Studies on arbuscular mycorrhiza (AM) in the Alentejo (Portugal) using pea mutants resistant to AM fungi as a control tool for field conditions

The utilization and management of arbuscular mycorrhiza (AM) symbiosis may improve production and sustainability of the cropping system. For this purpose, native AM fungi (AMF) were sought and tested for their efficiency to increase plant growth by enhanced P uptake and by alleviation of drought stress. Pot experiments with safflower (Carthamus tinctorius) and pea (Pisum sativum) in five soils (mostly sandy loamy Luvisols) and field experiments with peas were carried out during three years at four different sites. Host plants were grown in heated soils inoculated with AMF or the respective heat sterilized inoculum. In the case of peas, mutants resistant to AMF colonization were used as non-mycorrhizal controls. The mycorrhizal impact on yields and its components, transpiration, and P and N uptake was studied in several experiments, partly under varying P and N levels and water supply. Screening of native AMF by most probable number bioassays was not very meaningful. Soil monoliths were placed in the open to simulate field conditions. Inoculation with a native AMF mix improved grain yield, shoot and leaf growth variables as compared to control. Exposed to drought, higher soil water depletion of mycorrhizal plants resulted in a haying-off effect. The growth response to this inoculum could not be significantly reproduced in a subsequent open air pot experiment at two levels of irrigation and P fertilization, however, safflower grew better at higher P and water supply by multiples. The water use efficiency concerning biomass was improved by the AMF inoculum in the two experiments. Transpiration rates were not significantly affected by AM but as a tendency were higher in non-mycorrhizal safflower. A fundamental methodological problem in mycorrhiza field research is providing an appropriate (negative) control for the experimental factor arbuscular mycorrhiza. Soil sterilization or fungicide treatment have undesirable side effects in field and greenhouse settings. Furthermore, artificial rooting, temperature and light conditions in pot experiments may interfere with the interpretation of mycorrhiza effects. Therefore, the myc- pea mutant P2 was tested as a non-mycorrhizal control in a bioassay to evaluate AMF under field conditions in comparison to the symbiotic isogenetic wild type of var. FRISSON as a new integrative approach. However, mutant P2 is also of nod- phenotype and therefore unable to fix N2. A 3-factorial experiment was carried out in a climate chamber at high NPK fertilization to examine the two isolines under non-symbiotic and symbiotic conditions. P2 achieved the same (or higher) biomass as wild type both under good and poor water supply. However, inoculation with the AMF Glomus manihot did not improve plant growth. Differences of grain and straw yields in field trials were large (up to 80 per cent) between those isogenetic pea lines mainly due to higher P uptake under P and water limited conditions. The lacking N2 fixation in mutants was compensated for by high mineral N supply as indicated by the high N status of the pea mutant plants. This finding was corroborated by the results of a major field experiment at three sites with two levels of N fertilization. The higher N rate did not affect grain or straw yields of the non-fixing mutants. Very efficient AMF were detected in a Ferric Luvisol on pasture land as revealed by yield levels of the evaluation crop and by functional vital staining of highly colonized roots. Generally, levels of grain yield were low, at between 40 and 980 kg ha-1. An additional pot trial was carried out to elucidate the strong mycorrhizal effect in the Ferric Luvisol. A triplication of the plant equivalent field P fertilization was necessary to compensate for the mycorrhizal benefit which was with five times higher grain yield very similar to that found in the field experiment. However, the yield differences between the two isolines were not always plausible as the evaluation variable because they were also found in (small) field test trials with apparently sufficient P and N supply and in a soil of almost no AMF potential. This similarly occurred for pea lines of var. SPARKLE and its non-fixing mycorrhizal (E135) and non-symbiotic (R25) isomutants, which were tested in order to exclude experimentally undesirable benefits by N2 fixation. In contrast to var. FRISSON, SPARKLE was not a suitable variety for Mediterranean field conditions. This raises suspicion putative genetic defects other than symbiotic ones may be effective under field conditions, which would conflict with the concept of an appropriate control. It was concluded that AMF resistant plants may help to overcome fundamental problems of present research on arbuscular mycorrhiza, but may create new ones.

Causes of legume rotation induced yield increases in cereals grown on West African soils

This study was conducted to investigate soil biological and chemical factors that give rise to cereal yield enhancing effects of legume rotations on sandy, nutrient poor West African soils. The aim was not only to gain more information on the role of legume residues and microorganisms in the soil nutrient cycle. But the study aimed at evaluating if differences in substrate qualities (e.g. root residues) cause changes in the microbial community structure due to specific and highly complex microbe-root-soil interactions. Site and system specific reactions of microorganisms towards rewetting, simulating the onset of rainy season, were observed. Higher respiration rates, higher amounts of microbial biomass carbon (Cmic) and nitrogen (Nmic) as well as higher ergosterol, muramic acid, glucosamine and adenylate concentrations were measured in CL soils of Koukombo and in both soils from Fada. The immediate increase in ATP concentrations after rewetting was likely caused by rehydration of microbial cells where N was not immobilized and, thus, available for plants facilitating their rapid development. Legume root residues led only to slightly better plant performances compared to the control, while the application of cereal roots reduced seedling growth. In contrast to sorghum seedlings, the microbial community did not react to the mineral treatment. Thus the energy supply in form of organic amendments increased microbial indices compared to mineral P application and the control. The results of basal respiration rates, Cmic and Corg levels indicate that the microbial community in the soil from Koukombo is less efficient in substrate use compared to microorganisms in the soil from Fada. However, the continuous carbon input by legume root residues might have contributed to these differences in soil fertility. With the 33P isotopic exchange method a low buffering capacity was detected in both soils irrespective of treatments. Calculated E values (E1min to E1min-1d and E1d-3m) indicated a slowly release of P due to root turnover while applied mineral P is taken up by plants or fixed to the soil. Due to the fact that sorghum growth reacted mainly to the application of mineral P and the microorganisms solely to the organic inputs, the combination of both amendments seems to be the best approach to a sustainable increase of crop production on many nutrient-poor, sandy West African soils. In a pot experiment, were CC and CL soils from Fada and Koukombo were adjusted to the same level of P and N concentrations, crop growth was significantly higher on CL soils, compared to the respective treatments on CC soils. Mycorrhizal infection of roots was increased and the number of nematodes, predominantly free living nematodes, was almost halfed on rotation soils. In conclusion, increased nutrient availability (especially P and N) through the introduction of legumes is not the only reason for the observed yield increasing effects. Soil biological factors seem to also play an important role. In a root chamber experiment the pH gradient along the root-soil-interface was measured at three times using an antimony microelectrode. For Fada soils, pH values were higher on CL than CC soils while the opposite was true for the Koukombo soils. Site-specific differences between Fada and Koukombo soils in N content and microbial community structures might have created varying crop performances leading to the contrasting pH findings. However, the mechanisms involved in this highly complex microbe-root-soil interaction remain unclear.