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

Hada, Alkesh; Singh, Divya; Venkata Satyanarayana, Kranti Kavalipurapu Veera; Chatterjee, Madhurima; Phani, Victor; Rao, Uma

doi:10.21307/jofnem-2021-073

Figures & Tables

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

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

Figures & Tables

Figure 1:

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

List of primers used in this study_

Homology parameters of unannotated genes used in this study_

Paradigm

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