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Effect of fluensulfone on different functional genes of root-knot nematode Meloidogyne incognita

Alkesh Hada

Alkesh Hada

Division of Nematology, ICAR-Indian Agricultural Research InstituteNew Delhi, India
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Hada, Alkesh
, 
Divya Singh

Divya Singh

Division of Nematology, ICAR-Indian Agricultural Research InstituteNew Delhi, India
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Singh, Divya
, 
Kranti Kavalipurapu Veera Venkata Satyanarayana

Kranti Kavalipurapu Veera Venkata Satyanarayana

All India Coordinated Research Project on Nematodes, ICAR-Indian Agricultural Research InstituteNew Delhi, India
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Venkata Satyanarayana, Kranti Kavalipurapu Veera
, 
Madhurima Chatterjee

Madhurima Chatterjee

Division of Nematology, ICAR-Indian Agricultural Research InstituteNew Delhi, India
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Chatterjee, Madhurima
, 
Victor Phani

Victor Phani

Department of Agricultural Entomology, College of Agriculture, Uttar Banga Krishi ViswavidyalayaWest Bengal, India
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Phani, Victor
 oraz 
Uma Rao

Uma Rao

Division of Nematology, ICAR-Indian Agricultural Research InstituteNew Delhi, India
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Rao, Uma
  
17 sie 2021
Effect of fluensulfone on different functional genes of root-knot nematode Meloidogyne incognita's Cover Image
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Data publikacji: 17 sie 2021
Zakres stron: 1 - 14
DOI: https://doi.org/10.21307/jofnem-2021-073
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© 2021 Alkesh Hada et al., published by Sciendo.
This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1:

Relative fold-change expression of genes related to nematode chemosensation, GPCRs, parasitism, fatty acid metabolism, and neurotransmission in M. incognita J2s after treatment with 10, 50, and 100 ppm of fluensulfone for 5 and 10 hr. Each bar represents the log2-transformed mean ± SE (n = 3). Asterisk (*) indicates significant differential expression (*p < 0.05; **p < 0.01; ns = non-significant), analyzed by one-tailed t-test.
Relative fold-change expression of genes related to nematode chemosensation, GPCRs, parasitism, fatty acid metabolism, and neurotransmission in M. incognita J2s after treatment with 10, 50, and 100 ppm of fluensulfone for 5 and 10 hr. Each bar represents the log2-transformed mean ± SE (n = 3). Asterisk (*) indicates significant differential expression (*p < 0.05; **p < 0.01; ns = non-significant), analyzed by one-tailed t-test.

Assessment of mortality of M_ incognita J2s at variable concentrations of fluensulfone and time of exposure_

Nematode mortality (%)
Fluensulfone concentration (ppm) 1hr 2.5hr 5hr 7.5hr 10hr 24hr Remarks
100 0.86 3.47 8.62 18.51 27.78 100 Reduced motility and agility observed 0.5hr onward the drug treatment
75 0 1.74 4.35 8.62 18.18 100 Reduced motility and agility observed 1hr onward the drug treatment
50 0 1.74 8.08 13.79 13.79 100 Reduced motility and agility observed 1hr onward the drug treatment
25 0 0.86 2.59 8.62 11.01 90.74 Reduced motility and agility observed 4hr onward the drug treatment
10 0 0 0 2.59 6.03 75.92 Reduced motility and agility observed 4hr onward the drug treatment
5 0 0 0 1.74 1.74 43.48 Reduced motility and agility observed 7hr onward the drug treatment
1 0 0 0 0 1.74 29.57 Reduced motility and agility in few worms observed 7–8hr onward the drug treatment
Vehicle control (0.05% DMSO) 0 0 0 0 0 0 No change in motility and agility

List of primers used in this study_

Sl Primers name Gene name (accession number) Primers Sequences (5′–3′) Tm (°C) Purpose
1 RT miODR-1 F Mi-odr-1 (MG780832) GAACCACGCTCTTACGATTC 60°C qRT PCR
RT miODR-1 R CTCCAGAAGCGACCATGTA
2 RT miODR-3 F Mi-odr-3 (MG780833) CTGGCTATAGACCCACAGAA 60°C qRT PCR
RT miODR-3 R GAACGTTGTCCACCTACATC
3 RT miTAX-2 F Mi-tax-2 (MG780834) GAGGGAATATCCTGAAGCG 60°C qRT PCR
RT miTAX-2 R CCTGATTCCACTGTTCTGG
4 RT miTAX-4 F Mi-tax-4 (MG780835) GTGAAGTTCCTTGGCCTAT 60°C qRT PCR
RT miTAX-4 R CAGAGCTAGCAATCATACTC C
5 RT miOSM-9 F M. incognita osm-9 (MT676864) CATGCCTGAAGATTGGGAAG 60°C Cloning and qRT PCR
RT miOSM-9 R GGTCAGGATTAGCACCATAC
6 RT miGPC-1 F M. incognita gpc-1 (MT676857) CAACTTCGACAAGAGGCAA 60°C Cloning and qRT PCR
RT miGPC-1 R CCCTATTGGTCCGGTAATTAAG
7 RT mi GPA-11F M. incognita gpa-11 (MT676854) CGACTACGGGTATTTGTGAC 60°C Cloning and qRT PCR
RT miGPA-11 R GAAAGTTCGGCTACGAACAG
8 RT miGPA-13 F M. incognita gpa-13 (MT676855) GCTGAAACTTCAGAGGATGG 60°C Cloning and qRT PCR
RT miGPA-13 R GCTGGTATTCACGAGAACG
9 RT miGRK-2 F M. incognita grk-2 (MT676858) CTCAATTCTACGCAGCAGAG 60°C Cloning and qRT PCR
RT miGRK-2 R GGTGCCATATAGCCAACAG
10 RT miXYL-1 F Mi-xylanase (AF224342) GGGATTAGTTGCATACAGTTAAGATAAG 60°C qRT PCR
RT miXYL-1 R GCTGTTCGAGTAAGCAGTAGAG
11 RT miXYL-3 F Mi-xylanase (EU475876) GGAACTTCCGGTAAACCCTA 60°C qRT PCR
RT miXYL-3 R GACCGGACTGATGTGTTATC
12 RT miENG-1 F Mi-β-1,4-endoglucanase (AF100549) ACCGAGCAACTCACAAAC 60°C qRT PCR
RT miENG-1 R GGCATTGCTACCCGTATTT
13 RT miPEL F Mi-pectatelyase (AF527788) CGAATAACGACGAAGAGGAC 60°C qRT PCR
RT miPEL R AGAACCGACAACCCTACA
14 RT miMSP-20 F Mi-msp-20 (AY134439) TGGTGACGAACGCACACCTACATA 60°C qRT PCR
RT miMSP-20 R GCGCTGTCTTTGACCATTTGCTCT
15 RT miMSP-33 F Mi-msp-33 (AY142118) GTGGCCTCCTTTGCTTGGACATTT 60°C qRT PCR
RT miMSP-33 R CATCACCTCCAATTACTCCGGGTT
16 RT miFAT-6 F M. incognita fat-6 (MT676846) GAATTACTGCAGGTCCTCAC 60°C Cloning and qRT PCR
RT miFAT-6 R CATCAGTGTCAGTCCACTTG
17 RT miACS-2 F M. incognita acs-2 (MT676837) CCCACCAGAGGAAAGGATAA 60°C Cloning and qRT PCR
RT miACS-2 R CGCTCCTATACCAACGATCT
18 RT miMDT-15 F M. incognita mdt-15 (MT676859) CAGCTATGCCACCAGGTTT 60°C Cloning and qRT PCR
RT miMDT-15 R CCTAATCCTCCTCCACCATTTC
19 RT miNHR-49 F M. incognita nhr-49 (MT676861) GCCAATGAGGCAATTAGAG 60°C Cloning and qRT PCR
RT miNHR-49 R CCTCGTCTACCGGTTTAGAT
20 RT miELO-2 F M. incognita elo-2 (MT676842) CAAACGGATATTGGGTCTGG 60°C Cloning and qRT PCR
RT miELO-2 R GGCGTGATAGGGTAGGAATA
21 RT miECH-5 F M. incognita ech-5 (MT676839) CACTAGGTGGAGGACTAGAA 60°C Cloning and qRT PCR
RT miECH-5 R CTTGGGAGTCTTTGAGTTCC
22 RT miECH-6 F M. incognita ech-6 (MT676840) CGAGTCTTCTGGAAGGGATA 60°C Cloning and qRT PCR
RT miECH-6 R GCTTAGATGGCAAAGAGGTC
23 RT miFLP-12 F Mi-flp-12 (AY804187) TGAGGAAGCGGCCCGATAGTTCTT 60°C qRT PCR
RT miFLP-12 R GGATGAAGAAATGCTTGGACGAGT
24 RT miFLP-14 F Mi-flp-14 (AY907829) GCGAGTCCATGTGTAGCAGCTAAT 60°C qRT PCR
RT miFLP-14 R GGGAGATGAAGAACGTTTACTACTTTGCC
25 RT miFLP-16 F Mi-flp-16 (EU549831) GGCAATATTCACGAGACTGGCAAC 60°C qRT PCR
RT miFLP-16 R GGCCATTCAATGCCTGAAGAGGG
26 RT miFLP-18 F Mi-flp-18 (AY729022) AGGATGACTTATTGCGCCAGGA 60°C qRT PCR
RT miFLP-18 R TTCCTTTACCGAATCTGAGCACGC
27 RT miACE-1 F Mi-ace-1 (AF075718) CTCCTTGTTCTGAGGATTGTCT 60°C qRT PCR
RT miACE-1 R TTTATGGAGGTGGATTCTGGAG
28 RT miACE-2 F Mi-ace-2 (AF495588) AGATGTGGAATCCGCCTAATG 60°C qRT PCR
RT miACE-2 R TTTACTCGGGCTCTCCTTCT
29 RT miNLP-3 F Mi-nlp-3 (KY054882) TCCACAATTTGTGCCAGGTC 60°C qRT PCR
RT miNLP-3 R TTGTGGGGCTGATCGTTTTC
30 RT miNLP-12 F Mi-nlp-12 (KY054885) GATAGGCGAAAGCGGACTATT 60°C qRT PCR
RT miNLP-12 R CAAGGGCCTGTAATCTCTCTTT
31 RT mi18S rRNA F Mi-18S rRNA (HE667742) TCAACGTGCTTGTCCTACCCTGAA 60°C qRT PCR
RT mi18S rRNA R TGTGTACAAAGGGCAGGGACGTAA
32 RT mi actin F Mi-actin (BE225475) TGACTCTGGAGATGGTGTTACG 60°C qRT PCR
RT mi actin R GTGATGACTTGACCGTCAGGC

Homology parameters of unannotated genes used in this study_

Sl Primers name Gene name (accession number) Primers Sequences (5′–3′) Amplicon length (bp) Identity (%) Score E-value Purpose
1 RT miOSM-9 F M. incognita osm-9 homologue CATGCCTGAAGATTGGGAAG 150 100 124 3.6E‒62 qRT PCR
RT miOSM-9 R GGTCAGGATTAGCACCATAC
2 RT miGPC-1 F M. incognita gpc-1 homologue CAACTTCGACAAGAGGCAA 117 100 199 1.4E‒18 qRT PCR
RT miGPC-1 R CCCTATTGGTCCGGTAATTAAG
3 RT mi GPA-11F M. incognita gpa-11 homologue CGACTACGGGTATTTGTGAC 143 100 58 8.2E‒23 qRT PCR
RT miGPA-11 R GAAAGTTCGGCTACGAACAG
4 RT miGPA-13 F M. incognita gpa-13 homologue GCTGAAACTTCAGAGGATGG 112 100 194 1.1E‒17 qRT PCR
RT miGPA-13 R GCTGGTATTCACGAGAACG
5 RT miGRK-2 F M. incognita grk-2 homologue CTCAATTCTACGCAGCAGAG 124 100 202 4.4E‒19 qRT PCR
RT miGRK-2 R GGTGCCATATAGCCAACAG
6 RT miFAT-6 F M. incognita fat-6 homologue GAATTACTGCAGGTCCTCAC 162 100 306 1.9E‒33 qRT PCR
RT miFAT-6 R CATCAGTGTCAGTCCACTTG
7 RT miACS-2 F M. incognita acs-2 homologue CCCACCAGAGGAAAGGATAA 200 100 356 4.2E‒40 qRT PCR
RT miACS-2 R CGCTCCTATACCAACGATCT
8 RT miMDT-15 F M. incognita mdt-15 homologue CAGCTATGCCACCAGGTTT 100 100 105 2.5E‒5 qRT PCR
RT miMDT-15 R CCTAATCCTCCTCCACCATTTC
9 RT miNHR-49 F M. incognita nhr-49 homologue GCCAATGAGGCAATTAGAG 179 100 294 9.4E‒32 qRT PCR
RT miNHR-49 R CCTCGTCTACCGGTTTAGAT
10 RT miELO-2 F M. incognita elo-2 homologue CAAACGGATATTGGGTCTGG 148 100 64 2.2E‒26 qRTPCR
RT miELO-2 R GGCGTGATAGGGTAGGAATA
11 RT miECH-5 F M. incognita ech-5 homologue CACTAGGTGGAGGACTAGAA 130 100 209 4.7E‒20 qRT PCR
RT miECH-5 R CTTGGGAGTCTTTGAGTTCC
12 RT miECH-6 F M. incognita ech-6 homologue CGAGTCTTCTGGAAGGGATA 193 100 342 1.3E‒36 qRT PCR
RT miECH-6 R GCTTAGATGGCAAAGAGGTC
Język:
Angielski
Częstotliwość wydawania:
1 razy w roku
Dziedziny czasopisma:
Nauki biologiczne, Nauki biologiczne, inne
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