Transfer of Meloidogyne incognita Resistance Using Marker-assisted Selection in Sorghum

Name	PI number	Reference/source	Sorghum type
Collier	PI 641862	Maunder (2000)	sweet
Dale	PI 651495	Broadhead and Coleman (1973)	sweet
Entry 22	–	University of Florida	forage
GT-IR7	PI 602445	Widstrom (1998)	grain
Top 76-6	PI 583832	Day et al. (1995)	sweet

Table 2.

Sorghum primer sequences of microsatellite markers used for confirming sorghum crosses in the M. incognita resistance quantitative trait locus region.

Marker	Forward sequence	Reverse sequence	Repeat motif	Expected amplicon (bp)a
RKNP17	GCAGTTTTTCAAGGAACGTG	GAGGAATGGGTGATGAAACAA	(TTA)106	422
RKNP135	GTTTCGTTTCAATCGGCTTC	GCGCCCCATCATATGTCTT	(AAG)21	197
RKNP194	TCATACTACCACAGCCGCTAGA	TGGTGTAGATGTGTGTGATTCAA	(AT)40	236
RKNP259	AGCTCTTCAGGCACATCGTT	TCTCCTTCCCACCCTGTATG	(GA)48	243
RKNP342	TTCCAACAGGCAAACAACAG	TCATGGCCTGTGTATCAAGC	(CT)25	205
RKNP402	TCAGCAAGATGGTTGGTTGA	ACGAGGCCGTTGAGATTATG	(TTA)22	213
RKNP465	TGACTGAGAGGGTCTACCTAACG	CAACCGGAAGTACGCTGATT	(AT)19	247
RKNP529	GCGAAATGGAGAAGAACAGG	CGTCATCAGCTTCCAGGAGT	(GA)25	199
RKNP638	CCACACCGGTTTCCTGTTAT	TAATAAGCCCCGCATGAAGA	(ATA)21, (TAT)6, (TAA)25	251
RKNP709	GCAAGCTGAAGTGGCCTAGT	CTACTCCCTCCGTCCCAAAT	(CT)9, (TA)18	324
RKNP821	CTCGGCAGCACCAAATAAAA	TCTCAACCGATGATTGTCCA	(TA)51	199
TRKN1	TGTACACTGCATGCCAACCT	GCCTCGTCTGGTTCATTGTT	(AT)14	246
TRKN3	GAAGAATTGCTCCAGGAACG	AAGCAGTATCCGGGGAAGAT	(TA)10	271
TRKN4	CGTAAATGGAGGTGGCTACA	CCCGGTTGGTACAACATAGA	(AT)29	232
TRKN5	ACTGTTATGTCGGCTGGTCA	AGTGTTACTGCCTGGCCAAA	(TC)11	196

1 Note: ^aExpected amplicon size was obtained from the genomic sequence of sorghum genotype BTx623.

Table S1.

Genotyping information for the creation of sorghum BC₁F₆ lines.

Date of PCR	Sorghum seedling(s) with confirmed cross	Microsatelite markers used
7/17/2015	Honey Drip x Collier	TRKN1, TRKN3, TRKN4
8/4/2015	Honey Drip x GT-IR7	TRKN1, TRKN3, TRKN4, TRKN5
8/4/2015	Honey Drip x Entry 22	TRKN1, TRKN3, TRKN4, TRKN5
8/4/2015	Honey Drip x Entry 22	TRKN1, TRKN3, TRKN4, TRKN5
8/25/2015	Honey Drip x Dale	TRKN1, TRKN3, TRKN4, TRKN5
11/30/2015	Honey Drip x Top 76-6	RKNP402, RKNP342
11/30/2015	Entry 22 x (Honey Drip x Entry 22)	RKNP402, RKNP342
12/16/2015	Honey Drip x Top 76-6	RKNP194, RKNP259, RKNP402, RKNP529
1/26/2016	Dale x (Honey Drip x Dale)	RKNP194, RKNP259, RKNP465, RKNP529
2/19/2016	GT-IR7 x (Honey Drip x GT-IR7)	TRKN4, TRKN3, RKNP529, RKNP638
2/29/2016	Collier x (Honey Drip x Collier)	RKNP529, RKNP638
3/9/2016	GT-IR7 x (Honey Drip x GT-IR7)	RKNP529, TRKN3, RKNP638
3/14/2016	GT-IR7 x (Honey Drip x GT-IR7)	RKNP638, RKNP194, RKNP465
3/15/2016	GT-IR7 x (Honey Drip x GT-IR7)	RKNP638, RKNP194, RKNP465
3/28/2016	Entry 22 x (Honey Drip x Entry 22) F₂ homozygous	RKNP342, RKNP402, RKNP529
3/28/2016	Collier x (Honey Drip x Collier)	RKNP342, RKNP402, RKNP529
4/20/2016	Top 76-6 x (Honey Drip x Top 76-6)	RKNP342, RKNP402, RKNP529
4/20/2016	Entry 22 x (Honey Drip x Entry 22) F₂ homozygous	RKNP342, RKNP402, RKNP529
4/21/2016	Entry 22 x (Honey Drip x Entry 22) F₂ homozygous	RKNP342, RKNP402, RKNP529
5/6/2016	Dale x (Honey Drip x Dale) F₂ homozygous	RKNP342, RKNP402, RKNP529
5/11/2016	Dale x (Honey Drip x Dale) F₂ homozygous	RKNP342, RKNP402, RKNP529
6/1/2016	Dale x (Honey Drip x Dale) F₂ homozygous	RKNP342, RKNP402, RKNP529
6/8/2016	Dale x (Honey Drip x Dale) F₂ homozygous	RKNP342, RKNP402, RKNP529
6/9/2016	GT-IR7 x (Honey Drip x GT-IR7) F₂ homozygous	RKNP342, RKNP402, RKNP529
6/22/2016	Collier x (Honey Drip x Collier) F₂ homozygous	RKNP342, RKNP402, RKNP529
8/30/2016	Top76-6 x (Honey Drip x Top 76-6) F₂ homozygous	RKNP529, RKNP638, RKNP709, RKNP821
6/15/2017	Genotyping of BC₁F₆ lines to determine the size of the Honey Drip crossover in the RKN region	RKNP17, RKNP135, RKNP342, RKNP402, RKNP529

1 Note: Many seedlings were created for each cross. At the F₂ homozygous stage (where the F₂ plant is homozygous for Honey Drip in the Meloidogyne incognita resistance QTL region), a single seedling was selected from each backcross and advanced to the F₆ stage.

Table 3.

Presence of the introgression from ‘Honey Drip’ in the QTL-Sb.RKN.3.1 region of each backcross (BC₁F₆) line.

	Allele Sizes (bp)
	RKNP17^a	RKNP135	RKNP342	RKNP402	RKNP529
NC_012872 (bp)b	(52,744,750)	(53,190,542)	(53,974,058)	(54,199,591)	(54,678,311)
Genotype
Honey Drip	120, 120	153, 153	240, 240	173, 173	222, 222
Entry 22	198, 198	210, 210	222, 222	204, 204	185, 185
Entry 22-BC1F6	120, 120	153, 153	240, 240	173, 173	222, 222
Collier	198, 198	200, 200	220, 220	253, 253	116, 116
Collier-BC1F6	120, 120	153, 153	240, 240	173, 173	222, 222
GT-IR7	198, 245	210, 210	220, 220	265, 265	116, 232
GT-IR7-BC1F6	120, 120	153, 153	240, 240	173, 173	222 , 232
Dale	198, 198	206, 206	222, 222	234, 234	185, 185
Dale-BC1F6	120, 120	153, 153	240, 240	173, 173	222, 222
Topper	198, 198	202, 202	220, 220	255, 255	116, 232
Top 76-6-BC1F6	120, 120	153, 153	240, 240	173, 173	222 , 232

1 Notes: ^aRKNP17, RKNP135, RKNP342, RKNP402, and RKNP529 are microsatellite markers in the QTL-Sb.RKN.3.1 region.

2b The base pair position is the start site of the forward primer on NC_012872, the sorghum chromosome 3 genomic sequence. Allele sizes in bold italics indicate this allele is from ‘Honey Drip’.

Polyacrylamide gel image of microsatellite marker RKNP135 from the sorghum donor line ‘Honey Drip’, the recurrent parents ‘Collier’, GT-IR7, ‘Dale’, and ‘Top 76-6’, and the BC₁F₆ progeny lines. A 50-350 bp sizing standard (LI-COR Biosciences, Lincoln, NE) was loaded in the first and last lanes.

Table 4.

Fresh root weights of parental sorghum genotypes and their BC₁F₆ progeny.

	Trial 1		Trial 2
Genotype	Root weight (g)a		Root weight (g)a
Top 76-6	63.3	A^b	97.8	A^b
Top 76-6-BC1F6	56.6	AB	69.2	B
Honey Drip	43.9	BC	52.6	BCD
Dale-BC1F6	41.4	BCD	53.7	BC
Entry 22-BC1F6	41.1	CD	43.3	CDE
Collier	40.9	CD	32.9	DE
Collier-BC1F6	40.1	CD	31.5	E
GT-IR7	38.5	CD	27.9	E
GT-IR7-BC1F6	38.3	CD	30.6	E
Dale	37.1	CD	39.5	CDE
Entry 22	26.6	D	24.0	E

1 Notes: ^aRoot weights are from plants 8 weeks after infection with M. incognita.

2b Means within a column followed by the same letter are not statistically different (P ≤ 0.05).

Table 5.

Total M. incognita eggs and eggs/gram of root of parental sorghum genotypes and their BC₁F₆ progeny that have the M. incognita resistance QTL QTL-Sb.RKN.3.1.

Genotype	Total eggs^a		Eggs/g root^a
Dale	425775	A^b	11141	A^b
Top 76-6	297250	A	3977	A
Collier	206325	A	5856	A
GT-IR7	170125	A	5584	A
Entry 22	115808	A	4661	A
Honey Drip	2300	B	48	B
Collier-BC1F6	1175	B	36	B
Entry 22-BC1F6	975	B	20	B
Dale-BC1F6	875	B	19	B
Top 76-6-BC1F6	750	B	18	B
GT-IR7-BC1F6	625	B	16	B

1 Notes: ^aCombined data from two trials. Data was collected 8 weeks after infection with M. incognita. Statistical analysis performed using log₁₀ transformed data, however, untransformed numbers are presented in the table.

2b Means within a column followed by the same letter are not statistically different (P ≤ 0.05).

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DOI: https://doi.org/10.21307/jofnem-2021-087 | Journal eISSN: 2640-396X | Journal ISSN: 0022-300X

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Language: English

Page range: 1 - 10

Published on: Nov 11, 2021

Published by: Society of Nematologists, Inc.

In partnership with: Paradigm Publishing Services

Publication frequency: 1 issue per year

Keywords:

Breeding,

Markers,

Nematode resistance,

QTL,

Sorghum bicolor,

Southern root-knot nematode

Related subjects:

Life sciences,

Life sciences, other

© 2021 Richard F. Davis, Karen R. Harris-Shultz, Joseph E. Knoll, Hongliang Wang, published by Society of Nematologists, Inc.
This work is licensed under the Creative Commons Attribution 4.0 License.

Volume 53 (2021): Issue 1 (January 2021)

Transfer of Meloidogyne incognita Resistance Using Marker-assisted Selection in Sorghum

Figures & Tables

Table 1.

Table 2.

Table S1.

Table 3.

Figure 1:

Table 4.

Table 5.

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