Effects of preplant inoculation of apple (Malus domestica Borkh.) with arbuscular mycorrhizal fungi on population growth of the root-lesion nematode,Pratylenchus penetrans

Six species of arbuscular mycorrhizal (AM) fungi (Glomus aggregatum, G. clarum, G. etunicatum, G. intraradices, G. mosseae and G. versiforme) were evaluated, in three greenhouse experiments, for their effects on reproduction of the root-lesion nematode, Pratylenchus penetrans, and growth of Ottawa 3 apple rootstock. Glomus mosseae increased total dry weights of nematode-inoculated and non-inoculated rootstock in all three greenhouse experiments, and G. intraradices increased dry weights in two of three greenhouse experiments. Plants inoculated with G. mosseae generally supported fewer P. penetrans per gram of root than plants inoculated with other AM fungi, but did not differ significantly from the controls in any greenhouse experiment. Colonization of roots by AM fungi was reduced by P. penetrans at initial inoculum densities greater than 250 nematodes/L soil. In field trials, preplant inoculation with either G. intraradices or G. mosseae increased rootstock growth and leaf concentrations of P, Mg, Zn and Cu in fumigated plots but not in non-fumigated plots, indicating that colonization by native AM fungi in non-fumigated plots may have been sufficient for adequate nutrient acquisition. The abundance of vesicles and arbuscules was greater in roots of plants inoculated with AM fungi before planting than in roots of non-inoculated plants, in both fumigated and non-fumigated plots. P. penetrans per gram of root and per 50 ml soil were significantly lower for G. mosseae- inoculated plants than for non-inoculated plants in fumigated soil but not in non-fumigated soil.     

Pseudomonas chlororaphis Strain Sm3, Bacterial Antagonist of Pratylenchus penetrans

The interaction of Pseudomonas chlororaphis strain Sm3 and the root-lesion nematode Pratylenchus penetrans was investigated in three separate greenhouse experiments with soils from southern British Columbia, Canada. The bacteria were applied to the roots of strawberry plants and planted in unpasteurized field soils, with natural or supplemented infestation of P. penetrans. Nematode suppression in roots was evident after 6 or 10 weeks in all experiments. Root or shoot growth were increased after 10 weeks in two experiments. Population dynamics of P. chlororaphis Sm3 in the rhizosphere was followed using an antibiotic-resistant mutant of P. chlororaphis Sm3. There was no apparent correlation between bacterial density in the rhizosphere and P. penetrans suppression in strawberry roots and rhizosphere soil, although the soil with the highest nematode reduction also had the largest P. chlororaphis Sm3 population in the rhizosphere.     

SRAP polymorphisms associated with superior freezing tolerance in alfalfa (Medicago sativa spp. sativa)

Sequence-related amplified polymorphism (SRAP) analysis was used to uncover genetic polymorphisms among alfalfa populations recurrently selected for superior tolerance to freezing (TF populations). Bulk DNA samples (45 plants/bulk) from each of the cultivar Apica (ATF0), and populations ATF2, ATF4, ATF5, and ATF6 were evaluated with 42 different SRAP primer pairs. Several polymorphisms that progressively intensified or decreased with the number of recurrent cycles were identified. Four positive polymorphisms (F10-R14, Me4-R8, F10-R8 and F11-R9) that, respectively, yielded 540-, 359-, 213-, and 180-bp fragments were selected for further analysis. SRAP amplifications with genotypes within ATF populations confirmed that the polymorphisms identified with bulk DNA samples were reflecting changes in the frequency of their occurrence in response to selection. In addition, the number of genotypes cumulating multiple polymorphisms markedly increased in response to recurrent selection. Independent segregation of the four SRAP polymorphisms suggests location at unlinked loci. Homology search gave matches with BAC clones from syntenic Medicago truncatula for the four SRAP fragments. Analysis of the relationship with low temperature tolerance showed that multiple SRAP polymorphisms are more frequent in genotypes that maintain superior regrowth after freezing. These results show that SRAP analysis of bulk DNA samples from recurrent selections is an effective approach for the identification of genetic polymorphisms associated with quantitative traits in allogamous species. These polymorphisms could be useful tools for indirect selection of freezing tolerance in alfalfa.     

Embryogenic response of Mexican alfalfa (Medicago sativa) varieties

The in vitro embryogenic response of nine varieties of alfalfa (Medicago sativa L.) grown in México (five Mexican varieties: Puebla 76, Inia 76, Bajío 76, Sintético I and Sintético II and four foreign or introduced varieties: Moapa 69, San Joaquín II, Hairy Peruvian and Valenciana) were tested. We screened 25 genotypes from each variety in four tissue culture protocols. All the varieties, except San Joaquín II, gave a positive response in one or more of the protocols tested. The response in each variety was low; this was also observed in a wider screening performed with the varieties Moapa 69, Hairy Peruvian, Sintético I and Sintético II. Two plants from Moapa 69 were regenerated and appeared normal.     

Suppression of Pratylenchus penetrans Populations in Potato and Tomato using African Marigolds

Current strategies for management of Pratylenchus penetrans in both white potato and tomato consist of the use of fumigant or non-fumigant nematicides or crop rotation. The objective of this study was to determine if double-cropping African marigolds (Tagetes erecta) with potatoes or tomatoes could reduce P. penetrans populations. Plots were 10 m × 3 m arranged in a randomized complete block design with four replications. Treatments included marigolds, potatoes or tomatoes, and natural weedy fallow followed by either potatoes or tomatoes. Nematode populations were sampled before spring planting, between crops in August and after harvest in November. During the 3 years of the study, P. penetrans soil population density declined by an average of 93% from the pre-plant level when marigold was grown in rotation with potato and by 98% when marigold was grown.in rotation with tomato. Weedy fallow preceding potato resulted in an average decline in P. penetrans soil population density of 38%, and a similar decrease (37%) was seen when fallow preceded tomato. There was a significant reduction in the number of P. penetrans found in both potato and tomato roots when the crops followed marigolds. These results suggest that P. penetrans population density may be significantly reduced when marigolds are double-cropped with potatoes or tomatoes.     

Impact of rhizobacterial inoculants on plant growth and enzyme activities in soil treated with contaminated bottom sediments

The impact of contaminated bottom sediments on plant growth and soil enzyme activities was evaluated in a greenhouse pot study. The sediments were moderately contaminated with zinc and heavily contaminated with polycyclic aromatic hydrocarbons and polychlorinated dibenzo-p-dioxins and furans. The sediments were mixed with soil and planted with either Festuca arundinacea or Tagetes patula. The capacity of two rhizobacterial strains (Massilia niastensis P87 and Streptomyces costaricanus RP92), previously isolated from contaminated soils, to improve plant growth under the chemical stress was tested. Application of sediments to soil was severely phytotoxic to T. patula and mildly to F. arundinacea. On the other hand, the addition of sediments enhanced the soil enzymatic activity. Inoculation with both bacterial strains significantly increased shoot (up to 2.4-fold) and root (up to 3.4-fold) biomass of T. patula. The study revealed that the selected plant growth-promoting bacterial strains were able to alleviate phytotoxicity of bottom sediments to T. patula resulting from the complex character of the contamination.     

Suppression of Rhizoctonia solani diseases of sugar beet by antagonistic and plant growth-promoting yeasts

AIMS: Isolates of Candida valida, Rhodotorula glutinis and Trichosporon asahii from the rhizosphere of sugar beet in Egypt were examined for their ability to colonize roots, to promote plant growth and to protect sugar beet from Rhizoctonia solani AG-2-2 diseases, under glasshouse conditions. METHODS AND RESULTS: Root colonization abilities of the three yeast species were tested using the root colonization plate assay and the sand-tube method. In the root colonization plate assay, C. valida and T. asahii colonized 95% of roots after 6 days, whilst Rhod. glutinis colonized 90% of roots after 8 days. Root-colonization abilities of the three yeast species tested by the sand-tube method showed that roots and soils attached to roots of sugar beet seedlings were colonized to different degrees. Population densities showed that the three yeast species were found at all depths of the rhizosphere soil adhering to taproots up to 10 cm, but population densities were significantly (P < 0.05) greater in the first 4 cm of the root system compared with other root depths. The three yeast species, applied individually or in combination, significantly (P < 0.05) promoted plant growth and reduced damping off, crown and root rots of sugar beet in glasshouse trials. The combination of the three yeasts (which were not inhibitory to each other) resulted in significantly (P < 0.05) better biocontrol of diseases and plant growth promotion than plants exposed to individual species. CONCLUSIONS: Isolates of C. valida, Rhod. glutinis and T. asahii were capable of colonizing sugar beet roots, promoting growth of sugar beet and protecting the seedlings and mature plants from R. solani diseases. This is the first successful attempt to use yeasts as biocontrol agents against R. solani which causes root diseases. SIGNIFICANCE AND IMPACT OF THE STUDY: Yeasts were shown to provide significant protection to sugar beet roots against R. solani, a serious soil-borne root pathogen. Yeasts also have the potential to be used as biological fertilizers.     

Identification of common root-lesion nematode (Pratylenchus thornei Sher et Allen) loci in bread wheat.

Plant parasitic nematodes are a major biotic cause of wheat-yield loss in temperate wheat-growing regions. A major strategy to develop resistance to root-lesion nematodes (RLNs) in wheat is to assess and then exploit their natural genetic variation. This study examines RLN (Pratylenchus thornei) resistance in 1 Middle Eastern landrace (AUS4930 7.2) and 1 synthetic hexaploid wheat, CROC_1/AE.SQUARROSA (224)//OPATA (CROC), using F2 and F9 populations generated by crossing AUS4930 7.2 and CROC with the susceptible cultivar Pastor, and inoculating these crosses with P. thornei in greenhouse trials. Wheat microsatellite markers linked to previously identified quantitative trait loci (QTLs) for resistance to P. thornei and P. neglectus were used to screen both populations. In the AUS4930 7.2 x Pastor population, resistance loci on chromosomes 1B, 2B, and 6D were detected. Similarly, in the CROC x Pastor population, 2 resistance loci, located on chromosomes 1B and 3B, were identified. Interestingly, a resistance locus located on chromosome 6D was not detected. More detailed mapping is required in these 2 populations, developed using new RLN resistance sources, to determine whether the QTLs identified on these chromosomes are the same, are allelic, or are linked to different resistance loci from those previously identified, and to determine whether these 2 sources contain other novel resistance loci.     

Colonization behaviour of Pseudomonas fluorescens and Sinorhizobium meliloti in the alfalfa (Medicago sativa) rhizosphere

The colonization ability of Pseudomonas fluorescens F113rif in alfalfa rhizosphere and its interactions with the alfalfa microsymbiont Sinorhizobium meliloti EFB1 has been analyzed. Both strains efficiently colonize the alfalfa rhizosphere in gnotobiotic systems and soil microcosms. Colonization dynamics of F113rif on alfalfa were similar to other plant systems previously studied but it is displaced by S. meliloti EFB1, lowering its population by one order of magnitude in co-inoculation experiments. GFP tagged strains used to study the colonization patterns by both strains indicated that P. fluorescens F113rif did not colonize root hairs while S. meliloti EFB1 extensively colonized this niche. Inoculation of F113rif had a deleterious effect on plants grown in gnotobiotic systems, possibly because of the production of HCN and the high populations reached in these systems. This effect was reversed by co-inoculation. Pseudomonas fluorescens F113 derivatives with biocontrol and bioremediation abilities have been developed in recent years. The results obtained support the possibility of using this bacterium in conjunction with alfalfa for biocontrol or rhizoremediation technologies.     

Evaluation of Soil Streptomyces spp. for the Biological Control of Fusarium Wilt Disease and Growth Promotion in Tomato and Banana

Fusarium oxysporum f. sp. lycopersici (Fol) and Fusarium oxysporum f. sp. cubense (Foc), are the causal agent of Fusarium wilt disease of tomato and banana, respectively, and cause significant yield losses worldwide. A cost-effective measure, such as biological control agents, was used as an alternative method to control these pathogens. Therefore, in this study, six isolates of the Streptomyces-like colony were isolated from soils and their antagonistic activity against phytopathogenic fungi and plant growth-promoting (PGP) activity were assessed. The results showed that these isolates could inhibit the mycelial growth of Fol and Foc. Among them, isolate STRM304 showed the highest percentage of mycelial growth reduction and broad-spectrum antagonistic activity against all tested fungi. In the pot experiment study, the culture filtrate of isolates STRM103 and STRM104 significantly decreased disease severity and symptoms in Fol inoculated plants. Similarly, the culture filtrate of the STRM304 isolate significantly reduced the severity of the disease and symptoms of the disease in Foc inoculated plants. The PGP activity test presents PGP activities, such as indole acetic acid production, phosphate solubilization, starch hydrolysis, lignin hydrolysis, and cellulase activity. Interestingly, the application of the culture filtrate from all isolates increased the percentage of tomato seed germination and stimulated the growth of tomato plants and banana seedlings, increasing the elongation of the shoot and the root and shoot and root weight compared to the control treatment. Therefore, the isolate STRM103 and STRM104, and STRM304 could be used as biocontrol and PGP agents for tomato and banana, respectively, in sustainable agriculture.     

Suppression of fusarium wilts by fluorescent pseudomonads: Mechanisms and applications

Fluorescent pseudomonads are involved in the natural suppressiveness of some soils to fusarium wilts. These bacteria have been applied successfully to suppress fusarium wilts of various plant species grown in conducive soils and growing substrates. Suppression of fusarium wilts by fluorescent pseudomonads can be ascribed to direct and indirect effects against pathogenic Fusarium oxysporum. Direct effects are expressed by a reduction of the saprophytic growth of the pathogen leading to delay and reduction of root infections. This antagonism was demonstrated to be related to siderophore‐mediated iron competition. Iron competition was also shown to enhance the antagonistic effect of non‐pathogenic F. oxysporum by making the pathogen more susceptible to fungal competition for carbon. Indirect effects of fluorescent pseudomonads against pathogenic F. oxysporum are mainly related to the induction of host plant resistance which can be associated with the bacterial lipopolysaccharides. Suppression of fusarium wilts by some fluorescent pseudomonads could also be related to their ability to detoxify fungal metabolites such as fusaric acid. Association of different mechanisms of suppression increases the efficacy and consistency of the biological control under various experimental conditions. Increased knowledge of mechanisms of suppression now enables management of the environment, to some extent, to favour expression of the beneficial activities. These activities are only expressed if the bacterial density is sufficiently high.     

Ontogeny and ultrastructure of spontaneous nodules in alfalfa (Medicago sativa)

Summary The development of spontaneous nodules, formed in the absence ofRhizobium and combined nitrogen, on alfalfa (Medicago sativa L. cv. Vernal) was investigated at the light and electron microscopic level and compared to that ofRhizobium-induced normal nodules. Spontaneous nodules were initiated from cortical cell divisions in the inner cortex next to the endodermis, i.e., the site of normal nodule development. These nodules, on uninoculated roots, were white multilobed structures, histologically composed of nodule meristems, cortex, endodermis, central zone and vascular strands. Nodules were devoid of intercellular or intracellular bacteria confirming microbiological tests. Early development of spontaneous nodules was initiated by series of anticlinal followed by periclinal divisions of dedifferentiated cells in the inner cortex of the root. These cells formed the nodular meristem from which the nodule developed. The cells in the nodule meristems divided unequally and differentiated into two distinct cell types, one larger type being filled with numerous membrane-bound starch grains, and the other smaller type with very few starch grains. There were no infection threads or bacteria in the spontaneous nodules at any stage of development. This size differentiation is suggestive of the different cell sizes seen inRhizobium-induced nodules, where the larger cell type harbours the invading bacteria and the smaller type is essential in supportive metabolic roles. The ontogenic studies further support the claim that these structures are nodules rather than aberrant lateral roots, and that plant possess all the genetic information needed to develop a nodule with distinct cell types. Our results suggest that bacteria and therefore theirnod genes are not necessarily involved in the ontogeny and morphogenesis of spontaneous and normal nodules in alfalfa.Abbreviations EH smallest emergent root hair - EM electron microscope - enod2 early nodulin2 gene - RT root tip - RER rough endoplasmic reticulum - YEMG yeast extract-mannitol-gluconate     

Establishment of Peristenus digoneutis and P. relictus (Hymenoptera: Braconidae) in California for the control of Lygus spp. (Heteroptera: Miridae)

Lygus hesperus Knight is native to the western United States and is a perennial pest of numerous crops in California. It is responsible for triggering the early season application of insecticides on cotton, Gossypium hirsutum L., and strawberries, Fragaria L. Despite several surveys conducted in alfalfa (Medicago sativa L.) grown in central California, nymphal parasitoids associated with L. hesperus and L. elisus have not been found. Two exotic parasitoids were released into California beginning in 1998. Peristenus relictus (Ruhte), formerly P. stygicus Loan, and P. digoneutis Loan were collected from several locations in southern Europe and released at up to six locations over a 6-year period. At the original release site in Sacramento, a 0.25-ha plot of alfalfa, parasitism by P. digoneutis and P. relictus combined increased from zero to 90%, 3 years after the last releases were made. Parasitoids have been recovered from vacant fields of weedy annuals within 2 km of this site. Recoveries at more southerly release sites in central California have been poor.     

水分对苜蓿叶片光合特性的影响

采用田间试验, 对每茬灌水3次(W₃)、2次(W₂)、1次(W₁)和不灌水(W₀)四种条件下的土壤水分, 苜蓿(Medicago sativa)叶片的叶绿素荧光参数、气孔导度(G_s)、净光合速率(P_n)和蒸腾速率(T_r)进行测定。结果表明, 灌水提高了苜蓿叶片的原初光能转换效率(F_v/F_m)、P_n和T_r, 并随着灌水量的增加而增加。苜蓿叶片的F_v/F_m、P_n和T_r的日均值与土壤含水量均呈极显著正相关关系。苜蓿叶片的P_n与F_v/F_m和光合有效辐射(PAR)的乘积呈正相关关系。灌水还改变了苜蓿叶片P_n的日变化格局。灌水较多的处理(W₃和W₂), 苜蓿叶片没有出现光合“午休”现象,P_n的日变化趋势呈现“单峰”型。而灌水较少和不灌水的处理(W₁和W₀), 苜蓿叶片出现了明显的光合“午休”现象, 其P_n的日变化进程呈现“双峰”型。在相同的水分条件下, 初花期苜蓿叶片的P_n高于再生期的, T_r则相反。