Meloidogyne spp. Plant parasitic nematode (sedentary endo-parasitic) Wide host range (>2,000 plant species)

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2 Meloidogyne spp. Plant parasitic nematode (sedentary endo-parasitic) Wide host range (>2,000 plant species)

3 Southern root-knot (M. incognita) (Mi) Javanese root-knot (M. javanica) (Mj) Peanut root-knot (M. arenaria) (Ma) Yield losses in the United States were 9.6 million bushels in 2008 Riggs & Schmitt, 1987; Wrather & Koenning, 2008; Contributed by Roger Boerma

4 Nematode Soybean Cotton Peanut Corn Southern root-knot Host Host Non-host Host Peanut root-knot Host Non-host Host Host Javanese root-knot Host Non-host Non-host Host Reniform Host Host Non-host Non-host Soybean cyst Host Non-host Non-host Non-host

5 Contributed by Roger Boerma & Jennifer Yates

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7 S-100 CNS Tokyo FC33243 Lee D R Lee74 Dyer Bragg Pickett71 Forrest Govan Gregg Coker338 Gordon Pharaoh Hartwig Narow Doles Maxcy Manokin Walters PI Volstate Palmetto Jackson Gasoy17 Coker488 Wright Hutcheson Coker6738 Benning Cobb Centennial Braxton Hutton Sharkey Perrin Cook Lamer Haskell Hampton Young Hagood Dillon Ransom N Johnston Ha et al., Crop Sci. 44

8 S-100 CNS Tokyo FC33243 Lee D R Lee74 Dyer Bragg Pickett71 Forrest Govan Gregg Coker338 Gordon Pharaoh Hartwig Narow Doles Maxcy Manokin Walters PI Volstate Palmetto Jackson Gasoy17 Coker488 Wright Hutcheson Coker6738 Benning Cobb Centennial Braxton Hutton Sharkey Perrin Cook Lamer Haskell Hampton Young Hagood Dillon Ransom N Johnston Ha et al., Crop Sci. 44

9 Southern root-knot (Mi) PI96354 (Luzzi et al., 1987, Crop Sci. 27; Harris et al., 2003, Crop Sci. 43) Peanut root-knot (Ma) PI PI594427C PI594651B (Luzzi et al., 1987, Crop Sci. 27; Harris et al., 2003, Crop Sci. 43; Yates et al., 2010, Crop Sci. 50) Javanese root-knot (Mj) PI (Luzzi et al., 1987, Crop Sci. 27; Harris et al., 2003, Crop Sci. 43) G (Luzzi et al., 1996, Crop Sci.36) G (Luzzi et al., 1996, Crop Sci.36) G (Luzzi et al., 1997, Crop Sci.37)

10 Gm10 (Lg-O) Satt358 Mi Satt492 Satt445 (R 2 =56%) Gm13 (Lg-F) A757-2 Satt334 B212-1 Satt510 Ma Mj PI96354 Mi (h 2 =69%) PI Ma (h 2 =55%) PI Mj (h 2 =55%) (R 2 =32%) (R 2 =46%) Gm18 (Lg-G) Satt199 Mi Satt012 Satt503 (R 2 =18%) Gm01 (Lg-D1a) Sat_353 A725-2 Mj R013-2 (R 2 =13%) Gm15 (Lg-E) B212-2 Ma (R 2 =16%) A111-2 Satt553 Tamulonis et al.,1997, TAG 95 & Crop Sci. 37 ; Li et al., 2001, TAG103

11 110 F 2 PI96354 Bossier No. of galls QTL on LG-O/G; +/+ - / - Adapted from Li et al., 2001,TAG 103

12 92 F 6 Cook (R, 200bp) N7001 (S, 192bp) Nematode reaction Satt358 homoz. 200 bp Satt358 homoz. 192 bp Resistant lines 40 0 Susceptible lines 1 51

13 a) Satt358 containing genomic DNA clone on Gm10 PI ATTATAAATGCTATCCTTTAATTCTTAGCTATGCGCTTTATGTAACAATACGATTTCTAT 12 0 BOSSIER 61 ATTATAAATGCTATCCTTTAATACTTAGCTGTGCGCTTTATGTAACAATACGATTTCTAT 12 0 ********************** ******* ***************************** PI TATTATTATTATTATTATTATTATTATTATTATATTTTTTCCTATTTTTGGAAATATATT 21 0 BOSSIER 151 TATTATTATTATTATTATT ATTATATTTTTGCCTATTTTTGGAAATATATT 20 1 ******************* *********** ******************** b) PI96354 Satt TTTATATTTCTGTTTTCTCTTAAGAAAAAAAAAACTAAAACAAAATGTCGACAGTAGTTG containing genomic DNA clone on Gm BOSSIER 61 TTTATATTTCTGTTTTCTCTTAA--AAAAAAAAACTAAAACAAAATGTCCACAGTAGTTG 11 8 *********************** ************************ ********** PI ATTATTATTATTATTATTATTATTATTATTAATATTAGACTGTAAACGTTTTAATTACTA 23 0 BOSSIER 169 ATTATTA TTATTATTATTATTATTAGGCTGTAAACGTTTTAATTACTA 21 6 ******* ************ ****** ********************* Ha et al., 2007, Crop Plant Sci. Genome 47S

14 SNP199 A/G./TATTAGGCTGTAAACGTTTTAATTACTACTACTTTTTTACTTTATGAATTTTGTGAGTAAGTTGATGATGGACC/. 5 3 P AATCCGACATTTGCAAAATTAATGATGATGAAAAA GAAATACTTAAAACACTCATTCAACTACTACCTGG Sensor probe Anchor probe R F HybProbe P 3 5 ATAATCCGACATTTGCAAAATTAATGATGATGAAA Q F SimpleProbe Ha et al., 2008, J. Crop Sci. Biotech.11

15 SNP199 HybProbe SNP358 PI Hetero Bossier PI Bossier SNP199 SimpleProbe SNP358 PI Bossier PI Bossier Ha et al., 2008, J. Crop Sci. Biotech.11

16 US patent application No: Title : Methods and compositions for selecting soybean plants resistant to southern root knot nematode Inventors: Narvel, James; Concibido, Vergel; Cerny, Liesa; Tamulonis, John; Hancock, Floyd; Dougherty, Richard; Boerma, Henry Roger; Ha, Bo-Keun Application Filed: Aug 6, 2008 Publication Date: Mar 5, 2009

17 Satt 510 Gm13 (Lg-F) OPN 11 PI PI594427C PI594651B PI (Mj) 3 cm 83I9 98F7 S R S S S R R L20a (R 2 =41%) SoyHsp176 Yates, 2006, Ph.D.Diss.

18 188 F 2 PI594427C x CNS Ma resistant Ma susceptible Ma heterozygous Yates et al., unpublished

19 Greenhouse screening Planting + Inoculation + Gall counting = 35 days $/data $ 5.00 LightCycler 480 (SNP) DNA (1 week) PCR setup PCR/ melting curve Genotype calling $ 0.23 ~ 0.27 ABI3730 (SSR) DNA (1 week) PCR setup PCR ABI3730 $ 0.35 ~ 0.42 Time(hours) Boerma & Hussey, 1992, J Nemato. 24; Ha et al., 2008, J. Crop Sci. Biotech.11

20 Two backcross populations BenningRR(5) x G BoggsRR(6) x G BenningRR and BoggsRR = tawny G = gray Pubescence color Gm06 (Lg-C2) TT, or Tt = tawny tt = gray Phenotypic selection for high Mi resistance after each cycle of backcrossing Lines should have TAWNY Some of them were GRAY! (Woodworth, 1921)

21 Lines Entries (No.) Mean (eggs plant -1 ) Range (eggs plant -1 ) Bossier n/a G n/a Seed yield (kg ha -1 ) BenningRR a BenningRR(5) x G Tawny lines 8 170a b Gray lines 6 42b ab Bossier n/a G n/a BoggsRR 1 30 n/a 3101a BoggsRR(6) x G Tawny lines 5 63a a Gray lines 5 6b a LSD (0.05) LSD (0.01) Shearin et al., 2009, Crop Sci. 49

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