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.     

Evaluation of non-chemical seed treatment methods for the control of Phoma valerianellae on lamb’s lettuce seeds

Journal of Plant Diseases and Protection - The aim of the present study was to identify seed treatment methods for eradicating Phoma valerianellae from lamb`s lettuce seeds in organic vegetable...     

New subspecies-specific polymerase chain reaction-based assay for the detection of Acidovorax avenae subsp. citrulli

New subspecies-specific primers for the detection of Acidovorax avenae subsp. citrulli ( Aac ), the seedborne bacterium which causes bacterial fruit blotch of cucurbits, were designed based on PCR fragments obtained in ERIC- and BOX-PCR profiles of Aac strains. PCR with both primer sets identified 30 strains from different locations and hosts, but did not amplify DNA from other closely related species and subspecies assessed, with the exception of DNA of one strain of A. avenae subsp. avenae , which was amplified by one of the primer sets, named BX-S. This primer set, based on a BOX-PCR fragment, performed under high-stringency conditions without losing its detection sensitivity. The primers were also evaluated for their ability to detect the pathogen in contaminated watermelon and melon seed samples. The BX-S primers facilitated the detection of the pathogen from washings of 5000-seed samples with 0·02% infestation. This primer set was also assessed for detection using immunomagnetic separation polymerase chain reaction (IMS-PCR) and was shown to be as sensitive as a previously described primer set (AACF2/R3), detecting 0·02% infestation in seed samples. This highly specific and sensitive primer set could be used to improve PCR-based detection of this important pathogen.     

A semiselective medium for the isolation of Acidovorax valerianellae from soil and plant debris

Bacterial black spot of corn-salad ( Valerianella locusta ) caused by Acidovorax valerianellae appeared in France in the beginning of the 1990s. The disease is now widespread in the main area of corn-salad production and is responsible for high depreciation and significant economic losses every year. A semiselective medium is described for the detection of the bacterium in various environments and to identify primary sources of inoculum. The medium TSAV (tryptic soy broth, 3 g L⁻¹, agar 15 g L⁻¹, 5-fluorouracyl 5 mg L⁻¹, novobiocin 5 mg L⁻¹, propiconazole 5 mg L⁻¹) increased chances of obtaining cultures of A. valerianellae from naturally infested soil and from root-debris. Using TSAV, the pathogen was detected in root-debris up to 39 days after the harvest of a diseased crop. As only a few days usually separates harvest from the next sowing date, soil is likely to be an important source of inoculum for the next crop.     

Modelling the spread of Xanthomonas campestris pv. campestris in module-raised brassica transplants

A series of experiments was performed to quantify the rate of dispersal of Xanthomonas campestris pv. campestris in module-raised brassica transplants, in a simulated commercial plant-raising system. Seeds were sown in '308' module seed trays and set out in blocks in the glasshouse. Primary inoculum was introduced as inoculated seeds sown in one or more cells. Trays were watered via an overhead-gantry irrigation system, hand-watered or capillary-watered. Disease symptoms were monitored visually and the presence of the pathogen on samples of plants was monitored by leaf washing, dilution and plating on selective medium. Spread of symptoms was greatest in the gantry-watered trays, was very limited in hand-watered trays and was almost non-existent in capillary-watered trays. Dispersal of bacteria followed a similar pattern, but the proportion of plants contaminated was much greater than the proportion showing symptoms, and approached 100% after six weeks in the gantry-watered trays within 50 plants distance from a single primary infector. Models relating the proportion of plants with symptoms, or contaminated, to the distance from primary infector and time since sowing were fitted to the data. Predictions of the proportions of plants contaminated in commercial-scale blocks of transplants suggested that high levels of disease in the field could be explained by rapid rates of pathogen spread during plant-raising, and that the widely-used tolerance standard for seed health testing (0·01%) should be revised to 0·004%. In addition to seed health testing, control should focus on raising transplants under conditions that minimise the rates of disease spread and pathogen dispersal.     

Studies on the biology of black mould (Aspergillus niger) on temperate and tropical onions. 2. The effect of treatments on the control of seedborne A. niger

Treatment of A. niger -inoculated onion seed with benomyl dust (1 g a.i./kg), or a foliar spray of thiram (0·4% a.i./ha) applied to plants grown from inoculated seed under temperate (UK) conditions reduced the incidence of A. niger in harvested crops. Treatment of naturally contaminated Sudanese onion seed with a benomyl + thiram mixture at a rate of 2·5 + 2·5 g a.i./kg or soaking the seed in hot water (15 min at 60°C) reduced the incidence of black mould on bulbs grown in the Sudan in field soil that had not previously been used for onion production. The seed treatments were less effective in crops produced in fields regularly used for onion production. However, incubation of harvested bulbs in moist chambers showed that damage to the internal storage tissue of onion bulbs caused by A. niger was reduced by seed treatment.     

种子引发处理对无籽西瓜幼苗生长的影响和对细菌性果斑病菌消毒的效果

 种子出苗率低和传带细菌性果斑病菌是制约三倍体无籽西瓜生产的主要原因。本试验利用KMnO₄、CuSO₄和ZnSO₄的不同浓度溶液对无籽西瓜种子进行不同时间引发处理, 通过滤纸发芽试验, 观测其对种子发芽和幼苗生长的促进作用;借助平板法测定引发溶液对细菌性果斑病菌FC247的抑制作用, 免疫凝聚试纸条和传统PCR检测引发处理对种子人工接菌FC247的消毒效果。结果表明:0.1% CuSO₄溶液引发处理4 h和0.2% ZnSO₄溶液引发处理24 h, 分别比未引发处理的无籽西瓜种子发芽率提高71.1%和73.3%, 并能显著提高发芽整齐度和幼苗素质;同时, 引发溶液对细菌性果斑病菌有显著抑制作用, 并对人工接菌种子表现出-定程度的消毒效果。     

种衣剂对玉米田双斑长跗萤叶甲的防效评价

为评价种衣剂防治玉米田双斑长跗萤叶甲的可行性,通过选用内吸性强、持效期长的种衣剂,设置常规剂量对玉米种子进行包衣处理,采取田间罩网小区试验,明确种衣剂对双斑长跗萤叶甲的防效及对玉米出苗、生长的安全性。结果表明,30%噻虫嗪FS、10%氟虫腈FSC、38%噻虫胺FSC、40%噻虫嗪·溴氰虫酰胺FS、30%噻虫嗪·氟虫腈FSC、30%噻虫胺·氟虫腈FSC、20%噻虫胺·氟啶虫酰胺FSC等7种种衣剂对玉米出苗、生长均安全;38%噻虫胺FSC有效成分用量7.6 g/kg种子处理对双斑长跗萤叶甲的防治效果为70.6%～78.5%,防效最好,显著高于除30%噻虫胺·氟虫腈FSC 7.5 g/kg处理外的其他种衣剂处理(P<0.05),并且可使双斑长跗萤叶甲成虫羽化出土始现期、出土高峰期推迟5～10 d,可以使成虫高峰期避开玉米吐丝期。玉米田双斑长跗萤叶甲防控前移推荐使用38%噻虫胺FSC,有效成分剂量为7.6 g/kg种子。     

Comparison of extraction procedures and determination of the detection threshold for Clavibacter michiganensis ssp. michiganensis in tomato seeds

Several seed extraction procedures, used for detection of Clavibacter michiganensis ssp. michiganensis ( Cmm ) in naturally infected and artificially infested tomato seed lots were evaluated. Extraction methods that included grinding the seeds were significantly better at detecting the pathogen in three different seed lots than methods that used only soaking. The detection threshold of Cmm in relation to seed sample size was determined by adding naturally infected seeds into samples of three different sizes. Cmm was detected by agar plating assay, on three media (CNS, mSCM, D₂ANX), and by direct PCR from seeds and Bio-PCR (bacteria cultured on agar media prior to PCR). In samples of 10 000 seeds containing one infected seed, Cmm could be detected only by Bio-PCR and in only one replicate out of five. In samples containing five or 10 infected seeds per 10 000 seeds, three of five and five of five replicates, respectively, were detected by the three detection methods. In samples of 5000 seeds, one infected seed could be detected in all five replicates only after adding a concentration step. A high correlation ( R ² = 0·9448) between artificially infested seeds and the disease incidence was found. Seed lots infested with less than 58 colony-forming units (CFU) per g did not cause disease under glasshouse conditions, whereas lots with about 1000 CFU g⁻¹ caused disease in 78 plants out of 2000.     

哈蜜瓜种子带细菌性果斑病菌检测技术的研究

哈蜜瓜细菌性果斑病是世界性的检疫对象之一,主要以种子带菌进行远距离传播,本文通过温室育苗、室内分离和PCR(聚合酶链式反应)技术对哈蜜瓜种子带菌情况和主要带菌部位进行了检验,结果表明:3种方法都可以对种子带菌情况进行检测,分离检测和PCR检测还可以确定种子的带菌部位,PCR技术具有特异性强、灵敏度高、检测时间短等特点.另外检测结果也表明种子带菌以种皮为主,种仁的带菌量较少.     

Location of Acidovorax citrulli in infested watermelon seeds is influenced by the pathway of bacterial invasion

Watermelon seeds can become infested by Acidovorax citrulli, the causal agent of bacterial fruit blotch (BFB) of cucurbits via penetration of the ovary pericarp or by invasion of the pistil. This study investigated the effect of these invasion pathways on A. citrulli localization in seeds. Seed samples (n = 20 or 50 seeds/lot) from pistil- and pericarp-inoculated lots were dissected into testa, perisperm-endosperm (PE) layer, and embryo tissues and tested for A. citrulli by species-specific polymerase chain reaction (PCR) and by plating on semiselective media. Less than 8% of the testa samples were A. citrulli-positive regardless of the method of seed inoculation. Additionally, the difference in percentages of contaminated testae between the two seed lot types was not significant (P = 0.64). The percentage of A. citrulli-positive PE layer samples as determined by real-time PCR assay was significantly greater for seeds from pistil-inoculated lots (97%) than for seeds from pericarp-inoculated lots (80.3%). The mean percentage of A. citrulli-positive embryo samples was significantly greater for seeds from pistil-inoculated lots (94%) than for seeds from pericarp-inoculated lots (≈8.8%) (P = 0.0001). Removal of PE layers and testae resulted in a significant reduction in BFB seed-to-seedling transmission percentage for seeds from pericarp-inoculated lots (14.8%) relative to those from pistil-inoculated lots (72%). Additionally, using immunofluorescence microscopy, A. citrulli cells were observed in the PE layers and the cotyledons of pistil-inoculated seeds but only in the PE layers of pericarp-inoculated seeds. These results suggest that pericarp invasion results in superficial contamination of the testae and PE layers while pistil invasion results in the deposition of A. citrulli in seed embryos.     

Bio-PCR方法检测葫芦科作物种子携带 西瓜嗜酸菌的特异性引物筛选

 由西瓜嗜酸菌(Acidovorax citrulli)引起的细菌性果斑病是一种毁灭性的种传病害,可为害多种葫芦科作物并造成重大经济损失。该病原菌为检疫性有害生物,种子带菌是田间病害发生的最重要初侵染来源,因此,种子健康检测成为病害综合防控过程中的重要环节。Bio-PCR是当前种子携带细菌检测的常用方法,而特异性引物的选择和使用是检测的关键。本研究使用已报道的7对引物对17株西瓜嗜酸菌、10株嗜酸菌属其它种的菌株和6株其它属的植物病原细菌进行了Bio-PCR检测,筛选出对西瓜嗜酸菌特异性最好的引物为SEQID4^m/SEQID5。研究表明:使用该引物对西瓜嗜酸菌MH21纯菌菌悬液的检测限度为10² CFU·mL^-1;在人工添加菌悬液的模拟带菌西瓜种子中,使用ASCM和EBBA两种半选择性培养基结合引物SEQID4^m/SEQID5进行Bio-PCR检测,ASCM对种子中带菌量的检测限度可达到0.01 CFU·g^-1,EBBA对种子中带菌量的检测限度为0.1 CFU·g^-1。     

玉米疯顶病种子传播研究

连续3a对不同来源的玉米种子,利用显微镜镜检和试种观察植株发病显症的方法,检测其携带玉米疯顶病病菌状况。结果表明,病株病穗籽粒、病株健穗籽粒、病田健株籽粒都有较高的带菌率。因此,病菌随种子调运远距离传播是造成各地病害突发的主要原因。     

Novel in vivo use of a polyvalent Streptomyces phage to disinfest Streptomyces scabies-infected seed potatoes

A highly virulent and polyvalent Streptomyces phage was isolated from a potato field near Albany, Western Australia. The efficacy of the isolated phage to disinfest seed potato tubers artificially inoculated with a common scab-causing streptomycete was evaluated. The phage suspension was prepared in a mini-bioreactor. Diseased potatoes were bathed in a phage suspension (1 × 10⁹ plaque-forming units per mL) for 24 h. The suspension was constantly circulated within a novel 25 L phage bath by means of an air-sparging pipe driven from an air compressor. Phage-treated scab-affected seed potatoes planted into free-draining polystyrene boxes containing steam-pasteurized field soil produced tuber progeny with significantly ( P  < 0·05) reduced levels of surface lesions of scab (1·2%) compared with tubers harvested from nonphage-treated tubers (23%). The number of scab lesions was also significantly reduced ( P  < 0·05) by phage treatment of mother tubers. No significant differences were recorded in weight, size or number of harvested tubers from phage-treated or nontreated mother tubers. This is the first in vivo study that has used Streptomyces phage to significantly disinfest seed potatoes of Streptomyces scabies and thereby reduce contamination of soil from seed-tuber-borne inoculum and reduce infection of daughter tubers.     

Germination strategies of annual and short-lived perennial species in the Arabian Desert

Germination timing is highly regulated in short-lived plant species since it strongly influences recruitment success of vegetation. In deserts, the spatiotemporal distribution of plant-available water is highly episodic and unpredictable, making winter months more favorable for seed germination when other abiotic conditions co-occur. We hypothesized that changes in photoperiod and thermoperiod would impact germination more in seeds that had undergone in situ storage. We assessed 21 annual and short-lived perennial species in the Arabian Desert to find (1) if seeds were dormant at maturity, (2) if in situ seed storage increased germination percentage compared with no storage, (3) if photoperiod and thermoperiod germination requirements were influenced by in situ storage, and (4) if a phylogenetic association in seed germination could be observed. Seeds of each species collected in early 2017 were divided into two batches. One was tested for germination within one week (fresh seeds). The other was stored in situ at the maternal location (stored seeds) until October 2017 and tested for seed germination in the first week of November. Seed germination was conducted in incubators at two thermoperiods (15°C/20°C and 20°C/30°C; 12 h/12 h), and two photoperiods (12 and 0 h light per day). Results indicated that seed germination percentages of 13 species were significantly enhanced by in situ storage. A thermoperiod response was exhibited by stored, but not fresh seeds. Light exposure increased germination of fresh seeds but had only a minimal effect on stored seeds. Germination traits exhibited no phylogenetic correlation. This result indicated that selection pressure for germination strategy was stronger than that for taxonomic traits of these desert species.     

Long‐term survival of Acidovorax citrulli in citron melon (Citrullus lanatus var. citroides) seeds

Long‐term survival of Acidovorax citrulli in citron melon (Citrullus lanatus var. citroides) seeds was investigated. Citron melon seed lots infected with A. citrulli were generated in the field by inoculating either the pistils (stigma) or pericarps (ovary wall) of the female blossoms. Seventeen A. citrulli isolates from 14 different haplotypes belonging to two different groups (group I and II) were used for inoculation. After confirming that 100% of seed lots were infected, they were stored at 4°C and 50% RH for 7 years. After storage, the viability of A. citrulli cells from individual lots was determined by plating macerated seeds on semiselective medium as well as growing seeds for 14 days and scoring for bacterial fruit blotch symptoms. The type of A. citrulli isolate (group I or group II) used did not significantly influence bacterial survival. However, A. citrulli survival was significantly greater in seed lots generated via pistil inoculation (52·9 and 29·4%) than via pericarp inoculation (23·5 and 17·6%). Repetitive extragenic palindrome (rep)‐PCR on A. citrulli isolated from citron melon seed lots after storage displayed similar fingerprinting patterns to those of the reference strains originally used for blossom inoculation, indicating that cross‐contamination did not occur. The results indicate that A. citrulli may survive/overwinter in citron melon seeds for at least 7 years and bacterial survival in seed was influenced more by method of blossom inoculation than by the type of bacterial isolate.     

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.     

Identification, pathogenicity and distribution of Penicillium spp. isolated from garlic in two regions in Argentina

A total of 147 samples of garlic ( Allium sativum ) bulbs affected by blue mould were obtained from a variety of agroclimatic districts between December 1999 and February 2000. Penicillium species were identified using both morphological and chemotaxonomic characteristics. Penicillium allii was the predominant species isolated (81·8%) in this survey and the only species proven to be pathogenic on garlic. Other species were isolated much less frequently: P. chrysogenum (13·7%), P. brevicompactum (2·8%), P. phoeniceum (0·9%), P. aurantiogriseum (0·6%) and P. flavigenum (0·2%). Colonies of P. allii could be classified into four morphotypes and their distribution seemed to be influenced by seed trade and agricultural practices. Penicillium allii isolates were grouped into three aggressiveness phenotypes (low, medium and high) based on their ability to cause disease during field trials on susceptible (Fuego INTA) and less susceptible (Castaño INTA) garlic cultivars. The number of surviving plants at 191 days after planting and postharvest bulb weight contributed the most towards aggressiveness modelling.     

New weed management strategy using subterranean clover reseeding under different tillage systems: Numerical experiments with the subterranean clover-tillage dynamics model

Subterranean clover ( Trifolium subterraneum L.) appears to be a suitable winter legume cover crop for Japan because the subterranean clover stands maintain themselves by reseeding in autumn and the large seeds facilitate establishment and enable early fall production. However, there is little information on the relationship between reseeding and tillage systems for subterranean clover under Japanese climatic conditions. A weed-tillage population dynamics model was developed to investigate the effects of the tillage method and timing on the reseeding of subterranean clover. The field experiments were conducted in a silage corn and subterranean clover rotation system. The life cycle of subterranean clover was modeled to describe the seed production competing with weed growth, seasonal changes in the buried seeds' viability, and the movement of seeds by tillage treatments. In the numerical simulation, the effect of the tillage method on the seedling population of reseeded subterranean clover was investigated and the simulation results showed good agreement with the experimental results. Rotary tilling immediately after subterranean clover seed maturation successfully produced a good subterranean clover stand the following spring. However, rotary tillage conducted 2 months after seed maturation killed the emerging subterranean clover seedlings and the field was dominated by winter weeds. These simulation results suggest that a suitable tillage system can maintain successful subterranean clover re-establishment from year to year.     

SEED GERMINATION OF AGROSTIS GIGANTEA ROTH

Summary. To assess the role of seed production and seedling behaviour in the biology of the perennial grass weed Agrostis gigantea Roth, seed germination and seedling emergence of the species were investigated in the glasshouse and in the laboratory. These studies showed that the species may produce a large number of seeds: panicles collected from a cereal field in 1971 and 1972 contained about a thousand viable seeds in both years. Mature seeds of the species are mostly non-dormant and germinate readily on moist soil surfaces. Freshly-harvested seeds require light and alternating tempratures for germination, but older seeds can germinate in constant temparature in the dark. Covering seeds with 0·6 cm soil halved the percentage emergence and few seedlings emerged from seeds sown deeper than 3±8 cm, but many seeds survived under enforced dormancy. A. gigantea seeds may also persist long in frequently cultivated shallow soil. The species flowers late but its seeds become viable quickly—in 1969 a third of the number that were finally viable became so within a week of flowering.
Germination des semenes d' Agrostis gigantean Roth.