Immunolocalization of Pepper mild mottle virus in Capsicum annuum seeds

To clarify the mechanism of seed transmission of Pepper mild mottle virus (PMMoV), the virus was immunolocalized in Capsicum annuum seeds using fluorescence microscopy. Two distinct patterns were observed: In the first, PMMoV was present in the epidermis and parenchyma but not in the endosperm or embryo; in the second, the virus was restricted to the surface of the epidermis and parenchyma. These findings shed light on the fundamental mechanisms of seed transmission of tobamoviruses and may aid in the design of new methods to prevent the spread of seedborne virus diseases.     

Differential Interactions of aColletotrichum gloeosporioides isolate with green and red pepper fruits

Differential interactions ofColletotrichum gloeosporioides isolate KG 13 with green and red pepper fruits (Capsicum annuum were found when it was inoculated on unwounded and wounded fruits. The isolate produced the typically necrotic, sunken anthracnose symptom on unwounded and wounded green fruits, and wounded red ones, but not on unwounded red ones. Appressorial formation of the fungus on the surfaces of compatible green fruits was higher than on incompatible red ones up to 12 h after inoculation. More and longer infection pegs from appressoria were produced on green than on red fruits. When cuticular wax layers of green and red fruits were removed by dipping in chloroform, red ones only produced larger lesions and more conidia than water-dipped controls did. However, differences in lesion diameter and conidial production were not observed between green and red fruits wounded by pin-pricking. In addition, concentrations of wax extracted from the surface of green and red fruits affected conidial germination and appressorial formation of the fungus. These findings suggest that the isolate KG 13 ofC. gloeosporioides may react differentially to green and red pepper fruits, probably due to the physical and chemical differences in cuticular layers of the fruits.     

Molecular and Pathogenicity Characteristics of Phytophthora nicotianae Responsible for Root Necrosis and Wilting of Pepper (Capsicum annuum L.) in Tunisia

Isolates of Phytophthora from pepper, produced in Tunisia, were characterised according to molecular and pathogenicity criteria. Polymerase chain reaction amplification of the ITS1 region in the ribosomal DNA resulted in different sized fragments. The pepper isolates and P. nicotianae yielded a fragment of 310bp that distinguished it from P. capsici with a fragment of 270bp. The ribosomal RNA gene amplicons of both internal transcribed spacers and the 5.8 S of the pepper Phytophthora and P. nicotianae were digested with 8 endonucleases. The patterns generated, with the 2 enzymes that cut, were identical for both taxa. This molecular analysis corroborated the morphological and biological characteristics and suggests strongly that the isolates of Phytophthora from pepper belong to the species P. nicotianae. Inoculation of pepper, tomato, eggplant and tobacco plants with the isolates of P. nicotianae from pepper showed they were highly pathogenic on pepper but not on tobacco, while their pathogenicity was weak on tomato and eggplant and was associated with atypical symptoms not observed in the field. These pathogenicity tests suggest that pepper isolates of P. nicotianae are particularly adapted to their host and may thus constitute a forma specialis of P. nicotianae.     

Identification and development of molecular markers linked to Phytophthora root rot resistance in pepper (Capsicum annuum L.)

Phytopthora root rot in pepper (C. annuum) is caused by Phytophthora capsici L., which exhibits a high level of pathogenic diversity. Resistance to this disease is conditioned by a number of quantitative trait loci. Pyramiding resistance alleles is desirable and could be simplified by the use of molecular markers tightly linked to the resistance genes. The purpose of this study was development of molecular markers linked to Phytophthora root rot resistance. An F₈ recombinant inbred line (RIL) population derived from a cross between YCM334 and a susceptible cultivar ‘Tean’ was used in combination with bulk segregant analysis utilizing RAPD and conversion of AFLP markers linked to Phytophtora root rot resistance into sequence-characterized amplified region (SCAR) markers. In conversion: one marker was successfully converted into a co-dominant SCAR marker SA133_4 linked to the trait. In bulked segregant analysis (BSA): three RAPD primers (UBC484, 504, and 553) produced polymorphisms between DNA pools among 400 primers screened. Genetic linkage analysis showed that the SCAR and RAPD markers were located on chromosome 5 of pepper. Quantitative trait locus (QTL) analysis showed that the SA133_4 and UBC553 were linked to Phytophtora root rot resistance. These markers were correctly identified as resistant or susceptible in nine promising commercial pepper varieties. These markers will be beneficial for marker-assisted selection in pepper breeding.     

Selection forMeloidogyne incognita virulence against resistance genes from tomato and pepper and specificity of the virulence/resistance determinants

Experiments were designed to analyze the relationships between the root-knot nematodeMeloidogyne incognita and resistant tomato and pepper genotypes. From a natural avirulent isolate, near-isogenic nematode lineages were selected with virulence either against the tomatoMi resistance gene or the pepperMe3 resistance gene. Despite the drastic selection pressure used, nematodes appeared unable to overcome the pepperMe1 gene, therefore suggesting some differences in the resistance conferred byMe1 andMe3 in this species. Nematodes virulent onMi-resistant tomatoes were not able to reproduce onMe1-resistant nor onMe3-resistant peppers, and nematodes virulent onMe3-resistant peppers were not able to reproduce onMi-resistant tomatoes nor onMe1-resistant peppers. These results clearly demonstrate the specificity ofM. incognita virulence against resistance genes from both tomato and pepper, and indirectly suggest that gene-for-gene relationships could occur between these two solanaceous crops and the nematode.     

Critical period for weed control in direct seeded red pepper (Capsicum annum L.)

Direct seeded red pepper is a cash crop in Kahramanmara? province of Turkey as well as some other nearby provinces. Weeds are a major constraint in red pepper production. Field studies were conducted to determine critical period for weed control (CPWC) in direct seeded spice pepper in Kahramanmara?, in 2008 and 2009. The CPWC in red pepper based on a 2.5%, 5% and 10% acceptable yield loss (AYL) was calculated by fitting logistic and Gompertz equations to relative yield data. The CPWC in red pepper was determined from 0 to 1087 growing degree days (GDD) in 2008 and from 109 to 796 GDD in 2009 for 10% AYL after crop emergence in red pepper. For 2.5–5% AYL, the CPWC starts with germination and lasts until harvest. Direct seeded red pepper is very vulnerable to weed competition and weed control programs for direct seeded spice pepper in Turkey should include pre‐emergence and residual herbicides.     

Possibilities of biological and chemical control of Verticillium wilt in pepper

In pathogen populations in Serbia, the incidence, pathogenic and morphological characters ofVerticillium spp. were studied. Biological and chemical control ofVerticillium was investigated in pepper plants (Capsicum annuum L. cv. ‘Soroksari’) with the biofungicide Polyversum^® (Pythium oligandrum) and the conventional fungicides benomyl and propamocarb-hydrochloride. On the basis of macroscopic and microscopic characters of the isolates originating from eight localities in Serbia, it was established that they apparently belong to the speciesVerticillum dahliae. The isolates differed in their pathogenic characters. However, all of them caused marked wilting symptoms on pepper plants 40 days after inoculation, conducted when there were more than nine fully developed leaves on the primary stem. The fungicides were applied either before or after inoculation. Benomyl was the most efficient fungicide in wilt control (88.2% when applied after inoculation and 94.6% when applied before inoculation). Polyversum proved more efficient (66.6%) when applied before rather than after inoculation. Propamocarb-hydrochloride provided sufficient Verticillium wilt control; its efficacy and that of Polyversum were similar, and less efficient than benomyl, but still significantly different from the disease control.     

Nematicidal fluorescent pseudomonads for the in vitro and in vivo suppression of root knot (Meloidogyne incognita) of Capsicum annuum L

BACKGROUND: Considerable attention has been paid to plant‐growth‐promoting rhizobacteria (PGPR), especially the fluorescent group of Pseudomonas species, as the best alternatives to chemicals for facilitating ecofriendly biological control of soil‐ and seedborne microorganisms. On the basis of their novel plant‐growth‐promoting attributes, two rhizobacteria Pseudomonas aeruginosa VP1 and VP2 selected out of over 63 isolates from the rhizosphere of chilli (Capsicum annuum) were identified as potential candidates for biocontrol of the root‐knot nematode Meloidogyne incognita on chilli. RESULTS: The nematicidal activity of both strains was evaluated in vitro and in vivo for their efficacy against M. incognita. P. aeruginosa VP2 exhibited strong nematicidal activity in comparison with VP1, based on the in vitro killing of the second‐stage juveniles (J2) of M. incognita. Seed bacterisation with both strains VP1 and VP2 was able to manage root‐knot M. incognita on chilli (C. annuum) in a pot trial study. Increase in root and shoot length and in fresh and dry weight of root and shoot and reduction in the root‐knot index over the control were attained. In overall performance, VP2 was 29.5% more effective than VP1, and about 30% more effective than the control (non‐bacterised). CONCLUSION: The application of P. aeruginosa VP1 and P. aeruginosa VP2 controls the development of M. incognita in C. annuum, and hence they are recommended as efficient plant growth promotors and biocontrolling agents for raising healthy crop of C. annuum that can promote the growth of plants and reduce the nematode (M. incognita) population. Copyright © 2012 Society of Chemical Industry     

Bicarbonate solutions control powdery mildew (Leveillula taurica) on sweet red pepper and reduce the development of postharvest fruit rotting

Disease severity of powdery mildew (causal agentLeveillula taurica (Lév.) Arn.) on pepper (Capsicum annuum), defoliation and fruit sunscald rating were significantly reduced (P=0.05) by foliar applications of sodium or potassium bicarbonate solutions, compared with water or penconazole (Ophir). Preharvest application of bicarbonate solutions significantly reduced (P=0.05) postharvest decay development on sweet red pepper fruits.     

Reduction in aggressiveness among hybrids between host-specific pathotypes of Magnaporthe oryzae is caused by reduced ability to overcome adult resistance at the level of penetration

A genetic cross between a Triticum isolate (pathogenic on wheat) and a Setaria isolate (pathogenic on foxtail millet) of Magnaporthe oryzae yielded several F₁ cultures that were virulent on both wheat and foxtail millet at the primary leaf stage. To estimate whether these cultures survive in nature, they were sprayed onto 1-, 2-, and 3-week-old wheat and foxtail millet. As the age of the inoculated plants increased, the lesion number and size were greatly reduced. The F₁ cultures were almost nonpathogenic on both wheat and foxtail millet at the 3-week-old stage. Cytological analysis revealed that the low pathogenicity of the F₁ cultures on older plants was primarily associated with a reduced ability to penetrate the cuticle. When placed on wounded leaf surfaces, the F₁ cultures produced large lesions on 4-week-old wheat and foxtail millet. These results indicate that hybridization between species-specific pathotypes results in a reduction of aggressiveness. We therefore suggest that, even if such hybrids were produced in nature, they might not survive in the natural environment.     

Simultaneous detection of <Emphasis Type="Italic">Japanese yam mosaic virus</Emphasis> and <Emphasis Type="Italic">Yam mild mosaic virus</Emphasis> from yam leaves using a tube capture reverse transcription-polymerase chain reaction assay

To detect Japanese yam mosaic virus (JYMV) and Yam mild mosaic virus (YMMV) in yam plants in Japan, we developed a duplex RT-PCR assay consisting of a tube-capture procedure followed by one-step RT-PCR with two primer pairs. A 241-bp fragment of the coat protein region of JYMV and a 174-bp fragment of the nuclear inclusion protein b region of YMMV were amplified, thus identifying the two viruses from yam plants cultivated in Yamaguchi Prefecture in 2007. All water yam plants examined were infected with YMMV alone. All the Japanese yam and Chinese yam plants were infected with either JYMV alone or both JYMV and YMMV, suggesting that YMMV and JYMV are prevalent among field-grown yam plants.     

Timing and extent of hypersensitive response are critical to restrict local and systemic spread of Pepper mild mottle virus in pepper containing the L 3 gene

Amino acid changes in Pepper mild mottle virus (PMMoV) coat protein (CP) that enhance, decrease, or nullify the resistance-inducing activity in Capsicum plants carrying the L ³ gene have been identified. In this study, molecular events underlying the L ³ -gene-mediated resistance were analyzed through the expression of hypersensitive response (HR)-related genes, HSR203J-Cc and HIN1-Cc, and defense-related genes, PR1-Cc and PR4b-Cc, upon infection with PMMoV CP mutants. The expression kinetics of the genes correlated with the degree of restriction of virus distribution in the inoculated leaves. The results suggest that the timing and extent of HR are critical factors to restrict virus spread both locally and systemically in L ³ -gene-mediated resistance.The nucleotide sequence data reported are available in the DDBJ/GenBank/EMBL databases under accession numbers AB162220 (HSR203J-Cc), AB162221 (HIN1-Cc), AB162222 (PR1-Cc), and AB162223 (PR4b-Cc)     

Tree Mortality by the Bark Beetle Ips typographus (L.) in storm-disturbed stands

To maintain biodiversity in forests more wind-felled trees must be left. However, there is concern among forest owners that this may result in higher tree mortality caused by the spruce bark beetle (SBB). Data from six studies on number of trees killed by the SBB in storm-disturbed stands was analysed. The studies include 16 areas, 2–346 ha in size, where wind-felled spruce trees were left. In one of the studies also stands where all wind-felled trees were removed directly after the storm were included. Few trees were killed in the first summer following the storm disturbance. In most stands the tree mortality peaked already in the second or third summer following the storm. But in five of the stands tree mortality peaked later. The number of trees killed per ha by the SBB varies much between the studied stands. There was a trend that the larger areas experienced on average higher tree mortality per ha than the smaller areas. One of the studies, including six stands, demonstrated an almost perfect correlation between the number of colonised wind-felled trees and the number of killed trees in a 4-year period following the storm disturbance. In the same study the removal of wind-felled trees resulted in a 50% reduction in number of trees killed per ha in the 4-years following the storm.     

Application of pathogen surveys,disease nurseries and varietal resistance characteristics in an IPM approach for the control of wheat yellow rust

The present paper presents the rationale for the use of pathogen surveys, inoculated and non-inoculated disease nurseries and varietal resistance characteristics in an integrated approach to control wheat yellow rust in Denmark. The non-inoculated disease observation plots, which gave valuable information about yellow rust at the year, site and variety level, served as the primary sample source for the pathogen survey revealing pathogen virulence dynamics. This survey was also the main source for isolates of new pathotypes, a prerequisite for the assessment of the resistance characteristics of varieties and breeding lines in inoculated nurseries, and the postulation of race-specific resistance genes. A simple grouping of varieties into four categories with respect to resistance to the current yellow rust population proved robust, and this grouping was used as a determinant in a web-based decision support system for pesticide applications in cereals, Crop Protection On-line (CPO). The interplay between the different research and survey activities in the integrated pest management (IPM) approach demonstrated the need for a coherent and long-term involvement at all stages from plant breeding to the official variety approval system, extension service and research in disease epidemiology and resistance genetics.     

Validation of a QTL for resistance to ascochyta blight linked to resistance to fusarium wilt race 5 in chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.)

Ascochyta blight caused by Ascochyta rabiei and fusarium wilt caused by Fusarium oxysporum. f. sp. ciceris are the two most serious diseases of chickpea (Cicer arietinum). Quantitative trait loci (QTL) or genes for ascochyta blight resistance and a cluster of resistance genes for several fusarium wilt races (foc1, foc3, foc4 and foc5) located on LG2 of the chickpea map have been reported independently. In order to validate these results and study the linkage relationship between the loci that confer resistance to blight and wilt, an intraspecific chickpea recombinant inbred lines (RIL) population that segregates for resistance to both diseases was studied. A new LG2 was established using sequence tagged microsatellite sites (STMS) markers selected from other chickpea maps. Resistance to race 5 of F. oxysporum (foc5) was inherited as a single gene and mapped to LG2, flanked by the STMS markers TA110 (6.5 cM apart) and TA59 (8.9 cM apart). A QTL for resistance to ascochyta blight (QTL_AR3) was also detected on LG2 using evaluation data obtained separately in two cropping seasons. This genomic region, where QTL_AR3 is located, was highly saturated with STMS markers. STMS TA194 appeared tightly linked to QTL_AR3 and was flanked by the STMS markers TR58 and TS82 (6.5 cM apart). The genetic distance between foc5 and QTL_AR3 peak was around 24 cM including six markers within this interval. The markers linked to both loci could facilitate the pyramiding of resistance genes for both diseases through MAS.     

Corynespora blight of sweet pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis>) caused by <Emphasis Type="Italic">Corynespora cassiicola</Emphasis> (Berk. &; Curt.) Wei

In 2004, Corynesopra cassiicola was isolated from dark brown spots on leaves and fruits and from black blights on stems of sweet pepper plants in Kochi Prefecture, Japan. The isolated fungus was then used to inoculate sweet pepper plants and subsequently reisolated from the plants with dark brown spots and black blights, showing that C. cassiicola is a new pathogen causing Corynespora blight on sweet pepper plants. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases as accession numbers AB366649 (TS-C11), AB366650 (TS-C21), AB366651 (TI-C32) and AB366652 (TI-C51)     

芹菜APX基因家族鉴定及其表达分析

抗坏血酸过氧化物酶（Ascorbate peroxidase, APX）是植物活性氧代谢中重要的抗氧化酶之一,在植物抵抗氧化胁迫方面发挥重要作用。利用生物信息学方法对芹菜基因组中的APX基因家族成员进行鉴定和分析,并通过实时荧光定量PCR（quantitative real\|time PCR, qRT-PCR）验证分析AgAPXs在高温胁迫下的表达情况,为开展芹菜APX基因参与高温胁迫调控机制提供依据。结果表明：芹菜基因组中共有9个APX基因,随机分布在5个染色体上,并出现了基因片段复制现象;大多数基因被定位在细胞质中。系统发育分析表明,AgAPX基因家族可分为3个亚族,同一亚族中的成员具有相似的基因结构和基序。启动子顺式元件分析表明,大多数AgAPX基因含有多种与生长发育、植物激素和逆境胁迫相关的顺式元件。高温胁迫下,芹菜APX活性提高。qRT-PCR分析表明,AgAPXs在不同时间的高温处理下表达具有显著差异,并与转录组表达丰度相一致,AgAPX2、AgAPX3、AgAPX4、AgAPX5、AgAPX7的表达量和APX活性具有显著相关性,推测AgAPXs可能参与了芹菜抵御高温的调控过程。本研究初步鉴定并提供了芹菜APX基因家族成员信息,为今后进一步探索芹菜APX基因功能提供了重要的研究基础。     

Molecular basis for resistance to tribenuron in shepherd’s purse (Capsella bursa-pastoris (L.) Medik.)

Capsella bursa-pastoris, a winter annual weed in the mustard family, can not be controlled by tribenuron after the herbicide has been continuously used for several years. The resistant biotype Lz-R was the generation of a population collected from Liangzhu, a place where tribenuron had been used for more than 15 consecutive years. To confirm and characterize the resistance of C. bursa-pastoris to tribenuron, whole-plant bioassays were conducted in the greenhouse. The results of whole-plant bioassays revealed that Lz-R was highly resistant to tribenuron with the resistance index (GR₅₀ Lz-R)/(GR₅₀ Lz-S) up to 236.6. To investigate the molecular basis of resistance in C. bursa-pastoris, the acetolactate synthase (ALS) genes were sequenced and compared between susceptible and resistant biotypes. Analysis of the nucleotide and deduced amino acid sequences between the biotypes indicated that one substitution had occurred in Domain A, cytosine by thymine (CCT to TCT) at position 197, that led to a change of the amino acid proline in the susceptible to serine in the Lz-R.