Effect of two vesicular-arbuscular mycorrhizal fungi on the growth of micropropagated potato plantlets and on the extent of disease caused by <Emphasis Type="Italic">Rhizoctonia</Emphasis><Emphasis Type="Italic">solani</Emphasis>

Two micropropagated potato cultivars, Goldrush and LP89221, were inoculated into sowing trays with either Glomus etunicatum or G. intraradices in a greenhouse. After 2 weeks, plantlets were transplanted into pots and roots were challenged 7 days later with Rhizoctonia solani. At different times after R. solani infection, disease severity, mortality rate, root colonization levels, various growth parameters, and shoot mineral content were evaluated. In Goldrush, only inoculation with G. etunicatum led to a significant reduction in disease severity, ranging between 60.2% and 71.2%, on both shoot and crown. This decrease was not observed in LP89221. Compared with the control plantlets, inoculation of Goldrush with G. etunicatum or G. intraradices reduced significantly the mortality rate by 77% and 26%, respectively, whereas vesicular-arbuscular mycorrhizal (VAM) fungi did not significantly influence the mortality rate in LP89221. In Goldrush, inoculation with G. etunicatum significantly increased shoot fresh weight, root dry weight and the number of tubers produced per plant, whereas G. intraradices only significantly increased the number of tubers. Tuber and root fresh weights of both potato cultivars were significantly reduced by R. solani infection. However, R. solani-infected plantlets of both Goldrush and LP89221, inoculated with G. etunicatum, produced significantly greater tuber fresh weight than non-VAM plantlets. In R. solani-infected plantlets of Goldrush but not LP89221, G. etunicatum and G. intraradices increased root fresh weight by approximately 140.3% and 76.5%, respectively, compared with non-VAM plants. The potato cultivars Goldrush and LP89221 responded differently to VAM fungal inoculation and to R. solani infection in terms of shoot mineral content.     

Changes in root growth patterns of (Picea abies) spruce roots by inoculation with an ectomycorrhizal fungusPisolithus tinctorius and jasmonic acid treatment

Symbiosis between fungi and plant roots forming a mycorrhiza involves extensive interactions at the molecular level between both partners. The role of plant hormones in the regulation of mycorrhizal infection is not known to involve jasmonates. Their endogenous levels increase during pathogen attack; however, little has been done on their involvement in mycorrhizae. In our recent work, root growth patterns of 2-month-old spruce seedlings after inoculation withPisolithus tinctorius and/or jasmonic acid (JA) treatment were studied using a paper-sandwich technique. Changes in root length, the degree of branching, presence and length of root hairs, and infection parameters were followed using a stereomicroscope. The first mycorrhizal contact of hyphae with roots was significantly accelerated upon treatment with 0.5 M JA. Interactions between root hairs and fungal hyphae were seen by scanning electron microscopy. The multiplication of root hairs of non-mycorrhized seedlings treated with 5.0 M JA and changes of the root surface were observed by the same technique.     

Infection of Potato Tubers with the Common Scab Pathogen Streptomyces scabiei in a Soil‐less System

A novel soil‐less method was developed to define susceptibility of developing potato tubers accurately to infection with Streptomyces scabiei the causal agent of common scab disease. Hydroponic production enabled precise identification of individual tuber development. Direct inoculation of tubers with a spore suspension of S. scabiei resulted in disease development, demonstrating that infection could be initiated in a soil‐less media. Tubers were most susceptible to infection between 3 and 20 days after tuber initiation, confirming that this early period of tuber formation is critical to disease development.     

Comparative studies of light leafspot (Pyrenopeziza brassicae) epidemics on the growth and yield of winter oilseed rape

Comparisons of epidemics of light leafspot of differing duration and time of initiation were made in two experiments using a single cultivar of Brassica napus. Fungicide was applied before introduction of disease to prevent infection or some time after inoculation to stop further disease development. In the first experiment, substantial reductions in green leaf area and total plant dry-matter were found at flowering when disease was introduced in the autumn or in January. Plant dry weight at maturity was also greatly reduced in these treatments. The detrimental effect of an epidemic initiated in the autumn was avoided to a large extent if fungicide application began in February. Epidemics initiated in March had only small effects on final dry-matter yield. Seed yield was negatively correlated with the length of the epidemic. In a second experiment, early epidemics initiated in the autumn were halted after different time intervals. Commencing fungicide application even as early as December failed to prevent some loss of dry weight at flowering. At maturity, however, dry weight and seed yield were reduced significantly when fungicide application was delayed until February. Failure to control the disease resulted in a 46% loss of seed yield.     

Optimizing Controlled Environment Assessment of Levels of Resistance to Yam Anthracnose Disease Using Tissue Culture-derived Whole Plants

Studies were conducted to determine the timing and frequency of disease assessment required to effectively identify levels of resistance to yam anthracnose using tissue culture‐derived whole plant inoculation assay. The effects of inoculation methods (paint brush and spray), and disease scoring methods [individual leaf area (ILA) and whole plant area (WPA)] were also assessed. Spray inoculation resulted in rapid infection and higher variations among yam genotypes, leading to earlier discrimination of genotypes than with the paintbrush method. Both the ILA and WPA scoring methods showed variation among yam genotypes, and association between the two methods gave a high positive correlation (r > 0.90). However, the WPA was faster and had the advantage of detecting differences in reactions of yam genotypes to less aggressive pathogen isolates to which the ILA method showed no variation. A single disease evaluation at 7 days after inoculation was as good as the area under the disease progress curve (AUDPC) and the disease progress rate (R_d) derived from multiple evaluations. However, a significant time–genotype interaction, suggests a need for more than a single assessment for effective comparison of genotypes. AUDPC derived from two assessments (5 and 7 DAI) was better than AUDPC from three assessments (5, 7 and 9 DAI) in separating genotypes reactions to a less aggressive pathogen isolate. This study showed that the use of spray inoculation method, the WPA scoring method, and AUDPC derived from two assessments (5 and 7 DAI) provided best conditions for evaluating yam genotypes for levels of anthracnose resistance with the tissue culture‐derived whole plant assay.     

Root, shoot tissues of Brassica juncea and Cereal secale promote potato health

Brassica species are increasingly being used as cover crops to suppress soil-borne diseases in potato cropping systems. Experiments were conducted in controlled environments and in the field to evaluate the effects of cover crop root or shoot or a combination of root and shoot tissues on potato root and tuber health. In a lab assay we examined the extent to which volatile compounds released from tissues of two cover crop species, rye (Cereale secale L.) and oriental mustard (Brassica juncea L.), could inhibit mycelium growth of two important potato diseases, Rhizoctonia solani and Pythium ultimum. Twenty-four hours into the lab assay, volatile compounds from all residues suppressed fungal growth. After 48 h, marked suppression of hyphal growth continued in the presence of mustard residues but not in the presence of rye tissues or the control without tissues. A 75 L volume container experiment evaluated the effect of incorporating different quantities of mustard shoot and root tissues (none, comparable to field level and fourfold field level) into R. solani and P. ultimum infested soil on potato growth, root health and tuber disease. In the container study, incorporating mustard shoots at the highest dose increased potato yield by 54% and reduced disease rating to 2.3 compared to a severe rating of 4.4 in the control. In the field trial, potato growth, root health and tuber disease levels were evaluated in plots where disease management involved either incorporation of mustard or rye cover crop roots, shoots and whole plants (roots plus shoots) or standard farmer practice of a fumigated fallow as a control. White root tissue was used as a health indicator, and averaged 58 and 78% in the fumigated control and mustard cover crop treatments, respectively. The highest healthy root tissue status (91%) was recorded where whole plants of mustard were incorporated. In contrast to the visual assessment of root and tuber health, tuber yield in the field was not influenced by cover crop treatment. Across experiments, the incorporation of or exposure to whole mustard plants was consistently effective at suppressing soil-borne fungi and promoting healthy roots and tubers, especially at higher rates of biomass. Mustard should be managed so as to maximize incorporated biomass for effective biofumigation. Multipurpose management requiring removal of mustard shoots is incompatible with promoting potato rhizosphere health.     

Comparative Analysis of Tuber Development in Six Sweet Potato (Ipomoea batatas (L.) Lam) Cultivars: 1. Tuber initiation, tuber growth and partition of assimilate

Differences in tuber initiation, tuber growth and partitionof assimilate during development were investigated in six localsweet potato cultivars. Study of the quantitative morphogenesisof root types in the sweet potato root system indicated thattuber initiation was completed by 8 weeks after planting inmost cultivars and frustrated thereafter. Subsequent differencesin patterns of tuber development were not obviously relatedto final tuber yield. Thus, high yield resulted either froma short period of rapid tuber growth or a longer period of slowertuber growth. Maximum shoot growth was achieved by week 12 inall except one cultivar, and the cessation of shoot growth resultedin either increases or decreases in tuber growth rate or hadno affect on the rate of tuber growth (cv. A28/7). The partitionof assimilate to tubers at final harvest was the parameter mostclosely related to the yield of the six cultivars studied, butthere was some evidence that such partition was related to totaldry weight. It is suggested that limiting factors in sweet potatotuber yield, e.g. assimilate production and transport or capacityfor tuber growth varied with the cultivar studied.     

Basal fertiliser application method, tuber initiation nitrogen, foliar NPK and the tolerance of potatoes to infection by the potato cyst nematodes Globodera rostochiensis and G. pallida

The effects of broadcast granular, placed liquid and foliar fertilisers on the tolerance of potatoes to infection by potato cyst nematodes were investigated. The tolerance of the potato cv. Pentland Dell was not significantly improved by fertiliser application type but placed liquid fertiliser, with or without foliar applications, increased the concentrations of N, P and K measured in whole plant dry matter of PCN infected plants. The tolerance of the potato cv. Sante was not statistically improved by altering the balance of fertiliser nitrogen applications between planting and tuber initiation or by applying foliar nitrogen. Nitrogen applications of 120 kg N ha^-1 at planting and a further 120 kg N ha^-1 at tuber initiation supplemented with foliar N, however, achieved a larger tuber yield than the same nitrogen programme without foliar N and gave a significantly greater yield than the application of 240 kg N ha^-1 at planting plus foliar N. The emergence of both cultivars was delayed in the absence of oxamyl. N, P and K concentrations within whole plant dry matter were significantly higher in plants from oxamyl treated plots and both N and K concentrations were significantly increased by increasing the quantity of N at planting, at 56 DAP. Splitting the fertiliser N between planting and tuber initiation appears to be important in maintaining the availability of this nutrient to PCN infected plants throughout the season.     

Efficacy of fungicide combinations,phosphoric acid and plant extract from stinging nettle on potato late blight management and tuber yield

Late blight caused by Phytophthora infestans is a major constraint to potato production. Inadequate control of the disease has often resulted in potato yield losses. We assessed the efficacy of fungicides, phosphoric acid and stinging nettle extract combinations for late blight control at two locations in Kenya. Disease severity, relative area under disease progress curves (RAUDPC), pathogen lesions and tuber yield were quantified during the 2008 and 2009 cropping cycles. The application of metalaxyl alternated with phosphate resulted in the greatest suppressive effects on late blight. The average late blight severity ranged from 3.5 to 34% in 2008 and 4.7 to 50% in 2009 at Tigoni location. RAUDPC for the same location ranged from 5 to 40% and 5 to 50% in 2008 and 2009, respectively. Similar levels of late blight severity were recorded at Marimba location in both years. Lesion growth and pathogen lesion numbers on potato plants differed significantly (p < 0.05) among treatments. Fungicides, phosphoric acid and stinging nettle extract varied in late blight control. Potato tuber yield varied among treatments. Phosphoric acid treatment had significantly (p < 0.05) greater tuber yield compared to metalaxyl at both locations. Field plots treated with plant extracts from stinging nettle resulted in the lowest tuber yield compared to other treatments with the exception of the untreated control. Fungicides, phosphoric acid, stinging nettle extract and their combinations can be readily effective in the suppression of late blight severity and pathogen lesions with moderate increases in tuber yield.     

The phenology of skin spot (Oospora pustulans Owen &Wakef.) and other fungal diseases of potato tubers

Underground stems and roots of the potato varieties King Edward and Majestic became more severely infected by Oospora pustulans (skin spot) as the growing season advanced. Tubers became infected at their initiation in June and July but the proportion of infected eyes usually increased during both the growth of the crop and bulk storage. Some buds on tubers in bulk stores died in December, but few were killed until after January, when attacks increased until by March about a quarter of Majestic eyes were usually dead. Boxing (chitting) not only prevented most eyes from dying but also prevented much skin spotting on tubers. At High Mowthorpe, Majestic seed unboxed or boxed in March yielded, respectively, 3 and 1 ton/acre less than seed boxed by January. Early boxing increased the yield of seed–sized tubers but did not affect the infection of progeny tubers. Dipping Majestic seed tubers in organo–mercurial fungicide and then boxing soon after lifting decreased their infection by O. pustulans; it also lessened infection on plants they produced and increased the yield of progeny seed–sized tubers. However, dipping, even in successive years, had little effect on the infection of progeny tubers by O. pustulans. Rhizoctonia solani and Helminihosporum atrovirens became increasingly prevalent on tubers during growth after July and during bulk storage. H. atrovirens was decreased by early boxing or dipping tubers in fungicide. Verticillate conidiophores often grew on incubated tuber plugs but their prevalence was affected by past storage. The results suggest that some of the benefit for chitting may be from disease control, it was not possible to measure the extent to which benefits from chitting depend on effects on disease or on the physiology of the tuber.     

Effect of moisture stress, plant population density and pathogen inoculation on charcoal stalk rot of sorghum

The effects of moisture deficit stress, plant population density and pathogen inoculation technique on charcoal stalk rot in the sorghum hybrid CSH 6 were studied in the 1980–81 and 1981–82 post-rainy seasons at three locations in India. Incidence and severity of charcoal rot caused by Macrophomina phaseolina were compared in three plant population densities, subjected to different moisture stress regimes created by withholding irrigation at various plant growth stages. Natural infections were compared to artificial inoculation with M. phaseolina. Combinations of moisture stress, plant population and inoculation treatments were compared to identify the combination most likely to develop maximum disease. Lodging, the first external symptom of charcoal rot, was significantly correlated with other disease symptoms used to measure charcoal rot, such as soft stalk, number of nodes crossed by M. phaseolina infection, root damage and plant senescence. In both seasons the highest incidence of lodging occurred when moisture stress was induced at the 'flag leaf visible in the whorl' growth stage. The greatest incidence of the disease was recorded in the highest plant population (266 700 plant ha^-) at all three locations. No significant differences were found between artificially and naturally inoculated treatments. The maximum number of lodged plants was found at a density of 266 700 plants ha^-1 when moisture stress was induced at the 'flag leaf visible in the whorl' growth stage.     

Effects of low root zone temperatures on the early stages of symbiosis establishment between soybean [Glycine max (L.) Merr.] and Bradyrhizobium japonicum

It is known that low root zone temperatures (RZT) have moreeffect on infection and early nodule development than on nitrogenfixation by soybean [Glycine max (L.) Merr.]. However, therehave been no studies regarding how the low RZT inhibit the infectionstages of soybean. Two controlled environment experiments wereconducted to examine the effect of low RZT on bacterial attachmentto, and infection thread penetration of, soybean root hairs.The experimental designs were (1) plants maintained at 25, 17.5or 15C RZT, or transferred from 25 or 17.5 to 15C RZT at either0.5, 1, 2, or 7d after inoculation (DAI), (2) early symbioticestablishment between soybean and Bradyrhizobium japonicum wasexamined microscopically under three RZT (15, 17.5 and 25C).These results indicated that (1) keeping plants at 25C only0.5 DAI prior to transfer to a 15C RZT accelerates the onsetof N₂ fixation at 15C RZT by 6 d, (2) at RZT between 25 and17.5C the infection processes were progressively delayed astemperature declined, (3) RZT less than 17C strongly inhibitedinfection steps, such that when RZT dropped 8.5C from 25 to17.5C infection initiation was delayed 1 d, while when RZTdropped only 2.5C from 17.5 to 15C, infection initiation wasdelayed another 2 d. Key words: Bradyrhizobium japonicum, low temperature, nodulation, soybean     

Lipopolysaccharides of Rhizobium etli strain G12 act in potato roots as an inducing agent of systemic resistance to infection by the cyst nematode Globodera pallida

Recent studies have shown that living and heat-killed cells of the rhizobacterium Rhizobium etli strain G12 induce in potato roots systemic resistance to infection by the potato cyst nematode Globodera pallida. To better understand the mechanisms of induced resistance, we focused on identifying the inducing agent. Since heat-stable bacterial surface carbohydrates such as exopolysaccharides (EPS) and lipopolysaccharides (LPS) are essential for recognition in the symbiotic interaction between Rhizobium and legumes, their role in the R. etli-potato interaction was studied. EPS and LPS were extracted from bacterial cultures, applied to potato roots, and tested for activity as an inducer of plant resistance to the plant-parasitic nematode. Whereas EPS did not affect G. pallida infection, LPS reduced nematode infection significantly in concentrations as low as 1 and 0.1 mg ml(-1). Split-root experiments, guaranteeing a spatial separation of inducing agent and challenging pathogen, showed that soil treatments of one half of the root system with LPS resulted in a highly significant (up to 37%) systemic induced reduction of G. pallida infection of potato roots in the other half. The results clearly showed that LPS of R. etli G12 act as the inducing agent of systemic resistance in potato roots.     

Production of potato intraspecific somatic hybrids with improved tolerance to PVY and Pythium aphanidermatum

Somatic hybridization can be an interesting alternative for the selection of heterozygous and vigorous potato plants through combination of dihaploid genomes. The resulting hybrids can harbour interesting characters and thus can be used in agriculture if they are in agreement with agronomic criteria.

In this report, we used an intraspecific somatic hybridization technique for the production of tetraploid potato lines. Two parental combinations were used in protoplast electrofusion procedure: Aminca-Cardinal and Cardinal-Nicola. The selection of somatic hybrids was based on in vitro plant vigour. Therefore, among the 75 regenerated plants obtained from Aminca-Cardinal fusion, 3 putative hybrids were retained and 2 plant lines were selected among the 54 regenerated from the Cardinal-Nicola fusion. Heterosis was observed in the larger hybrid tuber size compared to the parents’. Our results also showed a precocity in the in vitro tuberization for the hybrids. Moreover, all of the regenerated putative hybrids were tetraploid (2n=4x=48 chromosomes). Isocitrate dehydrogenase and malate dehydrogenase isoenzyme analyses confirmed the hybrid nature of these lines. A molecular characterization performed by PCR amplification of simple sequence repeats and inter-simple sequence repeats confirmed that all these lines were somatic hybrids.

The effect of potato virus Y infection on these hybrid lines was tested by mechanical inoculation of plants cultivated in a greenhouse. The majority displayed a reduction of infection rate associated with a delayed appearance of symptoms compared to the parents. Moreover, complete resistance was noted for one hybrid line (CN2). All hybrids also showed improved tolerance to Pythium aphanidermatum infection during tuber storage or after plant inoculation.     

Effects of liberibacter-infective Bactericera cockerelli (Hemiptera: Triozidae) density on zebra chip potato disease incidence, potato yield, and tuber processing quality

In plant pathosystems involving insect vectors, disease spread, incidence, and severity often depend on the density of the vector population and its rate of infectivity with the disease pathogen. The potato psyllid, Bactericera cockerelli (Sulc), has recently been associated with zebra chip (ZC), an emerging and economically important disease of potato in the United States, Mexico, Central America, and New Zealand. "Candidatus Liberibacter solanacearum," a previously undescribed species of liberibacter has been linked to the disease and is transmitted to potato by B. cockerelli. Experiments were conducted under laboratory and field conditions to determine the impact of B. cockerelli density on ZC incidence, potato yield, and tuber processing quality. Insect densities ranging from one to 25 liberibacter-infective psyllids per plant were used during the experiments. Results showed that a single adult potato psyllid was capable of inoculating liberibacter to potato and causing ZC disease after a 72-h inoculation access period and was as damaging as 25 psyllids per plant. In addition, ZC-diseased plants showed a sharp reduction in tuber yield but the disease response was independent of the density of psyllids. Furthermore, both glucose and sucrose were found to have highly elevated concentrations in ZC-diseased potato tubers compared with noninfected ones and psyllid density did not vary the response. The high reducing sugar concentrations found in ZC-infected potato tubers are believed to be responsible for browning and reduced quality in processed ZC-infected tubers. This information could help ZC-affected potato producers in making effective management decisions for this serious disease.     

Defining Plant Resistance to Phytophthora cinnamomi: A Standardized Approach to Assessment

Phytophthora cinnamomi is a soil‐borne plant pathogen that causes devastating disease in agricultural and natural systems worldwide. While a small number of species survive infection by the pathogen without producing disease symptoms, the nature of resistance, especially under controlled conditions, remains poorly understood. At present, there are no standardized criteria by which resistance or susceptibility to P. cinnamomi can be assessed, and we have used five parameters consisting of plant fresh weight, root growth, lesion length, relative chlorophyll content of leaves and pathogen colonization of roots to analyse responses to the pathogen. The parameters were tested using two plant species, Zea mays and Lupinus angustifolius, through a time course study of the interactions and resistance and susceptibility defined 7 days after inoculation. A scoring system was devised to enable differentiation of these responses. In the resistant interaction with Z. mays, there was no significant difference in fresh weight, root length and relative chlorophyll content in inoculated compared with control plants. Both lesion size and pathogen colonization of root tissues were limited to the site of inoculation. Following inoculation L. angustifolius showed a significant reduction in plant fresh weight and relative leaf chlorophyll content, cessation of root growth and increased lesion lengths and pathogen colonization. We propose that this technique provides a standardized method for plant–P. cinnamomi interactions that could be widely used to differentiate resistant from susceptible species.     

Growth and yield of potato plants inoculated with rhizosphere bacteria

Pot and field experiments on small plots were carried out to study the effect of isolates ofPseudomonas fluorescens-putida bacteria, applied to potato tubers, on the growth response of potato plants. Inoculation of tuber pieces with the isolates in pot experiments caused a better growth of young potato plant (111 % increase). The effect depended on the viable count in the bacterial suspension and on growth conditions of the plants. Inoculation of potato seed pieces before planting in field experiments caused a 4 –30 % improvement in plant growth and tuber yield. The complex action of inoculation with rhizobacteria apparently caused changes in the microflora colonizing the roots and stolons, which in turn brought about a better growth and yield.     

THE INFECTION OF PLANTS BY VIRUSES THROUGH ROOTS

Roots of young tomato plants became infected when inoculated with tomato bushy stunt, tobacco mosaic, and potato X viruses. Root infections also occurred when these viruses were added to soil or culture solutions in which plants were growing.
The viruses were sometimes localized around their initial entry points in roots; sometimes they invaded the root system but not the shoots, and sometimes they produced full systemic infection of roots and shoots. In some experiments, but not all, systemic infections were more frequent when the upper tap root or superficial roots were inoculated than when fibrous roots were inoculated.
In both tomato and potato, virus X spread from diseased to healthy plants sharing the same culture solution, if their roots were in contact, but not otherwise. Infection of the roots of potato plants by inoculation, produced only one plant with virus-infected haulms, although several had infected tubers.     

Relationship of inoculum ofVerticillium dahliae to disease in tomato plants

Summary Tomato (Lycopersicon esculentum Mill.) seedlings were inoculated by dipping roots in suspensions of conidia ofVerticillium dahliae Kleb. There was a linear relationship between inoculum concentration and infection at 21 days after inoculation in steam-disinfested soil. Thereafter the number of infections per plant increased. At higher concentrations data indicated a synergistic interaction between conidia. Synergism was more pronounced and was detected earlier in soil not disinfested before inoculation. When conidia from 3-, 7-, and 14-day-old cultures were used, the greatest response was from 3-day-old cultures. A lower total response and lower rate of response to older inoculum indicated a decrease in aggressiveness of conidia with age. Older plants were less affected by the pathogen when plants inoculated at the fourth and sixth leaf stages with minimum root disturbances were compared. There was also an increase in infection with an increase in the volume of root zone infested. In field experiments infection reduced stand when roots were disturbed at inoculation. With minimum root disturbance yields were reduced without a reduction in stand. Part of thesis to be submitted by the senior author to the University of Stellenbosch in partial fulfilment of the Ph.D. (Agric.) degree.