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.     

Analysis of gene inheritance and expression in hybrids between transgenic sugar beet and wild beets

Reciprocal gene exchange between cultivated sugar beet and wild beets in seed production areas is probably the reason for the occurence of weed beets in sugar beet production fields. Therefore, when releasing transgenic sugar beet plants into the environment, gene transfer to wild beets ( Beta vulgaris ssp. maritima ) has to be considered. In this study the transfer of BNYVV- (beet necrotic yellow vein virus) resistance and herbicide-tolerance genes from two transgenic sugar beet lines that were released in field experiments in 1993 and 1994 in Germany to different wild beet accessions was investigated. In order to evaluate the consequences of outcrossing, manual pollinations of emasculated wild beet plants with homozygous transgenic sugar beet plants were performed. In the resulting hybrids the transgenes were stably inherited according to Mendelian law. Gene expression in leaves and roots of the hybrids was in the same range as in the original transgenic sugar beet plants. Moreover, it was found that in one of the wild beet accessions, transfer and expression of the BNYVV resistance gene did considerably increase the level of virus resistance.     

Electron microscopical proof of the presence of sugar beet yellows virus particles in the roots of sugar beet

The presence of beet yellows virus (BYV) particles was electron microscopically proved in the roots of sugar beet. Specimens for the electron microscopical examination of root sap were prepared by differential centrifugation. It was proved that, contrary to expectations, examinations in spring showed most virus particles in the basal part of the root. At the same time it was found by experiment that the diagnostical BYV antiserum, for which the antigen was prepared from sugar beet leaves, did not react with a purificate of BYV containing virus particles.     

Adenylate patterns of autumn-sown sugar beet differ from spring-sown sugar beet. Implications for root quality

Sugar beet ( Beta vulgaris L) is generally cultivated using two different planting and harvest patterns. In northern zones, spring sugar beet is sown in spring and harvested in autumn, whereas in subtropical latitudes, autumn sugar beet is sown in autumn and harvested in summer. The industrial quality of the root is frequently higher in spring-sown sugar beet crops. In order to explore physiological changes associated with this fact, this study has been focused on the seasonal changes of adenosine 5'-triphosphate and adenosine 5'-diphosphate levels in the storage roots of sugar beet plants, as an index of its metabolic status. The results obtained correspond to a different metabolic status of spring and autumn sugar beet at the moment of harvest. The adenylate patterns of autumn beets suggested a functional and active respiratory system. On the contrary, the patterns shown by spring beets corresponded to those we would expect to see in plants becoming dormant. The proline and glucose contents, which decrease the industrial quality of the root, and the respiratory rate measured in autumn-sown sugar beets, were nearly twice those of spring-sown sugar beets. The combination of an active respiratory system, which allows the carbohydrate catabolism and the synthesis of stress molecules, with the environmental factors at the time of the harvest, could be the underlying physiological mechanism causing some of the differences between spring- and autumn-sown sugar beet crops.     

Genetic approaches to sustainable pest management in sugar beet (Beta vulgaris)

Sugar beet (Beta vulgaris) is an important arable crop, traditionally used for sugar extraction, but more recently, for biofuel production. A wide range of pests, including beet cyst nematode (Heterodera schachtii), root‐knot nematodes (Meloidogyne spp.), green peach aphids (Myzus persicae) and beet root maggot (Tetanops myopaeformis), infest the roots or leaves of sugar beet, which leads to yield loss directly or through transmission of beet pathogens such as viruses. Conventional pest control approaches based on chemical application have led to high economic costs. Development of pest‐resistant sugar beet varieties could play an important role towards sustainable crop production while minimising environmental impact. Intensive Beta germplasm screening has been fruitful, and genetic lines resistant to nematodes, aphids and root maggot have been identified and integrated into sugar beet breeding programmes. A small number of genes responding to pest attack have been cloned from sugar beet and wild Beta species. This trend will continue towards a detailed understanding of the molecular mechanism of insect–host plant interactions and host resistance. Molecular biotechnological techniques have shown promise in developing transgenic pest resistance varieties at an accelerated speed with high accuracy. The use of transgenic technology is discussed with regard to biodiversity and food safety.     

Proteome analysis of sugar beet (Beta vulgaris L.) elucidates constitutive adaptation during the first phase of salt stress

Salinity is one of the major stress factors responsible for growth reduction of most of the higher plants. In this study, the effect of salt stress on protein pattern in shoots and roots of sugar beet (Beta vulgaris L.) was examined. Sugar beet plants were grown in hydroponics under control and 125 mM salt treatments. A significant growth reduction of shoots and roots was observed. The changes in protein expression, caused by salinity, were monitored using two-dimensional gel-electrophoresis. Most of the detected proteins in sugar beet showed stability under salt stress. The statistical analysis of detected proteins showed that the expression of only six proteins from shoots and three proteins from roots were significantly altered. At this stage, the significantly changed protein expressions we detected could not be attributed to sugar beet adaptation under salt stress. However, unchanged membrane bound proteins under salt stress did reveal the constitutive adaptation of sugar beet to salt stress at the plasma membrane level.     

Detection of beet yellows virus in sugar beet leaves and roots by enzyme-linked immunosorbent assay (ELISA)

Using the enzyme-linked immunosorbent assay (ELISA) beet yellows virus (BYV) could be detected reliably in the leaves of sugar beet andTetragonia expansa Pall. and in the roots of sugar beet. Specifio γ-globulin of BYV antiserum was coupled to horse radish peroxidase by periodate oxidation. Optimum dilutions of antigen (extract from infected leaves) were1: 50 to 1: 200 for BYV detection in sugar beet andT. expansa leaves and1: 2 to 1: 5 for detection in sugar beet roots. Extracts from beet roots are not to be purified by ultracentrifugation, however, by the described method virus can be demonstrated only in 80–90% of naturally infected sugar beet roots. The method is specific, no increase of extinction values was found in healthy or beet western yellows virus infected plants. Presence of virus can be demonstrated by visual as well as photometric evaluation. Results confirmed the suitability of peroxidase application for detection of plant viruses by ELISA.     

天山北坡甜菜内生菌分离鉴定及其动态变化

对新疆昌吉和石河子两地种植的甜菜内生菌进行了分离、鉴定和分析,结果表明甜菜内生菌多属于细菌,其中假单胞菌 (Pseudomonas sp. )和芽孢菌类(Bacillus sp.)的分离频率分别在33.2%～59.2%和12.7%～28.1%,是甜菜植株中的优势内生菌群.16S rDNA 和 ITS 序列同源性比较和系统发育分析表明内生菌具有丰富的多样性.根中内生菌的多样性高于茎、叶,昌吉地区种植的甜菜中分离出的内生菌种类较多.从感病品种及生长不良甜菜植株中分离出的内生菌种类比较丰富.通过回接分离及利用扫描电镜观察内生菌在植物体内分布发现,内生菌能够定殖于甜菜块根.     

Antagonistic potential of certain legumes to Meloidgyne incognita on sugar beet

Under greenhouse conditions, a pot experiment was conducted to clarify the potential of using some legumes as intercropped plants for reducing the root-knot nematode Meloidogyne incognita infecting sugar beet (Beta vulgaris L.) cv. DS-9004 compared to non-legume plant, garlic and non-intercropped plants. The obtained results revealed that all legumes including chickpea, Egyptian clover, faba bean, fenugreek, lentil and lupin significantly (p ≤ 0.05) reduced nematode criteria on the roots of sugar beet at different degrees. Chickpea and Egyptian clover reduced the number of galls on the roots of sugar beet as the percentage of reductions were 54 and 50%, respectively, followed by lupin and fenugreek, while garlic achieved 72% reduction compared to non-intercropped plants. Lupin reduced the number of egg masses by 59% followed by Egyptian clover and fenugreek (32%), three months after the treatment. On the other hand, six months after the treatment, chickpea reduced the number of galls by 55.7% followed by lupin (53.4%) and Egyptian clover (52.3%) and the percentage of reduction of egg masses behaved the same trend. Also, the treatments improved plant growth criteria of sugar beet, weight of roots (tubers) and the percentage of total soluble solids (TSS).     

Pectolytic clostridia isolated from sugar beet pulp silages in Italy

Twenty-eight strains of pectolytic clostridia were isolated from sugar beet pulp silages. Seventeen non-pigmented strains were presumed to be Clostridium acetobutylicum ; the remaining 11 pigmented strains were similar to Cl. felsineum. The addition of molasses to sugar beet pulps favoured the growth of other bacteria, particularly lactic acid organisms, whereas pectolytic clostridia were only occasionally found. The pectolytic clostridia promoted the structure loss of simulated silages. The use of molasses in sugar beet pulp ensiling was suggested to prevent texture loss of the ensiled mass.     

An improved CARV process for bioethanol production from a mixture of sugar beet mash and potato mash

A mixed mash of sugar beet roots and potato tubers with a sugar concentration of 23.7% w/v was used as a feedstock for bioethanol production. Enzymatic digestion successfully reduced the viscosity of the mixture, enabling subsequent heat pretreatment for liquefaction/sterilization. An energy-consuming thick juice preparation from sugar beet for concentration and sterilization was omitted in this new process.     

Efficient dsRNA-mediated transgenic resistance to Beet necrotic yellow vein virus in sugar beets is not affected by other soilborne and aphid-transmitted viruses

Rhizomania caused by Beet necrotic yellow vein virus (BNYVV) is one of the most devastating sugar beet diseases. Sugar beet plants engineered to express a 0.4 kb inverted repeat construct based on the BNYVV replicase gene accumulated the transgene mRNA to similar levels in leaves and roots, whereas accumulation of the transgene-homologous siRNA was more pronounced in roots. The roots expressed high levels of resistance to BNYVV transmitted by the vector, Polymyxa betae. Resistance to BNYVV was not decreased following co-infection of the plants with Beet soil borne virus and Beet virus Q that share the same vector with BNYVV. Similarly, co-infection with the aphid-transmitted Beet mild yellowing virus, Beet yellows virus (BYV), or with all of the aforementioned viruses did not affect the resistance to BNYVV, while they accumulated in roots. These viruses are common in most of the sugar beet growing areas in Europe and world wide. However, there was a competitive interaction between BYV and BMYV in sugar beet leaves, as infection with BYV decreased the titres of BMYV. Other interactions between the viruses studied were not observed. The results suggest that the engineered resistance to BNYVV expressed in the sugar beets of this study is efficient in roots and not readily compromised following infection of the plants with heterologous viruses.     

An Investigation of Wound Healing in Sugar Beet Roots Using Light and Fluorescence Microscopy

Following impact, wound healing was investigated in roots ofsugar beet using fluorescence microscopy in conjunction witha conventional lignin test. Samples of sugar beet roots wereharvested at different stages of development and impacted inthe laboratory with a falling bolt delivering 1–4 Joules.A response in the form of deposition of brown material, presumedto be melanin, along the outer and inner walls of cells at thewound surface was observed within 3 d of impact. This materialeventually became granular in appearance. Formation of a ligno-suberizedboundary layer from cells present at the time of impact firstoccurred in 16-week-old roots 9 d after impact. Intensity ofcalcofluor fluorescence supported the findings made using lightmicroscopy. As wound healing progressed with time, the intensityof calcofluor fluorescence declined, demonstrating interferenceby wound healing products with calcofluor binding. Aggressiveharvest and subsequent storage dramatically reduced calcofluorfluorescence indicating that this dye may have potential valuein the assessment of tissue damage. Copyright 2001 Annals ofBotany Company Sugar beet, wound healing, impact damage, bruising, fluorescence, calcofluor, lignin     

Enhanced drought resistance in fructan-producing sugar beet

Fructans are soluble polymers of fructose that are produced by approximately 15 % of the flowering plant species. Production of bacterial fructans in tobacco has been shown previously to lead to improved biomass production under polyethylene glycol-mediated drought stress. Here, we used the same SacB gene from Bacillus subtilis to produce bacterial fructans in sugar beet (Beta vulgaris L.). The transgenic sugar beets accumulated fructans to low levels (max. 0.5 % of dry weight) in both roots and shoots. Two independent transgenic lines of fructan-producing sugar beets showed significantly better growth under drought stress than untransformed beets. Drought stressed fructan-producing plants attained higher total dry weights (+25–35 %) than wildtype sugar beet, due to higher biomass production of leaves (+30–33 %), storage roots (+16–33 %) and fibrous roots (+37–60 %). Under well-watered conditions, no significant differences were observed between the transgenic and wildtype beets. In conclusion, the introduction of fructan biosynthesis in transgenic plants is a promising approach to improve crop productivity under drought stress.     

Differences in salt tolerance of three sugar beet genotypes

The effect of increasing NaCl concentrations (up to 150 m M ) on growth and mineral composition of three genotypes of sugar beet ( Beta vulgaris L., MONOHILL, ADA and FIA) has been studied. Growth was stimulated or little affected in water culture by 50 m M NaCl in all 3 genotypes. Further increase in NaCl concentration depressed growth in ADA more than in MONOHILL, whereas in FIA growth did not significantly differ from the untreated control. In all 3 genotypes, particularly in FIA, increasing NaCl concentrations decreased potassium content in the shoots more than in the fibrous and storage roots. Simultaneously, the accumulation of sodium and chloride in the shoots was considerably higher in FIA than in ADA, where in contrast larger proportions of these ions were retained in the roots. The results demonstrate considerable genotypic differences in salt tolerance of sugar beet and indicate a positive correlation between salt tolerance and accumulation of sodium and chloride in the shoots. FIA but not ADA may be suited for a breeding programme of sugar beet for improved salt tolerance.     

Taproot promoters cause tissue specific gene expression within the storage root of sugar beet

The storage root (taproot) of sugar beet (Beta vulgaris L.) originates from hypocotyl and primary root and contains many different tissues such as central xylem, primary and secondary cambium, secondary xylem and phloem, and parenchyma. It was the aim of this work to characterize the promoters of three taproot-expressed genes with respect to their tissue specificity. To investigate this, promoters for the genes Tlp, His1-r, and Mll were cloned from sugar beet, linked to reporter genes and transformed into sugar beet and tobacco. Reporter gene expression analysis in transgenic sugar beet plants revealed that all three promoters are active in the storage root. Expression in storage root tissues is either restricted to the vascular zone (Tlp, His1-r) or is observed in the whole organ (Mll). The Mll gene is highly organ specific throughout different developmental stages of the sugar beet. In tobacco, the Tlp and Mll promoters drive reporter gene expression preferentially in hypocotyl and roots. The properties of the Mll promoter may be advantageous for the modification of sucrose metabolism in storage roots.