scholarly journals First report of root rot caused by (Pythium dissotocum) on hydroponically grown collard greens (Brassica oleracea var. acephala)

Plant Disease ◽  
2021 ◽  
Author(s):  
Noah Carr Luecke ◽  
Kerri Crawford ◽  
Hanane Stanghellini ◽  
Alyssa Burkhard ◽  
Steve Koike

Collards (Brassica oleracea var. acephala) are grown throughout the United States. Hydroponic greens are more common now due to technological advances lowering the cost and increasing the efficacy of production. In January 2021, a 325 m2 indoor hydroponic farm opened to provide fresh produce for a school in Los Angeles County, CA. Three week old collard seedlings were purchased from a local nursery, rinsed of their rooting media, and transplanted into deep water culture beds (1.2 m x 2.5 m x 0.3 m). Two weeks later, symptoms including plant stunting, chlorosis, leaf curling and wilting, and brown necrotic roots appeared. By and by 80-100% of usable plants were lost to disease. Symptomatic roots were plated on corn meal agar (CMA) amended with 2 ml of 25% lactic acid and CMA amended with pimaricin, ampicillin, rifampicin, and pentachloronitrobenzene (PARP) (Kannwischer et al. 1978). After 2 days a single colony type emerged on PARP but no growth occurred on acidified CMA. Representative isolates were transferred to CMA and to filtered (0.02 µm) soil extract solution with boiled grass blades (Martin 1992), both of which were incubated at 22 C and ambient light conditions. On CMA, isolates produced coenocytic mycelium with minimal aerial hyphae. After 24 h in soil extract, isolates developed filamentous sporangia, elongated discharge tubes with slightly inflated tips, and zoospores. Oospores were not observed. Pathogenicity was confirmed by soaking the roots of five day old collard seedlings in beakers containing zoospores (1 x 102 zoospores/ml) in filtered soil extract. Four isolates were tested on 15 seedlings each. After 24 h at 22 C in ambient light conditions, plants were transferred to new beakers with roots placed on filter paper at the bottom and saturated with sterile distilled water. Three days after this transfer, leaves on all plants turned chlorotic and roots developed brown lesions from which morphologically identical colonies were isolated. Control plants, soaked in filtered soil extract, developed no root or foliar symptoms. To molecularly identify the collard isolates, DNA was extracted from mycelial original and re-isolated isolates and was amplified by PCR using mitochondrial primers for the cytochrome oxidase I (COI) gene (Robideau et al. 2011) and the cytochrome oxidase II (COX2) gene (Martin 2000). The only species that matched both loci from the original and re-isolated isolates with a high percent identity was Pythium dissotocum. The COI locus from the original isolate (MZ027311) matched P. dissotocum with 99% identity and with 332/334 base pairs matching the isolate with Sequence ID MT981134.1. From the re-isolated isolate (MZ027313), the COIequence perfectly matched 657/657 base pairs of P. dissotocum (Sequence ID MT981147.1). The COX2 locus from the original isolate (MZ027312) matched P. dissotocum (Sequence ID MG719859.1) with a 99% identity and 517/518 matching base pairs and the re-isolated isolate (MZ027314) perfectly matched P. dissotocum (Sequence ID MG719859.1) with 515/515 matching base pairs. Based on these molecular and morphological data, the isolates were identified as Pythium dissotocum. To our knowledge, this is the first report of P. dissotocum causing root rot on collards. At this same facility, P. dissotocum was also confirmed as the cause of declining bean (Phaseolus vulgaris) plants. As hydroponics will be necessary to feed a growing population – especially in urban areas -- and because leafy greens are a main crop of the hydroponics industry, we anticipate this issue may become common. Hydroponic systems are highly conducive to the persistence of Oomycetes and a record of infection and plan of action will be necessary to preserve crop health.

Plant Disease ◽  
2000 ◽  
Vol 84 (5) ◽  
pp. 593-593 ◽  
Author(s):  
G. S. Karaoglanidis ◽  
D. A. Karadimos ◽  
K. Klonari

A severe rot of sugar beet roots was observed in the Amyndeon area of Greece during summer 1998. Infected plants initially showed a temporary wilt, which became permanent, and finally died. Slightly diseased roots showed necrotic spots toward the base, whereas more heavily diseased roots showed a more extensive wet rot that extended upward. Feeder roots also were infected and reduced in number because of decay. Rotted tissue was brown with a distinguishing black margin. In most of the isolations, carried out on potato dextrose agar (PDA), the pathogen obtained was identified as Phytophthora cryptogea Pethybr. & Lafferty Mycelium consisted of fairly uniform, fine hyphae that showed a slightly floral growth pattern. In autoclaved soil-extract medium, chains or clusters of hyphal swellings (average 12 µm diameter) formed. Sporangia were not produced on solid media but were abundant in soil-extract medium. Sporangia were oval to obpyriform in shape, nonpapillate with rounded bases, and varied in size (39 to 80 × 24 to 40 µm). Oospores were plerotic, thick-walled, and averaged 25 µm in diameter. The isolated pathogen, cultured on PDA, could not grow at all at 36°C. The closely related species P. drechsleri Tucker has been reported to cause similar root rot symptoms on sugar beet (3). However, P. drechsleri grows well at 36°C, while P. cryptogea cannot grow at this temperature; this is the major distinguishing feature that separates the two species (1). To test the pathogenicity of the organism, surface-sterilized sugar beet roots (cv. Rizor) were inoculated with 5-mm-diameter PDA plugs containing actively growing mycelium. Sterile PDA plugs were used to inoculate control sugar beet roots. Inoculated roots were kept at 27°C in the dark for 10 days. Extensive decay of inoculated roots developed, similar to decay observed in the field, whereas control roots showed no decay. P. cryptogea was reisolated from rotted tissues. This pathogen has been recognized previously as a cause of root rot of sugar beet in Japan (1) and Wyoming (2). This is the first report of Phytophthora root rot of sugar beet in Greece. References: (1) D. C. Erwin and O. K. Ribeiro. 1996. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN. (2) P. C. Vincelli et. al. Plant Dis. 74:614, 1990. (3) E. D. Whitnew and J. E. Duffus, eds. 1986. Compendium of Beet Diseases and Insects. The American Phytopathological Society, St. Paul, MN.


Plant Disease ◽  
2010 ◽  
Vol 94 (7) ◽  
pp. 917-917
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
M. L. Gullino

Edgeworthia papyrifera, Oriental paperbush, is a deciduous flowering shrub becoming increasingly popular because of its clove-like perfumed flowers appearing in late winter-early spring. During August of 2009 in a commercial nursery close to Maggiore Lake (Verbano-Cusio-Ossola Province) in northwest Italy, 2-year-old plants of E. papyrifera showed extensive chlorosis and root rot. Twigs wilted and died, dropping leaves in some cases. Most frequently, wilted leaves persisted on stems. At the soil level, dark brown-to-black water-soaked lesions formed and coalesced, girdling the stem. All of the crown and root system was affected. Infected plants died within 14 days of the appearance of symptoms. Disease was widespread and severe, affecting 90 of the 100 plants present. After disinfestation for 1 min in a solution containing 1% NaOCl, rotting root and collar pieces of E. papyrifera consistently produced a Phytophthora-like organism when plated on a medium selective for oomycetes (3). The pathogen was identified morphologically as Phytophthora nicotianae (= P. parasitica) (2). On V8 agar, coenocytic hyphae, 4 to 8 μm in diameter, formed fluffy, aerial colonies and spherical, intercalary chlamydospores, 21.0 to 36.5 (average 26.7) μm in diameter. Colonies grew well at 35°C and stopped growing at 40°C. Sporangia were produced by growing a pure hyphal-tip culture in a diluted, sterilized soil-extract. Sporangia were borne singly, laterally attached to the sporangiophore, were noncaducous, spherical to ovoid, papillate, and measured 28.6 to 55.2 × 22.4 to 45.1 (average 42.4 × 34.6) μm, length/breadth ratio (1.1:1)-1.2:1-(1.3:1). Papillae measured 3.1 to 7.6 (average 4.6) μm. The internal transcribed spacer (ITS) region of rDNA of a single isolate was amplified with primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 839-bp segment showed 99% homology with the sequence of P. nicotianae (No. AJ854296). The sequence has been assigned the GenBank No. GU353341. Pathogenicity of isolates Edg.1 and Edg.2 obtained, respectively, from the root and collar of an infected plant was confirmed by inoculating 1-year-old plants of E. papyrifera. Both strains were grown for 15 days on a mixture of 70:30 wheat/hemp kernels, and 4 g/liter of the inoculum was mixed into a substrate containing sphagnum peat moss/pumice/pine bark/clay (50:20:20:10 vol/vol). One plant per 3-liter pot was transplanted into the substrate and constituted the experimental unit. Five plants were used for each test strain and noninoculated control treatment; the trial was repeated once. All plants were kept in a greenhouse at 25 to 28°C. Plants inoculated with Edg.1 and Edg.2 developed chlorosis and root rot 18 and 14 days after the inoculation, respectively, and wilt rapidly followed. Control plants remained symptomless. P. nicotianae was consistently reisolated from inoculated plants. To our knowledge, this is the first report of P. nicotianae on E. papyrifera in Italy as well as worldwide. The current economic importance of the disease is minor due to the limited number of farms that grow this crop in Italy, although spread could increase as the popularity of plantings expand. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phtytopathological Society, St Paul, MN, 1996. (3) H. Masago et al. Phytopathology 67:425, 1977.


Plant Disease ◽  
1998 ◽  
Vol 82 (11) ◽  
pp. 1282-1282 ◽  
Author(s):  
B. J. Aegerter ◽  
R. M. Davis

Damping-off of broccoli (Brassica oleracea var. italica) and cauliflower (B. oleracea var. botrytis) seedlings occurred in several greenhouses in Fresno, CA, in 1997. Symptoms included wilting and root and stem rot. Pythium polymastum was consistently isolated from symptomatic tissues placed on corn meal agar amended with 10 ppm pimaricin, 250 ppm ampicillin, 10 ppm rifampicin, and 25 ppm pentachloronitro-benzene. On grass leaves in water, the fungus produced numerous aplerotic oospores in oogonia 43 to 50 μm in diameter (average 46 μm) with spines about 7 μm long. Spherical sporangia were only rarely observed. In the greenhouse, 4-week-old broccoli and cauliflower seedlings were transplanted into potting mix amended with a colonized vermiculite/rye/V8 juice medium to produce approximately 2,500 CFUs per gram of potting medium. Control plants were transplanted into noninfested potting mix. There were six replicate pots per treatment and three plants per pot. After 12 days, the potting mix was gently washed from the roots and the seedlings were dried and weighed. Symptoms on inoculated plants included wilting, severe root rot, black streaks on the lower stems, and death. The fungus was recovered from symptomatic tissues. There were no symptoms on the control plants. Infection by P. polymastum reduced dry weights of surviving broccoli and cauliflower seedlings by 82 and 58%, respectively. Similar results were obtained in a second experiment. This fungus was previously characterized as a pathogen of both cultivated and wild crucifers in Canada (1). This is the first report of P. polymastum in California. Reference: (1) T. C. Vanterpool. Can. J. Bot. 52:1205, 1974.


Plant Disease ◽  
2021 ◽  
Author(s):  
PengYing MEI ◽  
Xuhong Song ◽  
Zhiyu Zhu ◽  
Longyun Li

Chongqing coptis (Coptis chinensis Franchet) industry produces more than 60% of the Chinese coptis crop, and has been exported to many countries and regions. Since 2008, root rot has become a serious and widespread disease on coptis plants in Shizhu county with an average incidence of 40%, and yield losses up to 67%. Symptomatic coptis plants showed stunted growth, with the fibrous roots and main roots having brown or black, rotten, necrotic lesions. To our knowledge, Fusarium solani, F. carminascens, F. oxysporum and F. tricinctum have been previously reported as pathogens of coptis root rot (Luo et al. 2014; Cheng et al. 2020; Wu et al. 2020), but non Fusarium pathogens has not been reported yet. In order to identify new pathogens, 33 diseased roots were collected from Shizhu (30°18'N, 108°30'E) in October 2019. Small samples (0.5 cm in length) were cut from the border between diseased and healthy tissue, and then put on PDA after surface sterilization. Cultures were incubated at 25°C in dark until fungal colonies were observed. After subculturing for 3 times, 3 out of 21 isolates yielded a similar type of fungal colony. White, aerial, fluffy mycelium were formed and reached 8.3 cm diameter within 7 days, and dark pigmentation developed in the centre. Colonies turned to gray with age, and abundant dark brown pycnidia and black stromata were formed at maturity. Alpha conidia were aseptate, hyaline, fusiform to ellipsoidal, often biguttulate, measuring (6.0-8.5)×(2.0-3.0) μm. Beta conidia were aseptate, hyaline, linear to hooked, measuring (18-30)×(1.0-1.5) μm (Figure S1). For further identification, a multigene phylogenetic analysis was carried out. The internal transcribed spacer (ITS), translation elongation factor 1ɑ (tef1-ɑ), histone H3 (his3), calmodulin (cal), and β-tubulin (tub2) gene regions were amplified with ITS1/ITS4, EF1-728F/EF1-986R, CYLH3F/H3-1b, CAL228F/CAL737R, T1/Bt2b (White et al. 1990; Glass and Donaldson 1995; Carbone and Kohn 1999; Crous et al. 2004). GenBank accession numbers of isolate H13 were MT463391 for the ITS region, MT975573 for tef1-ɑ, MT975574 for his3, MT975575 for cal, and MT975576 for tub2. BLAST results showed the ITS, tef1-ɑ, his3, cal and tub2 sequences revealed 99.82% (553/554 base pairs), 100% (347/347 base pairs), 100% (474/474 base pairs), 99.39% (486/489 base pairs), and 99.14% (803/810 base pairs) homology respectively with those of Diaporthe eres (MN816416.1, KU557616.1, KC343564.1, KU557595.1, and KY569366.1). Thus, H13 were identified as D. eres based on its morphological and molecular characteristics. Pathogenicity of D. eres in coptis was investigated using the H13 isolate (1 of the 3 isolates). The roots of 10 healthy 2-year-old coptis plants were individually inoculated with 5 ml of a 106 conidia/mL conidial suspension and sterilized water was used to mock inoculate. Thirty days after inoculation, most of the inoculated coptis roots showed dark brown and rotten root, similar to those observed in the field, whereas mock inoculated roots showed healthy. D. eres was recovered from symptomatic roots and identified based on morphology. To our knowledge, this is the first report of D. eres causing root rot of coptis not only in China but anywhere in the world.


2019 ◽  
Vol 18 (3) ◽  
pp. 209
Author(s):  
Muhammad Dailami ◽  
Darma Santi ◽  
. Murtihapsari ◽  
Hermawaty Abubakar ◽  
Abdul Hamid A. Toha

Cirrhilabrus cf. ryukyuensis from Cenderawasih Bay and Raja Ampat have different colormorphs with Cirrhilabrus ryukyuensis. This study was conducted to determine the phylogenetic relationship, evolution and genetic diversity of Cirrhilabrus cf. ryukyuensis based on Cytochrome oxidase sub unit 1 (COI) gene. Mitochondrial DNA extracted by using chelex 10%. FISH-BCH and BCL primers were used to amplify COI gene fragment and sequencing. Data analysis conducted by using software Sequencher 4.1 and MEGA 5. COI genes from six samples of C. cf rykyuensis from Cenderawasih Bay and Raja Ampat have total 613 base pairs. The average compositions of nucleotides are T 32.8%, C 26.3%, A 23.2% and G 17.7%. There is six positions of polymorphism that caused by transition. Conversion of nucleotides to amino acid sequence resulted in 204 amino acid with the same sequence. The phylogenetic tree from six sample showed that all in one clade, but different clade from Cirrhilabrus cyanopleura from GenBankAbstrakCirrhilabrus cf. ryukyuensis asal Teluk Cenderawasih dan Raja Ampat secara morfologi memiliki perbedaan warna dengan Cirrhilabrus ryukyuensis. Penelitian ini bertujuan menentukan hubungan filogenetik, evolusi, dan keragaman genetik  Cirrhilabrus cf. ryukyuensis berdasarkan gen Sitokrom Oksidase sub Unit I (COI). Genom DNA mitokondria diekstrak menggunakan larutan chelex 10%. Primer FISH-BCH dan BCL digunakan untuk mengamplifikasi fragmen Gen COI dan sekuensing. Analisis data dilakukan dengan menggunakan perangkat lunak Sequencher 4.1 dan MEGA 5. Gen COI dari enam sampel C. cf. rykyuensis asal Teluk Cenderawasih dan Raja Ampat, memiliki panjang basa men-capai 613 bp. Komposisi rata-rata nukleotida T 32,8%, C 26,3%, A 23,2% dan G 17,7%. Terdapat enam posisi poli-morfisme yang kesemuanya disebabkan oleh mutasi transition. Hasil penerjemahan nukleotida, diperoleh asam amino dengan panjang 204 asam amino dengan urutan yang sama. Pohon filogenetik menunjukkan keenam sampel berada dalam satu clade yang sama dan berbeda dari clade Cirrhilabrus cyanopleura data Gene Bank.


Plant Disease ◽  
2021 ◽  
Author(s):  
Jun-shan Qi ◽  
Bo Zhang ◽  
Li-guo Ma ◽  
Guoping Ma ◽  
Shu-jun Qin ◽  
...  

Head lettuce (Lactuca sativa L.) is an important crop for fresh consumption in China. In Shandong Province, head lettuce is planted in spring and in autumn each year. Because of the on-and-off rain for three weeks, head lettuce plants planted directly into the field in Jiyang City, in July 2017, 20% of the plants rapidly showed symptoms of rotting, water-soaked lesions on roots and stem bases, and then death. The diseased plants first appeared in low-lying areas prone to water accumulation. One-millimeter pieces were excised from water-soaked roots and stem bases, dipped in a 0.2% calcium hypochlorite solution for 10 min, then placed on V8 medium, and incubated in the dark at 28°C for 5 d. Two Pythium-like strains were isolated from the roots and stems. The isolates transferred to CMA and grown for 7 d, and the morphological characteristics of the two isolates on corn meal agar (CMA) were white with dense, cottony, aerial and well-branched mycelia. The two isolates produced sporangia, oogonia, antheridia and oospores. Most of the sporangia were lobate. The oogonia were smooth, nearly globose and terminal. Oospores were globose, smooth and aplerotic. The average dimensions of 50 oogonia and oospores respectively ranged from 19.5 to 25.2 (av. 23.1) µm and 17.8 to 22.3 (av. 19.9) µm. The antheridia were broadly sac-shaped. The isolates morphological characteristics were consistent with P. aphanidermatum (van der Plaats-Niterink, 1981). The COI gene and ITS region of the rDNA were amplified and sequenced using primers FM55/FM52R (Long et al. 2012) and ITS1/ITS4 (White et al. 1990), respectively. The two aligned COI sequences were identical for both isolates, as were the two ITS sequences. BLASTn analysis of the 1,133-bp COI sequence (accession no. MT952703) resulted in a 100% identity with accession number AY129164 from Lactuca sativa, which belongs to P. aphanidermatum, and the 808-bp ITS sequence (accession no. MT921597) showed a 99% identity with Genbank accession number HQ643442 belonging to P. aphanidermatum. Koch’s postulates were conducted by first soaking corn kernels for 24 h in water, and then autoclaving for 2 h at 121˚C. Isolate SDHL-1 was grown on CMA for 10 days, after which agar plugs were transferred to the sterilized corn kernels and incubated at 28℃ for approximately 15 d, until the corn kernels were covered in white hyphae. Ten healthy head lettuce plants were transplanted into a sterilized loam potting soil artificially infested with the corn inoculum (3 g inoculum per 100 g loam mixture). Inoculated plants and noninoculated controls were maintained in a greenhouse at 28°C and 100% relative humidity with a 12-h photoperiod; the experiment was repeated once. All twenty inoculated plants exhibited symptoms within one week similar to those observed. Pythium aphanidermatum was recovered only from the water-soaked roots and stem bases of inoculated plants and the re-isolated cultures again identified based on morphological characteristics and sequencing of the ITS and COI genes. No symptoms were observed on the control plants. Sclerotinia sclerotiorum is reported to cause stem base rot of L. sativa in China (Zhou et al. 2011). To our knowledge, however, this is the first report of root rot of head lettuce caused by Pythium aphanidermatum. Identification of the pathogen will assist in devising strategies to reduce yield loss.


2019 ◽  
Vol 21 (1) ◽  
Author(s):  
DEWI IMELDA ROESMA ◽  
DJONG HON TJONG ◽  
DYTA RABBANI AIDIL

Abstract. Roesma DI, Tjong DH, Aidil DR. 2020. Phylogenetic analysis of transparent gobies in three Sumatran lakes, inferred from mitochondrial Cytochrome Oxidase I (COI) gene. Biodiversitas 21: 43-48. The transparent gobies fish found in three lakes in Sumatra island is known as Rinuak fish (in Maninjau Lake and Singkarak Lake, West Sumatra, Indonesia) or Badar fish (in Siais Lake, North Sumatra, Indonesia), and are morphologically very similar to the Gobiopterus brachypterus. The phylogenetic study was carried out by analyzing 619 base pairs of the mitochondrial DNA cytochrome oxidase subunit I (COI) gene in 12 fish individuals from the three lakes. Rinuak and Badar fish in three populations have four haplotypes. The sequence divergences in and between populations are very low (0.0-0.5%). This value indicates that Rinuak and Badar fish are the same species with low genetic diversity. The phylogenetic tree illustrates that this fish belongs to the group of Gobiidae and a sister taxon from G. brachypterus.


Plant Disease ◽  
2012 ◽  
Vol 96 (4) ◽  
pp. 591-591
Author(s):  
M. Vargas ◽  
C. Loyola ◽  
N. Zapata ◽  
V. Rivera ◽  
G. Secor ◽  
...  

Chicory (Cichorium intybus L. var sativum Bisch.), a relatively new high-value crop in Chile, was introduced for commercial production of inulin. Inulins are polysaccharides extracted from chicory tap roots that are used in processed foods because of their beneficial gastrointestinal properties. Approximately 3,000 ha of chicory are grown for local processing in the BioBio Region near Chillan in south central Chile. Recently, a severe rot of 1 to 3% of mature roots in the field and after harvest has been observed in most fields, which caused yield and quality losses. Typical symptoms include a brown discoloration and a soft, watery decay of the root. Tissue pieces from symptomatic roots were placed on water agar and clarified V8 juice agar medium amended with antibiotics (1) for isolation of the causal pathogen. A Phytopthora sp. had been consistently isolated from root lesions, and axenic cultures were obtained using single-hypha transfers. The species was provisionally identified as Phytopthora cryptogea (Pethybridge and Lafferty, 1919) on the basis of morphological and cultural characteristics (1). Mycelia grew between 5 and 30°C with optimal growth at 20 to 25°C and no growth at 35°C. All isolates produced hyphal swellings and nonpapillate, persistent, internally proliferating, and ovoid to obpyriform sporangia with mean dimensions of 45 × 31 μm in sterile soil extract. The isolates were of A1 mating type because they produced oospores only when paired with reference isolates of P. cinnamomi A2 on clarified V8 juice agar amended with thiamine, tryptophan, and β-sitosterol (1) after 20 days at 20°C in the dark. On the basis of morphological and sequence data from cytochrome c oxidase subunit 1 and 2, internal transcribed spacer 2, and β-tubulin (GenBank Accession Nos. JQ037796 to JQ037798, respectively), the pathogen was identified as P. cryptogea. Pathogenicity tests were conducted using three isolates of P. cryptogea by placing a 7-mm-diameter disk from a 1-week-old V8 agar culture on 10 wounded and nonwounded healthy chicory roots (2). Control roots were mock inoculated with agar plugs. The inoculated roots were incubated at 20°C in a moist chamber. Root rot symptoms, identical to those observed both in field and storage, developed after 4 to 6 days only on wounded sites inoculated with the pathogen, and P. cryptogea was reisolated from these inoculated plants. Mock-inoculated roots remained healthy. This experiment was completed twice and similar results were obtained. To our knowledge, this is the first report of Phytophthora root rot of chicory caused by P. cryptogea in Chile. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (2) M. E. Stanghellini and W. C. Kronland. Plant Dis. 66:262, 1982.


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