Colonization of citrus seed coats by 'Candidatus Liberibacter asiaticus': implications for seed transmission of the bacterium

Huanglongbing is an economically damaging disease of citrus associated with infection by 'Candidatus Liberibacter asiaticus'. Transmission of the organism via infection of seeds has not been demonstrated but is a concern since some citrus varieties, particularly those used as rootstocks in commercial plantings are propagated from seed. We compared the incidence of detection of 'Ca. Liberibacter asiaticus' DNA in individual fruit peduncles, seed coats, seeds, and in germinated seedlings from 'Sanguenelli' sweet orange and 'Conners' grapefruit fruits sampled from infected trees. Using real-time quantitative PCR (qPCR) we detected pathogen DNA in nucleic acid extracts of 36 and 100% of peduncles from 'Sanguenelli' and from 'Conners' fruits, respectively. We also detected pathogen DNA in extracts of 37 and 98% of seed coats and in 1.6 and 4% of extracts from the corresponding seeds of 'Sanguenelli' and 'Conners', respectively. Small amounts of pathogen DNA were detected in 10% of 'Sanguenelli' seedlings grown in the greenhouse, but in none of 204 extracts from 'Conners' seedlings. Pathogen DNA was detected in 4.9% and in 89% of seed coats peeled from seeds of 'Sanguenelli' and 'Conners' which were germinated on agar, and in 5% of 'Sanguenelli' but in none of 164 'Conners' seedlings which grew from these seeds on agar. No pathogen DNA was detected in 'Ridge Pineapple' tissue at 3 months post-grafting onto 'Sanguenelli' seedlings, even when pathogen DNA had been detected initially in the 'Sanguenelli' seedling. Though the apparent colonization of 'Conners' seeds was more extensive and nearly uniform compared with 'Sanguenelli' seeds, no pathogen DNA was detected in 'Conners' seedlings grown from these seeds. For either variety, no association was established between the presence of pathogen DNA in fruit peduncles and seed coats and in seedlings.     

Transmission of rice blast from seeds to adult plants in a non‐systemic way

Rice blast, caused by the fungal pathogen Magnaporthe oryzae, is a serious threat to rice production worldwide. In temperate regions, where rice is not cultivated for several months each year, little is known about the initial onset of the disease in the field. The main overwintering and primary inoculum sources reported are infested residues and seeds, but the subsequent steps of the disease cycle are largely unknown, even though a systemic infection has been proposed but not demonstrated. The present work follows rice blast progression in infected seeds from germination to seedling stage, with direct and detailed microscopic observations under both aerobic conditions and water seeding. With the use of GFP‐marked M. oryzae strains, it was shown that spores are produced from contaminated seeds, infect emerging seedling tissues (coleoptile and primary root) and produce mycelium that colonizes the newly formed primary leaf and secondary roots. Using different rice cultivars exhibiting distinct levels of resistance/susceptibility to M. oryzae at the 2/4‐leaf stage, it was observed that resistance or susceptibility of a considered genotype is already established at the seedling stage. The results also showed that when plants are inoculated either at ripening stage (mature panicles), heading stage (flowering/immature panicles) or even before heading (flag leaf fully developed), they produce infested seeds. These seeds produce contaminated seedlings that mostly die and serve as an inoculum source for healthy neighbouring plants, which gradually develop disease symptoms on leaves. The possible rice blast disease cycle was reconstructed on irrigated rice in temperate regions.     

Factors Affecting the Spread of Plum pox virus Strain M in Peach Orchards Subjected to Roguing in France

ABSTRACT We evaluated the impact of roguing on the spread and persistence of the aggressive Plum pox virus strain M (PPV-M) in 19 peach orchard blocks in Southern France. During a 7- to 10-year period, orchards were visually inspected for PPV symptoms, and symptomatic trees were removed every year. Disease incidence was low in all orchards at disease discovery and was <1% in 16 of the 19 orchard blocks. The spread of Sharka disease was limited in all 19 blocks, with an annual disease incidence between 2 and 6%. However, new symptomatic trees were continuously detected, even after 7 to 10 years of uninterrupted control measures. An extended Cox model was developed to evaluate to what extent tree location, orchard characteristics, environment, and disease status within the vicinity influenced the risk of infection through time. Eleven variables with potential effect on tree survival (i.e., maintenance of a tree in a disease- free status through time) were selected from survey data and databases created using a geographical information system. Area of the orchard, density of planting, distance of a tree from the edge of the orchard block sharing a boundary with another diseased orchard, and distance to the nearest previously detected symptomatic tree had a significant effect on the risk for a tree to become infected through time. The combined results of this study suggest that new PPV-M infections within orchards subjected to roguing resulted from exogenous sources of inoculum, disease development of latent infected trees, as well as infected trees overlooked within the orchards during visual surveys. A revision of the survey and the roguing procedures used for more effective removal of potential sources of inoculum within the orchards and in the vicinity of the orchards would improve disease control suppression of PPV.     

Detection of lethal yellowing phytoplasma in embryos from coconut palms infected with Cape St Paul wilt disease in Ghana

This study investigated the potential of seed transmission of Cape St. Paul wilt disease (CSPWD) in coconuts. PCR amplification was used to assess the distribution of phytoplasmas in parts of West African Tall (WAT) palms infected with CSPWD. Employing phytoplasma universal primer pair P1/P7 in standard PCR, or followed with a nested PCR using CSPWD–specific primer pair G813f/AwkaSR, phytoplasma infection was detected in the trunks, peduncles, spikelets, male and female flowers of four infected WAT coconut palms. Through nested PCR, phytoplasma was also detected in four of 19 embryo DNA samples extracted individually from fruits harvested from three of the four infected palms and was confirmed as CSPWD by cloning and sequencing. Subsequently, CSPWD phytoplasma was again detected in five of 33 embryos from nine infected palms, and in one of eight fruits from two symptomless palms. Fruits from infected palms recorded higher percentage germinations in two field nurseries (average of 71·0%) compared to fruits from healthy palms (average of 57·6%), and matured fruits that had dropped from infected palms showed the same levels of germination as those harvested directly from the palms. This indicates that infected fruits retain the ability to germinate whether harvested or dropped. No phytoplasmas were detected in any of the resulting seedlings and plantlets obtained through embryo in-vitro culture. Therefore, although phytoplasma DNA can be detected in embryos, there is as yet no evidence that the pathogen is seed transmitted through to the seedling to cause disease in progeny palms.     

Transformation of soil microbial community structure and rhizoctonia-suppressive potential in response to apple roots

ABSTRACT Changes in the composition of soil microbial communities and relative disease-suppressive ability of resident microflora in response to apple cultivation were assessed in orchard soils from a site possessing trees established for 1 to 5 years. The fungal community from roots of apple seedlings grown in noncultivated orchard soil was dominated by isolates from genera commonly considered saprophytic. Plant-pathogenic fungi in the genera Phytophthora, Pythium, and Rhizoctonia constituted an increasing proportion of the fungal community isolated from seedling roots with increasing orchard block age. Bacillus megaterium and Burkholderia cepacia dominated the bacterial communities recovered from noncultivated soil and the rhizosphere of apple seedlings grown in orchard soil, respectively. Populations of the two bacteria in their respective habitats declined dramatically with increasing orchard block age. Lesion nematode populations did not differ among soil and root samples from orchard blocks of different ages. Similar changes in microbial communities were observed in response to planting noncultivated orchard soil to five successive cycles of 'Gala' apple seedlings. Pasteurization of soil had no effect on apple growth in noncultivated soil but significantly enhanced apple growth in third-year orchard block soil. Seedlings grown in pasteurized soil from the third-year orchard block were equal in size to those grown in noncultivated soil, demonstrating that suppression of plant growth resulted from changes in the composition of the soil microbial community. Rhizoctonia solani anastomosis group 5 (AG 5) had no effect on growth of apple trees in noncultivated soil but significantly reduced the growth of apple trees in soil from third-year orchard soil. Changes in the ability of the resident soil microflora to suppress R. solani AG 5 were associated with reductions in the relative populations of Burkholderia cepacia and Pseudomonas putida in the rhizosphere of apple.     

Sanitation practices that inhibit reproduction of Monosporascus cannonballus in melon roots left in the field after crop termination

Ascospores of Monosporascus cannonballus function as primary inoculum for infection of melon roots. Previous studies demonstrated that pathogen reproduction (i.e. ascospore production) occurs on infected melon roots primarily after the crop has been terminated. Therefore, the key to maintaining low soil population densities of the pathogen is to destroy the hyphae of the pathogen in infected roots as soon as possible after crop termination, thereby inhibiting ascospore production. Results from a 3-year field study demonstrated that, relative to the nontreated controls, an immediate postharvest application of metam sodium (applied via the drip irrigation system at 187 L ha⁻¹) or cultivation (which lifts roots onto the surface of the soil for rapid desiccation) significantly inhibited pathogen reproduction in infected melon roots, as shown by the number of roots subsequently bearing perithecia. Additionally, ascospore populations in plots that received either the metam sodium or cultivation treatment were significantly lower ( P  < 0·05) than populations in the nontreated control plots at the end of the 3-year study. These results demonstrated the efficacy of these postharvest treatments in the inhibition of pathogen reproduction and retardation of inoculum build-up in soil.     

Seed and pollen transmission of <Emphasis Type="Italic">Apple latent spherical virus</Emphasis> in apple

To examine whether Apple latent spherical virus (ALSV) has spread among apple trees in an orchard, we surveyed 21 apple trees surrounding two ALSV-infected trees for virus infection using a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). None of the 21 trees were infected, indicating that ALSV has not spread from the infected trees to the neighboring apple trees since it was first detected in 1984. We analyzed seed embryos and seedlings derived from infected trees and detected ALSV in 10 of 223 seed embryos (4.5%) and 10 of 227 seedlings (4.4%). From these results, we conclude that ALSV is seed-transmitted at a rate of ca. 4.5% in apple. We also analyzed seed embryos and seedlings from uninfected apple trees that were hand-pollinated with pollen from infected trees. We detected ALSV in only 1 of 260 seed embryos and in none of the 227 apple seedlings. This result indicated that the seed transmission rate via infected pollen is only 0–0.38%. In situ hybridization analysis of ALSV-infected apple flower buds showed that ALSV was present inside almost all pollen grains and in all ovary and ovule tissues, including the embryo sac and inner integument.     

Seed transmission of maize downy mildew (Peronosclerospora sorghi) in Nigeria

In an area of Nigeria where downy mildew of maize is present, histological assessment of maize seed revealed the presence of mycelium and oospores of Peronosclerospora sorghi in the kernels. Seed transmission of downy mildew of maize was demonstrated when grain purchased at local markets gave mean seedling infection rates of 12·3% (untreated seeds) and 10·0% (in metalaxyl-treated seeds) within 7 days of emergence, after storage in a desiccator for 30 days. When untreated seeds taken from nubbin ears of systemically infected plants from four states in southern Nigeria were planted at 9 days (17–22% moisture content) and 27 days (9–22% moisture content) after harvest, 20·0% infected seedlings resulted in both trials. Seeds from Borno state in northern Nigeria had 26·6% systemic seedling infection after 9 months of storage at 11% moisture content. When seeds harvested from maize plants inoculated with P. sorghi through silks were examined histologically, hyphae of P. sorghi were observed mostly in the scutellum of the embryo. Transmission of disease to seedlings was observed when the silk-inoculated seeds (9% moisture content) were planted in pots in a greenhouse; however, no disease transmission was observed when such seeds were planted in the field. The epidemiological significance of seed transmission is discussed with particular reference to survival of inoculum and development of epidemics. Also noteworthy is the overall significance of seed transmission in Nigeria, where the major source of seed is that saved by farmers from their grain crop, occasionally supplemented by seed bought from the local market.     

Transmission of <Emphasis Type="Italic">Colletotrichum coccodes via</Emphasis> tomato seeds

Anthracnose of tomato caused by Colletotrichum coccodes is a devastating disease of ripe fruits. This pathogen may also infect tomato roots, stems and leaves. In the present study, C. coccodes is shown to be capable of contaminating seeds collected from artificially inoculated tomato fruits. Seedlings germinating from these infected seeds exhibited disease symptoms and therefore may transmit the pathogen to the next crop. The proportion of infected seeds ranged between 20% and 63% in all C. coccodes isolates tested and correlated with the aggressiveness of the isolates to tomato fruits. Fungicidal treatment of the collected seeds reduced, but did not eliminate, seed infection. A transgenic C. coccodes isolate expressing green fluorescent protein was used to visualize the pathogen. Mycelium was observed both on surfaces of the seed coat and within 1% of the embryos.     

A major gene for resistance to white pine blister rust in Western white pine from the Western cascade range

ABSTRACT A dominant gene for resistance to white pine blister was indicated by Mendelian segregation in full-sib families of western white pine parent trees selected for phenotypic resistance in six heavily infected stands in the Western Cascades of Oregon and Washington. Seedlings were artificially inoculated three times between 1959 and 1964 and observed for development of stem infection. Segregation at this locus (Cr2) occurred in only two of the six parent populations sampled: one a natural stand, Champion Mine (CM), and the other a plantation of unknown seed origin. At CM, reduced penetrance of this gene was expressed by altered Mendelian ratios (mostly less-than-expected resistant phenotypes) in families of specific combinations of certain parents, indicating the presence of modifier genes with effects that ranged from mild to almost complete suppression of Cr2. Between 1968 and 1994, an apparent shift in virulence at CM caused all of the resistant selections to become infected and die. Recent inoculations of many of the same or related families from these parents, made from grafted ramets in a seed orchard, showed that Cr2 conditions a classical hypersensitive reaction (HR) in needle tissues, the primary infection courts. In the latter tests, seedlings were challenged with wild-type and four other sources of inoculum at and near CM that were also suspected of having wider virulence than wild type. No seedlings segregating for HR that were inoculated with wild type subsequently developed stem symptoms, but the other inocula induced both susceptible and HR needle spots on Cr2- genotypes, and many of these seedlings did develop stem infections. This implied that spore genotypes with specific virulence to Cr2 are carried in these inocula.     

Factors affecting mummification and sporulation of pome fruit infected by Monilinia fructigena in Dutch orchards

A 2-year field experiment (1997–98, 1998–99) was conducted to study mummification and subsequent sporulation in spring of apple (cvs James Grieve, Golden Delicious) and pear (cv . Conference) fruits infected by Monilinia fructigena . Most mummified fruits were found in James Grieve and Conference, whereas in late-infected Golden Delicious, fruits were still soft when examined in April. In the first year, these late-infected fruits had a significantly higher sporulation intensity per sporulating fruit ( P  = 0·05) compared with Golden Delicious fruits infected 9 and 5 weeks before harvest maturity, which were partly mummified. It was concluded that early- and late-infected fruits contributed to primary inoculum in the next season. In a postinfection regime of 25°C and 65–75% relative humidity under controlled conditions, the number of Conference fruits sporulating decreased rapidly, and after 12 weeks' incubation sporulation had completely ceased. After 8 weeks' incubation, sporulation intensity in the postinfection regime at 10°C was significantly higher than that at 20 and 25°C in a first experiment with inoculated unripe fruit ( P  = 0·05). Results of a second experiment with ripe fruit were less clear. These results are discussed in relation to orchard disease management.     

东北地区番茄细菌性溃疡病的发生和病原鉴定研究

 1988~1989年从北京市、辽宁和黑龙江省等地调查采集的患溃疡病的病株和病果上分离到17个细菌菌株,接种番茄幼苗、果实及叶片,均能产生典型的溃疡病症状,各菌株致病力无明显差异。各菌株经细菌染色反应,形态特征,培养性状,生理生化反应,血清学反应(ELISA法)及蛋白质凝聚丙烯酰胺胶电泳等鉴定,认为番茄溃疡病的病原细菌是:密执安棒杆菌密执安亚种(Clavibacter michiganense subsp.michiganense (Smith) Davies et al.)。试验表明病原细菌可经种子带菌传染,病残体和病土壤可能是初侵染菌源。抗病性测定45个番茄品种均属感病品种。寄主范围除番茄外,尚能侵染茄子、龙葵和心叶烟。     

Field studies of anthracnose symptoms and pathogen infection in different phases of the persimmon growing season

Anthracnose is the main disease of persimmon and is caused by Colletotrichum spp. The study of field epidemiology is essential for the development of efficient management of this disease. In this study, we investigated infection by Colletotrichum spp. throughout the persimmon growing season to understand the host–pathogen interactions better. We observed the production of primary inoculum of persimmon anthracnose and described how epidemics progress from secondary infections during the fruit crop season. The field study was carried out in an organic orchard with three susceptible persimmon cultivars, Fuyu, Kakimel and Jiro, for three consecutive seasons. Our results indicate that the pathogen survives in 1-year-old shoots, which are the sources of primary inoculum. Later that growing season, the inoculum reaches flowers and new shoots, developing symptoms and producing the secondary inoculum. Fruit drop was also observed, with or without symptoms of anthracnose, throughout the plant cycle. In some of the symptomless fruit, collected from the plant and from the ground where they had fallen, latent infections of Colletotrichum spp. were detected. Shoots, flowers, immature and ripened fruit remained infected throughout the growing season, producing conidia that could lead to new secondary infections within and among plants. The incidence of anthracnose in fruit at harvest and postharvest proved to be less relevant for disease management. Practices for chemical and cultural control of the disease throughout the persimmon growing season are discussed.     

Inoculum availability and seasonal survival of Potebniamyces pyri in pear orchards

Potebniamyces pyri (anamorph Phacidiopycnis piri) is the causal agent of Phacidiopycnis rot, a postharvest disease of pear fruit (Pyrus communis). Infections of pear fruit by P. pyri occur in the orchard, and symptoms develop after harvest during storage or in the market. P. pyri also is the cause of a canker and twig dieback disease of pear trees. To determine inoculum availability of P. pyri, dead bark and dead fruit spurs were periodically collected in two commercial ‘d’Anjou’ pear orchards and examined for the presence and viability of fruiting bodies of P. pyri. To determine seasonal survival of P. pyri, 2-year-old twigs of ‘d’Anjou’ pear in a research orchard were inoculated approximately monthly over 2 years with P. pyri and monitored for canker development. Inoculated twigs were removed from the trees 6 months post inoculation and examined for formation, viability of pycnidia of P. pyri, and reisolation of the pathogen. In both commercial orchards, all sampled trees were infected by P. pyri; viable pycnidia of P. pyri were observed on 42–78 % of the sampled bark and 5–32 % of the sampled fruit spurs; and viable conidia were observed at all sampling times during the fruit growing season. Apothecia of P. pyri also were observed on sampled dead bark and fruit spurs, but at a frequency ranging from 0 % to 19 %. P. pyri was recovered from inoculated twigs 6 months after inoculation at all sampling times during the 2-year study, but recovery frequency varied. P. pyri formed pycnidia on most cold-injured and inoculated twigs. The results suggest that: i) the conidial state of P. pyri is the main type of inoculum in pear orchards in the region; ii) viable inoculum of P. pyri for potential fruit infections is available during the pear fruit-growing season; iii) P. pyri can form pycnidia on cankers of twigs infected by the fungus at different seasons during the year; and iv) P. pyri can survive as mycelium in diseased pear twigs year round in the orchard.     

Phytophthora infestans Produces Oospores in Fruits and Seeds of Tomato

ABSTRACT Tomato fruits at the mature green stage coinoculated with A1 + A2 sporangia of Phytophthora infestans, the late blight causal fungus, showed abundant oospores in the vascular tissues, pericarp, columella, and placenta. Oospores were also formed on the surface of fruits kept in moisture-saturated atmosphere. Occasionally, oospores were enclosed between the epidermal hairs of the seed coat. In a few seeds, oospores were detected inside the embryo. The data suggest that blighted tomato fruits may carry a large number of oospores, thus making them a threatening source of blight inoculum. Such fruits may also release airborne oosporic inoculum that may introduce recombinant genotypes within a growing season. Although Phytophthora infestans is seedborne in tomato, to our knowledge, this is the first report on the occurrence of oospores in tomato seeds. Whether such tomato seeds produce blighted seedlings remains to be shown.     

玉米丝黑穗病菌侵染的研究

 玉米丝黑穗菌,对玉米幼根、幼芽都能侵染,但以幼芽侵染较多。在幼芽的芽鞘和根茎部位,侵染程度没有明显差异。     

Spread of seed-borne <Emphasis Type="Italic">Erwinia rhapontici</Emphasis> in bean,pea and wheat

Studies were conducted in the laboratory and greenhouse to determine the distribution of Erwinia rhapontici in plants arising from naturally infected seeds of pea or artificially inoculated seeds of bean and wheat, and whether the pathogen is transmitted to the subsequent generation of seeds. Infected seeds were planted in pots of Cornell mix in the greenhouse, and sampled at specified intervals throughout the plant growth cycle (seedling stage, elongation stage, flowering stage, seed formation stage, and maturity). Plating of surface sterilized lateral roots, tap roots, basal stems, mid-stems, apical stems, petioles, pods, and seeds of pea and bean, and of lateral roots, sub-crown internodes, basal stems, mid-stems, apical stems, peduncles, glumes, and seeds of wheat revealed that the bacterial pathogen spread from infected seeds to the lower parts of the plant tissues, but failed to spread further to the seeds produced on these plants. The study concludes that E. rhapontici did not establish systemic infection throughout the plants. Possible mechanisms of infection of seeds are discussed.     

Studies on the spread of the olive scab pathogen,Spilocaea oleagina1

Investigations were carried out to study the occurrence and effectiveness of wind dispersal of conidia of the olive spot pathogen Spilocaea oleagina. Dissemination in absence of rain was confirmed both by collecting conidia at various distances from an inoculum source and by using trap plants. The dispersal gradient of conidia was very steep: 3–10% of leaves of olive seedlings placed 20 m from diseased trees became infected.     

Effects of the mycoparasite Verticillium biguttatum on barley stunt disease caused by Rhizoctonia solani anastomosis group 8 in a model system

The height of barley stunted by Rhizoctonia solani anastomosis group 8 was significantly increased by up to 72·8% after incubation for 8 days at 20°C in seedling tray tests following application of the mycoparasite Verticillium biguttatum. The pathogen and mycoparasite were applied at the rate of 1% Perlite maizemeal inoculum (w/w potting mixture) resulting in propagule densities of approximately 24·0 and 6·6 × 10⁵ colony-forming units (cfu) per g potting mixture, respectively. Sieving (2 mm) R. solani inoculum prior to dilution in potting mixture increased the recovery of propagules from 1·2 × 2·1 × 10³ cfu per g inoculum compared with recovery when inoculum was sieved after dilution. Applications of a V. biguttatum isolate from the UK (vbl) and a Dutch isolate (M73) reduced stunting to a similar extent but did not stimulate the growth of healthy plants. The height of stunted plants was significantly increased after application of V. biguttatum inoculum after 6 days if inoculated trays were preincubated for 1 day prior to planting but a similar increase was only detected after 7 days if seeds were planted immediately. The number of stunted plants which emerged after 4 days was significantly increased by treatment with V. biguttatum but preincubation had no additional effect. These results suggested that control of R. solani was effected both before and after the initiation of disease.     

苹果短粗根病的发生及病原线虫种的鉴定

 本文首次报道了苹果上的新病害——短粗根病:根系团状,侧根丛生,黑褐色,短、粗,尖端钝或稍膨大,无根毛,无吸收功能与活力。地上部生长衰退,枝条生长缓慢,产量低,重者枯死。该病发生在北京市南口农场,经诊断为巴基斯坦毛刺线虫(Trichodorus pakiatanensis Siddiqi,1962)所致。病原线虫主要形态特征是雄虫具3个颈乳突、三个腹乳突和一对尾乳突,交合刺有环纹;雌虫有一对体侧孔。