Can a chitin‐synthesis‐inhibiting turfgrass fungicide enhance black cutworm susceptibility to a baculovirus?

BACKGROUND: Developmental resistance, i.e. reduced virulence and speed of kill of late instars, is a limiting factor in the use of baculoviruses for caterpillar control. Agrotis ipsilon multicapsid nucleopolyhedrovirus (AgipMNPV) is highly infective to young black cutworms, Agrotis ipsilon, but too slow‐acting against late instars for effective curative control on golf courses or sports fields. Chitin‐synthesis‐inhibiting fungicides containing the active ingredient polyoxin‐d are used to control fungal diseases in turfgrass, and similar compounds have been shown in the laboratory to synergize baculoviruses by disrupting peritrophic membrane function. This study tested whether applying the virus together with such a fungicide can synergize AgipMNPV activity against A. ipsilon in turfgrass. RESULTS: The addition of a chitin synthesis inhibitor failed to increase AgipMNPV infectivity to A. ipsilon in the field. Rather, delayed and slightly reduced mortality from viral infection was seen when larvae fed on fungicide/virus‐treated grasses as opposed to virus‐only treatments. Choice tests revealed the fungicide residues to be a mild feeding deterrent. CONCLUSION: Because polyoxin‐d does not deactivate AgipMNPV, the two substances are compatible. However, combination applications of polyoxin‐d and Agip MNPV on turfgrass might interfere with larval ingestion of a lethal virus dose, resulting in prolonged larval feeding in the field. Copyright © 2011 Society of Chemical Industry     

近50 a中国≥10 ℃有效积温时空演变

根据全国583个气象站点近50 a（1961-2010年）日均气温资料,以ArcGIS为技术平台,采用反距离插值和Mann Kendall趋势性检验等方法,研究了我国≥10 ℃有效积温等值线中心和典型有效积温带面积的变化规律,以及≥10 ℃有效积温、起始时间和持续时间的时空分布特征。结果表明：我国≥10 ℃有效积温的时空分布在1985年前后差异性显著。≥10 ℃有效积温整体上呈上升趋势,其变化率在0~10 ℃•d•a^-1,≥10 ℃典型有效积温带整体北移显著。≥10 ℃有效积温为0~3 400 ℃•d的区域面积显著减小,而 ≥10 ℃有效积温为3 400~8 000 ℃•d的区域面积明显增加,增速达2.0×10⁴ km²•a^-1。1985年后我国≥10 ℃有效积温起始时间提前和持续时间增加的区域面积显著增加;有效积温的起始时间整体有所提前,提前幅度集中在0~4 d•(10a)^-1;有效积温持续时间均有所增加,增加梯度集中在0~6 d•(10a)^-1。     

Identification of a novel point mutation in the <Emphasis Type="Italic">β</Emphasis>-tubulin gene of <Emphasis Type="Italic">Botrytis cinerea</Emphasis> and detection of benzimidazole resistance by a diagnostic PCR-RFLP assay

The molecular basis of resistance to benzimidazole fungicides with laboratory and field mutant isolates of Botrytis cinerea was investigated. After chemical mutagenesis with N-methyl-N-nitrosogouanidine (NMNG) two different benzimidazole-resistant phenotypes were isolated on media containing carbendazim or a mixture of carbendazim and diethofencarb. The mutant isolates from the fungicide-mixture-containing medium were moderately resistant to carbendazim with wild-type tolerance to diethofencarb while mutant isolates from carbendazim-containing medium were highly resistant to carbendazim but sensitive to diethofencarb. The studied field isolates were highly resistant to benzimidazoles and sensitive to diethofencarb. Study of fitness characteristics of benzimidazole highly-resistant isolates showed that the resistance mutation(s) had no apparent effect on fitness-determining parameters. Contrary to this, the moderately benzimidazole-resistant strains, with no increased diethofencarb sensitivity, had a significant reduction in certain ecological fitness-determining characteristics. Analysis of the sequence of the β-tubulin gene revealed two amino acid replacements in the highly benzimidazole-resistant mutants compared to that of the wild-type parent strain. One was the glutamic acid (GAG) to alanine (GCG) change at position 198 (E198A), identified in both laboratory and field highly benzimidazole-resistant isolates, a mutation previously implicated in benzimidazole resistance. The second was a novel benzimidazole resistance mutation of glutamic acid (GAG) to glycine (GGG) substitution at the same position 198 (E198G), identified in a highly benzimidazole-resistant laboratory mutant strain. Molecular analysis of the moderately benzimidazole-resistant strains revealed no mutations at the β-tubulin gene. A novel diagnostic PCR-RFLP assay utilising a BsaI restriction site present in the benzimidazole-sensitive (E198) but absent in both resistant genotypes (E198G and E198A) was developed for the detection of both amino acid replacements at the β-tubulin gene.     

<Emphasis Type="Italic">Amaranthus</Emphasis> spp.: a new host of “<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma aurantifolia”

Mexico is considered to be one of the centers of origin of grain amaranth species. Recently, plants with abnormal anatomical features were observed in experimental fields established in Central Mexico. The most noticeable symptoms, which consisted of excessive stem and bud proliferation, mosaics and unusual coloration, suggested that they might be phytoplasma-induced disorders. Thus, different accessions of grain amaranth (Amaranthus hypochondriacus and A. cruentus) plants were subjected to polymerase chain reaction (PCR) analysis specifically designed to detect these pathogens. Two universal phytoplasma-specific primer pairs were tested in a nested PCR assay, with primer pair P1/tint (followed by primer pair R16F2/R16R2). Further DNA sequencing analysis of the resulting amplicons indicated that these phytoplasmas may be related to others already affecting important agricultural crops in Mexico, such as soybean. Data are presented that disclose the etiology of these syndromes by the use of molecular techniques. To the best of our knowledge, this finding constitutes the first report of a phytoplasma-related disease in grain amaranth.     

East Adriatic—a reservoir region of severe <Emphasis Type="Italic">Citrus tristeza virus</Emphasis> strains

Citrus tristeza virus (CTV) represents one of the major threats to citrus production worldwide. In the East Adriatic region, CTV symptoms are mostly absent due to traditional citrus grafting on trifoliate orange (Poncirus trifoliata), a CTV-tolerant rootstock. Therefore, the virus has been continuously spreading by the propagation of infected material. The genetic variability of CTV was studied on nineteen citrus samples, collected from orchards in the coastal region of Croatia, Montenegro and Albania, that previously tested positive by ELISA and immunocapture RT-PCR. Single-strand conformation polymorphism of the amplified coat protein gene demonstrated the presence of different CTV variants in each amplicon, while sequence analysis of cloned CP gene variants confirmed their clustering into six out of the seven phylogenetic groups so far delineated. Four of these groups include sequences of severe quick decline, seedling yellows and stem-pitting (SP) isolates, thought to be found only rarely in the Mediterranean region. Regardless of the lack of symptoms in the field, CTV isolates from the East Adriatic displayed high genetic variability and pathogenic potential, additionally confirmed by biological characterisation. The high percentage of mixed infections suggest the potential for further diversification and a greater risk of severe variants spreading into new areas.     

<Emphasis Type="Italic">Spiraea salicifolia:</Emphasis> a new plant host of “<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma ziziphi”-related phytoplasma

Spiraea salicifolia is widely grown in China as an ornamental plant, and its roots and young leaves have many medical uses. On the campus of Northwest A&F University, we observed diseased S. salicifolia plants that had yellowed, dwarfed, deformed leaves and other symptoms resembling diseases caused by phytoplasma. This study was aimed at determining the causal agent of the disease. On the basis of phytoplasma-specific DNA amplification by PCR, a phytoplasma infection of S. salicifolia was confirmed. The phytoplasma was related to “Ca. Phytoplasma ziziphi” according to RFLP and phylogenetic analyses. This report is the first of phytoplasma infection of S. salicifolia in China.     

Reduced BYDV–PAV transmission by the grain aphid in a <Emphasis Type="Italic">Triticum monococcum</Emphasis> line

The luteovirus Barley yellow dwarf virus–PAV (BYDV–PAV) and its vector, the aphid Sitobion avenae are two major sources of yield losses in cereal crops. We report in this paper the effects of a Triticum monococcum line (TM44), resistant by antibiosis to S. avenae, on the different steps of transmission of one BYDV–PAV isolate by the aphids. First, it was shown that TM44 is strongly resistant to BYDV–PAV transmission, but exclusively when S. avenae is the vector. Second, that TM44 is resistant (1) to BYDV–PAV acquisition by S. avenae and (2) to its inoculation, whatever the respective duration of these two periods. Third, that both resistances have partially additive effects. In the discussion, several lines of evidence are given to support the hypothesis that resistance of TM44 to PAV transmission is due to the same disturbances to S. avenae feeding behaviour that are involved in its antibiosis against this aphid species. Reasons for caution in releasing resistant material in the field are presented.     

Detection of the fire blight biocontrol agent <Emphasis Type="Italic">Bacillus subtilis</Emphasis> BD170 (Biopro<Superscript>®</Superscript>) in a Swiss apple orchard

Fire blight, caused by Erwinia amylovora, is a major disease threat to apple, pear and other pome fruit worldwide. The disease is widespread in Europe and has recently become established in Switzerland. Antibiotics are the most effective controls used in North America but these are not permitted for agricultural use in most European countries. A newly registered biological control product Biopro^®, based on the antagonist Bacillus subtilis strain BD170, is being used as an alternative strategy for fire blight management. A specific molecular marker was developed for monitoring the spread of this agent on blossoms after Biopro^® spray application in a Swiss apple orchard throughout the bloom period for 2years. Direct spraying resulted in efficient primary colonisation of pistils in flowers that were open at the time of treatment. Subsequent bacterial dissemination (secondary colonisation) of flowers that were closed or at bud stage at the time of treatment was observed but was found to be dependent on the timing of treatments relative to bloom stage in the orchard. Foraging honeybees were shown to be disseminators of Biopro^®. We also report detection of the biocontrol agent in honey collected from hives where bees were exposed by placing Biopro^® at the entrance or in the hatching nest and from hives that were simply placed in sprayed orchards.     

近50 a新疆≥0 ℃持续日数和积温时空变化

利用新疆95个气象站1961-2010年的逐日平均气温资料,使用线性趋势分析、累积距平和[WTBX]t[WTBZ]检验以及基于ArcGIS的混合插值法,对新疆近50 a日平均气温稳定≥0 ℃的初日、终日、持续日数和活动积温的时空变化进行分析。结果表明：≥0 ℃的初日在空间分布上南疆早,北疆晚;平原和盆地早,山区晚的格局。≥0 ℃终日的空间分布与≥0 ℃初日大体相反。≥0 ℃持续日数和≥0 ℃积温的空间分布表现为南疆多,北疆少;盆地多,山区少的格局。在全球变暖背景下,1961-2010年新疆≥0 ℃初日总体呈显著提早趋势,≥0 ℃终日呈极显著推迟趋势,≥0 ℃的持续日数和积温分别以2.58 d·（10a）^-1和66.26 ℃·d·（10a）^-1的倾向率呈极显著的增多趋势,并且分别于1997年或1994年发生了突变。但突变后较突变前各要素的变化量具有明显的区域性差异,总体来说,≥0 ℃初日提前幅度的空间分布呈南疆大,北疆小的格局。≥0 ℃终日的推迟幅度呈北疆大,南疆小;山区大,平原和盆地小的特点,≥0 ℃持续日数的延长幅度表现为山区大,平原和盆地小的格局,≥0 ℃积温的增加幅度呈平原和盆地多,山区少的空间分布格局。     

Diversity of defence mechanisms in plant–oomycete interactions: a case study of <Emphasis Type="Italic">Lactuca</Emphasis> spp. and <Emphasis Type="Italic">Bremia lactucae</Emphasis>

Plant pathogenic oomycetes, including biotrophic downy mildews and hemibiotrophs/necrotrophs such as Phytophthora and Pythium, cause enormous economic losses on cultivated crops. Lettuce breeders and growers face the threat of Bremia lactucae, the causal agent of lettuce downy mildew. This pathogen damages leaf tissues and lettuce heads and is also frequent on wild Asteraceae plants. The interactions of Lactuca spp. with B. lactucae (abbr. lettuce–Bremia) display extreme variability, due to a long co-evolutionary history. For this reason, during the last 30 years, the lettuce–Bremia pathosystem has been used as a model for many studies at the population, individual, organ, tissue, cellular, physiological and molecular levels, as well as on genetic variability and the genetics of host–parasite interactions. The first part of this review summarizes recent data on host–parasite specificity, host variability, resistance mechanisms and genetics of lettuce–Bremia interactions. The second part focuses on the development infection structures. Phenotypic expression of infection, behaviour of B. lactucae on leaf surfaces, the process of penetration, development of primary infection structures, hyphae and haustoria are discussed in relation to different resistance mechanisms. In the third part, the components of host resistance and the variability of defence responses are analysed. The role of reactive oxygen species (ROS), antioxidant enzymes, nitric oxide (NO), phenolic compounds, reorganization of cytoskeleton, electrolyte leakage, membrane damage, cell wall disruption, hypersensitive reaction and plant energetics are discussed in relation to defence responses. In general, the extreme variability of interactions between lettuce and Bremia, and their phenotypic expression, results from diversity of the genetic background. Different mechanisms of resistance are conditioned by an orchestra of defence responses at the tissue, cell, and molecular levels. The various events responsible for defence involve a complex interaction of the processes and reactions mentioned above. This review also provides an overview on the timing of pathogen development, host pathological anatomy, cytology and physiology of lettuce–Bremia associations. The significance of these factors on the expression of different resistance mechanisms (non-host and host resistance, race-specific and race non-specific resistance, field resistance) is discussed.     

β-aminobutyric acid protects <Emphasis Type="Italic">Brassica napus</Emphasis> plants from infection by <Emphasis Type="Italic">Leptosphaeria maculans.</Emphasis> Resistance induction or a direct antifungal effect?

Resistance to infection in plants can be induced by treatment with various chemicals. One such compound is β-aminobutyric acid (BABA). Its positive effect on disease resistance has been noted in several pathosystems. Here we demonstrate that treatment with BABA protects Brassica napus plants from infection by the fungal pathogen Leptosphaeria maculans. Surprisingly, BABA also displayes in vitro antifungal activity against L. maculans with EC₅₀ similar to the fungicide tebuconazole. Both spore germination and hyphal growth were affected. The toxic effect can be reverted by addition of trypton to the culture medium. We hypothesised that BABA might inhibit inorganic nitrogen assimilation. Suppression of disease progression in plants and antifungal activity in vitro was weaker for α-aminobutyric acid and negligible for γ-aminobutyric acid. In contrast to a resistance inducer benzothiadiazole, the effect of BABA on disease development was nearly independent of the timing of treatment, indicating possible antifungal activity in planta. On the other hand, quantification of multiple hormones and an expression analysis have shown that treatment with BABA induces a synthesis of salicylic acid (SA) and expression of SA marker gene PR-1, but no evidence was observed for priming of SA responses to L. maculans. While we have not conclusively demonstrated how BABA suppresses the disease progression, our results do indicate that antifungal activity is another mechanism by which BABA can protect plants from infection.     

Is there a relationship between the intrinsic rate of propagation and <Emphasis Type="Italic">in-vitro</Emphasis> migration and virulence of the pinewood nematode, <Emphasis Type="Italic">Bursaphelenchus xylophilus</Emphasis>?

We investigated if rates of propagation and migration were related with the level of virulence in the pinewood nematode Bursaphelenchus xylophilus using 17 offspring lines from the F₂ crosses between virulent and avirulent isolates. Virulence was tested by inoculating seedlings of Pinus thunbergii with the nematodes. The proportion of dead seedlings ranged from 13.3% to 77.8%, 20 weeks after inoculation. Migration rate of the nematodes was estimated by measuring their migration distance per unit time in an artificial substrate that imitated pathways in pine trees. Migration rate varied from 0.85 to 3.53 mm min⁻¹. Propagation rate was determined based on population growth on the fungus Botrytis cinerea, and it ranged between 10^3.88 and 10^4.99 per 12 days. Statistical analyses revealed that virulence was not correlated with migration rate, but was negatively correlated with propagation rate on Botrytis cinerea, suggesting that the nematodes paid some cost for virulence. Also, there was no relationship between rates of migration and propagation. Cluster analysis showed that the biological parameters varied between crossbred lines, with no kinship bias, suggesting the absence of sex-linked inheritance in virulence and rates of propagation and migration.     

Susceptibility of eight Algerian olive cultivars to <Emphasis Type="Italic">Bactrocera oleae</Emphasis> infestation – a pomological and nutritional quality perspective

The olive fruit fly, Bactrocera oleae (Rossi), is the major olive pest in the Mediterranean Basin, being responsible for high losses in olive production. The objective of this work was to study the influence of B. oleae infestation on the pomological parameters and nutritive value (lipids, proteins, carbohydrates and ash) of eight olive cultivars (Abani, Aellah, Blanquette de Guelma, Chemlal, Ferkani, Limli, Rougette de Mitidja and Souidi) from Algeria. The results displayed different degrees of susceptibility of the studied cultivars to this pest. Fruit size (weight) and the Fruit weight / stone weight ratio are significantly correlated with the attack. The statistical treatment of the results (PCA and HAC) highlighted different behaviors from the cultivars, concerning the attack. There are some cultivars more susceptible, being the nutritional value also affected. The lipid and protein contents were positively correlated with the attack and the carbohydrates negatively, suggesting a careful choice by the B. oleae. Ovipositional preference of olive fruit fly females and the success of larval development in different olive cultivars are crucial to establishing new approaches in Integrated Pest Management (IPM) against this pest.     

Is zinniol a true phytotoxin? Evaluation of its activity at the cellular level against <Emphasis Type="Italic">Tagetes erecta</Emphasis>

Zinniol, a non-host selective phytotoxin commonly produced by fungi of the Alternaria genus, has been reported as the metabolite responsible for the phytotoxicity of the lipophilic fraction of A. tagetica. While both the lipophilic fraction and zinniol have been shown to produce necrosis on leaves of susceptible marigold (Tagetes erecta) plants, the true role of zinniol in the infectious process remains uncertain. Using marigold cell cultures as a model, we evaluated the effects of zinniol and the lipophilic fraction at the cellular level and showed that pure zinniol is not markedly phytotoxic at concentrations known to induce necrosis in leaves of T. erecta. Moreover, the effects of zinniol on cell membranes and DNA fragmentation are less intense than those caused by the lipophilic fraction. These results suggest that zinniol may not play a significant role in the A. tagetica–T. erecta interaction and, consequently, its classification as a non-host selective phytotoxin is questionable.     

Detection and identification of a ‘<Emphasis Type="Italic">Candidatus</Emphasis> phytoplasma aurantifolia’-related strain (16SrII-A subgroup) associated with <Emphasis Type="Italic">corchorus aestuans</Emphasis> phyllody in China

In September 2015, a phyllody that is typical of phytoplasma infection was observed on Corchorus aestuans plants in Haikou, Hainan Province, China. Total DNA from symptomatic and asymptomatic plants was extracted for molecular diagnosis. On the basis of sequence analysis and phylogenetic trees based on 16S rDNA and rp genes, the phyllody phytoplasma was ascertained to be related to ‘Candidatus phytoplasma aurantifolia’. To the best of our knowledge, this is the first report of a phytoplasma infecting C. aestuans in the world.     

Enhanced protection against two major fungal pathogens of groundnut, <Emphasis Type="Italic">Cercospora arachidicola</Emphasis> and <Emphasis Type="Italic">Aspergillus flavus</Emphasis> in transgenic groundnut over-expressing a tobacco β 1–3 glucanase

Groundnut is an important oilseed crop of the Indian subcontinent. Yield losses due to fungal diseases are enormous in the cultivation of this crop. Over-expression of PR proteins leads to increased resistance to pathogenic fungi in several crops. The PR protein glucanase hydrolyses a major cell-wall component, glucan, of pathogenic fungi and acts as a plant defense barrier. We report in this paper, overexpression of a tobacco glucanase in transgenic groundnut and its resistance towards Cercospora arachidicola and Aspergillus flavus. PCR, Southern and Northern hybridization confirmed stable integration and expression of the glucanase gene in groundnut transgenics. When screened for resistance against Cercospora arachidicola the transgenics showed not only reduction in the number of spots but also delay in the onset of disease. Resistance was also demonstrated against one another important pathogen of groundnut, Aspergillus flavus. The transgenics not only resisted hyphal spread but also did not accumulate aflatoxin in the seeds. The results demonstrate the potential of a PR protein from a heterologous source in developing fungal disease resistant groundnut.     

Molecular typing of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma mali’ and epidemic history tracing by a combined T-RFLP/VNTR analysis approach

Apple proliferation caused by ‘Candidatus Phytoplasma mali’ is a disease of apple trees gaining increasing importance in Europe. The present study describes a high-throughput method for simultaneous typing of ‘Ca. P. mali’ at two genetic loci. This novel approach combines terminal restriction fragment length polymorphism (T-RFLP) analysis of a putative rhodanese-like protein gene and the analysis of the variable number of tandem repeats (VNTR) of the ribosomal protein L22 gene. The typing approach was applied to analyse a collection of DNA isolates from 310 apple trees tested positive for ‘Ca. P. mali’. Samples were taken between 2002 and 2010 in South Tyrol (Northern Italy). In addition, 15 samples of Cacopsylla melanoneura and 19 of C. picta were typed. Seven combined genetic profiles were found in the samples of infected apple trees: AT-2/rpX-A (81.0%), AT-1/rpX-D (8.4%), AT-1/rpX-E (4.2%), AT-1/rpX-A (3.2%), AT-1/rpX-B (1.6%), AT-1/rpX-C (1.0%) and AP/rpX-A (0.3%), and one mixed infection AP + AT-1/rpX-A + rpX-D (0.3%). Subtype rpX-E was discovered for the first time. In C. melanoneura samples the most frequent subtype was AT-1/rpX-E, followed by AT-1/rpX-D and AT-1/rpX-C. All C. picta samples displayed subtype AT-2/rpX-A. Analysis of the temporal distribution of subtype frequencies in apple trees revealed that exclusively subtype AT-1 in combination with four rpX subtypes was present in South Tyrol in the period from 2002 to 2004. From 2006 onwards subtype AT-2/rpX-A became dominant with an average frequency of 90%. The data obtained suggest that there may be a co-adaptation of particular ‘Ca. P. mali’ subtypes with different insect vector species.     

Detection of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma brasiliense’ in a new geographic region and existence of two genetically distinct populations’

‘Candidatus Phytoplasma brasiliense’, a phytoplasma taxon associated with hibiscus witches’ broom disease was first described in 2001 in Brazil. In September 2007, a peach tree (Prunus persica) displaying yellowing symptoms reminiscent of phytoplasma infection was sampled in Guba region of Azerbaijan. A phytoplasma was detected in the diseased peach tree by nested PCR amplification of its 16S rDNA with universal primers for phytoplasmas. Phylogenetical analyses of the amplified 16S rDNA showed that the phytoplasma infecting the peach tree corresponded to ‘Ca. P. brasiliense’, a species never reported in Euro-Mediterranean area. To set up a detection assay, cloning of a ‘Ca. P. brasiliense’ DNA fragment was undertaken by comparative RAPD. The amplified dnaK-dnaJ genetic locus was used to design a nested PCR assay able to amplify all ‘Ca. P. brasiliense’ isolates of the subgroup 16SrXV-A without amplifying the related members of the group 16SrII. This assay also allowed confirming the first detection of ‘Ca. P. brasiliense’ in diseased basil collected in south Lebanon.