Comparison and differentiation of Wheat Yellow Mosaic Virus(WYMV), Wheat Spindle Streak Mosaic Virus (WSSMV) and Barley Yellow Mosaic Virus (BaYMV) isolates using WYMV monoclonal antibodies

Twelve monoclonal antibodies (MAbs) were obtained by immunizing mice with a French isolate (F1) of wheat yellow mosaic virus (WYMV). Three of these (3D12, 2C1, 6C3) belong to the IgM class and the nine others to the IgG class (3D8, 3H1, 2B8, 1F2, 3C10, 4F12, 3H9, 1G5, 54). In antigen-coated plate (ACP) ELISA and indirect double antibody sandwich (IDAS) ELISA, all MAbs recognize the WYMV (F1) both in the form of purified particles and in wheat leaf extract. The analysis of numerous French isolates of WYMV shows a variable reactivity with MAbs 3D8, 3H1, 2B8, 3C10, 3H9 and 1G5 in IDAS — and ACP-ELISA. The Japanese isolate of WYMV and United States isolates of wheat spindle streak mosaic virus (WSSMV) were detected in IDAS- and ACP-ELISA by ten of the MAbs tested showing that the wheat bymoviruses originating from the three locations share a high epitopic homology. French isolates of barley yellow mosaic virus (BaYMV; pathotypes 1 and 2) were only detected in ACP-ELISA with MAbs 6C3, 3D8, 3H1 and 2B8 whereas the two Japanese strains (I-1, II-1) of MaYMV were recognized with these and also with that of 3C10. In IDAS-ELISA, the two Japanese strains were clearly detected by MAbs, 6C3, 3D8, 3H1, 1F2, 3C10 and 1G5 and the British and Belgian (pathotype 2) isolates only by that of 6C3. Only the Japanese strain of BaYMV, 1-1 could be detected with MAb 3H9 in this ELISA system.     

Occurrence of Honeysuckle Yellow Vein Virus (HYVV) containing a monopartite DNA-A genome in Korea

Two newly emerged begomoviruses were isolated from naturally infected tomato (Solanum lycopersicum) plants grown in greenhouses at Jeju Island and Dangjin in Korea and their genomes were characterized. These viruses-infected plants had very small leaves that curled upward, yellow margins and a leathery appearance, and a bushy and stunted appearance with short internodes. Nucleotide (nt) sequence analysis of their genomes showed that they have a DNA-A component of a monopartite begomovirus. Their genomes comprised 2763 and 2764 nucleotides with six open reading frames. The results of nt sequence similarity analysis of DNA-A genome between the two Korean isolates and isolates of Tobacco leaf curl Japan virus (TbLCJV), Honeysuckle yellow vein virus (HYVV), Honeysuckle yellow vein mosaic virus (HYVMV), and Eupatorium yellow vein virus in Japan (EpYVV) showed that they are likely similar to HYVV-[Masuda] (89.4–92.8% nt identity). Consequently, we tentatively propose the two isolates’ names as HYVV-Jeju and -DJ according to the ICTV geminivirus rules. Phylogenetic relationship analysis of 33 DNA-A genome sequences using PAUP* 4.0b10 and MrBayes revealed that HYVV-Jeju and -DJ belong to the Far East Asian begomovirus species complex. Within the Far East Asian begomovirus species complex, HYVV-Jeju and -DJ are distantly related to EpYVV, HYVMV, and TbLCJV groups. Based on the presence of a recombination fragment spanning the C3 ORF, a recombinant origin was suggested for both HYVV-Jeju and –DJ, with parents close to Japanese isolates HYVMV-[SP1:00] and Eupatorium yellow vein virus (EpYVV)-[Suya]. In addition, the presence of a further recombination fragment spanning the IR suggested the parents of HYVV-DJ were close to HYVV-Jeju and EpYVV-[Suya].     

Comparison of Resistance to Tomato Leaf Curl Virus (India) and Tomato Yellow Leaf Curl Virus (Israel) among Lycopersicon Wild Species, Breeding Lines and Hybrids

The objective of this study was to screen wild and domesticated tomatoes for resistance to Tomato yellow leaf curl virus, Israel (TYLCV-Is) and Tomato leaf curl virus from Bangalore isolate 4, India (ToLCV-[Ban4]) to find sources of resistance to both viruses. A total of 34 tomato genotypes resistant/tolerant to TYLCV-Is were screened for resistance to ToLCV-[Ban4] under glasshouse and field conditions at the University of Agricultural Sciences, Bangalore, India. Resistance was assessed by criteria like disease incidence, symptom severity and squash-blot hybridization. All the tomato genotypes inoculated with ToLCV-[Ban4] by the whitefly vector Bemisia tabaci (Gennadius) produced disease symptoms. In some plants of the lines 902 and 910, however, the virus was not detected by hybridization. The tomato genotypes susceptible to ToLCV-[Ban4] by whitefly-mediated inoculation were also found susceptible to the virus under field conditions. However, there were substantial differences between genotypes in disease incidence, spread, symptom severity and crop yield. Despite early disease incidence, many genotypes produced substantially higher yields than the local hybrid, Avinash-2. Sixteen tomato genotypes from India resistant/tolerant to ToLCV-[Ban4] were also tested for TYLCV-Is resistance at the Hebrew University of Jerusalem, Rehovot, Israel. Accessions of wild species, Lycopersicon hirsutum LA 1777 and PI 390659 were the best sources of resistance to both viruses. Lines 902 and 910, which were, resistant to TYLCV-Is were only tolerant to ToLCV-[Ban4] and accession Lycopersicon peruvianum CMV Sel. INRA, resistant to ToLCV-[Ban4], was only tolerant to TYLCV-Is. Implications of using the resistant lines in breeding programme is discussed.     

Immunocytochemical Evidence that Secretion of Pectin Occurs During Gel (Gum) and Tylosis Formation in Trees

ABSTRACT During gel (gum) formation in angiosperm trees, fibrillar material accumulated in protective layers of xylem parenchyma cells before being secreted across half-bordered pit membranes into vessel elements. Immunogold labeling demonstrated that this fibrillar material was mainly composed of partially esterified pectic polysaccharides. The primary wall of expanding tyloses, an extension of the parenchyma protective layer, secreted similar pectic substances to completely block vessel elements. In most studies, these occluding structures were reported to be formed in response to causative factors such as aging processes, injuries, or infections. Current observations support the view that partial to complete embolism, which almost always accompanies these factors, might be the main cause triggering the formation of vessel occlusions. Whereas pectin seems to be the basic component of gels (gums) and of the external layer of tyloses, other substances, such as phenols, were also detected either as a part of these plugs or as accumulations beside them in vessels. Finally, it is proposed that the term 'gel' instead of 'gum' be used in future studies to describe the occluding material secreted by ray and paratracheal parenchyma cells.     

我国大麦条纹花叶病毒的研究Ⅰ.用ELISA方法检测麦种中的大麦条纹花叶病毒

 用ELISA检测麦种中的大麦条纹花叶病毒(Barlcy stripe mosaicvirus BSMV)方法简便,灵敏度高,检测抗原的最高灵敏度可达10毫微克/毫升。一粒麦芽稀释1000倍还有阳性反应,也能测出带毒麦粒的种皮、胚和胚芽等部位中的BSMV。另外,还检测了国内外的部分麦种,其中来自新疆、北京、江苏和丹麦的小麦品种中存在BSMV。     

Occurrence of Cucurbit Yellow Stunting Disorder Virus (CYSDV) and Beet Pseudo-yellows Virus in Cucurbit Crops in Spain and Transmission of CYSDV by Two Biotypes of Bemisia tabaci

The relative occurrence in Spain of two whitefly transmitted closteroviruses causing similar yellowing diseases in melon and cucumber greenhouse crops was studied. Based on a RT-PCR assay, a 1994–1997 survey of Spanish greenhouses showed that the recently described Bemisia tabaci-transmitted cucurbit yellow stunting disorder virus (CYSDV) has displaced the Trialeurodes vaporariorum-transmitted beet pseudo-yellows virus (BPYV), a virus that was present in the area since the late 1970s. The CYSDV transmission rates by each of the two biotypes of B. tabaci present in Spain were compared. The results showed that the ubiquitous B biotype and the resident Q biotype (found in Spain and Portugal) were able to transmit CYSDV with similar efficiency.     

Modulation of carbohydrate metabolism and chloroplast structure in sugarcane leaves which were infected by Sugarcane Yellow Leaf Virus (SCYLV)

Non-symptomatic sugarcane plants infected with Sugarcane Yellow Leaf Virus showed starch in mesophyll and bundle sheath cells. In situ-hybridization of mRNAs of sucrose-phosphate phosphatase and ADP-glucose pyrophosphorylase revealed that infected leaves contained SPPase and AGPase in mesophyll cells, Kranz cells and bundle sheath cells. In contrast virus-free leaves contained SPPase only in Kranz cells and AGPase only in bundle sheath cells. Infected leaves exhibited ultrastructural changes in Kranz cell chloroplasts and a shift of the chlorophyll a/b ratio. No obstruction of plasmodesmata was observed. The results indicate that SCYLV-infected plants, even when visually non-symptomatic, underwent strong metabolic and ultrastructural changes.     

Variation in Pathogenicity and Multiplication of Beet Necrotic Yellow Vein Virus (BNYVV) in Relation to the Resistance of Sugar-beet Cultivars

Some partially resistant cultivars varied in their response to beet necrotic yellow vein virus (BNYW), which could be due to the occurrence of different pathotypes. In the past three different types of BNYVV could be identified. Since in the field no consistent cultivar×virus source interaction could be detected, greenhouse trials were carried out under more standardised conditions, starting with a similar initial density of BNYVV. Cultivars with different degrees of resistance varied in their response to various types of beet necrotic yellow vein virus (BNYVV). The B type appeared to be less damaging than the A and P types. The virus content in the tap roots and the ratio of the virus content in tap roots to that in lateral roots were both higher in P type than in A or B type infections indicating that the P type moves more rapidly in the plants than the two other BNYVV types. The percentage of plants in which the virus reaches only a low concentration (less than 56ng/ml of sap) is much lower in P type than in A or B type infections. Frequency distribution diagrams of individual plants showing different BNYVV levels reveal considerable differences between various cultivars.     

Evidence that a Leaf-Disk Test Allows Assessment of Isolate-Specific Resistance in Brassica oleracea Crops Against Downy Mildew (Peronospora parasitica)

A rapid resistance/susceptibility test for Peronospora parasitica (downy mildew) was established by inoculating leaf-disks of four Brassica oleracea accessions. Several conditions were tested: disk disinfection or not, agar medium with or without nutrients and with 50 or 100 ppm of benzimidazole. Using disinfected disks placed on agar (no nutrient and benzimidazole at 50 or 100 ppm), the responses of leaf-disks to four isolates were similar to those obtained using the classical cotyledon test, whereas undesired contaminations occurred in all other conditions. The possible effect of the particular leaf used for obtaining the disks was also studied. In each incompatible interaction tested, disks were resistant whatever the leaf used. In compatible interactions, susceptible phenotypes were observed on disks derived from the six lowest leaves, but disks from upper leaves were resistant. The genetic basis of resistance in a F1 hybrid broccoli was assessed, by testing six isolates on an F2 population derived from this hybrid. The cotyledon test only allows inoculation of two isolates per seedling, whereas many isolates can be tested on each plant by using leaf-disks. The segregation of the resistance to each of the six isolates was analysed: two dominant genes (tightly linked) control resistance to all isolates (one to five isolates; the other to only one isolate).     

The Effect of Different Levels of Beet Cyst Nematodes (Heterodera schachtii) and Beet Necrotic Yellow Vein Virus (BNYVV) on Single and Double Resistant Sugar Beet Cultivars

Beet cyst nematode-resistant cultivars, which were introduced recently, originated from the homozygous inbred line B883. This translocation product was unstable and the transmission of resistance when crossed with a susceptible cultivar did not exceed 94%. Tests with the resistant cultivars in climate cabinets showed a wide variety of resistance against Heterodera schachtii and beet necrotic yellow vein virus (BNYVV), expressed as average numbers of infective units per plant and percentages of resistant plants. In a series of field trials at different levels of infection of H. schachtii, their multiplication rates on all resistant cultivars depended on the initial density, which was caused by the presence of small numbers of susceptible plants. Since tolerance to wilting was also incorporated in B883, reasonable yields were obtained in the presence of H. schachtii. However, at increasing initial densities of H. schachtii, yields decreased considerably, since penetrating juveniles cause a hypersensitivity reaction in resistant plants. Based upon the results of three series of field trials, it was concluded that resistant cultivars should preferably be applied at population densities between 500 and 2000 eggs and juveniles of H. schachtii per 100ml of soil. Cultivars with double resistance against H. schachtii and BNYVV behaved like those with H. schachtii resistance in soils infected with beet cyst nematodes, but not with BNYVV. In soils with a combined infection of H. schachtii and BNYVV double resistant cultivars were far superior to single resistant ones, since damage caused by BNYVV was far more serious than damage caused by H. schachtii. No substantial interaction between soil pathogens nor types of resistance could be detected.     

Evidence of cork barrier formation as a resistance mechanism to berry disease (Colletotrichum coffeanum) in arabica coffee

The histology of resistance to coffee berry disease (CBD) was studied in artificially inoculated berries and hypocotyls of 6-week old seedlings of a number of varieties ofCoffea arabica L. In resistant varieties a phellogen was quickly formed some cell layers below the site of infection and progress of the fungal invasion was effectively blocked by a complete barrier of suberized cells. Such barriers were absent or incompletely developed in CBD susceptible varieties. A highly significant correlation (r=0.87) was found between the frequencies of complete barrier formation in berries and in hypocotyls of young seedlings, while both were also highly correlated to observed mature plant resistance (r>-0.93). Resistance to CBD in arabica coffee may to an important extent be based on the formation of cork barriers. These cork barriers confine the pathogen to the small volume of tissue external to the barrier so that its growth is severely restricted. Such a resistance mechanism is likely to be stable (race-nonspecific). The almost identical response to infection observed in berries and hypocotyls provides further evidence that plants with resistance to CBD can be reliably preselected by the hypocotyl inoculation test.     

一株莱氏绿僵菌的筛选及其对草地贪夜蛾的毒力

开展草地贪夜蛾病原真菌的筛选和毒力测定可为草地贪夜蛾的绿色防控提供新的生物制剂.本研究从广西南宁田间采集的草地贪夜蛾幼虫僵虫上分离获得一株病原真菌,采用形态学和ITS-rDNA序列分析相结合的方法对该菌株进行了鉴定,并采用浸虫法测定了该菌株对草地贪夜蛾的致病力.结果表明新分离的菌株为莱氏绿僵菌Metarhizium r...     

Molecular and Biological Characterization of Lettuce mosaic virus (LMV) Isolates Reveals a Distinct and Widespread Type of Resistance-Breaking Isolate: LMV-Most

ABSTRACT Lettuce mosaic virus (LMV) causes an economically important seedborne and aphid-transmitted disease of lettuce and ornamental crops worldwide. The genetic diversity among 73 LMV isolates was examined based on a 216-nucleotide sequence at the variable region encoding the NIb-coat protein junction. Three clusters of LMV isolates were distinguished: LMV-Yar, LMV-Greek, and LMV-RoW. In the latter cluster, two subgroups of isolates, LMV-Common and LMV-Most, accounted for a large proportion of the LMV isolates analyzed. These two subgroups included the seedborne isolates, consistent with this property contributing a selective advantage and resulting in widespread distribution. In addition to being seedborne, LMV-Most isolates overcome the two resistance genes commonly used in lettuce, mo1(1) and mo1(2), and thus represent a potential threat to lettuce cultivation. The complete sequence of an LMV-Most isolate (LMV-AF199) was determined, allowing a better definition of the genetic relationships among LMV-Most, LMV-Common, and an additional isolate of the LMV-RoW cluster.     

球孢白僵菌对斜纹夜蛾高毒菌株筛选与制剂的研发

斜纹夜蛾Spodoptera litura是一种危害极大的农业害虫,为了探索1条可持续、无污染的新型防治方法,我们通过标准喷塔法从12株球孢白僵菌Beauveria bassiana中筛选出1株对斜纹夜蛾2龄幼虫高毒力的菌株--球孢白僵菌Bb2860菌株.在喷雾浓度为(2592±275)孢子·mm-2的高浓度下,菌株Bb2860对斜纹夜蛾2龄幼虫在第7d的校正死亡率达82.43%±6.03%.不同制剂的室内生测试验表明,孢子水悬液和2种油剂(分别为不含稳定剂和含有稳定剂)在中浓度319～529孢子·mm-2的喷雾浓度下,7d后对斜纹夜蛾2龄幼虫的累计死亡率分别为54.8%、70.0%和76.7%;菌药协同作用为86.8%.另外,于2009年9月在安徽马鞍山进行自然条件下对花椰菜叶上斜纹夜蛾幼虫的田间小区药效试验.处理9d后,纯菌剂的相对防效为77.64%;低剂量和常规用量5%氟啶脲(抑太保)的相对防效分别为49.02%和89.70%;菌药混用的相对防效为91.13%.本研究显示出利用球孢白僵菌制剂及其与低剂量抑太保混用防治斜纹夜蛾的良好应用前景.     

Initial report of glufosinate and paraquat multiple resistance that evolved in a biotype of goosegrass (Eleusine indica) in Malaysia

Field and glasshouse studies have confirmed the presence of a glufosinate‐ and paraquat‐resistant goosegrass biotype that has infested a bitter gourd field in Air Kuning, Perak, Malaysia. Glufosinate and paraquat had been applied at least six times per year to the affected fields (originally a rubber plantation) for more than four consecutive years. Paraquat had been used since 1970 for weed control in the rubber plantation. An on‐site field trial revealed that the control of the goosegrass plants, measuring 20–35 cm in height, ranged from 20 to 35% 3 weeks after being treated with each herbicide at twice the recommended rate. Dose–response tests were conducted in the glasshouse, using seedlings at the three‐to‐four‐leaf stage that had been obtained from the plants that had received repeated exposure to these herbicides and a biotype with no history of any herbicide resistance. The comparison of the GR₅₀ (the herbicide rate that is required to reduce the shoot fresh weight by 50%) of the seedlings indicated that the resistant biotype of goosegrass is 3.4‐fold and 3.6‐fold more resistant than the susceptible biotype following treatment with glufosinate and paraquat, respectively. This study has demonstrated the world's first field‐evolved instance of multiple resistance in goosegrass to two non‐selective herbicides, glufosinate and paraquat.     

Characterization of a potyvirus from eggplant (Solanum melongena) as a strain of potato virus Y by N-terminal serology and sequence relationships

A potyvirus (eggplant mottle virus, EMoV) causing mosaic mottling in eggplant ( Solanum melongena ) was characterized on the basis of biological, serological and partial nucleotide sequence properties. EMoV infected Chenopodium amaranticolor and members of the Solanaceae. Polyclonal antiserum against EMoV showed antigenic relationship with henbane mosaic potyvirus (HMV) and potato Y potyvirus (PVY). Virus-specific antibodies directed to the N-terminal region of EMoV cross-reacted only with PVY. Determination and comparison of nucleotide sequence of the coat protein (CP) and the 3'-untranslated region (UTR) of EMoV with other potyviruses showed that the level of homology was highest with PVY isolates. Comparative sequence analyses of the CP amino acid and 3'-UTR sequences with distinct PVY isolates placed EMoV within the PVY^O subgroup.     

Risks and consequences of gene flow from herbicide-resistant crops: canola (Brassica napus L) as a case study

Data from the literature and recent experiments with herbicide-resistant (HR) canola (Brassica napus L) repeatedly confirm that genes and transgenes will flow and hybrids will form if certain conditions are met. These include sympatry with a compatible relative (weedy, wild or crop), synchrony of flowering, successful fertilization and viable offspring. The chance of these events occurring is real; however, it is generally low and varies with species and circumstances. Plants of the same species (non-transgenic or with a different HR transgene) in neighbouring fields may inherit the new HR gene, potentially generating plants with single and multiple HR. For canola, seed losses at harvest and secondary dormancy ensures the persistence over time of the HR trait(s) in the seed bank, and the potential presence of crop volunteers in subsequent crops. Although canola has many wild/weedy relatives, the risk of gene flow is quite low for most of these species, except with Brassica rapa L. Introgression of genes and transgenes in B rapa populations occurs with apparently little or no fitness costs. Consequences of HR canola gene flow for the agro-ecosystem include contamination of seed lots, potentially more complex and costly control strategy, and limitations in cropping system design. Consequences for non-agricultural habitats may be minor but appear largely undocumented.     

Mapping the spread of apricot chlorotic leaf roll (ACLR) in southern France and implication of Cacopsylla pruni as a vector of European stone fruit yellows (ESFY) phytoplasmas

An epidemiological study on European stone fruit yellows (ESFY) phytoplasmas infecting Prunus fruit trees was carried out from 1994 to 2000 in Languedoc-Roussillon (southern France). The spread of the disease was monitored for 7 years by visual observation of symptoms and by PCR detection of the phytoplasma in an experimental orchard planted with apricot hybrid seedlings. This indicated that aerial vectors were responsible for disease spread, and that transmission rates were low at the beginning of the spread. Seventy thousand homopteran insects were captured within and in the surroundings of highly ESFY-infected apricot orchards, of which about 10 000 were used in PCR and nested-PCR assays with universal ribosomal and ESFY-specific nonribosomal primers to detect ESFY phytoplasmas. The other insects were confined in cages for trials of transmission to test plants. ESFY phytoplasmas could not be detected by PCR in any of the leafhopper species captured but could be detected in the psyllid Cacopsylla pruni caught on Prunus domestica and Prunus cerasifera rootstock suckers of apricot trees and on Prunus spinosa . Nested PCR revealed ESFY phytoplasmas in one individual of the deltocephalid Synophropsis lauri captured on an apricot tree. Transmission trials confirmed the role of Cacopsylla pruni as the ESFY phytoplasma vector in France. When apricot seedlings were used as bait plants from April to November during two consecutive years, no natural transmission could be demonstrated. However, one out of 50 apricot seedlings left for the whole year in the orchard became infected. An early spring ESFY infection is in agreement with both the natural transmission results and the life cycle of Cacopsylla pruni .