Seed transmission of Sweet potato leaf curl virus in sweet potato (Ipomoea batatas)

Sweet potato leaf curl virus (SPLCV) infects sweet potato and is a member of the family Geminiviridae (genus Begomovirus). SPLCV transmission occurs from plant to plant mostly via vegetative propagation as well as by the insect vector Bemisia tabaci. When sweet potato seeds were planted and cultivated in a whitefly‐free greenhouse, some sweet potato plants started to show SPLCV‐specific symptoms. SPLCV was detected by PCR from all leaves and floral tissues that showed leaf curl disease symptoms. More than 70% of the seeds harvested from SPLCV‐infected sweet potato plants tested positive for SPLCV. SPLCV was also identified from dissected endosperm and embryos. The transmission level of SPLCV from seeds to seedlings was up to 15%. Southern blot hybridization showed SPLCV‐specific single‐ and double‐stranded DNAs in seedlings germinated from SPLCV‐infected seeds. Taken altogether, the results show that SPLCV in plants of the tested sweet potato cultivars can be transmitted via seeds and SPLCV DNA can replicate in developing seedlings. This is the first seed transmission report of SPLCV in sweet potato plants and also, to the authors' knowledge, the first report of seed transmission for any geminivirus.     

Indian peanut clump virus (IPCV) infection on wheat and barley: symptoms, yield loss and transmission through seed

Wheat and barley crops were shown to be susceptible to Indian peanut clump virus (IPCV) under field conditions. In wheat, the Hyderabad isolate of IPCV (IPCV-H) induced symptoms resembling the rosette caused by soil-borne wheat mosaic virus, and these were apparent only three weeks after emergence. Early-infected plants were severely stunted and dark green, with chlorotic streaks on the youngest leaves, which turned necrotic as the plants aged; most of these plants died. Late-infected plants were also stunted and were conspicuous in the field because of their dark green appearance as a result of delayed maturity. The virus was detected by ELISA and nucleic acid hybridization in all plants with symptoms. These plants usually produced fewer tillers than healthy ones. Spikes were malformed, often did not emerge from the flag leaf, and they contained few, shrivelled seeds. Grain yield was decreased, on average, by 58%. In barley, IPCV-H caused severe stunting and general leaf chlorosis. As the plants aged, the leaves became necrotic and the few infected plants that reached maturity produced small spikes. IPCV-H antigens were detected by ELISA in every wheat seed from infected plants and the virus was transmitted through wheat seed at a frequency of 0.5–1.3%. Storage at 4°C for more than a year did not affect seed transmission frequency. The virus was detected in leaves and roots of seed-transmitted plants. Seed transmission was not detected in barley. The Durgapura isolate (IPCV-D) was detected in wheat crops (cv. RR-21) at 3 different locations in Rajasthan State, India. Infected plants showed reduced growth without any overt symptoms.     

Role of seed infection by the Ascochyta blight pathogen of dried pea (Mycosphaerella pinodes) in seedling emergence, early disease development and transmission of the disease to aerial plant parts

The role of infected seed in the epidemiology of Ascochyta blight of pea, caused by Mycosphaerella pinodes, was studied both under growth chamber and field conditions, using healthy seeds, naturally infected seeds and artificially infected seeds. Results suggest that infected seeds caused serious losses, as a result of poor germination and high transmission of the disease, to parts of the plants under soil level. Foot rot symptoms often caused the death of young seedlings. Losses were increased by low temperatures during the early stage of crop development. M. pinodes progressed from seeds to aerial parts of the plants, but no Ascochyta blight symptoms occurred, the disease remaining near to the basal parts of the plants as a foot rot symptom. This suggests that seeds cannot be regarded as a source of contamination in the epidemiology of the disease.     

The Relationship Between Wheat Seed Weight, Infection by Fusarium culmorum or Microdochium nivale, Germination and Seedling Disease

The distribution of seeds by weight for three lots of winter wheat cv. Avalon infected by Fusarium culmorum and three lots of winter wheat cv. Riband infected by Microdochium nivale was determined. The distribution of infected seeds within each seed lot was then determined by isolating F. culmorum from seeds on moist filter paper and M. nivale from seeds on potato dextrose agar. The distribution of M. nivale infected seeds between seeds of different weight was similar to that of the seed lot as a whole, whereas the distribution of F. culmorum was greater in light seeds than heavy seeds. The percentage germination of infected seeds decreased with seed weight. A similar situation was found with respect to seedling emergence in compost for F. culmorum infected seeds. However, with M. nivale infection, similar numbers of seedlings emerged from both light and heavy infected seeds. Seed treatment with guazatine increased seedling emergence for both light and heavy seed infected by M. nivale. However, seedling emergence from F. culmorum infected seed was poor even following treatment with guazatine. Poor emergence was most evident from light seed.     

A model system for plant-virus interaction—infectivity and seed transmission of <Emphasis Type="Italic">Cherry leaf roll virus</Emphasis> (CLRV) in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

The wide natural incidence of Cherry leaf roll virus (CLRV) in deciduous forest trees and nurseries in northern Europe is believed to have occurred, apart from occasional mechanical spread and transmission through grafting, mainly by seed transmission. The mode of the vertical transmission and its role in the epidemiology of the virus has not been investigated, basically due to the inconvenient host-pathogen combinations studied to date. With the aim of obtaining an appropriate system for identification of viral genes and products participating in infection processes and seed transmission of CLRV, we performed infection and seed transmissibility tests with CLRV in Arabidopsis thaliana plants. Two phylogenetically and serologically different CLRV isolates were tested. Both of them were found able to infect A. thaliana plants, exhibited clear symptoms of the infection and spread systemically in the plants. Infection of the seeds and of a remarkable number of seedlings generated from infected seeds was possible for two consecutive generations. These results, for first time, report seed transmission of CLRV in the model plant A. thaliana and allow the assumption to be made of embryo invasion during seed transmission. Furthermore, first indications are given that genetically diverse CLRV isolates exhibit different abilities for vertical transmission in A. thaliana. The CLRV-A. thaliana model system is suitable for investigating viral invasion of developing plant organs and meristematic tissue, a prerequisite for successful virus dissemination via vertical transmission through seed.     

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.     

<Emphasis Type="Italic">Indian citrus ringspot virus:</Emphasis> localization of virus in seed tissues and evidence for lack of seed transmission

Indian citrus ringspot disease is an important viral disease in kinnow mandarin orchards where disease incidence up to 100% has been recorded. The disease is caused by Indian citrus ringspot virus (ICRSV), a positive sense flexuous RNA virus. The transmission of ICRSV is generally through budwood. Association of ICRSV with pollens of naturally infected flowers from cv. ‘Kinnow’ mandarins has been shown previously and this study demonstrates the presence of ICRSV in seed tissues. DAC-ELISA revealed the presence of virus in seed coats but not in embryo and endosperm of seeds collected from the fruits of ICRSV-infected Kinnow plants. Of the infected seed coats, 18% were found to harbor the virus. The seedlings in the grow-out test did not show any symptom for 2 years and the virus could not be detected in seedlings by DAC-ELISA and RT-PCR. The present study indicated that ICRSV could be localized in the testa of seeds but its transmission to progeny was not observed.     

The question of seed transmissibility of Plum pox virus

For many years, Plum pox virus (PPV) was considered to be transmissible by seed, increasing the fear of long-distance spread of the disease. In the late 1970s, it was claimed on the basis of biological transmission of the virus to herbaceous indicator plants and the development of serological diagnosis based on polyclonal antibodies, that PPV was seed-transmitted, with a different infection rate according to the plant species and part of the seed which was tested. In the 1990s, PPV was characterized into four different types, and specific monoclonal antibodies were produced for them. These new and more sensitive diagnostic techniques, together with RT-PCR with different sets of specific primers, were used to approach once again the problem of PPV transmission through seeds. The virus was detected in seed coats and cotyledons, but embryonic tissue and seedlings obtained from germinated seeds never showed symptoms, and gave negative results for PPV with both ELISA and PCR assays. No PPV isolate is currently recognized to be seed transmitted, so vertical transmission of PPV from infected mother plants to their progeny does not occur. Hypothetically, the only possibility of seed transmission would arise from a mutation in the helper component of the virus, associated with high susceptibility of the infected Prunus cultivar.     

Detection of Diplodia sapinea in Corsican pine seeds

Diplodia sapinea is one of the major pathogens of pines worldwide. Despite the putative critical importance of seed infection in the epidemiology of the disease, this aspect of the biology of the fungus is poorly known. Here, biological and molecular methods were developed for the detection of the fungus and applied to assess D. sapinea infection in Corsican pine seeds. A buffered medium containing tannic acid and malt extract as a nutrient base was the most efficient and selective for D. sapinea recovery. A molecular method based on DNA extraction with a commercial kit and specific amplification, including an internal amplification control, was developed. A high percentage of infection (57% positive isolations) was observed in seeds obtained from fallen cones in a Corsican pine stand with no apparent symptoms of D. sapinea. Seeds collected from trees in a seed orchard showing severe symptoms of dieback caused by D. sapinea had comparatively lower infection (38%). Moreover, very low infection levels (1–5%) were observed after the standard treatment used for seed extraction, which included heating at 40°C. Diplodia sapinea was not recovered from seedlings grown from infected seed lots submitted to water stress. Overall, results suggest that the risk of disease transmission by commercial seeds is probably low, but could be further reduced by thermotherapy.     

Presence of Xylella fastidiosa in Sweet Orange Fruit and Seeds and Its Transmission to Seedlings

ABSTRACT Xylella fastidiosa, a xylem-limited bacterium, causes several economically important diseases in North, Central, and South America. These diseases are transmitted by sharpshooter insects, contaminated budwood, and natural root-grafts. X. fastidiosa extensively colonizes the xylem vessels of susceptible plants. Citrus fruit have a well-developed vascular system, which is continuous with the vascular system of the plant. Citrus seeds develop very prominent vascular bundles, which are attached through ovular and seed bundles to the xylem system of the fruit. Sweet orange (Citrus sinensis) fruit of cvs. Pera, Natal, and Valencia with characteristic symptoms of citrus variegated chlorosis disease were collected for analysis. X. fastidiosa was detected by polymerase chain reaction (PCR) in all main fruit vascular bundles, as well as in the seed and in dissected seed parts. No visual abnormalities were observed in seeds infected with the bacterium. However, the embryos of the infected seeds weighed 25% less than those of healthy seeds, and their germination rate was lower than uninfected seeds. There were about 2,500 cells of X. fastidiosa per infected seed of sweet orange, as quantified using real-time PCR techniques. The identification of X. fastidiosa in the infected seeds was confirmed by cloning and sequencing the specific amplification product, obtained by standard PCR with specific primers. X. fastidiosa was also detected in and recovered from seedlings by isolation in vitro. Our results show that X. fastidiosa can infect and colonize fruit tissues including the seed. We also have shown that X. fastidiosa can be transmitted from seeds to seedlings of sweet orange. To our knowledge, this is the first report of the presence of X. fastidiosa in seeds and its transmission to seedlings.     

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.     

Seed transmission of Melon necrotic spot virus and efficacy of seed-disinfection treatments

Rates of seed transmission of Melon necrotic spot virus (MNSV) were estimated in seedlings grown from commercial melon ( Cucumis melo ) cv. Galia F₁ seeds. Seedlings at the cotyledon stage and adult plants were assayed for MNSV by DAS-ELISA and RT-PCR. None of the seedling groups tested positive for MNSV by ELISA. The proportion of seedlings infected with MNSV was at least 7 and 8% in seed lots 05 and 06, respectively, as estimated from RT-PCR analysis of grouped seedlings. Fourteen and eight grouped samples (10 seedlings per group), of a total of 200 and 100 seedlings, respectively, grown from infected seeds were MNSV-positive in seed lots 05 and 06, respectively, corresponding to seed-to-seedling transmission rates of 11·3 and 14·8%, respectively. Several seed-disinfection treatments were evaluated for their ability to prevent seed transmission of MNSV. The results suggest that a treatment of 144 h at 70°C can be used to eradicate MNSV in melon seeds without hindering germination.     

Pathogenicity of isolates of Magnaporthe spp. from wheat and grasses infecting seedlings and mature wheat plants in Argentina

Wheat blast of wheat (Triticum aestivum), caused by Magnaporthe oryzae pathotype triticum (MoT; anamorph Pyricularia oryzae) is a destructive disease in the South American countries of Brazil, Paraguay and Bolivia. In Argentina, the fungus was recently recorded on wheat and barley plants in the northeast part of the country, Buenos Aires and Corrientes Provinces, with a potential for spreading. This work aimed to study, for the first time, the morphocultural and pathogenic characteristics of Magnaporthe isolates collected from wheat and other herbaceous species in Argentina and three neighbouring countries (Paraguay, Brazil and Bolivia) and determine their aggressiveness on wheat varieties. Statistical differences among isolates, culture media, and development conditions were found for conidia colour, growth rate, size and sporulation rate. Pathogenicity tests performed on seedlings with 19 isolates of Magnaporthe spp. under greenhouse conditions showed a maximum disease severity of 55.3% and 66.7% for varieties BIOINTA 3004 and Baguette 18, respectively. Weed and grass isolates were infectious on wheat, demonstrating their potential epidemiological role on the disease. Spike disease severity was 34.6% for the host × pathogen interaction of BIOINTA 3004 × PY22. Observed symptoms included partial or total spike bleaching, and glume and rachis discolouration. The 1000‐grain weight was significantly reduced to 38.5% and 63.1% for cultivars BIOINTA 3004 and Baguette 18, respectively. The disease affected grain germination, which fell to 65.9% for seeds infected with the PYAR22 isolate. Symptoms observed in infected grains were partial spotting, grain softening, and rot symptoms with the presence of a greyish mould.     

The importance of the infected seed tuber and soil inoculum in transmitting Potato mop‐top virus to potato plants

Potato mop‐top virus (PMTV), the cause of spraing in potato tubers, is transmitted by Spongospora subterranea, the cause of powdery scab, and by planting infected seed tubers. This study was undertaken to determine the relative importance of these sources of infection in seed potato production in Scotland. The transmission of PMTV from tested seed tubers to daughter plants was examined over 2 years and six cultivars. The development of foliar symptoms varied with year and cultivar. Infection of daughter tubers derived from PMTV‐infected seed tubers was more prevalent on plants affected by foliar symptoms than those without symptoms. The rate of transmission of PMTV from infected seed tubers to daughter tubers ranged from 18 to 54%. Transmission was affected by cultivar and by origin of seed tubers used for a cultivar, but not by a cultivar's sensitivity to PMTV infection. The incidence of PMTV in daughter tubers of cv. Cara grown from seed potatoes from one source (common origin) by more than 25 seed producers was examined over two successive generations. The incidence of PMTV in daughter tubers was not correlated with that in the seed tubers but appeared to be strongly associated with soil inoculum. The incidence of PMTV was correlated with powdery scab in those crops in which both were present. There was some evidence from soil tests conducted in 2006 using a tomato bait plant and real‐time RT‐PCR that planting PMTV‐infected seed potatoes could increase the risk of introducing the virus into land not infested by PMTV.     

Assessment of the loose smut fungi (Ustilago nuda and U. tritici) in tissues of barley and wheat by fluorescence microscopy and real-time PCR

Loose smut fungi of barley and wheat (Ustilago nuda and U. tritici, respectively) colonize the plant without causing obvious disease symptoms before heading. The availability of diagnostic methods to detect and follow the growth of these pathogens in the plant would therefore be highly advantageous for both resistance breeding and the development of effective seed treatments. Using seed lots of barley and wheat highly infected with loose smut, we studied the early establishment of the loose smut pathogens in the plant by fluorescence microscopy. In hand-cut sections stained with the fluorochrome Blankophor?, fungal hyphae were observed to invade the shoot apical meristem and leaf primordia during the first days after the onset of germination. At the first node stage the ear and leaf primordia were generally extensively colonized. Hyphae of U. nuda were also regularly observed in high density in the nodes. A protocol was developed for the specific amplification of U. nuda and U. tritici DNA extracted from infected plant tissue. PCR screening of U nuda in seedlings from infected and healthy seed lots was compared to ELISA, microscopy and ultimately head infection of mature plants derived from tillers of the tested seedlings. The results indicated that a prediction of loose smut infection by real-time PCR is possible at the second leaf stage, and that the assay is equally suited for use with spring and winter varieties of barley and wheat.     

大豆褐斑粒与大豆花叶病毒若干株系的关系

 近年来发生的大豆褐斑粒是由大豆感染大豆花叶病毒的黄斑株系和顶枯株系等所致。人工接种上述病毒证明了这一点。种子斑驳的严重度与植株发病的严重度有某种相关性。
病株所结种子并不一定全部为褐斑粒。褐斑粒或病株所结无病状种子都可以传病,带毒种子产生疫苗的多少视环境条件、大豆品种及病毒的株系而异。     

Temperature Dependent Seed Transmission of Stagonospora nodorum in Wheat

The transmission of Stagonospora nodorum from four naturally infected winter wheat seedlots was quantified in controlled environment germination chambers at 9, 13, 17, 21, and 25 °C. Seedlings were harvested when the second leaf began to emerge. Coleoptiles and first seedling leaves were examined for the presence of lesions caused by S. nodorum. First leaves were incubated on Bannon's medium for 2 weeks, after which they were examined for pycnidia of S. nodorum. Transmission to the coleoptile occurred at all temperatures, but decreased from 100% to 72% as temperature increased from 9 to 25 °C. Transmission to the first leaf was less, dropping from 37% to 2% as temperature increased from 9 to 25 °C. At least 44% of infected first leaves were symptomless at all temperatures, with 96% of infected leaves showing no symptoms at 25 °C. Transmission to seedling leaves occurred over a broad temperature range. Under the high densities at which wheat is sown, a significant number of infected seedlings per unit area may originate from relatively low initial seed infection levels and transmission efficiencies.