Humberto A. Leblanc

Detection of nitrogen transfer from N2-fixing shade trees to cacao saplings in 15N labelled soil: ecological and experimental considerations

Theobroma cacao seedlings were grown alone (TCA) or associated with saplings of N2-fixing shade trees Gliricidia sepium and Inga edulis in 200 l of 15N labelled soil within a physical root barrier for studying direct nitrogen transfer between the trees and cacao. Root:shoot partitioning ratio for sapling total N was lower than biomass root:shoot ratio in all species. Sapling total 15N was partitioned between root and shoot in about the same ratio as total N in cacao and inga but in gliricidia much higher proportion of 15N than total N was found in roots. Thus, whole plant harvesting should be used in 15N studies whenever possible. Average percentage of fixed N out of total tree N was 74 and 81% for inga estimated by a yield-independent and yield-dependent method, respectively, and 85% for gliricidia independently of estimation method. Strong isotopic evidence on direct N transfer from trees to cacao was observed in two cases out of ten with both tree species. Direct N transfer was not correlated with mycorrhizal colonisation of either donor or receiver plant roots. Direct N transfer from inga and gliricidia to cacao is conceivable but its prevalence and the transfer pathway via mycorrhizal connections or via reabsorption of N-rich legume root exudates by cacao require further study. Competition in the restricted soil space may also have limited the apparent transfer in this study because the trees accumulated more soil-derived N than cacao in spite of active N2 fixation.

Dinitrogen fixation by legume shade trees and direct transfer of fixed N to associated cacao in a tropical agroforestry system

Natural abundance of (15)N (δ (15)N) was determined in bulk soil, rhizospheric soil and vegetation in an organically managed cacao (Theobroma cacao L.) plantation with Inga edulis Mart. legume trees (inga) as the principal shade for studying the nitrogen (N) cycle in the system. Cacao without contact with legumes in an adjacent plantation was used as the reference for N2 fixation and direct N transfer calculations. Bulk and rhizospheric soils contained 72 and 20%, respectively, of whole- system N. No vegetation effect on δ (15)N in rhizospheric soil was detected, probably due to the high native soil N pool. Fine roots of the cacaos associated with inga contained ∼35% of N fixed from the atmosphere (Nf) out of the total N. Leaves of all species had significantly higher δ (15)N than fine roots. Twenty percent of system Nf was found in cacao suggesting direct N transfer from inga via a common mycelial network of mycorrhizal fungi or recycling of N-rich root exudates of inga. Inga had accumulated 98 kg [Nf] ha(-1) during the 14-year history of the plantation. The conservative estimate of current N2 fixation rate was 41 kg [Nf] ha(-1) year(-1) based on inga biomass only and 50 kg [Nf] ha(-1) year(-1) based on inga and associated trees.

Leaf and root litter of a legume tree as nitrogen sources for cacaos with different root colonisation by arbuscular mycorrhizae

Traditionally cacao (Theobroma cacao L.) is cultivated under legume shade trees, which produce N-rich litter that improves soil organic matter content, microbial activity, and recycles N to the crop. Arbuscular mycorrhiza forming fungi (AMF) are known to play an important role in plant nutrient uptake, yet their role in plant N uptake from organic residues in tropical agroforestry systems is not clear. We studied root and leaf litter of the legume shade tree Inga edulis Mart. as a source of N for cacao and the importance of AMF colonisation in the uptake of litter N under controlled conditions. Leaf and root litter of I. edulis enriched with 15N was added to cacao pots filled with field soil. Half of the cacao saplings were AMF-inoculated and the soil of non-inoculated saplings was treated with fungicide to suppress AMF. During the 10-week experiment, young cacao leaves were sampled for 15N analyses and at the end of the experiment whole plants were harvested. Microbial populations in the soil were determined using phospholipid fatty acid (PLFA) analysis, and AMF structures in the roots were quantified. Fungicide treatment decreased AMF structures in roots and increased bacterial populations, but did not affect the decomposition rate of either litter type. Inoculated and non-inoculated cacao saplings used 2.6 and 2.1%, respectively, of N added to the pots in leaf litter and 12.1 and 7.1% of N available in root litter indicating that root litter of I. edulis may be a more efficient N source than leaf litter for cacao. Although the fungicide treatment did not completely suppress AMF in non-inoculated pots, it created sufficient contrast in root AMF colonisation for concluding that AMF significantly enhanced cacao N use from both litter types. The role of root litter of shade trees as a N source in agroforestry should not be neglected.

Response of Theobroma cacao and Inga edulis seedlings to cross-inoculated populations of arbuscular mycorrhizal fungi

Response of Theobroma cacao L. (cacao) and the shade tree Inga edulis Mart. (inga) seedlings from an organically-grown cacao plantation to inoculation with native arbuscular mycorrhizae forming fungi (AMF) was studied in a cross-inoculation assay under greenhouse conditions. Seedlings of inga and cacao were grown in pots filled with heat-treated soil from the plantation. Control was heat-treated soil without inoculum and roots of cacao and inga from the plantation were applied as AMF inocula. Undisturbed soil blocks were used as a “positive control” of the inoculation potential of untreated soil and roots combined. No AMF structures were observed in the roots of either species in the heat-treated control. All inocula were infective in both hosts and the differences in the total AMF colonization percentage between the hosts were not significant but inga had significantly higher colonization by hyphal coils and arbuscules. Cacao roots but neither inga roots nor soil block inocula stimulated cacao growth. All inocula significantly increased growth of inga, which had higher relative mycorrhizal responsiveness than cacao. Thus, in spite of the strong infectivity of the inocula in both hosts, cacao and inga responded differently to the same AMF populations. The strong conspecific preference of cacao suggests that attention must be paid to the AMF inoculum used for this species. However, the strong response of inga to cacao root inoculum indicates that the two species may share same AMF symbionts, thus enabling positive interactions between them, including formation of common mycelial networks.