Thermal sensitivity of Phytophthora cinnamomi and long-term effectiveness of soil solarisation to control avocado root rot

Root rot caused by the fungus Phytophthora cinnamomi is a major disease of avocados worldwide. Heat sensitivity of a collection of P. cinnamomi isolates was determined by exposing agar discs containing mycelium or mycelium plus chlamydospores at various temperatures for different periods. Long‐term effectiveness of soil solarisation to control Phytophthora root rot was evaluated in two field trials. In the first, soil disinfestation by solarisation was applied in 1990 to a naturally infested plot before planting avocado (Persea americana) and viñatigo (Persea indica) seedlings. In the second trial, established avocado trees were solarised for four consecutive summers (1996–1999). Results for heat sensitivity showed that fungal mycelium was inactivated after 1–2 h at 38°C. However, 1–2 h at 40°C was needed to kill all propagules when chlamydospores were present. Fungal growth inhibition after thermal treatments was related to levels of time and temperature, and detrimental effects occurred as consequence of sublethal thermal doses. Soil solarisation presented long‐term positive effects when applied as a preplanting treatment. Five years after solarisation, disease severity (0–5 scale where 0 = healthy and 5 = dead plant) of avocado and viñatigo planted in solarised soil was 2.03 and 0.71, respectively, compared with 4.65 and 4.84 in controls. Eleven years after solarisation, the percentage of dead plants in solarised soil was 73% for avocado and 43% for viñatigo but 100% in controls. In contrast, an insufficient level of control was observed in established orchards, probably because of the lower temperature reached during solarisation under the shade of tree canopy. In this situation, maximum temperatures at 5‐cm depth were 10–13.7°C lower than under solar‐heated mulch, only exceeding 40°C in 1997.     

Cloning and expression of a gene encoding a root specific nitrate reductase in bean (Phaseolus vulgaris)

A structural gene encoding nitrate reductase (NR) in the legume Phaseolus vulgaris cv. Saxonia has been cloned and sequenced. The NR gene encodes a protein of 881 amino acids with a MW of 99.2 kDa. The coding sequence is interrupted by three introns, which are located in the molybdopterin cofactor binding domain. In the flanking regions the signals of a functional eukaryotic gene are present. The gene is the smallest NR gene so far identified in higher plants. Comparison to other NRs shows homology ranging from 65 to 74% at the amino acid level. The similarity is highest for the three functional domains, and lowest in the N-terminal end of the protein. mRNA studies demonstrate that the gene is nitrate inducible and expressed exclusively in the roots of bean. Southern blot analysis indicates the presence of a second NR gene in bean. This gene may encode a NR enzyme expressed in leaves.     

Effect of different soil and clove treatments in the control of white rot of garlic

Soil solarisation was consistently efficacious in reducing inoculum density to undetectable levels in a field naturally‐infested with Sclerotium cepivorum. This treatment delayed epidemic onset of white rot of garlic 2–3 months as compared with the untreated control or the inoculation of planting furrows with Glomus intraradices. Furthermore, significant reductions of disease incidence and of the standardised AUDPC were also observed in solarised plots, resulting in quantitative and qualitative yield improvement. Similar effects were observed in plots planted with tebuconazole‐treated cloves, confirming previous results, whereas Trichoderma harzianum was ineffective as a biocontrol agent, when applied to planting furrows. The inoculation of plots with G. intraradices before planting, in three consecutive years, was neither effective for disease control nor on the development of garlic, although the root systems of garlic plants from all the experimental treatments were heavily mycorrhizal at harvest, indicating the presence of native arbuscular mycorrhizal propagules in the soil and their survival after soil solarisation.     

Towards a better understanding of the genetic and physiological basis for nitrogen use efficiency in maize

To enhance our understanding of the genetic basis of nitrogen use efficiency in maize (Zea mays), we have developed a quantitative genetic approach by associating metabolic functions and agronomic traits to DNA markers. In this study, leaves of vegetative recombinant inbred lines of maize, already assessed for their agronomic performance, were analyzed for physiological traits such as nitrate content, nitrate reductase (NR), and glutamine synthetase (GS) activities. A significant genotypic variation was found for these traits and a positive correlation was observed between nitrate content, GS activity and yield, and its components. NR activity, on the other hand, was negatively correlated. These results suggest that increased productivity in maize genotypes was due to their ability to accumulate nitrate in their leaves during vegetative growth and to efficiently remobilize this stored nitrogen during grain filling. Quantitative trait loci (QTL) for various agronomic and physiological traits were searched for and located on the genetic map of maize. Coincidences of QTL for yield and its components with genes encoding cytosolic GS and the corresponding enzyme activity were detected. In particular, it appears that the GS locus on chromosome 5 is a good candidate gene that can, at least partially, explain variations in yield or kernel weight. Because at this locus coincidences of QTLs for grain yield, GS, NR activity, and nitrate content were also observed, we hypothesize that leaf nitrate accumulation and the reactions catalyzed by NR and GS are coregulated and represent key elements controlling nitrogen use efficiency in maize.     

Calcium and temperature effects on seedling exudation and root rot infection of common bean on an acid sandy soil

Soil born fungi such as Phytium ultimum, Fusarium ssp., and Rhizoctonia solani (Kühn) severely restrict stand establishment of common bean (Phaseolus vulgaris L.) on acid soils of the Tropics. Calcium application is known to alleviate fungal infection in many legumes but the causes are still unclear. To investigate environmental factors and physiological mechanisms involved, growth chamber experiments were conducted with an acid sandy soil from Mexico. Treatments were soil liming at a rate of 0.67 g Ca(OH)₂ kg^-1, gypsum application at 0.49 g CaSO₄ 2H₂O kg^-1 soil placed around the seed, and an untreated control. Beans were grown under three temperature regimes with constant night and one constant day vs. two sinusoidal day temperatures. To examine patterns of seed and seedling exudation at regular intervals leachates of germinating seeds were collected on filter paper soaked with equilibrium solutions from soils of the three treatments. The severity of root rot in the control treatment was highest when plants were stressed by temperature extremes. At a sinusoidal day temperature peaking at 40°C soil liming and gypsum application to the seed increased the number of healthy seedlings similarly by over 60%. However, only liming which effectively eliminated growth constraints by low pH and high aluminum concentrations led to an increase in hypocotyl elongation by 22% and in total root length by 8%. Both calcium amendments increased the calcium and potassium contents in the hypocotyl tissue. From seeds exposed to the equilibrium solution of unlimed soil with pH 3.7, 1 mM Ca, and 0.6 mM Al considerable amounts of amino acids and carbohydrates were leached. In contrast, exposure to the equilibrium solution from limed soil with pH 4.3, 3 mM Ca, and negligible concentrations of Al led to a net uptake of amino acids and decreased leaching of carbohydrates. Exposure to the equilibrium solution of the gypsum treatment with pH 3.6, 20 mM Ca, and 1.2 mM Al resulted in a somewhat smaller net uptake of amino acids compared to liming. During germination pH around the seeds steeply increased in the untreated control but significantly less with both amendments. The results indicate that pH and the Ca/Al ratio in the soil solution around bean seeds determine their pattern of exudation and solute uptake. For bean germination and early growth on acid soils locally placed application of small amounts of gypsum as seed pelleting seems as effective as soil liming in reducing the incidence of root rot. The results indicate that this may be accomplished by decreasing the amount of leachates available for fungal development.     

Towards an integrated linkage map of common bean. 4. Development of a core linkage map and alignment of RFLP maps

 Three RFLP maps, as well as several RAPD maps have been developed in common bean (Phaseolus vulgaris L.). In order to align these maps, a core linkage map was established in the recombinant inbred population BAT93×Jalo EEP558 (BJ). This map has a total length of 1226 cM and comprises 563 markers, including some 120 RFLP and 430 RAPD markers, in addition to a few isozyme and phenotypic marker loci. Among the RFLPs mapped were markers from the University of California, Davis (established in the F₂ of the BJ cross), University of Paris-Orsay, and University of Florida maps. These shared markers allowed us to establish a correspondence between the linkage groups of these three RFLP linkage maps. In total, the general map location (i.e., the linkage group membership and approximate location within linkage groups) has been determined for some 1070 markers. Approaches to align this core map with other current or future maps are discussed. Received: 10 March 1998 / Accepted: 22 April 1998     

Thermal time assessment of suitable areas for navy bean (Phaseolus vulgaris) production in the UK

In field trials at eight sites throughout the UK the mean thermal time requirement for navy beans from sowing to harvest for a standard cultivar, Marcus, was 2069 Ontario Heat Units (OHU). Low level plastic covers increased the range of warm environments at one site and gave a mean thermal time required of 2098 OHU. Analysis of daily air temperatures from six weather stations throughout the UK over a 29 year period, showed a 14 day possible planting period on the south coast of England, but gave a high probability of crop failure in Scotland. Maps of England and Wales indicating the probability of achieving 2000 OHU were produced from 5 km grid point temperatures. Less than 2% of the land had more than a 60% chance of receiving 2000 OHU under the present climate, however this area increased to 70% with a mean temperature rise of 1.5°C. Although 2000 OHU is often used as the thermal time requirement of navy beans, these trials showed that it may be more accurate to use the higher figure of 2087 OHU from sowing to harvest, and restrict the use of 2000 OHU to the period between emergence and harvest. When the map was redrawn using 2087 OHU and current climate, no parts of England and Wales showed a reasonable chance of growing the present cultivars of navy beans.     

A simple and efficient PCR method for the specific detection of Pseudomonas syringae pv phaseolicola in bean seeds

Using Southern blot hybridizations, it was found that the gene encoding the phaseolotoxin-insensitive ornithyl carbamoyl transferase (argK) was specific for Pseudomonas syringae pv. phaseolicola, the causal agent of the halo-blight disease. Based on these findings, a PCR protocol was developed for the specific detection of P.syringae. pv. phaseolicola in water-extracts of soaked bean seed. For this PCR protocol, two oligonucleotide primers were designed, based on the sequence of argK, which allowed the detection of a specific 1kb fragment. The protocol is simple since PCR was directly applied to bacterial suspensions, thus avoiding DNA extraction. The sensitivity of detection was increased by allowing the bacteria present in seed extracts to multiply in semi-selective media for 18h prior to PCR amplification. The detection threshold by visual detection using ethidium bromide staining was one naturally infected seed in lots of 400 to 600 seeds.     

Towards a better understanding of the Chenopodium album aggregate (Amaranthaceae) in the Middle East: A karyological,cytometric and morphometric investigation

The study of variation in nuclear genome size, especially when combined with common garden experiments, significantly contributes to disentangling interspecies relationships within taxonomically complicated plant groups. The Chenopodium album aggregate is among the morphologically most variable groups and consists of many weakly differentiated cosmopolitan entities. We analysed nuclear genome size variation in diploid and polyploid species of the aggregate from Iran using flow cytometry of 282 accessions from 88 populations of 7 species. To this end, we also determined chromosome numbers and performed a morphometric study to reveal the extent of intraspecific morphological variation. We found that Iranian species are exclusively diploid (C. vulvaria), tetraploid (C. novopokrovskyanum, C. strictum, C. sosnowskyi and C. chaldoranicum) or hexaploid (C. album subsp. album, C. album subsp. iranicum and C. opulifolium). Six homogeneous relative genome size groups were distinguished among the species studied. Our morphometric study surprisingly revealed that under similar ecological conditions Chenopodium species are morphologically stable and well distinguishable, exhibited very little morphological variation. Hence, immense variation in leaf shapes, branching and inflorescence organization seen in the field has not been repeated under greenhouse conditions. The only exception was C. album s. str. which exhibited numerous morphotypes, covering the variation of remaining species.     

Integrated control of root rot and wilt disease of faba bean by soil amendment with suppressive compost in combination with seed coating with an antagonistic yeast

For this study, 21 isolates of fungi belonging to Rhizoctonia and Fusarium genera were isolated from the diseased faba bean plants, obtained from the different localities in Assiut governorate, showing root rot and wilt symptoms. The isolates proved to be pathogenic on Masr 1 faba bean cultivar under greenhouse conditions. F. oxysporum isolates caused wilt disease; however, the isolates of R. solani and other Fusarium species caused root rot. The virulence of isolates on the tested faba bean cultivar was different. The highly pathogenic isolates of these fungi were employed in this study. The effect of soil amendment with Planta Rich and Rich Composts (CMs) alone or in combination with seed coating by the antagonistic yeast Pichia guilliermondi before sowing on the severity of Rhizoctonia and Fusarium root rot and Fusarium wilt of faba bean was tested under greenhouse and field conditions. The tested isolates of yeast proved to be highly antagonistic to the pathogen in vitro. The test rates of CMs were equivalent to 2, 7, 10 and 14 ton/feddan in the greenhouse and 7 and 10 t/feddan in the field conditions. Uncomposted soil was used as a control. The results showed that the tested CMs have a suppressive effect on the severity of root rot and wilt diseases of faba bean under greenhouse and field conditions. The application of CMs (Planta Rich and Rich) alone at the rates equivalent to 2, 7, 10 and 14 t/feddan in greenhouse and 7 and 10 t/feddan in the field conditions to the soil infested with the tested pathogens reduced percentage of the tested diseases compared with uncomposted soil. Combined CMs treatments with yeast seed treatment increased the suppressive effect of CMs on the disease severity.     

Variation in physical and chemical characteristics of common bean (Phaseolus vulgaris L.) grain along a domestication gradient

The objective of this study was to compare six samples of Mexican wild common bean (Phaseolus vulgaris L.) against three landraces and three improved cultivars with respect to physical and chemical attributes, and the culinary quality potential of their grain. A completely randomized experimental design was used to characterize the twelve genotypes. Data were analyzed by analysis of variance and pair-wise comparison of the treatment means by the Tukey test. In addition, correlation and principal-component analysis (PCA) were carried out using twelve characteristics of raw and four of cooked wild and domesticated grains. The results show a larger variability of the physical and chemical characteristics in wild than in domesticated beans. The PCA confirmed that grain gigantism was the main physical characteristic resulting of domestication, whereas the protein and tryptophan contents tended to be higher in wild than domesticated genotypes. Some wild samples from Chihuahua and Durango, Mexico, showed to be a genetic resource to improve food quality, because of their richness in minerals, protein, lysine, tryptophan, and dietary fibers.     

Development of a system simulating commercial production conditions for assessing the potential spread of Phytophthora cryptogea root rot of hardy nursery stock in recirculating irrigation water

A system was developed for monitoring the potential spread of Phytophthora cryptogea root rot of the hardy nursery stock (HNS) species Chamaecyparis lawsoniana, Erica x darlyensis and Calluna vulgaris in recirculating irrigation water on semi-commercial scale production beds. The spread of disease was monitored by measurements of symptoms and by baiting bioassays. A range of baits was tested and the most sensitive was found to be shoot segments of Lupinus angustifolius. The pathogen spread rapidly throughout the system from infected plants in the recirculating irrigation water. Although the percentage of plants infected was high (in Chamaecyparis 73% in 1994 and 93% in 1995), the percentage of infections actively sporulating at any one sample date was much less (47% in 1994 and 63% in 1995) and shoot symptoms were even less evident (in Chamaecyparis 22% in 1994 and 38% in 1995). In Chamaecyparis the incidence of shoot symptoms was less than expected from results of inoculations of similar plants not irrigated with recirculated water (80% infected, 60% with symptoms). The reason for this difference is not known but it poses a serious threat to the quality of HNS produced using recirculated irrigation water and underlines the need for reliable pathogen control methods.     

Histological investigation of walnut cotyledon fragments for a better understanding of in vitro adventitious root initiation

Summary Rapid formation of adventitious roots by walnut cotyledon fragments in vitro was traced by light microscopy. It was shown that this plant model is characterized by two major developmental processes: a) confined elongation of the cotyledon petiole caused by a limited number of cell divisions and b) formation of a morphogenetic zone around each initially wounded vascular bundle within 36 h after detachment of the embryonic axis. During the first phase of development, granular storage protein bodies dissolved, and starch grains were deposited mainly in the distal portion of the cotyledon fragments. Rapidly, new globular protein bodies were formed, and phenolic inclusions accumulated in the vacuoles of epidermal and subepidermal cells and of individual cells close to the vascular bundles. Each adventitious root was found to be in continuity with a single vascular bundle of the cotyledon petiole. A short auxin treatment suppressed the formation of large roots and induced numerous tiny rootlets dispersed all over the surface of the cotyledons.Abbreviations IBA indole-3-butyric acid     

Novel characteristics and regulation of a divergent cinnamate 4-hydroxylase (CYP73A15) from French bean: engineering expression in yeast

cDNAs showing high sequence similarity (>70%) over large stretches to plant CYP73A orthologues from other species were isolated from a cDNA library derived from mRNAs expressed in elicitor-treated suspension-cultured cells. These clones appear to code for a full-length 1554 bp open reading frame with a 78 bp 5-untranslated region and a 140 bp 3-untranslated region. The open reading frame, determined by sequence similarity, codes for a protein with a predicted Mr of 59 229 and a pI of 8.8. It contains the conserved cysteine haem-binding site found in all cytochrome P450s. The protein encoded by this cDNA diverges however from other CYP73As in its N- and C-terminus and in four domains internally, so that overall sequence similarity is in the range 58–66%. Many clones contained an identical intron, which may be associated with a novel regulatory mechanism. Sequence similarity is sufficient for it to be classified as CYP73A15, although it is the least similar member of this family classified so far. The cDNA was expressed in yeast. Successful expression of cinnamate 4-hydroxylase activity required removal of the intron. High-level expression also required modification of the N-terminus to that of CYP73A1. Yeast did not process the intron at all and the leader sequence for A15 was not as compatible as that of A1. The mRNA for CYP73A15 was shown to be rapidly induced by elicitor treatment of suspension-cultured cells of French bean but induction was more transient than that of phenylalanine ammonia-lyase (PAL). In contrast, induction in cells undergoing xylogenesis was much more coordinate with PAL. The cloned cDNA may represent a cinnamate 4-hydroxylase isoform, whose expression is more related to differentiation than the responses to stress in which the majority of CYP73As cloned so far are involved.     

Propyl gallate,a free radical scavenger,counteracts the benefits of exogenously applied salicylic acid and aggravates the deleterious effects of the southern bean mosaic virus in Rhizobium-nodulated Phaseolus vulgaris plants

Infection of Rhizobium-nodulated Phaseolus vulgaris by the southern bean mosaic virus (SBMV) markedly decreased the growth and nodulation of plants. Exogenous applications of salicylic acid (SA) at concentrations ≥10 μM further decreased growth and nodulation of virus-infected (V) plants. Only SA concentration of 5 μM in the solution improved the growth, nodulation, chlorophyll concentration and the catabolism of ureide in leaves of V plants. The spray of leaves with 2 mM propyl gallate (+Pg) decreased growth, nodulation as well as the chlorophyll and leaf ureide concentrations in V plants, regardless of the concentration of SA at which plants were grown. Ultrastructural damages in leaf cells caused by SBMV were also enhanced in V+Pg plants. The massive proliferation of virus particles and the presence of virus crystalline arrays within symbiosomes of nodules in V+Pg plants were attributed to proliferation of branched plasmodesmata in leaf vascular-parenchyma cell walls that facilitated virus movement. Virus particles were never observed in leaf and nodule tissues of V plants not sprayed with Pg. Exogenous applications of SA hindered while Pg increased the symbiotic performance of H plants, pointing out the complexity to be addressed in breeding for virus resistance in Rhizobium-nodulated beans.     

Oat residue and soil compaction influences on common root rot (Aphanomyes euteiches) of peas in a fine-textured soil

Management of common root rot (Aphanomyces euteiches Drechs.) in peas (Pisum sativum L.) is sought primarily by host crop avoidance for several years. Soil compaction is known to aggravate A. euteiches disease in peas but effects on infection and subsequent symptom development are not sufficiently known to assist in cultural control. Several isolated observations have noted that oat crop residues may suppress A. euteiches infection and disease in pea roots. The individual and combined influence (a factorial combination of two factors each at two levels) of a prior oat crop and soil compaction were studied for their effects on common root rot severity in processing peas grown in an A. euteiches disease nursery on a fine-textured soil in the northern Corn Belt of the USA. A previous crop of summer oats relative to prior-year peas significantly suppressed common root rot and increased pea fresh vine weight 210% at peak bloom stage. Both fresh vine weight and green pea yield were reduced as much as 63% by soil compaction and increased as much as 48% by a prior oat crop. Greater soil bulk density at the 10 to 25-cm depth identified wheel traffic compaction patterns in each year. A 10-fold reduction of saturated hydraulic conductivity in the 10 to 25-cm compacted zone and high soil-water potentials within the upper 60 cm both confirmed an impaired water drainage, especially during infiltration events. These observations support the use of a previous full season or summer oat crop jointly with chisel plowing, plus the prevention of excessive traffic during secondary tillage and planting, to reduce common root rot in a field infested with A. euteiches. Shallow incorporation of oat shoot and root residue by chiseling could be a crucial component of the cultural control of the disease. R Rodriguez Kabana Section editor     

Towards a better understanding of the respective effects of milk yield and body condition dynamics on reproduction in Holstein dairy cows

The overall reproductive performance has decreased over the last decades, involving changes in cyclicity, oestrous behaviour and fertility. High milk yield (MY), low body condition score (BCS) and large body condition (BC) loss have been identified as risk factors. However, these effects are often confounded, as high MY and body lipid reserve mobilization are correlated. The aim of this study was to evaluate the respective effects of MY and BC on post-partum ovarian cyclicity, oestrus and fertility of Holstein cows. This study provides novel information, as MY and BC change were uncoupled in the overall dataset that included 98 lactations and milk progesterone profiles. Cows were assigned to two feeding-level groups: high feed, which achieved high MY and moderate BC loss throughout lactation (8410 kg, -1.17 unit from calving to nadir BCS), and low feed, which limited MY and triggered a large BC loss (5719 kg, -1.54 unit). MY and BC had different effects at different stages of the reproductive process. Cyclicity as well as non-fertilization and early embryo mortality were mainly driven by body lipid reserves, whereas oestrous behaviour and late embryo mortality were related to MY. The results point to possible uncoupling between cyclicity, oestrus and early and late embryo survival allowing compensation along the reproductive process and leading to similar final reproductive performance. In compact calving systems, which require high pregnancy rates within a short period, higher MY strategies appear unsuitable even where BCS is maintained, owing to depressed oestrous behaviour and probably increased late embryo mortality, which delays rebreeding. Similarly, strategies that compromise cyclicity and fertility by excessively low BCS are unsuitable.