First Report of Black Spot Caused by Penicillium citreosulfuratum on Saffron in Chongming Island

  • Saffron (Crocus sativus L.) is a world-famous source of dye and spices and an important medicinal plant, which is cultivated on a large scale on Chongming Island (31.62° N, 121.39° E) in Shanghai, China. In August 2020, a survey of saffron was conducted in this area, and black spots were observed on about 10% of plants.
  • Characteristics of the disease were: The bottom of the corm was darkened in the groove and scattered black spots could be observed after peeling off the membranous scale leaves. The junction of lesions and healthy parts was light brown. As the lesions expanded, approximately 80% of the surface of the corm became dark brown to bluish gray.
  • The inside of the corm was also necrotic. In order to isolate the pathogen, ten diseased corms with typical symptoms were selected. All corms were first treated with 75% ethanol for 30 s, 0.1% HgCl2 for 5 min, and then rinsed with sterile water 5 times. Next, tissue pieces (5 mm × 5 mm) at the margin of lesions were cut out and placed on the potato dextrose agar (PDA) medium. After incubating at 28°C for 5 days, fungi were separated and purified by using the hyphal-tip technique.
  • A total of six pure cultures with different colony morphologies were obtained, of which only the isolate MF3 was present in ten diseased corms. The isolate MF3 was inoculated on the PDA and cultured at 28°C for 10 days and characteristics of the fungus colonies were: colonies sub-circular, reaching 28 to 30 mm diam, from above rough, dense, fluffy, blue-gray, with some white spots and central point raised, and outer margin form an irregular, narrow, white ring; from below, yellow with light-green. The hyphae were slender, with many septa.
  • The conidiophores were typically smooth walled, short, and slender and either monoverticillate with very short stipes or as irregularly biverticillate. Phialides were ampulliform, 5 to 10 per metula, 5 to 8 × 2 to 3 µm. Conidia were smooth and globose, and ranged in diameter from 1.4 to 1.7 µm (n=50).
  • Molecular identification of the fungus was made by PCR amplification of the internal transcribed spacer (ITS) region of rDNA and β-tubulin gene using primers ITS1/ITS4 (White et al. 1990), TUB2Fd/TUB4Rd (Aveskamp et al. 2009) respectively.
  • The ITS (MW881446) and β-tubulin (MW911464) sequences of the fungus were similar to the ITS (MN592912) and β-tubulin (KY469126) sequences of the epitype of Penicillium citreosulfuratum with 99.81% and 99.56% identity, respectively.
  • According to the morphological and molecular characterization, the isolate MF3 was identified as P. citreosulfuratum (Visagie et al. 2016). For pathogenicity testing, the fungus was grown on PDA and incubated at 28°C for 5 days.
  • Then mycelial plugs (5 mm diam.) were inoculated on the scalpel incision square wounds of surface-disinfected corms and mock-inoculated corms received only PDA plugs. Corms were placed in sterile plastic bottles and observed after culturing at 28°C for 21 days. Each treatment had three replicates and the experiment was repeated twice.
  • The results showed that corms inoculated with P. citreosulfuratum developed diseased with similar symptoms as in the field. No disease symptoms were observed on control corms. Re-isolations were performed from inoculated corms, and all re-isolated fungi were confirmed as P. citreosulfuratum, verifying the fungus as the pathogen based on Koch’s postulates.
  • To our knowledge, this is the first report of the pathogen causing black spot disease of saffron. Although the disease is not fatal to saffron, to a certain extent it will cause a reduction in the production of the crop. In addition, Empty Bottles Plastic this pathogen has not been reported to be pathogentic to other plant species.

First Report of Spot Blotch Caused by Bipolaris sorokiniana on Winter Rye in Kentucky.

Brown bottle
Brown bottle

The Kentucky distilling industry ranks as one of the state’s largest industries and continues to expand. In 2017, the Kentucky distilling industry was responsible for approximately $235 million in state and local tax revenues (Coomes and Kornstein, 2019). Rye (Secale cereale L.) grains are a vital component for production of some distilled spirits.

  1. Although winter rye is produced on relatively few hectares in Kentucky currently, a recent initiative has supported expanding production to help meet the growing demand of local distilleries.
  2. University of Kentucky winter rye research field trials were visited in Caldwell and Logan Counties, KY in May 2018, and in Fayette County, KY in May 2019. Leaves were collected that had dark brown, oval to irregular-shaped lesions with definite margins and yellow halos. Symptoms were present on approximately 50% to 80% of the flag leaves, with severity ranging from 5% to 30% of the flag leaf area affected.
  3. Leaves were surface-disinfested by soaking in a 2% NaOCl solution for 1 min and rinsed twice in sterilized water and then placed in a humidity chamber (plastic bag with moist paper towels) at room temperature (approximately 24°C) to induce fungal sporulation.
  4. Seventeen single-spore isolates were obtained and stored at -80°C in 15% glycerol solution. Isolates were grown on potato dextrose agar under 12 h cycles of white light/darkness for 5 days. Colonies were gray to black. Conidia that formed were mostly straight or slightly curved, dark olivaceous brown, 3-7 septate, and 41.0-90.4 × 15.2-29.3 µm.
  5. Based on the symptoms observed on the collected leaves and these morphological characteristics similar to those described by Chang and Hwang (2000) and Manamgoda et al. (2014), the fungus was tentatively identified as Bipolaris sorokiniana (Sorokin) Shoemaker. The sequence of internal transcribed spacer regions (ITS) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were used to identify three isolates (18Bs004, 18Bs111 and 19Bs064) using primer ITS1/ITS4 (White et al. 1990) and GPD1/GPD2 (Berbee et al. 1999), respectively.
  6. The sequences were deposited in GenBank with accession numbers MT457817, MT457818 and MZ066635 for ITS sequences and MZ073644 to MZ073646 for GAPDH sequences. BLAST searches with ITS and GAPDH sequences matched 100% identity (344/344 bp and 515/515 bp for ITS and GAPDH sequences, respectively) to B. sorokiniana (GenBank accession No. MT254731 and MH844813, respectively).
  7. To prove pathogenicity, a conidial suspension (1 × 105 conidia/ml) was used to inoculate 15-day-old cultivar ‘Serafino’ winter rye plants in the greenhouse. Leaves of 8 plants were inoculated with 50 ml of the conidial suspension using a spray bottle. Plants were covered with a transparent plastic bag for 48 h, and symptoms were observed 10 days after inoculation.
  8. Leaf lesions, similar to those described above, were present on all inoculated plants, but no symptoms were observed on non-inoculated control plants. Bipolaris sorokiniana was reisolated from symptomatic leaves and the identity of the pathogen was confirmed based on the morphology previously described.
  9. To our knowledge, this is the first report of spot blotch caused by B. sorokiniana on winter rye in Kentucky, but B. sorokiniana has been reported on rye in the neighboring state of Virginia (Roane 2009).
  10. Kentucky produces approximately 150,000 and 4,000 ha of winter wheat (Triticum aestivum) and winter barley (Hordeum vulgare) annually, respectively, which are both known hosts of B. sorokiniana (Kumar et al. 2002).
  11. An isolate of B. sorokiniana from rye was reported by Ghazvini and Tekauz (2007) to be less virulent on barley differential lines.

Further research is needed to better understand spot blotch distribution, susceptibility in winter rye cultivars, and potential yield and quality loss implications in winter rye production and end use. It is unknown how susceptible various winter rye cultivars grown in Kentucky are to spot blotch.

Nanoengineering of eco-friendly silver nanoparticles using five different plant extracts and development of cost-effective phenol nanosensor

The production of environmentally friendly silver nanoparticles (AgNPs) has aroused the interest of the scientific community due to their wide applications mainly in the field of environmental pollution detection and water quality monitoring. Here, for the first time, five plant leaf extracts were used for the synthesis of AgNPs such as Basil, Geranium, Eucalyptus, Melia, and Ruta by a simple and eco-friendly method.

  • Stable AgNPs were obtained by adding a silver nitrate (AgNO3) solution with the leaves extract as reducers, stabilizers and cappers. Only, within ten minutes of reaction, the yellow mixture changed to brown due to the reduction of Ag+ ions to Ag atoms. The optical, structural, and morphology characteristics of synthesized AgNPs were determined using a full technique like UV-visible spectroscopy, FTIR spectrum, XRD, EDX spectroscopy, and the SEM. Thus, Melia azedarach was found to exhibit smaller nanoparticles (AgNPs-M), which would be interesting for electrochemical application.
  • So, a highly sensitive electrochemical sensor based on AgNPs-M modified GCE for phenol determination in water samples was developed, indicating that the AgNPs-M displayed good electrocatalytic activity. The developed sensor showed good sensing performances: a high sensitivity, a low LOD of 0.42 µM and good stability with a lifetime of about one month, as well as a good selectivity towards BPA and CC (with a deviation less than 10%) especially for nanoplastics analysis in the water contained in plastics bottles.

Duran Bottles 100Ml Plastic coat

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Plastic bottles 250ml supply/waste

WAT4842 Scientific Laboratory Supplies PK2

Flystuff (59-194P) Droso-Plugs Cap for Plastic Bottles

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Disposable Meat Sample Plastic tubes with caps (5 ml) (1000/pk)

MST5-1000 Alpha Diagnostics 1 pk

Corning 1L Square Polycarbonate Storage Bottles with 45mm Caps

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Corning 150mL Square Polycarbonate Storage Bottles with 45mm Caps

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Corning 250mL Square Polycarbonate Storage Bottles with 45mm Caps

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Corning 250mL Easy Grip Polystyrene Storage Bottles with 45mm Caps

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Costar 1L Traditional Style Polystyrene Storage Bottles with 45mm Caps

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Costar 125mL Traditional Style Polystyrene Storage Bottles with 45mm Caps

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Pyrex WM Bottles with Cap and Ring - 250mL

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Pump Sampler Set with Glass Bottles

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LCP EZ-Cut films (50 pieces, with liners)

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Bottle Duran Plastic Coated 5L.

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Pump Sampler Set with 12 x PE Bottles 150ml

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Plastic Bottle, Wide Neck, 125ml, Natural

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Plastic Bottle, Wide Neck, 15ml, Natural

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Plastic Bottle, Wide Neck, 250ml, Natural

B003N-250 Bio Basic 10, 10UNIT

Plastic Bottle, Wide Neck, 30ml, Natural

B003N-30 Bio Basic 10, 10UNIT

Plastic Bottle, Wide Neck, 500ml, Natural

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Plastic Bottle, Wide Neck, Sterile, 125ml, Natural

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Plastic Bottle, Wide Neck, Sterile, 250ml, Natural

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Plastic Bottle, Wide Neck, Sterile, 500ml, Natural

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Plastic Bottle, Wide Neck, Sterile, 70ml, Natural

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Media bottles 500 ml petg

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Bottles Nalgene 500 Ml

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Duran Bottles 1000ml No Cap

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Bottles Cent Pc 500 Ml

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Flat One Piece Moulded Plastic Lid to suit 1 Litre Capacity Bath

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Flat One Piece Moulded Plastic Lid to suit 6 Litre Capacity Bath

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Flat One Piece Moulded Plastic Lid to suit 30 Litre Capacity Bath

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Bottles Amber Hdpe 125 Ml Nn

2004-0004 Scientific Laboratory Supplies PK12

Adapter 250 mL bottles high-cap

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BROWN LOOPED SCREW CAPS WITH "O" RINGS. FITS 0.5ML, 1.5ML AND 2.0ML TUBES

SCO-LP-BR CORNING 500/pk

Bottle Amber Lab Bottles 500ml

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Bottles Wash W/M Ldpe 500 Ml

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4oz Clear Bottles

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Adapter tubes/ bottles/ plates

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Rack For 9 Bottles 2000ml

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Rack For 16 Bottles 1000ml

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Bottles N/M Amber 30 Ml Nalgene

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Mycoprep 75Ml Bottles

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FiveGo sample bottles

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Corning Storage Bottles 250ml

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Corning Storage Bottles 500ml

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Rack For 20 Bottles 500ml

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Rack For 44 Bottles 100ml

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Solution Basin, 55 mL Capacity, White, 50 pieces/CS

42734050-1 Bio-WORLD 50 Piece (s)

Nalgene narrow-mouth boston round bottles HDPE with PP closure bulk pack

312002-0032 Scientific Laboratory Supplies PK50

Honey Jar 375ml with Plastic Cap Sterile

CON7774 Scientific Laboratory Supplies PK100

Pen for Azlon Bottles

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Bottles Boston PC 500mL

BOT0350 Scientific Laboratory Supplies PK12

Bottles Boston PC 1000mL

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Tube Culture With Svl Plastic

B99485 Scientific Laboratory Supplies PK40

Sample Bottles Undosed Sterile Green Cap 1000ml

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Spray bottle with air spray - 50 mL

DD670885 Scientific Laboratory Supplies EACH

60ml HDPE Nalgene Bottles

3121890002 Scientific Laboratory Supplies PK1000

Bottles Powder Clear 250ml

BP250C Scientific Laboratory Supplies PK42

Powder Bottles 30ml Clear

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Reagent Bottles Amber 500ml

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Droso-filler For Bottles

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Heat Extractor with Plastic Knob

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Staining Dish with Cover Plastic

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Pack of 2 suction bottles 100ml

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Sample Bottles Undosed Sterile Green Cap 250ml

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Sample Bottles Undosed Sterile Green Cap 500ml

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Rack For 48 Bottles 50ml

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Sample Bottles Non Sterile Undosed Red Cap 1000ml

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Nalge Nn Bottles ppr 4ltr

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Nalgene Square Media Bottles PETG with HDPE closures in shrink wrapped trays. sterile.

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Sample Bottles EDTA Dosed Sterile White Cap 1000ml

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Boston Bottles 8ml nm HDPE

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Bottles Sod Thio Dosed 18mg/L St. Blue Cap 1000ml

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Bottles Sod Thio Dosed 180mg/L St. Blue Cap 1000ml

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250ml Adaptors Conical Bottles

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Sample Bottles Non Sterile Undosed Red Cap 250ml

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Sample Bottles Non Sterile Undosed Red Cap 500ml

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Nalgene Bottle Rack for 8x500ml Bottles

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Sample Bottles EDTA Dosed Sterile White Cap 250ml

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Sample Bottles EDTA Dosed Sterile White Cap 500ml

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M-Vac Jr. PP Barbed Cap for 2L Bottles Red.

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BottleS Sod. Thio Dosed 18mg/L St. Blue Cap 250ml

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Bottles Sod Thio Dosed 180mg/L St. Blue Cap 500ml

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Flystuff (59-194) Droso-Plugs Cap for Bottles

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Amber N/M Bottles Ppr 7.5Ml

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Pouring Ring For Simax Bottles

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HDPE Bottles Narrow Mouth 30mL

312002-0001 Scientific Laboratory Supplies PK1000

HDPE Bottles Narrow Mouth 60mL

312002-0002 Scientific Laboratory Supplies PK1000

Biological Stain Kit, 5 Bottles

43320000-1 Bio-WORLD 1 PK(s)

Pallet Knife with Plastic Handle 100mm

SAM1325 Scientific Laboratory Supplies EACH

Pallet Knife with Plastic Handle 200mm

SAM1326 Scientific Laboratory Supplies EACH

Pallet Knife with Plastic Handle 300mm

SAM1327 Scientific Laboratory Supplies EACH

500 ml Glass Bottle with cap

B603 ABM Set of 5

Nalgene Rectangular Amber HDPE Bottles 500ml

2009-0016 Scientific Laboratory Supplies PK48

50 ml Tube Storage Boxes, Plastic

B619 ABM 5/pack

Rayon Balls X-large For Bottles

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Support stand with plastic convertor clamp

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Stuart Rotisserie for 7x40mm Bottles

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Stuart Rotisserie for 2x75mm Bottles

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Plastic tuberculine syringe with drain tube

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Bottles HDPE NM Low Particulate 1L

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Drechsel bottles heads Sinter 24/29

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Plastic syringe 30ml with tube and clip

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Bottles HDPE NM Low Particulate 250mL

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Bottles HDPE NM Low Particulate 500mL

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South American FBS Serum 500ml Bottles

35-079-CV Scientific Laboratory Supplies 500ML

DURAN(R) PRESSURE PLUS BOTTLES VOLUME

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Duran Stirred Reactor for 250mL Bottles

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Water Sampling Bottles HDPE 500ml Dosed

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Wide mouth wash bottles safety labeled

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Water Sampling Bottles HDPE 250ml Undosed

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Water Sampling Bottles HDPE 500ml Undosed

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Bismuth Pieces, 99.9999%

GX4649-250 Glentham Life Sciences 250

Bismuth Pieces, 99.9999%

GX4649-50 Glentham Life Sciences 50

Gallium Pieces, 99.99999%

GX9105-10 Glentham Life Sciences 10

Gallium Pieces, 99.99999%

GX9105-100 Glentham Life Sciences 100

Gallium Pieces, 99.99999%

GX9105-50 Glentham Life Sciences 50

Gallium Pieces, 99.9999%

GX1351-10 Glentham Life Sciences 10

Gallium Pieces, 99.9999%

GX1351-100 Glentham Life Sciences 100

Gallium Pieces, 99.9999%

GX1351-50 Glentham Life Sciences 50

Antimony Pieces, 99.9999%

GX8733-25 Glentham Life Sciences 25

Antimony Pieces, 99.9999%

GX8733-250 Glentham Life Sciences 250

Antimony Pieces, 99.9999%

GX8733-50 Glentham Life Sciences 50

Safety Labeled Wash Bottles, Saline Solution

43320027-1 Bio-WORLD 1 PK (s)

Germanium Pieces max. 2 cm (50 Ohm cm)

GX2202-GX2202100g Glentham Life Sciences GX2202-100g

Germanium Pieces max. 2 cm (50 Ohm cm)

GX2202-GX220210g Glentham Life Sciences GX2202-10g

Germanium Pieces max. 2 cm (50 Ohm cm)

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Indium Pieces, 99.999%

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Indium Pieces, 99.999%

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hybex Media Storage Bottle 1000mL w/ Assorted Caps

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Bismuth Pieces, 99.999%

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GX2132-250 Glentham Life Sciences 250

Bismuth Pieces, 99.999%

GX2132-50 Glentham Life Sciences 50

Antimony Pieces, 99.999%

GX7845-100 Glentham Life Sciences 100

Antimony Pieces, 99.999%

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GX7845-500 Glentham Life Sciences 500

ELISA Microplate Washer Automated (#MPW-30) bottle set (3 bottles and cap assembly)

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Gallium Pieces,

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Water Sampling Bottles HDPE 250ml Dosed Water Sampling Bottle

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5650P-100 CORNING 1/pk

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5650P-25 CORNING 1/pk

VOL FLASK,250ML,PLASTIC,CLASS A,PFA, WITH GL-25 SCREW CAP,1/5

5650P-250 CORNING 1/pk

VOL FLASK,50ML,PLASTIC,CLASS A,PFA, WITH GL-18 SCREW CAP,1/6

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VOL FLASK,500ML,PLASTIC,CLASS A,PFA, WITH GL-25 SCREW CAP,1/4

5650P-500 CORNING 1/pk

Wash bottle carrier pp holds up to 6 8 oz or 16oz bottles

Z408514-1EA Scientific Laboratory Supplies EACH

Strontium Pieces max. 20 mm, under argon, coated with oil, Ba max. 1%, 99%

GX0588-100 Glentham Life Sciences 100

Duran Stirred Reactor for 500mL and 1000mL Bottles

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Bottle 750ml with GL45 Cap

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Bottle 150ml with GL45 Cap

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vented caps for bottle falcon biocoat 25 cm

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Square bottle 500 mL sterile with septum cap Nalgene

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Slide Rack (24 Slides), Plastic Basket with Metal Handle

IMS002 ScyTek Laboratories 1 ea.

Semivolatile Calibration Kit - 5 Pack of 5.5mL Bottles

ECS-KN-050 Scientific Laboratory Supplies 5.5ML

Terbium Pieces, 99.9%

GX6615-10 Glentham Life Sciences 10

Terbium Pieces, 99.9%

GX6615-2 Glentham Life Sciences 2

Gallium Pieces, 99.99%

GX7573-10 Glentham Life Sciences 10

Gallium Pieces, 99.99%

GX7573-100 Glentham Life Sciences 100

Gallium Pieces, 99.99%

GX7573-50 Glentham Life Sciences 50

Holmium Pieces, 99.9%

GX2124-25 Glentham Life Sciences 25

Holmium Pieces, 99.9%

GX2124-5 Glentham Life Sciences 5

Hafnium Pieces, 99.9%

GX4121-10 Glentham Life Sciences 10

Hafnium Pieces, 99.9%

GX4121-100 Glentham Life Sciences 100

Hafnium Pieces, 99.9%

GX4121-25 Glentham Life Sciences 25

Hafnium Pieces, 99.9%

GX4121-250 Glentham Life Sciences 250

Yttrium pieces, 99.9%

GX0115-10 Glentham Life Sciences 10

Yttrium pieces, 99.9%

GX0115-50 Glentham Life Sciences 50

The obtained results are repeatable and reproducible with RSDs of 5.49% and 3.18% respectively. Besides, our developed sensor was successfully applied for the determination of phenol in tap and mineral water samples. The proposed new approach is highly recommended to develop a simple, cost effective, ecofriendly, and highly sensitive sensor for the electrochemical detection of phenol which can further broaden the applications of green silver NPs.

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