legumes wherein examination for the presence of nodules on the roots becomes normally d iffic u lt. The details regarding occurrance and position of nodules on the roots and their description v i z size, shape, interio r and exterior colour and surface are given in Table 3. Table 2 : New reports of leguminous species for nodulation, along with their taxonomic position, habit, and place of collection. Sr. Plant Species Subfamily Tribe Habit Place of No. Subtribe of the Collection plant 1. Cassia kleinii W. 5 A. Caesalpinioideae Cassieae Shrub Kam ala h i l l Dist. Raigad. 2. Alysicarpus pubescens Papilionoideae Headysareae Shrub Maishej Ghat Law . Desmodiinae 3. Atylosia sericea Phaseoleae Shrub Varandha Ghat Benth. Cajaninae 4. Butea monospemia (Lam) Phasealeae Tree Kamala hill Taub. Erythrininae Dist. Raigad. 5. Crotalaria albida Genisteae Shrub Mulshi Heyne. Crotalariinae Dist. Pune 6. Crotalaria calycina Genisteae Shrub Sinhagad Schranko Cotalariinae Dist. Pune 7. Crotalaria leptostachya Genisteae Shrub Bhimashankar Benth. Crotalariinae Dist. Pune Crotalaria triquetra Genisteae Shrub Shivneri h i l l Dalz. Crotalriinae Dist. Pune. 9. Dalbergia lanceolaria Dalbergieae Tree Katraj Ghat L. Lonchocarpinae Dist. Pune 10. Erythrina stricta Phaseoleae Tree Pune Roxbo phaseolinae 11. Erythrina suberosa Phaseoleae Tree Parvati hill Roxb. phaseolinae Pune 12. Indigofera cassioides Galgeae Shrub Mahabaleshwar Rottler ex. Dc. Indigoferinae Dist. Satara 13. Mucuna monosperma Phasealeae Climber Malkapur Erythrininae Dist. Kolhapur 14. Priotropsis cystisoides Genisteae Slirub Pune ITFX Crotalariinae 15. Rhynchosia rothii Phaseoleae Shrub Kliandala BenthoEx.Ait. Cajaninae Dist. Pune Sr. Plant Species Subfamily Tribe § Habit Place of NOo Subtribe of the Collection plant 16. Smithia salsuginea Papilionoideae Headysareae Shrub Amboli Ghat Hance Aeschynominae Dist. Kolhapur 17. Tephrosia pauciflora Galegeae Shrub Dapoli Grab, Tephrosiinae Dist. R a tn a g iri. 18. Tephrosia tinctoria Galegeae Shrub Bhimashankar Pers. Tephrosiinae Dist. Pune 19. Acacia tomentosa Mimosoideae Acacieae Tree Paud WTT3 Dist. Pune. <D U tO " rj 4J m o t'O tiO o o o o DOTJ o a o o o o D O :3 o o o F3 e e a o o CO cr: cy;) cii P::: oo CO CO cc 3 : <D p: rC: r>^ r-^ i/ r o c: CU3 O c C c •H ■ H •H }h PW K-q Xi a. (X :3 o r—H O (3) CO r d r C u n:3 •r-< to to to r-S^. • H p : ^ c f i: •H c ^ c •H p; U •M ; : : f-^'- '2 - ^ x: :s r3 • H O ClQ o Jh 0 c 0 ^ o bO o r. 0 w> :3 t-H r C •H o i j- ( • rH ■H W •H •H •H o r-P k J 4:) Q ^ CL, .- 4 ^ ^ ,£ 2 Pl, ^ X >< X r : EEE X X E (D EEEE n E E E E N t o t^) E E E E E E E E E E LO g ^ e E E CO r-H E E (N1 1 n r - j t o t^') -=f fNl r-l .o to CM to rH •-.-I fU o O <D T 3 (U 0) I r-X^ +-J r *H •M • 1 p . 0 0 0 1 +-> c:3 ■4 c3 I «' o » rt cd <u (/> to to to 03 O -r:i c e u QJ rH u 0) rC to 0 0 0 0 0 L-O 0 0 v-t to ;/> 1—1 bo ^ J-4 t—< U C/) 0 o . C C 3 03 :3 P .p uf o3 0 0 0 0 P 0 C P. +-> +-> C 0 r H r-i rH r H o ‘H 03 rH o rC 0 .- 4 oj 03 CT3 03 O O.H s •r+- CO a 0 0 '-H (T. CD o 0 0 > " p q o P m c : ■M 4-> 0 0 0 r-H 0 r H •r+ 0 ) 0 0 03 0 o3 03 03 rt C rt 4 J ^ Jh to f - to U !/> to to rt fn to :3 • H +-) ■p (U •!-> o ; 4-> 0 ) +-> O 4-> <D + J 0 4J o to 0 P.P. • P 0 (X 4-> 0 i-> o ■M O +-J O CU <M O c l 0 c i 0 03 03 03 0 03 03 0 o cd o 03 O 03 03 O C l. 0 H H ►-) H H ^ u ►-J ^ E—' 1^ tiO to 1 O') I V ) CJ U 0 ) cd ^ 03 ^ r H r : r H + J ^ QJ ;> H 0 ) •M cu D p : 0 ) ,Q 5 CD C c n 0 03 -C J E Mh P u S c u 03 T—1 w 0 0 0 p • H (D 'J o ::3 • H <D :3 c J“ « r-: E C >> X) 'm E C CM u c3 P ^ E 03 ^ E D 0 U Mh u C CD 0 (D P O C 0 <D :3 0 0 CO < k—( - P > P . P CO h-< 4J Pm G < to rHo O a . 03 w :1 cz o (D E •H o3 E U }••. p . ,rl fC - C j u 0 2 d U U) 0 cd +-> 0> r ; •H H tn o3 0 'J c c } } •M E c : >v 4J CO rH H r-v 4-J C j c j Cd a t::! r -< 0 +-> u rH c3 >•• . 0 , t o rH 4-J Zj 0 O h < P 'i Jl u I o rH X) , 03 U O CO ?!: 47 It could be seen from the table that position of nodules was on tap root in 4 species, on lateral roots in 11 species and on both tap and lateral roots in 4 species. The root nodules of the 19 species exhibited different shapes, sizes and colours. As fcr the out side colour, all the nodules showed a variety of colours ranging from light brown to dark brown except for flucuna monosperma which had black coloured nodules. It may be mentioned here that Dolichos lablab, Centrosema pubescens and Macroptilium atropurpureum have been reported to be having black coloured nodules (Cloonan 1963, Brockwell 1980}. When the nodules were cut, following 13 species viz. A3ysicarpus~ pubescens, Atylosia sericea, Crotalaria albida, C.calycina, C.leptostachya, C.triqetra, Dalbergia lanceolaria, Erythrina suberosa, Indigofera cassioides, Priotropsis cystisoides, Rhynchosia ro th ii, Smithia salsuginea and Tephrosia pauciflora had pink coloured interior. The remaining six species v iz . Cassia k le in ii, Butea monosperma, Erythrina stricta, Mucuna monosperma, Tephrosia tinetoria and Acacia tbmentosa, on the other hand, had reddish to brown colouration of the in te rio r. As for the shape of the nodules, Cassia kle in ii, Alysicarpus pubescens, Atylosia sericea, Butea monosperma, Pelbergia lanceolaria, Erythrina stricta, F.suberosa, Indigofera 48 cassioides and Acacia tomentosa had globose shape (F ig .2) Crotalaria albida, Rhynchosia rothii, Tephrosia pauciflora and T .tinctoria had elongate shape (Fig. 3), Crotalaria leptostachya , C . tr iquetra and Priotropsis cys 11 soi,des had bifurcate shape (Fig.4) C.albida, C.calycina and C.triquetra had fan shaped nodules (F ig .5) C.leptostachya and P.cystiso ides had Coralloid nodules (F ig .6] and Smith1 a salsucinea had siamese nodules (F ig .7). Tlie nodules o ^ Mucuna monosperma were irregular in shape (F ig .8). According to Corby (1971) and Lim and Ng (1977) nodule shapes are characteristics of legume species and independent of Rhi zobium strain s. As for the size of the nodules, it ranged between 2 mm to 18 mm, the smallest being shown by Smithia salsuginea and largest by Crotalaria leptostachya. As for the surface of the nodules, i: varied from smooth to hard and woody. In case of Frythrina suberosa the hard surface of nodule also showed whitish streaks. The data thus show that nodulation on 19 wild leguminous species is a first report. 5.2 Isolation and Identification of root nodule bacteria The details of the growth of the isolates from root nodules of 19 leguminous spp. on YB>! agar are given in Table 4, together with the results of Ketolactose test, growth in 0) vj- u.D’ U'J u (/} rC Cj • H rt X X rC ■fj 4-> rC 4-> rC rC 4-> u-» X J-J a) ciO":3 tO 'O o o t O cj)nd CO O •#-> 'H D o :3 ^ o o o ;n. o ::3 -*--> o o o rt e G c E O CTj o E cy^ 1/1 c^: a:: CO lO q : CO Cr: cx in rC q; tn (/) T3 ■M P •H •H •H c •H rP ^ 'T^ in t/) o P r: r:J C C q P •T3 o P •ri f~.