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.

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

MST5-1000 Alpha Diagnostics 1 pk

SCREW CAPS WITH "O" RINGS. BROWN

SCO-BR CORNING 500/pk

Disposable Meat Sample Plastic tubes with caps (5 ml) (250/pk)

MST5-250 Alpha Diagnostics 1 pk

ERLENMEYER FLASK, 50 ML, PP

4985P-50 CORNING 6/pk

ERLENMEYER FLASK, 50 ML, PMP

4990P-50 CORNING 6/pk

0.2mL PCR tubes, individual, with flat caps, 1000/pk

P3030 MTC Bio 1000/pack

0.2mL PCR tubes, individual, with domed caps, 1000/pk

P3040 MTC Bio 1000/pack

PLASTIC CLAMP, 125 ML

480149 CORNING 1/pk

PLASTIC CLAMP, 250 ML

480150 CORNING 1/pk

PLASTIC CLAMP, 500 ML

480151 CORNING 1/pk

Bottles 2L Nalgene with Quick Connect

BTLN52 Next Advance 2pack

50 ml Stripette individually paper/plastic wrapped serological pipettes sterile

GR238023 Genorise Scientific Case of 100

Tubes (empty) pack of 1000 with caps and sealing ring

D1031-T20 Benchmark Scientific 1 PC

PBS Tablets (1000 ml)

2129-10 Biovision

PBS Tablets (1000 ml)

2129-100 Biovision

Plastic Columns

C-50 ABTBeads 50/pk

0.2 ml Individual PCR Tubes with attached Flat Caps

GP101 GenDepot 1,000 tubes

0.2 ml Individual PCR Tubes with attached Domed Caps

GP102 GenDepot 1,000 tubes

0.5 ml Individual PCR Tubes with attached Flat Caps

GP103 GenDepot 1,000 tubes

0.5 ml Individual PCR Tubes with attached Domed Caps

GP104 GenDepot 1,000 tubes

TruStrip Sample Transfer Strips, 50-ul, 50/Pk (with sample tracking dye)

STSD-50-50 Alpha Diagnostics 1 Pk

Tubes (empty) pack of 50 with caps and sealing ring

D1031-T21 Benchmark Scientific 1 PC

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

SCO-LP-BR CORNING 500/pk

ELISA PLATE READER : MICRO READ 1000

1000 Global Diagnostics B 1 unit

0.2 ml 8-Strip PCR Tube with Individually Attached Caps

GP900 GenDepot 120 strips

Plastic XL Columns

CXL-50 ABTBeads 50/pk

Plastic XXL Columns

CXXL-50 ABTBeads 50/pk

Plastic Columns Frits

FC-50 ABTBeads 50/pk

D(+)Glucose 20 % 1000 ml

12-9165-5 Medicago 1000 ml

Sodium Chloride tablets 1000 ml

09-9104-10 Medicago 1000 ml

Sodium Chloride tablets 1000 ml

09-9104-100 Medicago 1000 ml

pepsine cleaning solution 1000 ml

B232 Consort ea

Heat Extractor with Plastic Knob

IMS006 ScyTek Laboratories 1 ea.

Antigen Retrieval Solution, Citrate pH 6.0 (50 ml, 100X)

ARS6-50 Alpha Diagnostics 5 L

Antigen Retrieval Solution, EDTA pH 8.0 (50 ml, 100X)

ARS8-50 Alpha Diagnostics 5 L

Human Calcyphosine (CAPS) ELISA Kit

DLR-CAPS-Hu-48T DL Develop 48T

Human Calcyphosine (CAPS) ELISA Kit

DLR-CAPS-Hu-96T DL Develop 96T

Human Calcyphosine (CAPS) ELISA Kit

RD-CAPS-Hu-48Tests Reddot Biotech 48 Tests

Human Calcyphosine (CAPS) ELISA Kit

RD-CAPS-Hu-96Tests Reddot Biotech 96 Tests

Human Calcyphosine (CAPS) ELISA Kit

RDR-CAPS-Hu-48Tests Reddot Biotech 48 Tests

Human Calcyphosine (CAPS) ELISA Kit

RDR-CAPS-Hu-96Tests Reddot Biotech 96 Tests

AlbuVoid Buffer Kit - 1000

AVBK-1000 Biotech Support Group 1000 mL

HemoVoid Buffer Kit - 1000

HVBK-1000 Biotech Support Group 1000 mL

UPCK Buffer Kit - 1000

UPBK-1000 Biotech Support Group 1000 mL

AXYPREP MAG DYECLEAN KIT- 50 ML - 5000 PREPS

MAG-DYECL-50 CORNING 1/pk

RESERVOIR, POLYPROPYLENE, 50 ML, V-BOTTOM, NONSTERILE, BULK

RES-V-50 CORNING 100/pk

Plastic XL Columns Frits

FCXL-50 ABTBeads 50/pk

Plastic XXL Columns Frits

FCXXL-50 ABTBeads 50/pk

TC-G 1000

B7730-50 ApexBio 50 mg

Antigen Retrieval Solution, Tris-EDTA pH 9.0 (50 ml, 100X)

ARS9-50 Alpha Diagnostics 5 L

50 ml Reagent Reservoir

GR238016 Genorise Scientific Pack of 200

Disposable Plastic Pestles, Conical, 210mm, 50/Bag

MGR113 Bio Basic 1PK, 50UNIT

Disposable Plastic Pestles, Round, 205mm, 50/Bag

MGR114 Bio Basic 1PK, 50UNIT

Disposable Plastic Pestles, Conical, 70mm, 50/Bag

MGR115 Bio Basic 1PK, 50UNIT

EDTA buffer pH 8.0, 1000 ml

12-9160-5 Medicago 1000 ml

Potassium chloride 1 M, 1000 ml

12-9175-10 Medicago 1000 ml

Potassium chloride 3 M, 1000 ml

12-9176-5 Medicago 1000 ml

Sodium chloride 3 M, 1000 ml

12-9190-5 Medicago 1000 ml

Sodium chloride 5 M, 1000 ml

12-9191-5 Medicago 1000 ml

Tris buffer pH 7.4, 1000 ml

12-9198-10 Medicago 1000 ml

Tris buffer pH 8.0, 1000 ml

12-9199-10 Medicago 1000 ml

Tris buffer pH 8.3, 1000 ml

12-9200-10 Medicago 1000 ml

PBS-Tween® Tablets (1000 ml)

2130-10 Biovision

PBS-Tween® Tablets (1000 ml)

2130-100 Biovision

AXYPREP FRAGMENTSELECT-I KIT- 50 ML - 380 PREPS

MAG-FRAG-I-50 CORNING 1/pk

RESERVOIR, POLYPROPYLENE, 50 ML, V-BOTTOM, STERILE, BULK

RES-V-50-S CORNING 5/pk

MINI BIOREACTOR, 50 ML CENTRIFUGE TUBE WITH VENT CAP, STERILE

431720 CORNING 25/pk

Phosphate Buffered Saline with Tween 20 (PBS-T) pH 7.4, 1000 ml

09-9410-100 Medicago 1000 ml

Phosphate Buffered Saline with Tween 20 (PBS-T) pH 7.4, 1000 ml

09-9410-30 Medicago 1000 ml

SCREW CAPS WITH "O" RINGS

SCO-R CORNING 500/pk

SCREW CAPS WITH "O" RINGS

SCO-W CORNING 500/pk

SCREW CAPS WITH "O" RINGS

SCO-Y CORNING 500/pk

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

5650P-50 CORNING 1/pk

AXYPREP MAG PCR NORMALIZER KIT- 50 ML - 384 PREPS

MAG-PCR-NM-50 CORNING 1/pk

RESERVOIR, POLYPROPYLENE, 50 ML, V-BOTTOM, STERILE, INDIVIDUALLY PACKAGED

RES-V-50-SI CORNING 100/pk

Disposable Plastic Pestles, Conical, 210mm, 50/Bag, Sterile

MGR113-S Bio Basic 1PK, 50UNIT

Disposable Plastic Pestles, Round, 205mm, 50/Bag, Sterile

MGR114-S Bio Basic 1PK, 50UNIT

Disposable Plastic Pestles, Conical, 70mm, 50/Bag, Sterile

MGR115-S Bio Basic 1PK, 50UNIT

Chloramphenicol Solution (50 mg/ml)

C090-10ML TOKU-E 10 mL

Chloramphenicol Solution (50 mg/ml)

C090-10x10ML TOKU-E 10 x 10 mL

PBS solution pH 7.4, 1000 ml/bottle

10-9402-10 Medicago 1000 ml/bottle

Tris-Glycine buffer pH 8.3, 1000 ml

12-9122-10 Medicago 1000 ml

Sodium hydroxide powder 5 M, 1000 ml

12-9183-5 Medicago 1000 ml

Sodium acetate buffer pH 7.0, 1000 ml

12-9188-5 Medicago 1000 ml

1000 ml leak-resistant HDPE bottle, colorless

GR238010 Genorise Scientific Pack of 6

PCR-Tubes 0,2 ml in strips; 8 x 250 tubes with domed caps

A23232 GeneOn 8x250 tubes&caps

Meat sample spatulas (individually wrapped) 1000/pk

MSSW-1000 Alpha Diagnostics 1 pk

SCREW CAPS WITH "O" RINGS. BLUE

SCO-B CORNING 500/pk

SCREW CAPS WITH "O" RINGS. CLEAR

SCO-C CORNING 500/pk

SCREW CAPS WITH "O" RINGS. GREEN

SCO-G CORNING 500/pk

AXYPREP MAG PCR CLEAN UP KIT- 50 ML - 1110 PREPS

MAG-PCR-CL-50 CORNING 1/pk

50 ML AMPHOTERICIN B, LIQUID 250 µG/ML SOLUBILIZED

30-003-CF CORNING 50 mL/pk

10 ML GENTAMICIN SULFATE, LIQUID 50 MG/ML SOLUTION

30-005-CR CORNING 10 mL/pk

50 ML KANAMYCIN SULFATE, LIQUID, 5,000 µG/ML SOLUTION

30-006-CF CORNING 50 mL/pk

Dressed Gold Streptavidin at 50 O.D./mL, 1 mL

DSTAL-B001 BioAssayWorks 1 mL

Dressed Gold Streptavidin at 50 O.D./mL, 9 mL

DSTAL-B009 BioAssayWorks 9 mL

Dressed Gold NeutrAvidin at 50 O.D./mL, 1 mL

DNTAL-B001 BioAssayWorks 1 mL

Dressed Gold NeutrAvidin at 50 O.D./mL, 9 mL

DNTAL-B009 BioAssayWorks 9 mL

ML 228

A4508-50 ApexBio 50 mg

ML-204

9448-50 Biovision

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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