The combined effect of 1‐methylcyclopropene and citral suppressed postharvest grey mould of tomato fruit by inhibiting the growth of Botrytis cinerea

1‐Methylcyclopropene (1‐MCP, 1 μl/L) and 1 × minimum fungicidal concentration (MFC) citral alone and in combination were used to treat on postharvest tomato fruits to investigate their influence on disease incidence and postharvest quality during fruit storage, which were stored at 90%–95% relative humidity and 25 ± 2°C. Weight loss, pH, hue angle (Hue^°), total soluble solid (TSS), ascorbic acid content, firmness and antioxidant enzyme activities were evaluated after each storage period. 1 μl/L 1‐MCP or 1 × MFC citral reduced weight loss, retarded peel colour changes and retained postharvest fruit quality. 1 μl/L 1‐MCP + 1 × MFC citral could better maintain firmness and ascorbic acid content and increase antioxidant enzyme activities, compared to other treatments. Disease incidence of tomato fruit was significantly decreased, and spore germination and mycelia growth of Botrytis cinerea were suppressed by the combined treatment with 1 μl/L 1‐MCP and 1 × MFC citral. These results indicate that the combined treatment could effectively delay postharvest tomato fruits senescence and inhibit postharvest pathogens in vitro.     

The effect of chemical and biological treatment on root and crown rot disease caused by Phytophthora cryptogea Pethybr. & Lafferty in Gerbera

Phytophthora root and crown rot (Phytophthora cryptogea) on gerbera is difficult to manage because most gerbera cultivars are susceptible to P. cryptogea. This study was conducted in order to determine the in vivo (pot experiment) efficacy of some fungicides and biofungicides. In pot experiments, fungicides were applied 7 days after inoculation with P. cryptogea, while biofungicide was applied 7 days before inoculation. In this study, soil drenches of five fungicides were tested. “Ametoctradin+dimethomorph (100 ml/day),” “mandipropamid+difenoconazole (60 ml/day),” “propamocarb+fosetyl‐Al (200 ml/day),” “mancozeb+metalaxyl‐M (250 g/day)” and “azoxystrobin+difenoconazole (100 ml/day)” active substances were used. Similarly, one biofungicide Bacillus amyloliquefaciens syn. MBI 600 (50 g/100 L) was applied by soil drenching. Efficacy of treatments was assessed according to the percentage of the root system which was visibly rotten at the end of the experiment. Root and crown rot severity was rated on a scale of 0 = 0% root system necrotic, 1 = 1%‐25% necrotic, 2 = 26%‐50% necrotic, 3 = 51%‐75% necrotic and 4 = 76%‐100% necrotic from 12 to 21 days. In this experiment, “azoxystrobin 200 g/L + difenoconazole 125 g/L” exhibited the highest efficacy against P. cryptogea with a ratio of 43.75%. The other fungicides and biofungicides ametoctradin 300 g/L + dimethomorph 225 g/L, mandipropamid 250 g/L + difenoconazole 250 g/L, propamocarb 530 g/L + fosety‐Al 310 g/L, mancozeb 64%+metalaxyl‐M 4% and Bacillus amyloliquefaciens syn. MBI 600 11% were ineffective. Importance should be given to management strategies of P. cryptogea of and more experiments should be carried out for a better understanding of the use of registered fungicides and biofungicides.     

Immunoassay‐based Techniques for Early and Specific Detection of Latent Postharvest Anthracnose in Mango

The postharvest anthracnose pathogen Colletotrichum gloeosporioides inciting latent or quiescent infection of mango was detected in early stages using immunoassay methods. Twenty‐five pathotypes isolated from different agroclimatic zones of Tamil Nadu, Karnataka and Pondicherry, India, revealed the variation in protein profile analysis (SDS‐PAGE). The polyclonal antibodies (PCA) were raised against the unfractioned mycelial protein (UMP) and a 40‐kDa polypeptide present in all pathotypes. Standardization of antigen and antiserum dilutions revealed that an antigen dilution of 1 : 200 (protein concentration of 20 μg/ml) and antiserum dilution of 1 : 100 (protein concentration of 40 μg/ml raised against UMP) and 1 : 200 (protein concentration of 20 μg/ml raised against 40 kDa polypeptide) was found to be optimum for the detection of anthracnose pathogen. Both antisera detected the C. gloeosporioides antigen in enzyme‐linked immunosorbent assays (ELISAs), dot immunobinding assays (DIBAs) and Western blots. The specificity in reaction was compared by isolating other Colletotrichum spp. from various hosts viz., C. lindemuthianum (beans), C. falcatum (sugarcane), C. musae (banana), C. capsici (chillies) and Botryodiplodia theobromae (mango). The antisera generated against UMP revealed the cross‐reaction with other host isolates and mango stem end rot pathogen (B. theobromae). The PCA raised against 40‐kDa polypeptide exhibited the specific reaction with C. gloeosporioides isolates in all the immunoassay techniques. By utilizing both PCA, the presence of latent infection was observed in healthy‐looking leaves, flowers and fruits in orchard conditions. The fruit tissues recorded high absorbance values followed by flowers and leaves in all the detection methods. The ELISA technique was also useful in assessing the pathogen inoculum at various biocontrol formulations sprayed mango trees under field conditions. The fluorescent pseudomonad strains mixture (KFP1 + FP7) amended with chitin sprayed at 30‐day intervals revealed the significant reduction in pathogen load than other formulations and unsprayed control.     

Identification and Pathogenicity of Lasiodiplodia Species from Eucalyptus urophylla × grandis,Polyscias balfouriana and Bougainvillea spectabilis in Southern China

Species of Lasiodiplodia are important pathogens of a wide variety of plants covering a wide geographical distribution. These fungi can be associated with different symptoms such as stem cankers, shoot blights, fruit rots, dieback and gummosis. Diseases caused by Lasiodiplodia were surveyed on Eucalyptus urophylla × grandis, Polyscias balfouriana and Bougainvillea spectabilis in a nursery in southern China. Based on morphology characteristics and phylogenetic analyses of ITS rDNA sequences and translation elongation factor 1‐alpha (TEF‐1α) gene regions, four species of Lasiodiplodia were identified. Lasiodiplodia theobromae was identified from E. urophylla × grandis, P. balfouriana and B. spectabilis. L. hormozganensis, L. iraniensis and L. pseudotheobromae were identified from B. spectabilis. To our knowledge, with the exception of L. theobromae on E. urophylla × grandis, this study represents the first report of these fungi on the host plants. Pathogenicity tests showed that all Lasiodiplodia spp. obtained in this study are virulent to E. urophylla × grandis and B. spectabilis, and L. theobromae was virulent to P. balfouriana.     

First report of Lasiodiplodia theobromae causing spadix rot in Anthurium andraeanum

Spadix rot was reported in Anthurium andraeanum cultivation farms in southern Thailand. The pathogen was identified as Lasiodiplodia theobromae, based on morphology and the nucleotide sequences of the internal transcribed spacer (ITS) and translation elongation factor 1-α (EF1-α) genes. Pathogenicity test proved Koch's postulates, supporting that L. theobromae caused spadix rot in A. andraeanum. To our knowledge, this is the first report of L. theobromae causing spadix rot in A. andraeanum, in Thailand and elsewhere.     

Physical postharvest treatments in the control of stem‐end rot of mango

Botryosphaeria dothidea is the major pathogen of mango in Brazil, causing stem‐end rot, which causes significant losses during transportation and storage. The current strategy to control this particular disease using synthetic fungicides has been ineffective, leaving residues in the fruit. The objective of the research was to study the effect of physical treatments, with hot water rinse brushing (HWRB) and ultraviolet C irradiation (UV‐C), individually and in combination, to control stem‐end rot of mango. Physicochemical parameters, respiration and resistance induction of the fruit were also analysed. The in vitro trials demonstrated that B. dothidea is a thermoresistant fungus. The individual treatments with HWRB at 65°C for 15 s and 2.5 kJ/m² of UV‐C presented the best results, showing less symptoms of the disease during 18 days of storage. The combination of HWRB with UV‐C did not improve the control of the disease when compared to the treatments applied individually. The physicochemical parameters and the consumer acceptance evaluation showed that both physical treatments preserved the appearance of the fruit and delayed the ripening–senescence process. The induction of defence‐related enzymes revealed that induced resistance was an important mechanism involved in the control of stem‐end rot of mango.     

Effect of cold storage on larval and adult Anastrepha ludens (Diptera: Tephritidae) viability in commercially ripe, artificially infested Persea americana 'Hass'

Commercially ripe 'Hass' avocados, Persea americana Mill, artificially exposed to wild Anastrepha ludens (Loew) (Diptera: Tephritidae) females 24 h after harvest were placed in a cold storage facility to determine the effect of low temperature on larval survival and adult viability. Fruit were left for 3, 6, 9, and 12 d in a cold room at 5 degrees C followed by a 20-25-d period at ambient temperature to allow for larval development and pupation. Hass avocados and grapefruit, Citrus paradisi Macfadyen, maintained at ambient temperature served as controls. Overall, only 0.23% of the Hass avocados and 19.30% of the grapefruit were infested. The number of infested fruit increased with decreasing exposure time to cold. Puparia from cold-treated Hass avocados were significantly smaller than those stemming from cold-treated grapefruit. Hass avocados exposed for 12 d to 5 degrees C yielded no puparia, and those exposed for 6 and 9 d yielded 22 and two puparia, respectively, but no adults. Although Hass avocados exposed to cold temperature for 3 d yielded adults that reached sexual maturity (N = 16), females laid inviable eggs. Grapefruit exposed to cold for 12 d yielded normal-sized puparia (but no adults), whereas those exposed over 9 d yielded females able to lay viable eggs. We conclude that exposing fruit to cold storage after packing and during transport represents an effective risk-mitigating procedure in the highly improbable event that a gravid A. ludens female might lay eggs in a commercially ripe Hass avocado that had been left unprotected in a packinghouse.     

Evaluation of Clove Essential Oil as a Mycobiocide Against Rhizopus stolonifer and Fusarium solani,Tuber Rot Causing Fungi in Yam (Dioscorea rotundata Poir.)

To exploit natural products for plant disease control, the essential oil of Syzygium aromaticum (L.) Merr. & Perr. (clove) was investigated for its antifungal activity against Rhizopus stolonifer and Fusarium solani, the postharvested yam tuber rot pathogens. The essential oil was obtained by hydrodistillation using a Clevenger‐type apparatus. The chemical composition of the oil was determined by gas chromatography and gas chromatography coupled with mass spectrometry. Antifungal activities of the oil were tested in vitro against mycelia growth and spores germination. In situ tests were conducted on healthy yam tubers, and necrosis symptoms were assessed. Results showed that eugenol (79.4%), eugenylacetate (9.2%) and isocaryophyllene (7.0%) were the major components. The oil exerted antifungal activities with total inhibition (TI) of the mycelia growth of R. stolonifer and F. solani was recorded at 200 and 300 ppm, respectively, while TI of spores germination was recorded at 31.25 and 250 ppm, respectively. For the standard fungicide (Ridomil^®), TI value of mycelia growth was 1600 ppm for the both pathogens, while TI of spores germination were 200 ppm and 1600 ppm, respectively, for Rhizopus and Fusarium. In situ tests showed complete inhibition of yam tuber rot when the essential oil was applied at 2000 ppm for preventive tests. This oil also reduced significantly (P ≤ 0.05) necrosis development on yam tuber for curative test at the same concentration. Total inhibition of the necrosis by Ridomil (3000 ppm) was observed only for Rhizopus on preventive test. There were positive correlations between the oil concentration and the reduction of necrosis cause by R. stolonifer and F. solani. These findings showed that clove essential oil may serve as environmental friendly bio‐fungicide for the management of postharvest yam tuber rot.     

Effect of an Antagonistic Yeast in Combination with Microwave Treatment on Postharvest Blue Mould Rot of Jujube Fruit

The potential of using an antagonistic yeast alone or in combination with microwave treatment for controlling blue mould rot of jujube fruit, and its effect on postharvest quality of fruit, was investigated. The results showed that the growth of Penicillium citrinum was completely inhibited by a 2450‐MHz microwave heating for 2 or more minutes in vitro. The population density of P. citrinum in surface wounds of fruit treated with microwave treatment for 2–3 min was significantly lower than that of controls. When tested on jujube fruit, antagonistic yeast or microwave treatment, as stand‐alone treatment, the disease incidence of infected wounds was reduced from 100% to 45.0% and 36.0%, and lesion diameters were reduced from 1.92 cm to 1.50 cm and 1.38 cm, respectively. However, in fruit treated with a combination of Metschnikowia pulcherrima and microwave treatment, the disease incidence of infected wounds and lesion diameters was only 21.0% and 1.00 cm, respectively. The natural decay incidence on jujube fruit treated with the combination of microwave treatment and M. pulcherrima was 6.2% after storage at 2 ± 1°C for 45 days and at 22°C for 7 days. None of the treatments impaired quality parameters of fruits. Thus, the combination of microwave treatment and M. pulcherrima could provide an alternative to synthetic fungicides for controlling postharvest blue mould rot of jujube fruit.     

Characterization of blackleg and soft rot from potato in northern Thailand

Blackleg and soft rot of potato cause economic loss through reduced yield and quality. The causal agents of bacterial blackleg and soft rot of potato were identified based on biological data and sequence analyses of the 16S rDNA gene. Between 2016 and 2018, diseased potato stems and tubers were collected in Chai Prakan District, Chiang Mai Province, and Chiang Khum District, Pa Yao Province. The symptoms included black stem lesions, soft rot on tubers, wilting, break down of the stem vascular ring and foliar yellowing. Of 13 bacterial isolates, five were identified as Pectobacterium carotovorum subsp. brasiliense, four‐Dickeya dadantii, two‐Pseudomonas putida and two‐Bacillus altitudinis. Pathogenicity tests of P. carotovorum subsp. brasiliense and D. dadantii resulted in lower leaves turning yellow and wilting followed by blackleg symptoms on lower stems and maceration of tuber tissue. Symptoms caused by P. putida were yellowing and wilting of leaves. B. altitudinis caused yellowing of the lower leaves and wilting followed by drying of leaf tissue. This is a first report of these bacterial pathogens causing blackleg and soft rot of potato in Thailand.     

Postharvest Fruit rot of Persimmon (Diospyros kaki) in Spain Caused by Lasiodiplodia theobromae and Neofusicoccum spp.

Three different fungi (isolates IVIA QCV‐1, IVIA QCV‐3 and IVIA QCV‐4) were isolated as potential causal agents of postharvest decay losses observed on sweet persimmons (Diospyros kaki L.) cv. ‘Rojo Brillante’ from commercial packinghouses in the Valencia area (Spain). Disease symptoms were irregular brownish and soft lesions mainly located under and surrounding the fruit calyx (stem‐end) that expanded rapidly at room temperature and turned to dark brown or black colour producing apparent and in some cases abundant white to grey mycelium. The identification of Lasiodiplodia theobromae, Neofusicoccum mediterraneum and Neofusicoccum luteum by macroscopic and microscopic morphological observations was confirmed with the amplification and subsequent sequencing of the ribosomal DNA ITS2 region. Representative nucleotide sequences were deposited in GenBank. Pathogenicity of all three isolates was demonstrated by fulfilling Koch's postulates.     

Interactions Between Rhizoctonia Root Rot and the Foliar Common Bean Diseases Anthracnose and Rust

The effects of co‐inoculation of Rhizoctonia solani and Colletotrichum lindemuthianum or Uromyces appendiculatus at different inoculum levels were studied on the disease dynamics and on the growth of bean plants under greenhouse conditions. Bean seeds were sown in R. solani‐infested soil. Additional experiments in which seedlings were transplanted to infested soil were also carried out. Conidial suspensions of C. lindemuthianum or uredospores of U. appendiculatus were inoculated onto leaves at plant developmental stages V2 and V3, respectively. Interactions between root rot and the aerial diseases were observed depending on the inoculum levels and on the timing of R. solani inoculation. Anthracnose severity tended to be higher on R. solani‐infected plants. Conversely, R. solani infection significantly reduced diameter of pustules and rust severity. When seedlings were transplanted to soil infested with low levels of R. solani, root rot severity and density of R. solani in the soil were magnified at high levels of C. lindemuthianum or U. appendiculatus. In these experiments, a synergistic interaction between root rot and anthracnose was observed to affect the plant dry weight. Antagonistic effects on the plant dry weight were found for the combination root rot/rust only when seeds were sown in infested soil.     

Pythium aphanidermatum Causing Pythium Rot on Lampranthus spectabilis in Korea

Pythium aphanidermatum was found to be associated with rotting of leaves and stems in Lampranthus spectabilis in the summers of 2013 and 2014. Infected plants were initially characterized by water‐soaked and discoloured tissue, which are soon covered with cottony aerial hyphae. Subsequently, infected tissues wilted, leaves and stems appeared desiccated, and infected plants died. Pathogenicity of a representative isolate was proved, fulfilling Koch's postulates. Sequence of the internal transcribed spacer region of ribosomal DNA from the isolate shared 100% identity with that of P. aphanidermatum. To our knowledge, this is the first report of P. aphanidermatum causing leaf and stem rot on L. spectabilis in Korea.     

Carvacrol and eugenol effectively inhibit Rhizopus stolonifer and control postharvest soft rot decay in peaches

Aims

This study aimed to investigate the antifungal mechanism of carvacrol and eugenol to inhibit Rhizopus stolonifer and the control of postharvest soft rot decay in peaches.

Methods and Results

To investigate the antifungal mechanism, the effects of carvacrol and eugenol on the mycelium growth, leakages of cytoplasmic contents, mycelium morphology, cell membrane and membrane composition of R. stolonifer were studied. Carvacrol and eugenol both exhibited dose‐dependent antifungal activity against R. stolonifer, carvacrol at a concentration of 2 μl per plant and eugenol at a concentration of 4 μl per plant inhibited fungal growth completely. The two essential oils (EOs) increased cell membrane penetrability and caused the leakage of cytoplasm, nucleic acid and protein content. The observation using scanning electron microscopy and fluorescent microscopy showed modification of the hyphal morphology and breakage of the cell plasma membrane. Decreased ergosterol contents confirmed that the two EOs could destroy the membrane of R. stolonifer. For the in vivo test, the inhibition of soft rot disease and the induction of defence‐related enzymes were investigated. Carvacrol and eugenol significantly reduced the incidence and severity of soft rot decay in inoculated peaches. The best treatments for controlling soft rot decay were obtained at 0·5 μl l^?1 for carvacrol and 1 μl l^?1 for eugenol. The activities of defence‐related enzymes in peaches were also enhanced by fumigation with two EOs.

Conclusion

This study showed that carvacrol and eugenol could effectively inhibit the growth of R. stolonifer in vitro and successfully control the incidence of soft rot decay in honey peaches.

Significance and Impact of the Study

The above findings may be the main antifungal mechanism of carvacrol and eugenol on R. stolonifer. Furthermore, carvacrol and eugenol are helpful for their commercial application on the preservation of fresh fruit.     

Baseline sensitivity and control efficacy of fluazinam against Sclerotinia sclerotiorum in Henan Province,China

Sclerotinia stem rot, caused by Sclerotinia sclerotiorum, is a devastating disease in Henan Province, of the main rapeseed production areas in China. Fluazinam belongs to the broad‐spectrum phenylpyridinamine fungicides, which have high activity in inhibiting the mycelial growth of S. sclerotiorum. In this study, 191 field isolates were obtained from different oilseed rape fields in Henan Province, before being exposed to fluazinam in 2015. The baseline sensitivity of S. sclerotiorum to fluazinam was established. The effective concentration for 50% inhibition of mycelial growth (EC₅₀) ranged from 0.0019 to 0.0337 μg/ml, and the mean EC₅₀ value was 0.0084 ± 0.0055 μg/ml. The range of the frequency distribution was narrow. The results of a cross‐resistance assay revealed no cross‐resistance between fluazinam and carbendazim, dimethachlone, boscalid or fludioxonil. Field efficacy tests showed that the control efficacies of fluazinam (50% WG) applied at 150, 225 and 300 g ai ha^?1 were 67%, 73% and 88%, respectively. In contrast, the control efficacies of boscalid (50% WG) and carbendazim (50% WP) applied at 225 and 1,500 g ai ha^?1 were 71% and 52%, respectively.     

Bacterial iturins mediate biocontrol activity of Bacillus sp. against postharvest pear fruit‐rotting fungi

A potential antagonist, Bacillus sp. LYLB4 isolated from pear fruits, was tested for its antifungal activity against postharvest pear pathogens. LYLB4 had a remarkable antifungal effect on Botryosphaeria dothidea. Although it showed a weak inhibition effect on the growth of Rhizopus stolonifer on potato dextrose agar (PDA) plates, LYLB4 almost completely destroyed R. stolonifer during direct contact in potato dextrose broth (PDB). LYLB4 treatment was able to significantly reduce disease incidence (by 68.9% and 100%, respectively) and lesion diameter (by 68.7% and 100%, respectively) of ring rot caused by B. dothidea and soft rot caused by R. stolonifer in pears. LYLB4 also suppressed several other phytopathogens in vitro, suggesting a broad‐spectrum antagonistic activity against fungal pathogens. 16S rRNA and gyrA sequence analysis indicated that LYLB4 is closely related to B. velezensis. Genome mining indicated that LYLB4 had 11 secondary metabolites encoding clusters, but that the surfactin and fengycin gene clusters may not be functional because of a large deletion. Matrix‐assisted laser desorption ionization‐time of flight mass spectra (MALDI‐TOF‐MS) demonstrated that iturins were the major lipopeptides, and that C₁₆/C₁₇ Bacillomycin D synthesis was stimulated when LYLB4 was co‐cultured with B. dothidea compared to the control. Overall, these results demonstrate that the main biocontrol mechanism adopted by LYLB4 could be through the production of toxic metabolites and direct contact with pathogens.     

Postharvest disease control in mangoes using high humidity hot air and fungicide treatments

The disease control efficacy of quarantine heat treatments developed for fruit fly disinfestation in mangoes cv. Kensington Pride was evaluated in this study. Heat was applied using high humidity (>95% r.h.) hot air (HHHA) at temperatures ranging from 47–49°C. Anthracnose, caused by Colletotrichum gloeosporioides, was well controlled in mangoes heated to a core temperature of 46°C, 47°C or 48°C for 24, 10 or 8 min respectively, prior to ripening at 23°C for 16 days. Stem end rot, caused by Dothiorella dominicana and Lasiodiplodia theobromae, was not satisfactorily controlled by these treatments. In a subsequent experiment, fruit were immersed in a hot benomyl (0.5 g a.i. litre“¹ at 52°C for 5 min) or unheated prochloraz (0.25 ml a.i. litre¹ at 28°C for 30 s) dip before or after the application of HHHA (core temperature of 47°C for 10 min). During storage at 23°C for 15 days, the incidence of stem end rot was reduced by HHHA alone, although immersion in hot benomyl either before or after HHHA treatment greatly improved stem end rot control. HHHA treatment (core temperature of 46.5°C for 10 min) alone reduced the incidence of anthracnose in mangoes stored at 13°C for 14 days prior to ripening at 22°C, although a combination treatment consisting of HHHA and either hot benomyl or unheated prochloraz gave complete control of anthracnose under these storage conditions. HHHA treatment alone gave no control of stem end rot in mangoes stored at 13°C prior to ripening at 22°C. A supplementary hot benomyl treatment was required for acceptable control of this disease in cool-stored mangoes. The development of yellow skin colour in fruit was accelerated by HHHA treatment.     

The Lactoperoxidase System: a Natural Biochemical Biocontrol Agent for Pre‐ and Postharvest Applications

Controlling pests in pre‐ and postharvest crops using natural and low‐impact products is a major challenge. The lactoperoxidase system is an enzymatic system that exists in all external secretions in mammals and is part of the non‐immune system. We tested its efficacy in in vitro microplates on Phytophthora infestans, Penicillium digitatum, Penicillium italicum, Penicillium expansum and Botrytis cinerea to determine the most suitable concentrations for use. Then, we verified its efficacy in planta under controlled conditions. Solutions prepared with 5.4 mm iodide and 1.2 mm thiocyanate and diluted threefold inhibited pathogen growth in vitro by 63–100%. Twofold‐diluted solutions protected potato plants against P. infestans by 60–74% under controlled conditions. Undiluted solution inhibited orange's and apple's postharvest pathogens in curative application with efficacy levels ranging between 84 and 95% in orange and between 63 and 74% in apple. 1.5‐fold concentrated solutions inhibited postharvest pathogens of apple in curative application with efficacy levels ranging between 84 and 92%. Our results also show that the oxidative stress response of fruit following wounding could interfere with ion efficiency. Our tests demonstrate for the first time that this biochemical method is as efficient as a conventional synthetic chemical method under controlled conditions.     

The complete chloroplast and mitochondrial genomes of the diatom Nitzschia palea (Bacillariophyceae) demonstrate high sequence similarity to the endosymbiont organelles of the dinotom Durinskia baltica

Nitzschia palea is a common freshwater diatom used as a bioindicator because of its tolerance of polluted waterways. There is also evidence it may be the tertiary endosymbiont within the “dinotom” dinoflagellate Durinskia baltica. A putative strain of N. palea was collected from a pond on the University of Virginia's College at Wise campus and cultured. For initial identification, three markers were sequenced—nuclear 18S rDNA, the chloroplast 23S rDNA, and rbcL. Morphological characteristics were determined using light and scanning electron microscopy; based on these observations the cells were identified as N. palea and named strain “Wise.” DNA from N. palea was deep sequenced and the chloroplast and mitochondrial genomes assembled. Single gene phylogenies grouped N. palea—Wise within a clearly defined N. palea clade and showed it was most closely related to the strain “SpainA3.” The chloroplast genome of N. palea is 119,447 bp with a quadripartite structure, 135 protein‐coding, 28 tRNA, and 3 rRNA genes. The mitochondrial genome is 37,754 bp with a single repeat region as found in other diatom chondriomes, 37 protein‐coding, 23 tRNA, and 2 rRNA genes. The chloroplast genomes of N. palea and D. baltica have identical gene content, synteny, and a 92.7% pair‐wise sequence similarity with most differences occurring in intergenic regions. The N. palea mitochondrial genome and D. baltica's endosymbiont mitochondrial genome also have identical gene content and order with a sequence similarity of 90.7%. Genome‐based phylogenies demonstrated that D. baltica is more similar to N. palea than any other diatom sequence currently available. These data provide the genome sequences of two organelles for a widespread diatom and show they are very similar to those of Durinskia baltica's endosymbiont.     

‘Cylindrocarpon’ and Ilyonectria Species Causing Root and Crown Rot Disease of Potted Laurustinus Plants in Italy

During the 2012 and 2013 growing seasons, a disease was detected on potted laurustinus (Viburnum tinus) plants in two nurseries located in the Catania province (eastern Sicily, Italy). ‘Cylindrocarpon’‐like species were consistently recovered from crown rot and stem rot tissues. Based on morphological characteristics, DNA sequencing and phylogenetic analysis of β‐tubulin (TUB), histone H3 (HIS3) and translation elongation factor‐1α (TEF‐1 α) gene sequences, the fungi associated with symptomatic tissues were identified as ‘Cylindrocarpon’ pauciseptatum, Ilyonectria novozelandica and I. torresensis. Koch's postulates were fulfilled by pathogenicity tests carried out on potted V. tinus cuttings. To our knowledge, this is the first report worldwide of ‘C.’ pauciseptatum, I. novozelandica and I. torresensis causing disease on V. tinus.