Current Distribution of Huanglongbing (citrus greening disease) in India as Diagnosed by Real‐Time PCR

The widespread occurrence of Huanglongbing (HLB) was recorded in sixteen citrus growing states of India using the real‐time quantitative PCR and the derived threshold cycle (Ct) value. All the commercially important citrus varieties of mandarin, sweet orange, lime and lemon, pummelo and Satkara were infected with ‘Candidatus Liberibacter asiaticus’, the bacterium associated with HLB. Ct values positive for HLB were found in all the states except Arunachal Pradesh. The primer–probe combination HLBas‐HLBr‐HLBp was found specific to Ca. L. asiaticus and do not exhibit any cross‐reactivity with other pathogenic residents of citrus.     

Effect of host deprivation on the foraging behavior of the Asian citrus psyllid parasitoid Tamarixia radiata: observations from the laboratory and the field

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is a significant citrus pest and the parasitoid Tamarixia radiata (Waterston) (Hymenoptera: Eulophidae) has been released in various citrus‐producing areas in classical biological control programs targeting D. citri. We investigated the effect of host deprivation on the foraging behavior and patch utilization by T. radiata. In the laboratory, females deprived of hosts for 3 days tended to leave patches of 12 nymphs without parasitizing hosts during the ca. 30 min they spent in the patch before leaving. Moreover, half of these females failed to host feed, and those that did host feed, on average, needed more than 15 min to complete feeding. Conversely, non‐host‐deprived females parasitized on average three nymphs before leaving patches without host feeding during the ca. 39 min they spent in the patch. These laboratory observations were compared to mass‐reared female T. radiata that were released onto colonies of D. citri nymphs infesting citrus in the field. Release vials were provisioned with honey and these females had no opportunities to host feed over the 1‐ to 2‐day containment period prior to release. When introduced onto D. citri colonies, 68% T. radiata females abandoned D. citri patches prior to probing hosts, in part, because Argentine ants, Linepithema humile (Mayr) (Hymenoptera: Formicidae), tending colonies disturbed searching parasitoids. These results from laboratory and field studies are discussed in the context of classical biological control, with the aim of understanding how to manipulate host availability and ant activity so establishment rates and impact of T. radiata can be improved.     

Potential of Periplaneta fuliginosa densovirus as a biocontrol agent for smoky-brown cockroach, P. fuliginosa

A densovirus, named Periplaneta fuliginosa densovirus (PfDNV), affects smoky-brown cockroach (P. fuliginosa), which is a major peridomestic cockroach in China, southeastern United States, Japan, and Southeast Asia. We reported here that PfDNV could be mass-produced using virus rescue technology. Pathology and ultrastructural studies revealed the presence of enlarged and dense nuclei in hindgut cells of infected smoky-brown cockroach nymphs and adults. Laboratory bioassays of the PfDNV against P. fuliginosa showed that it has the potential to be used as a biocontrol agent. High mortality was recorded in adult smoky-brown cockroaches treated with PfDNV baits at the dosage of 6 × 10⁴ virions per gram against P. fuliginosa in both no-choice tests and two-choice tests (LT₅₀’s were 13.37 and 13.50 days, respectively). PfDNV was also effective in large population chamber tests. LT₅₀ was 17.57 days and mortality achieved was 95.6% in 28 observation days.     

Roq1 mediates recognition of the Xanthomonas and Pseudomonas effector proteins XopQ and HopQ1

Xanthomonas spp. are phytopathogenic bacteria that can cause disease on a wide variety of plant species resulting in significant impacts on crop yields. Limited genetic resistance is available in most crop species and current control methods are often inadequate, particularly when environmental conditions favor disease. The plant Nicotiana benthamiana has been shown to be resistant to Xanthomonas and Pseudomonas due to an immune response triggered by the bacterial effector proteins XopQ and HopQ1, respectively. We used a reverse genetic screen to identify Recognition of XopQ 1 (Roq1), a nucleotide‐binding leucine‐rich repeat (NLR) protein with a Toll‐like interleukin‐1 receptor (TIR) domain, which mediates XopQ recognition in N. benthamiana. Roq1 orthologs appear to be present only in the Nicotiana genus. Expression of Roq1 was found to be sufficient for XopQ recognition in both the closely‐related Nicotiana sylvestris and the distantly‐related beet plant (Beta vulgaris). Roq1 was found to co‐immunoprecipitate with XopQ, suggesting a physical association between the two proteins. Roq1 is able to recognize XopQ alleles from various Xanthomonas species, as well as HopQ1 from Pseudomonas, demonstrating widespread potential application in protecting crop plants from these pathogens.     

Glutaminyl‐tRNA Synthetase from Pseudomonas aeruginosa: Characterization,structure, and development as a screening platform

Pseudomonas aeruginosa has a high potential for developing resistance to multiple antibiotics. The gene (glnS) encoding glutaminyl‐tRNA synthetase (GlnRS) from P. aeruginosa was cloned and the resulting protein characterized. GlnRS was kinetically evaluated and the K_M and k_cat^obs, governing interactions with tRNA, were 1.0 μM and 0.15 s^?1, respectively. The crystal structure of the α₂ form of P. aeruginosa GlnRS was solved to 1.9 Å resolution. The amino acid sequence and structure of P. aeruginosa GlnRS were analyzed and compared to that of GlnRS from Escherichia coli. Amino acids that interact with ATP, glutamine, and tRNA are well conserved and structure overlays indicate that both GlnRS proteins conform to a similar three‐dimensional structure. GlnRS was developed into a screening platform using scintillation proximity assay technology and used to screen ~2,000 chemical compounds. Three inhibitory compounds were identified and analyzed for enzymatic inhibition as well as minimum inhibitory concentrations against clinically relevant bacterial strains. Two of the compounds, BM02E04 and BM04H03, were selected for further studies. These compounds displayed broad‐spectrum antibacterial activity and exhibited moderate inhibitory activity against mutant efflux deficient strains of P. aeruginosa and E. coli. Growth of wild‐type strains was unaffected, indicating that efflux was likely responsible for the lack of sensitivity. The global mode of action was determined using time‐kill kinetics. BM04H03 did not inhibit the growth of human cell cultures at any concentration and BM02E04 only inhibit cultures at the highest concentration tested (400 μg/ml). In conclusion, GlnRS from P. aeruginosa is shown to have a structure similar to that of E. coli GlnRS and two natural product compounds were identified as inhibitors of P. aeruginosa GlnRS with the potential for utility as lead candidates in antibacterial drug development in a time of increased antibiotic resistance.     

Bioprospecting of Saprobe Fungi from the Semi‐Arid North‐East of Brazil for the Control of Anthracnose on Sorghum

Anthracnose, caused by the hemiotrophic fungus Colletotrichum sublineolum, is one of the most important diseases affecting sorghum production worldwide. The main goal of this study was to select saprobe fungi from the semi‐arid north‐east of Brazil that could increase sorghum resistance to anthracnose and investigate this increased resistance at both physiological and biochemical levels. Plants were sprayed with Curvularia inaequalis, Gonytrichum macroladum, Memnoniella levispora, Pithomyces chartarum, Periconia hispidula, Phaeoisaria clematidia, Dictyochaeta heteroderae, Sarcopodium circinatum, Periconia byssoides, Moorella speciosa, Stachybotrys chartarum, Pseudobotrytis terrestres, Memnoniella echinata, Stachybotrys globosa and Gonytrichum clamydosporium 24 h before inoculation with C. sublineolum. Plants sprayed with water served as the control treatment. The area under the anthracnose progress curve was significantly reduced in comparison with the control treatment only for plants sprayed with C. inaequalis. Therefore, C. inaequalis was selected for physiological and biochemical evaluations. The peroxidases, chitinases and β‐1,3‐glucanases activities were significantly higher for plants sprayed with C. inaequalis and inoculated with C. sublineolum than for plants not sprayed with C. inaequalis and inoculated with C. sublineolum. There was no apparent decrease in the values of net carbon assimilation rate, stomatal conductance to water vapour or transpiration rate for plants sprayed with C. inaequalis and infected by C. sublineolum in comparison with plants not sprayed with C. inaequalis and infected by C. sublineolum. In conclusion, sorghum resistance against C. sublineolum infection was greatly potentiated by C. inaequalis without any apparent change in the photosynthetic capacity of the infected plants.     

Evaluation of Fopius arisanus as a biological control agent for the olive fruit fly in California

1 The egg‐prepupal parasitoid Fopius arisanus (Hymenoptera: Braconidae) was evaluated in quarantine facilities as a potential biological control agent for the olive fruit fly Bactrocera oleae (Diptera: Tephritidae) in California, U.S.A. 2 Nontarget testing of two weed biological control agents confirmed that F. arisanus will not attack Tephritidae that feed in inflorescences or galls. It may, however, pose risks to native Tephritidae that feed in fruit. 3 Females preferentially oviposited in eggs, although first‐instar B. oleae were also attacked. Low lifetime reproductive potential and high rates of direct mortality inflicted on host eggs indicate that rearing on B. oleae may prove difficult. 4 In multiparasitized B. oleae, F. arisanus prevailed in competition against two species of larval–pupal parasitoids, Diachasmimorpha kraussii and Psyttalia concolor (both Hymenoptera: Braconidae). 5 The broad host‐range of F. arisanus with respect to fruit‐feeding Tephritidae may preclude its introduction to California, as may its low fecundity and its intrinsic competitive superiority over larva l–pupal parasitoids, which include specialists on B. oleae that are currently being introduced to California. High rates of direct mortality, however, point to potential uses in augmentative biological control. Whether or not F. arisanus is released in California, its biology as a parasitoid of B. oleae has been little studied to date and the results herein may be applied in other regions worldwide where B. oleae is a problem.     

A petal‐specific InMYB1 promoter from Japanese morning glory: a useful tool for molecular breeding of floricultural crops

Production of novel transgenic floricultural crops with altered petal properties requires transgenes that confer a useful trait and petal‐specific promoters. Several promoters have been shown to control transgenes in petals. However, all suffer from inherent drawbacks such as low petal specificity and restricted activity during the flowering stage. In addition, the promoters were not examined for their ability to confer petal‐specific expression in a wide range of plant species. Here, we report the promoter of InMYB1 from Japanese morning glory as a novel petal‐specific promoter for molecular breeding of floricultural crops. First, we produced stable InMYB1_1kb::GUS transgenic Arabidopsis and Eustoma plants and characterized spatial and temporal expression patterns under the control of the InMYB1 promoter by histochemical β‐glucuronidase (GUS) staining. GUS staining patterns were observed only in petals. This result showed that the InMYB1 promoter functions as a petal‐specific promoter. Second, we transiently introduced the InMYB1_1 kb::GUS construct into Eustoma, chrysanthemum, carnation, Japanese gentian, stock, rose, dendrobium and lily petals by particle bombardment. GUS staining spots were observed in Eustoma, chrysanthemum, carnation, Japanese gentian and stock. These results showed that the InMYB1 promoter functions in most dicots. Third, to show the InMYB1 promoter utility in molecular breeding, a MIXTA‐like gene function was suppressed or enhanced under the control of InMYB1 promoter in Arabidopsis. The transgenic plant showed a conspicuous morphological change only in the form of wrinkled petals. Based on these results, the InMYB1 promoter can be used as a petal‐specific promoter in molecular breeding of floricultural crops.     

Effects of local vegetation and plantation age for the parasitoid Asecodes mento– a biocontrol agent in organic strawberry fields

Abstract The parasitoid Asecodes mento (Walker, 1839) (Hymenoptera: Eulophidae) is the most important biocontrol agent of the strawberry leaf beetle Galerucella tenella (L.) (Coleoptera: Chrysomelidae) in northern Europe. Here, I investigated whether natural parasitism in organic strawberry plantations was affected by the presence of the alternative host plant meadowsweet (Filipendula ulmaria), and whether parasitism rates differed between plantations of different ages (6 to 79 years). I also investigated whether parasitoid brood size, body size and sex ratio differed between the two host plants in the field. Parasitism was very low (0%) in newly established plantations and increased to a plateau (～40%) in fields where strawberries had been grown for approximately 20 years or longer. Such an extended colonization process is unacceptable for commercial growers. It would thus be desirable to find a method to catalyze parasitoid population buildup in young plantations. Parasitoid brood sizes were larger in beetles collected from meadowsweet, while body size and sex ratio did not differ between parasitoids collected from the two plants. These findings suggest that meadowsweet can export parasitoids to neighboring strawberry fields. Although this is a possibility, I did not find any significant differences in parasitism rates between isolated strawberry fields and fields adjacent to meadowsweet stands, indicating that effects of local vegetation are small on parasitism rates. Releasing parasitoids in newly established strawberry plantations may be a better strategy for quickly obtaining high parasitism than intercropping with meadowsweet.     

Intercropping oilseed rape as a potential relay crop for enhancing the biological control of green peach aphids and aphid‐transmitted virus diseases

Tobacco viruses transmitted by green peach aphids, Myzus persicae (Sulzer) (Hemiptera: Aphididae), cause severe disease in flue‐cured tobacco, Nicotiana tabacum L. (Solanaceae), in China and throughout the world. Field experiments were conducted in 2016 and 2017 in Longyan City, Fujian Province, China, to determine whether M. persicae and aphid‐transmitted virus diseases are affected by intercropping of oilseed rape, Brassica napus L. (Brassicaceae), in tobacco fields. The results showed that, compared with those in monocultured fields, the densities of M. persicae and winged aphids in intercropped fields significantly decreased in both 2016 and 2017. In particular, the appearance of winged aphids was delayed by ca. 7 days. Moreover, the densities of Aphidius gifuensis Ashmead (Hymenoptera: Aphidiidae), a parasitoid of the aphid, significantly increased in 2016 and 2017. Accordingly, the incidence rates of aphid‐transmitted virus diseases (those caused by the cucumber mosaic virus, potato virus Y, and tobacco etch virus) significantly decreased in the intercropped fields in 2016 and 2017. Tobacco yields and monetary value significantly increased in 2016 (by 10–25 and 14–29%, respectively) and 2017 (by 17–22 and 22–34%, respectively). Consequently, our results suggest that intercropping oilseed rape in tobacco fields is a good approach to regulating and controlling aphids and tobacco mosaic viruses, for example potyvirus, and this intercropping can help control aphid‐transmitted virus diseases in tobacco.     

1‐Phenyl‐3‐pentanone,a Volatile Compound from the Edible Mushroom Mycoleptodonoides aitchisonii Active Against Some Phytopathogenic Fungi

Volatiles produced by mycelia of mushrooms with aromatic odour were investigated for their antifungal activity against plant‐pathogenic fungi. The results of the screening of 23 species of basidiomycetes revealed that volatile substances from mycelia of Mycoleptodonoides aitchisonii (TUFC10099), an edible mushroom, strongly inhibited the mycelial growth, spore germination and lesion formation on host leaves of some plant‐pathogenic fungi including Alternaria alternata, A. brassicicola, A. brassicae, Colletotrichum orbiculare and Corynespora cassiicola. The volatile compounds were isolated from the culture filtrate of M. aitchisonii, and 1‐phenyl‐3‐pentanone was identified as a major antifungal volatile. The compound had significantly inhibitory activity against plant‐pathogenic fungi at 35 ppm. This is the first report that the volatile compound produced by mycelia of M. aitchisonii has antifungal activity against plant‐pathogenic fungi.     

Preliminary studies on Ramularia crupinae sp. nov. as a potential biological control agent for common crupina (Crupina vulgaris) in the USA

Common crupina, Crupina vulgaris (Asteraceae), is an alien weed of pastures and disturbed noncrop lands in the western USA. Research is underway in Europe to search for the natural enemies to control the weed. Among various natural enemies attacking C vulgaris, a new fungal species, Ramularia crupinae, has been found to cause serious damage to the weed. The fungus has been grown on various media and a modified V8 juice agar has proved to be suitable for both growth and sporulation. Greenhouse and field studies have shown that R. crupinae can kill infected leaves, thus reducing the plant growth, and in severe cases can kill the young plants. During preliminary host-range tests using various members of the Asteraceae, including artichoke and safflower, the fungus attacked only C. vulgaris.     

Escape of Sgs1 from Rad9 inhibition reduces the requirement for Sae2 and functional MRX in DNA end resection

Homologous recombination requires nucleolytic degradation (resection) of DNA double‐strand break (DSB) ends. In Saccharomyces cerevisiae, the MRX complex and Sae2 are involved in the onset of DSB resection, whereas extensive resection requires Exo1 and the concerted action of Dna2 and Sgs1. Here, we show that the checkpoint protein Rad9 limits the action of Sgs1/Dna2 in DSB resection by inhibiting Sgs1 binding/persistence at the DSB ends. When inhibition by Rad9 is abolished by the Sgs1‐ss mutant variant or by deletion of RAD9, the requirement for Sae2 and functional MRX in DSB resection is reduced. These results provide new insights into how early and long‐range resection is coordinated.     

用黄瓜花叶病毒卫星RNA生防制剂大面积防治麦茬辣椒病毒病

用黄瓜花叶病毒（ＣＭＶ）卫星ＲＮＡ生防制剂Ｓ＿（５２）免疫辣椒,能防治辣椒由ＣＭＶ引起的病毒病。用Ｓ＿（５２）免疫接种辣椒的大田对比试验,防效达５９．５－７１．０％,产量比对照增加２６．１－３２．２％,产值增加３３．５－４５．２％。小区对比试验,用Ｓ＿（５２）免疫接种５０天的辣椒,防效达７１．３－９２．９％,免疫接种８０天,防效达５５．９－７８．５％,产量比对照增加３６．０－６５．３％,产值增加４９．３－９３．５％,还有刺激生长,促进早熟和增强对真菌、细菌病害抵抗作用。     

Vermistella arctica n. sp. Nominates the Genus Vermistella as a Candidate for Taxon with Bipolar Distribution

A new amoebozoan species, Vermistella arctica n. sp., is described from marine habitats in the central part of Svalbard archipelago. This is the first report on Arctic amoebae belonging to the genus Vermistella Moran and Anderson, 2007, the type species of which was described from the opposite pole of the planet. Psychrophily proved in the new strains qualifies the genus Vermistella as a bipolar taxon. Molecular phylogenetic analyses based on 18S rDNA and actin sequences did not show any affinity of the genus Vermistella to Stygamoeba regulata ATCC^® 50892^? strain. A close phylogenetic relationship was found between Vermistella spp. and a sequence originating from an environmental sample from Cariaco basin, the largest marine permanently anoxic system in the world. Possible mechanisms of bipolar distribution are discussed.     

LjMOT1, a high‐affinity molybdate transporter from Lotus japonicus,is essential for molybdate uptake,but not for the delivery to nodules

Molybdenum (Mo) is an essential nutrient for plants, and is required for nitrogenase activity of legumes. However, the pathways of Mo uptake from soils and then delivery to the nodules have not been characterized in legumes. In this study, we characterized a high‐affinity Mo transporter (LjMOT1) from Lotus japonicus. Mo concentrations in an ethyl methanesulfonate–mutagenized line (ljmot1) decreased by 70–95% compared with wild‐type (WT). By comparing the DNA sequences of four AtMOT1 homologs between mutant and WT lines, one point mutation was found in LjMOT1, which altered Trp²⁹² to a stop codon; no mutation was found in the other homologous genes. The phenotype of Mo concentrations in F₂ progeny from ljmot1 and WT crosses were associated with genotypes of LjMOT1. Introduction of endogenous LjMOT1 to ljmot1 restored Mo accumulation to approximately 60–70% of the WT. Yeast expressing LjMOT1 exhibited high Mo uptake activity, and the K_m was 182 nm . LjMOT1 was expressed mainly in roots, and its expression was not affected by Mo supply or rhizobium inoculation. Although Mo accumulation in the nodules of ljmot1 was significantly lower than that of WT, it was still high enough for normal nodulation and nitrogenase activity, even for cotyledons‐removed ljmot1 plants grown under low Mo conditions, in this case the plant growth was significantly inhibited by Mo deficiency. Our results suggest that LjMOT1 is an essential Mo transporter in L. japonicus for Mo uptake from the soil and growth, but is not for Mo delivery to the nodules.     

Feeding range studies of Rodolia cardinalis (Mulsant), a candidate biological control agent of Icerya purchasi Maskell in the Galápagos islands

The immediate threat of the cottony cushion scale, Icerya purchasi Maskell (Homoptera: Margarodidae), to the conservation of endangered flora in the Galápagos islands prompted conservation groups to assess the risks associated with the introduction of its natural enemy, Rodolia cardinalis (Mulsant) (Coleoptera: Coccinellidae). Although R. cardinalis has been widely used for controlling this exotic pest, little information was found to confirm its presumed narrow feeding range. Consequently, studies were deemed necessary to determine whether the introduction of R. cardinalis would harm the island’s native invertebrate fauna, in particular rare or threatened species. Using no-choice trials, we tested neonate and third instar larvae of R. cardinalis against 16 and 11 potential prey species, respectively. Adults with prior feeding experience on I. purchasi were tested against eight non-target species and naïve adults (those that had not fed on I. purchasi) were tested against six. These trials included up to 35% of the Homoptera species of conservation value presumed to have the highest risk of being preyed upon by R. cardinalis. To maximize the range of species exposed to R. cardinalis, feeding trials were also carried out with some introduced species representative of groups containing potential non-target species that were not located for testing. R. cardinalis was unable to complete its life cycle on any of the test prey species and only fed on Margarodes similis Morrison (Homoptera: Margarodidae), a species closely related to the cottony cushion scale. M. similis, however, is subterranean and in its natural habitat is not at risk from foraging by R. cardinalis. Based on these trials, we believe that immature stages of R. cardinalis will have no impact on the non-target invertebrate fauna of the Galápagos islands because they specialize on Margarodidae. Although the limited nature of our testing prevents us from reaching a definitive conclusion about the prey range of R. cardinalis adults, our results indicate that it is also narrow. According to our field and laboratory studies, niche overlap with native predators of Homoptera will be minimal and intraguild predation should not occur.