Nodulated tree legumes and

Nodulated tree legumes and

their symbiotic

Bradyrhizobium in African

and South-American

Tropical Rainforests

Y. Prin, B. Dreyfus, C. LeRoux, G. Bena,

M. Diabaté, P. de Lajudie, A. Bâ,

S. M. de Faria, A. Munive & A. Galiana

Laboratoire des Symbioses Tropicales et

Méditerranéennes (LSTM), Campus

International de Baillarguet, 34398

Montpellier Cedex 5, France.

The Leguminosae family

• 650 genera, 18,000 species

• Three sub-families:

– Caesalpinioideae

(about 26% nodulated)

• 1,900 species (woody perennials, tropics)

– Mimosoideae

(about 95% nodulated)

• 2,700 species (woody perennials, tropics)

– Papilionoideae

(about 97% nodulated)

• 13,400 species (herbaceous to trees,

temperate to tropical)

A wide diversity of nodule types in tropical rainforest trees

from West Africa (Guinea) and South America (French Guyana)

Occcurence of nodulation in leguminous species of the

primary rain forest in Guinea (West Africa)

Sub-family / Tribe /Species Plant habit and height

Location Nodulation New species observed

New genus observed

CAESALPINIOIDEAE

CAESALPINIEAE

Bussea occidentalis TŹ; 25 m Ziama - yes yes

Chidlowia sanguinea T ; 25 m Ziama + Ņ Ņ

Delonix regia T ; 12 m Nimba - no no

Erythrophleum ivorense T ; 40 m Ziama + Ņ Ņ

Erythrophleum suaveolens T ; 40 m Bˇ ro + Ņ ŅŹ

Mezoneuron benthamianum S ; 5 m Diˇc kˇ - yes ŅŹ

CASSIEAE

Cassia aubrevillei TŹ; 15 m Nimba - yes no

Cassia sieberiana TŹ; 15 m Bˇ ro - Ņ Ņ

Chamaecrista mimosoides H ; 1.5 m Nimba + no Ņ

Dialium aubrevillei T ; 30 m Ziama - yes Ņ

Dialium dinklagei T ; 20 m Diˇc kˇ - ć Ņ

Dialium guineense T ; 15 m Bˇ ro - no Ņ

Dialium pobeguinii T ; 15 m Ziama - yes Ņ

Distemonanthus benthamianus

T ; 35 m Ziama - Ņ yes

Duparquetia orchidacea SŹ; 8 m Nimba - Ņ ŅŹ

Senna podocarpa S ; 5 m Nimba - Ņ no

Senna siamea T ; 20 m Bˇ ro - no Ņ

CERCIDEAE

Bauhinia thonninguii T ; 9 m Pic de Fon - yes no

Griffonia simplicifolia SŹ; 5 m NÕzˇ rˇk orˇ - Ņ yes

DETARIEAE

Afzelia africana T ; 30 m Bˇ ro - no no

Afzelia bella var.gracilior T ; 35 m Ziama - yes Ņ

Copaifera salikounda T ; 30 m Ziama - Ņ Ņ

Daniellia ogea T ; 30 m Bˇ ro - ŅŹ yes

Daniellia thurifera T ; 35 m Ziama - Ņ Ņ

Detarium heudelotianum T ; 20 m - Ņ Ņ

Detarium macrocarpum T ; 25 m Diˇc kˇ - Ņ Ņ

Detarium senegalense T ; 25 m Bˇ ro - ŅŹ Ņ

Guibourtia copallifera T ; 25 m Ziama - Ņ no

Guibourtia ehie T ; 30 m Ziama - Ņ Ņ

Guibourtia leonensis T ; 30 m Diˇc kˇ - Ņ Ņ

Tessmannia baikiaeoides T ; 10 m Nimba - Ņ yes

AMHERSTIEAE

Anthonotha crassifolia T ; 20 m Bˇ ro - yes no

Anthonotha fragrans T ; 30 m Ziama - no Ņ

Anthonotha macrophylla T ; 12 m Diˇc kˇ - yes Ņ

Cryptosepalum tetraphyllum T ; 25 m Ziama - Ņ Ņ

Gilbertiodendron bilineatum T ; 15 m Ziama - Ņ Ņ

Gilbertiodendron limba T ; 15 m Ziama - Ņ Ņ

Nodulation of leguminous species from Guinea

(continued)

Sub-family / Tribe /Species

Plant habit

and height

Location Nodulation

New

species

observed

New

genus

observed

MIMOSOIDEAE

PARKIEAE

Parkia bicolor

T ; 30 m

Nimba

-

yes

no

Parkia biglobosa

T ; 15 m

Bˇ ro

-

no

Ņ

Pentaclethra macrophylla

T ; 25 m

Ziama

+

Ņ

Ņ

MIMOSEAE

Adenopodia scelerata

V ; 30 m

Nimba

-

yes

yes

Aubrevillea kerstingii

T ; 30 m

Ziama

-

Ņ

Ņ

Aubrevillea platycarpa

TŹ; 30 m

Ziama

+

Ņ

Ņ

Calpocalyx aubrevillei

TŹ; 25 m

Ziama

+

Ņ

no

Calpocalyx brevibracteatus

T ; 25 m

Ziama

-

Ņ

Ņ

Dichrostachys cinerea

S ; 8 m

Bˇ ro

+

no

ŅŹ

Entada africana

TŹ; 12 m

Bˇ ro

+

yes

Ņ

Entada gigas

VŹ; 30 m

Diˇc kˇ

+

ŅŹ

Ņ

Entada mannii

S ; 15 m

Ziama

+

Ņ

Ņ

Mimosa invisa

S ; 0.5 m

Diˇc kˇ

+

no

Ņ

Newtonia aubrevillei

T ; 30 m

Ziama

-

yes

Ņ

Newtonia duparquetiana

TŹ; 25 m

Nimba

-

Ņ

Ņ

Piptadeniastrum africanum

TŹ; 40 m

Ziama

+

no

no

Tetrapleura tetraptera

T ; 20 m

Ziama

+

yes

yes

Xylia evansii

T ; 25 m

Diˇc kˇ

+

Ņ

no

INGEAE

Albizia adianthifolia

T ; 20 m

Diˇc kˇ

+

no

no

Albizia altissima

T ; 20 m

Ziama

+

yes

Ņ

Albizia ferruginea

T ; 20 m

Bˇ ro

+

no

Ņ

Albizia glaberrima

T ; 15 m

Nimba

+

Ņ

Ņ

Albizia zygia

T ; 20 m

Ziama

+

Ņ

Ņ

Samanea dinklagei

T ; 20 m

Ziama

+

yes

Ņ

Tetrapleura tetraptera T ; 20 m Ziama + yes yes

Aubrevillea platycarpa T ; 30 m Ziama + yes yes

Nodulation of leguminous species from Guinea

(continued)

Sub-family / Tribe /Species Plant habit

and height Location Nodulation New speciesobserved New genusobserved

PAPILIONOIDEAE

SWARTZIEAE

Bobgunnia fistuloides T ; 15 m Diˇc kˇ - yes no

SOPHOREAE

Amphimas pterocarpoides T ; 25 m Ziama + yes yes

Angylocalyx oligophyllus S ; 4 m Nimba - Ņ Ņ

Baphia nitida T ; 10 m Nimba + no no

Baphia capparidifolia T ; 8 m Diˇc kˇ + yes Ņ

DALBERGIEAE

Dalbergia afzeliana V ; 35 m Nimba - yes no

Dalbergia bignonae V ; 25 m Bˇ ro + Ņ Ņ

Dalbergia hostilis V ; 15 m Ziama + Ņ Ņ

Dalbergia oblongifolia S ; V ; 10 m Diˇc kˇ - Ņ Ņ

Dalbergia saxatilis S ; V ; 20 m Nimba - Ņ Ņ

Pterocarpus erinaceus TŹ; 15 m Bˇ ro + no Ņ

Pterocarpus mildbraedii T ; 20 m Ziama + yes Ņ

Pterocarpus santalinoides TŹ; 15 m Ziama - no Ņ

MILLETTIEAE

Dalbergiella welwitschii S ; V ; 20 m Nimba - yes no

Leptoderris brachyptera S ; V ; 20 m Ziama + Ņ Ņ

Leptoderris fasciculata S ; V ; 20 m Nimba + Ņ Ņ

Lonchocarpus cyanescens S ; V ; 30 m Nimba - Ņ Ņ

Millettia dinklagei S ; V ; 15 m Nimba + Ņ Ņ

Millettia griffoniana T ; 15 m Nimba - Ņ Ņ

Millettia lucens S ; V ; 15 m Bˇ ro - Ņ Ņ

Millettia rhodantha T ; 12 m Ziama + Ņ Ņ

Millettia zechiana S ; 10 m Ziama + Ņ Ņ

Platysepalum hirsutum S ; V ; 20 m Nimba - Ņ yes

Tephrosia flexuosa S ; 1.5 m Nimba - Ņ no

DESMODIEAE

Droogmansia scaettaiana H ; 2 m Nimba + yes no

PHASEOLEAE

Dioclea reflexa S ; V ; 15 m Nimba - no no

Dolichos dinklagei S ; 1.5 m Nimba - yes Ņ

Dolichos tonkouiensis SŹ; 1. 5 m Nimba + Ņ Ņ

Erythrina milbraedii T ; 30 m Ziama + Ņ Ņ

Erythrina senegalensis S ; 8 m Diˇc kˇ + no Ņ

Mucuna flagellipes SŹ; VŹ; 5 m Bˇ ro - yes Ņ

Mucuna poggei S ; V ; 20 m Bˇ ro + no Ņ

Physostigma venenosum S ; V ; 6 m Bˇ ro - yes yes

AESCHYN OMENEAE

Kotschya lutea S ; 2 m Nimba + yes no

Kotschya ochreata S ; 2.5 m Nimba + Ņ Ņ

Ormocarpum megalophyllum S ; 1. 5 m Nimba - Ņ Ņ

CROTALARIEAE

Crotalaria lathyroides S ; 2 m Bˇ ro + no Ņ

Amphimas pterocarpoides + yes yes

Pterocarpus erinaceus +

Pterocarpus mildbraedii

+

-Occurence of nodulation in leguminous tree species

of the Amazonian bassin (French Guyana)

Sub-family / Tribe /Species Nodulation New species observed

New genus observed

CAESALPINIOIDEAE

CAESALPINIEAE

Dimorphandra polyandra + yes no

Tachigali melinonii + no Ņ

Tachigali myrmecophila + yes Ņ

CASSIEAE

Chamaecrista diphylla + no no

DETARIAE

Macrolobium bifolium - yes no

MIMOSOIDEAE

MIMOSEAE Stryphnodendron pulcherrimum + no no INGEAE Hydrochorea corymbosa + no no Inga disticha + Ņ Ņ

Inga sertulifera + yes Ņ

Zygia latifolia + no Ņ Zygia cataracteae + Ņ Ņ

PAPILIONOIDEAE

SWARTZIEAE Bocoa prouacensis - no no Swartzia polyphylla + Ņ Ņ SOPHOREAE Diplotropis purpurea + no no Ormosia coutinhoi + Ņ Ņ

Spirotropis longifolia + yes yes

DALBERGIEAE Andira inermis + no no Dalbergia monetaria + Ņ Ņ Hymenolobium flavum + Ņ Ņ Machaerium lunatum + Ņ Ņ Paramachaerium ormososioides + Ņ Ņ Pterocarpus officinalis + Ņ Ņ Vatairea erythrocarpa - Ņ Ņ BRONGNIARTIEAE Poecilanthe hostmannii + no no PHASEOLEAE Clitoria fairchildiana + no no

Erythrina amazonica + yes Ņ

Confirmation of nodulation status in nursery

Chidlowia sanguinea

Piptadeniastrum africanum

Tetrapleura tetraptera

Erythrophleum ivorense

a

b

g

Methylobacterium extorquens

Rickettsia rickettsii

Bradyrhizobium sp

Paracoccus denitrificans

Agrobacterium rhizogenes

Xanthobacter agilis

Azospirillum brasilense

Methylobacterium nodulans

Azoarcus indigens

Bradyrhizobium japonicum

Rhizobium leguminosarum

Sinorhizobium meliloti

Mesorhizobium loti

Azorhizobium caulinodans

Burkholderia

sp.

STM678

Burkholderia

sp.

STM815

Ralstonia taiwanensis

Neisseria meningitidis

Alcaligenes eutrophus

Burkholderia graminis

Burkholderia brasilense

Burkholderia cepacia

Bordetella pertusis

Nitrosovibrio tenuis

Spirillum volutans

0.01 changes

d

e

Phylogeny of 16S rDNA

Robinioids

Millettioids

Mirbelioids

Dalbergioids

Trifolium, Pisum, Vicia ... Trigonella, Melilotus, Medicago... Astragalus, Clianthus, Onobrychis ... Robinia, Gliricidia... Lotus, Anthyllis, Sesbania... Milletia, Tephrosia... Phaseolus, Macrotyloma, Vigna... Glycine, Pueraria... Indigofera Bossiaea, Daviesia... Pterocarpus, Stylosanthes, Arachis...

Aeschynomene

Dalbergia, Machaerium... Chaetocalyx, Nissolia Zornia... Amorpha, Dalea...

Phylogeny of Leguminosae based on parsimony analyses of matK gene sequences, adapted from Wojciechowski et al. (2004).

Lacking Inverted

Repeat Clade

Polygala californica Quillaja saponaria Vauquelinia californica Suriana maritima

Genistoids

Cladrastris

Papilionioids

Mimosoids

Caesalpinioids crown clade

Bradyrhizobium,Mesorhizobium, Rhizobium, Ochrobactrum Methylobacterium Genista, Lupinus, Cytisus Crotalaria Sophora... Bowdichia, Diplotropis, Hovea... Nodulated, by ? Vataira Uribea... Non-nodulated Cladastris Non-nodulated Myrospermum,

Dipteryx... Non-nodulated or no nodulation report Swartzia

Ateleia... Nodulated, by ?

Bradyrhizobium,Rhizobium, Sinorhizobium, Mesorhizobium, Allorhizobium Albizia, Calliandra, Acacia, Neptunia, Leucaena, Prosopis... Tetrapleura, Entada,

Xylia... Bradyrhizobium

,

nodulated by ?, or non-nodulated

Cercidium, Dinizia... Non-nodulated Non-nodulated or Bradyrhizobium Senna Chamaecrista, Caesalpinia... Gleditsia, Ceratonia... Non-nodulated

Non-nodulated or no nodulation report

Berlinia, Prioria, Tessmannia, Cercis...

Phylogeny of Leguminosae based on parsimony analyses of matK gene sequences, adapted from Wojciechowski et al. (2004).

STM726 Albizia zygia STM927 Millettia zechiana STM739 Albizia sassaSTM924 Albizia zygia STM928 Millettia zechianaSTM930 Millettia zechiana

STM929 Millettia zechiana STM724 Samanea dinklagei

STM741 Erythrophleum ivorensisSTM723 Samanea dinklagei STM788 Dalbergia monetariaSTM838 Dalbergia monetaria STM834 Dalbergia monetariaSTM845 Dalbergia monetaria STM826 Dalbergia monetariaSTM938 Andira inermis STM835 Dalbergia monetaria STM823 Dalbergia monetaria STM833 Dalbergia monetariaSTM756 Dalbergia monetaria STM827 Dalbergia monetaria STM757 STM803 Dalbergia monetariaAndira inermis

Bradyrhizobium japonicumT _{Genomic species I}

Bradyrhizobium spGenomic species V Bradyrhizobium spBradyrhizobiumGenomic species IVliaoningense

T_{Genomic species III} Group A Group B Group C Bradyrhizobiumsp Genomic species VI Bradyrhizobiumsp

Genomic species VII STM257 Albizia ferruginea

BradyrhizobiumspGenomic species VIII

Azorhizobium cluster

Mesorhizobium cluster BradyrhizobiumspGenomic species IX Bradyrhizobium elkaniiT_{Genomic species II}

STM759 Indigofera sp STM728 Chidlowia sanguinea STM841 Erythrina amazonica STM786 Erythrina amazonica STM751 Erythrina amazonica STM731 Chidlowia sanguinea STM727 Chidlowia sanguinea STM794 Chidlowia sanguinea

BradyrhizobiumspGenomic species XI

Group G

Group F Bradyrhizobiumsp Genomic species X

STM750 Erythrina amazonica

STM783 Andira inermis

STM749 STM758 Indigofera spAndira inermis

STM840 Indigofera sp STM745 Erythrina amazonica

STM744 Erythrina amazonica

Group E

ORS1812 Abrus pulchellus

Group D

ORS170 Faidherbia albida STM792 STM851 Piptadeniastrum africanumMillettia zechiana

STM790 Millettia zechiana STM795 Millettia zechiana STM725 STM931 Albizia ferrugineaMillettia rothonda STM934 STM916 Pterocarpus mildbraediiAlbizia adianthifolia

STM926 Millettia zechiana STM923 Albizia zygia STM922 Albizia ferruginea STM722 Pterocarpus mildbraedii STM933 Piptadeniastrum africanum STM932 Piptadeniastrum africanum STM732 Pterocarpus mildbraedii ITS phylogeny of Bradyrhizobium rDNA

Plant Genus species: Amazonian species

Inoculation trials in Republic of Guinea

- Inoculation in nursery (50 replicates)

-Transplantation to the field after 3 months

Effect of rhizobial inoculation on growth of Albizia

spp. and Samanea dinklagei

(after 4 months)

A. altissima x STM916

S. dinklagei x STM922

A. ferruginea x STM922

A. zigia x STM923

A. adianthifolia x STM916

Height

+75%

+20%

+50%

Effect of rhizobial inoculation on growth of

Millettia spp.

(after 4 months)

M. Zechiana x

STM926

M. rhodantha x

_STM931

M. zechiana x

_STM851

Height

+107%

+58%

+63%

Ctrl (-)

Effect of rhizobial inoculation on growth of

Erythrophleum spp.

(after 4 months

)

E. ivorensis x

STM934

E. guineensis x

STM934

Height

+22%

40%

Ctrl(-)

CONCLUSION

• Tropical rainforests = huge reservoirs of symbioses

• Predominance of the genus Bradyrhizobium

• Need for collaborative research between plant and

bacteria phylogenists

• Large potential of applications for reforestation with

endangered species

- IRAG, BP 1523, Conakry, République de Guinée

- Universita Autonoma del Puebla, Mexico

- Embrapa/Agrobiologia, BR465, Seropedica, Brazil

- LSTM, Montpellier, France