Identification and Characterization of Verticillium longisporum Lineage A1/D1 from Brassica Crops in Manitoba, Canada
Abstract
:1. Introduction
2. Results
2.1. Pathogen Isolation
2.2. Morphological Identification
2.3. PCR Characterisation
2.4. Disease Assessment
3. Discussion
4. Materials and Methods
4.1. Sample Collection and Fungal Pathogen Isolation
4.2. Morphological Identification of Isolates
4.3. DNA extraction, PCR Amplification and PCR-Restriction Fragment Length Polymorphism
4.4. Pathogenicity Tests of V. longisporum on Canola
4.5. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Inoculum. | Days Post-Inoculation (dpi) | ||||
---|---|---|---|---|---|
7 | 14 | 21 | 28 | 35 | |
Westar-water | 1.00(±0.00) 1a 2 | 1.00(±0.00)a | 1.00(±0.00)a | 1.85(±0.00)a | 2.15(±0.00)a |
Westar-VL-H1 | 1.10(±0.00)a | 2.05(±0.00)a | 3.65(±0.00)b | 4.85(±0.25)b | 5.95(±0.21)b |
Westar-VL-H2 | 1.00(±0.00)a | 2.25(±0.00)a | 2.95(±0.00)b | 4.75(±0.22)b | 6.15(±0.23)b |
Westar-VL-H3 | 1.25(±0.00)a | 2.15(±0.00)a | 3.25(±0.10)b | 4.20(±0.20)b | 5.25(±0.18)b |
Westar-VL-H4 | 1.15(±0.00)a | 2.00(±0.00)a | 3.75(±0.15)b | 4.45(±0.17)b | 5.75(±0.23)b |
Westar-VL-H5 | 1.05(±0.00)a | 2.25(±0.10)b | 4.05(±0.20)b | 4.95(±0.21)b | 5.75(±0.24)b |
Westar-VL-H6 | 1.00(±0.00)a | 2.35(±0.12)b | 3.85(±0.10)b | 4.75(±0.22)b | 6.05(±0.18)b |
Westar-VL-H7 | 1.10(±0.00)a | 2.00(±0.00)a | 3.45(±0.00)b | 4.95(±0.21)b | 5.75(±0.20)b |
Westar-VL-H8 | 1.15(±0.00)a | 2.05(±0.10)a | 3.35(±0.00)b | 4.50(±0.15)b | 5.75(±0.20)b |
Westar-VL-H9 | 1.25(±0.00)a | 2.00(±0.00)a | 3.65(±0.10)b | 4.70(±0.21)b | 5.65(±0.16)b |
Westar-VL-H10 | 1.20(±0.00)a | 2.15(±0.10)a | 3.75(±0.12)b | 4.95(±0.24)b | 5.85(±0.18)b |
Westar-VL-H11 | 1.10(±0.00)a | 3.05(±0.10)b | 3.95(±0.12)b | 4.65(±0.20)b | 5.25(±0.15)b |
Westar-VL-H12 | 1.05(±0.00)a | 2.15(±0.09)a | 3.75(±0.15)b | 4.80(±0.22)b | 5.75(±0.18)b |
Westar-VL-H13 | 1.25(±0.00)a | 2.25(±0.10)a | 3.95(±0.10)b | 5.15(±0.20)b | 6.15(±0.18)b |
Westar-VL-H14 | 1.00(±0.00)a | 2.00(±0.00)a | 3.45(±0.10)b | 4.75(±0.19)b | 5.75(±0.20)b |
Westar-VL-H15 | 1.15(±0.00)a | 2.25(±0.00)a | 3.70(±0.10)b | 4.95(±0.20)b | 5.90(±0.20)b |
Westar-VL-H16 | 1.00(±0.00)a | 1.95(±0.00)a | 3.45(±0.10)b | 4.50(±0.15)b | 5.65(±0.15)b |
Westar-VL-H17 | 1.10(±0.00)a | 2.25(±0.00)a | 3.50(±0.10)b | 4.75(±0.21)b | 6.05(±0.22)b |
Westar-VL-H18 | 1.20(±0.00)a | 2.00(±0.00)a | 3.50(±0.00)b | 4.90(±0.25)b | 5.85(±0.20)b |
Westar-VL-H19 | 1.00(±0.00)a | 2.15(±0.08)a | 3.45(±0.20)b | 4.75(±0.24)b | 5.75(±0.20)b |
Westar-VL-H20 | 1.20(±0.00)a | 2.25(±0.00)a | 3.75(±0.10)b | 5.05(±0.22)b | 5.45(±0.15)b |
Westar-VL-H21 | 1.05(±0.00)a | 2.45(±0.00)b | 3.80(±0.20)b | 4.95(±0.20)b | 5.70(±0.23)b |
Westar-VL-H22 | 1.20(±0.00)a | 2.00(±0.00)a | 3.50(±0.15)b | 4.85(±0.25)b | 5.85(±0.21)b |
Westar-VL-H23 | 1.00(±0.00)a | 2.05(±0.00)a | 3.55(±0.11)b | 4.45(±0.20)b | 5.40(±0.15)b |
Westar-VL-H24 | 1.50(±0.00)a | 2.55(±0.05)b | 3.75(±0.10)b | 4.95(±0.19)b | 6.15(±0.25)b |
Westar-VL-H25 | 1.15(±0.00)a | 2.25(±0.00)a | 3.55(±0.10)b | 4.55(±0.15)b | 5.65(±0.20)b |
Westar-VL-H26 | 1.15(±0.00)a | 2.15(±0.00)a | 3.50(±0.10)b | 4.95(±0.22)b | 5.80(±0.16)b |
Westar-VL-H27 | 1.20(±0.00)a | 2.05(±0.00)a | 3.75(±0.10)b | 4.75(±0.25)b | 5.40(±0.25)b |
Westar-VL-H28 | 1.00(±0.00)a | 2.00(±0.00)a | 3.25(±0.10)b | 4.50(±0.20)b | 5.05(±0.15)b |
Westar-VL-H29 | 1.20(±0.00)a | 2.15(±0.00)a | 3.45(±0.00)b | 4.80(±0.25)b | 5.25(±0.11)b |
Westar-VL-R 3 | 1.25(±0.00)a | 2.45(±0.00)a | 3.80(±0.11)b | 5.05(±0.21)b | 6.45(±0.25)b |
Westar-Vd-M 4 | 1.00(±0.00)a | 2.00(±0.00)a | 3.25(±0.10)b | 4.50(±0.21)b | 5.25(±0.14)b |
Primer. | Sequence | Amplicon/Locus | Reference |
---|---|---|---|
VlspF1 | AGCCTGAGTCACGAGAGATATGGG | 18S intron | Banno et al. 2011 [15] |
VlspR4 | CAAACCACGCCACTGCATTCTCGT | ||
VeruniF2 | TCGTAGTAGAAGCTCGGCCTCCGGTC | 18S rDNA | Ikeda et al. 2012 [9] |
VeruniR3 | TAAGAAGTCGGCGTACTACCGGGGT | ||
VdMAT1-1-1F01 | CAACTCCTGGACTCTCATCG | MAT1-1-1 | Banno et al. 2015 [16] |
VdMAT1-1-1R01 | AAATGGTCTGAGTCGGGGGT | ||
VdMAT1-2-1F02-2 | CTAAGGCTCCAAGTCAATCC | MAT1-2-1 | Banno et al. 2015 [16] |
VdMAT1-2-1R02-2 | TGCTGCCACTTGTTCAAACG | ||
D1f | CCCCGGCCTTGGTCTGAT | GPD gene1 | Inderbitzin et al. 2013 [26] |
AlfD1r | TGCCGGCATCGACCTTGG | ||
A1f | AAGTGGAGCCCCGTATCTTGAAT | EF-1α2 | Inderbitzin et al. 2013 [26] |
A1r | CAACTGGCAACAGGGCTTGAAT | ||
Df | CCGGTCCATCAGTCTCTCTG | ITS | Inderbitzin et al. 2013 [26] |
Dr | CTGTTGCCGCTTCACTCG | ||
NMS1 | CAGCAGTGAGGAATATTGGTCAATG | mtSSU-rDNA | Li et al. 1994 [27] |
NMS2 | GCGGATCATCGAATTAAATAACAT | ||
cobF1 | GTTGAATTTATCTGAGGAGG | Cytochrome b | Ikeda et al. 2012 [9] |
cobR1 | GGAGGAGTTTGCATTGGATTAGCC |
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Zou, Z.; Bisht, V.; Fernando, W.G.D. Identification and Characterization of Verticillium longisporum Lineage A1/D1 from Brassica Crops in Manitoba, Canada. Int. J. Mol. Sci. 2020, 21, 3499. https://doi.org/10.3390/ijms21103499
Zou Z, Bisht V, Fernando WGD. Identification and Characterization of Verticillium longisporum Lineage A1/D1 from Brassica Crops in Manitoba, Canada. International Journal of Molecular Sciences. 2020; 21(10):3499. https://doi.org/10.3390/ijms21103499
Chicago/Turabian StyleZou, Zhongwei, Vikram Bisht, and W. G. Dilantha Fernando. 2020. "Identification and Characterization of Verticillium longisporum Lineage A1/D1 from Brassica Crops in Manitoba, Canada" International Journal of Molecular Sciences 21, no. 10: 3499. https://doi.org/10.3390/ijms21103499