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Research Article

Screening of Cabbage (Brassica oleracea L.) Germplasm for Resistance to Black Rot

Khandker Shazia Afrin1, Md Abdur Rahim1,2, Jong-In Park1, Sathishkumar Natarajan1, Mehede Hassan Rubel1, Hoy-Taek Kim1, Ill-Sup Nou1,*
Plant Breeding and Biotechnology 2018;6(1):30-43.
Published online: March 1, 2018

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*Corresponding author: Ill-Sup Nou, nis@sunchon.ac.kr, Tel: +82-61-750-3249, Fax: +82-61-750-3208
• Received: January 20, 2018   • Revised: January 31, 2018   • Accepted: January 31, 2018

Copyright © 2018 The Korean Society of Breeding Science

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Screening of Cabbage (Brassica oleracea L.) Germplasm for Resistance to Black Rot
Plant Breed. Biotech.. 2018;6(1):30-43.   Published online March 1, 2018
DOI: https://doi.org/10.9787/PBB.2018.6.1.30
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Screening of Cabbage (Brassica oleracea L.) Germplasm for Resistance to Black Rot
Plant Breed. Biotech.. 2018;6(1):30-43.   Published online March 1, 2018
DOI: https://doi.org/10.9787/PBB.2018.6.1.30
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Screening of Cabbage (Brassica oleracea L.) Germplasm for Resistance to Black Rot
Image Image Image
Fig. 1 The disease reaction scoring criteria used in this study for black rot of cabbage. Scales: 0 = no visible symptoms; 1 = small necrosis or chlorosis near the inoculation point; 3 = typical small V-shaped lesion with black veins; 5 = typical lesion half way to the middle vein; 7 = typical lesion succeeding to the middle vein; and 9 = lesion reaching the middle vein (Vicente et al. 2002). 0, high resistant (HR); 1–3, resistant (R); 5–7, susceptible (S); 9, highly susceptible (HS).
Fig. 2 The resistant and susceptible cabbage lines against different races of Xanthomonas campestris pv. campestris (Xcc) at 2 weeks after inoculation. The letters a, b, c, d, e, f and g, represents Xcc race 1, 2, 3, 4, 5, 6 and 7, respectively.
Fig. 3 Chromosomes location of five reported polymorphic markers on Brassica oleracea.

Fig. 1

Fig. 2

Fig. 3

Screening of Cabbage (Brassica oleracea L.) Germplasm for Resistance to Black Rot

Xanthomonas campestris pv. campestris (Xcc) races used for bioassay-based screening of cabbage lines.

Sl. No. Bacterial race/strains Source Reference
1 Xanthomonas campestris pv. campestris Race 1 (B100) UK Vicente et al. 2001
2 Xanthomonas campestris pv. campestris Race 2 (3849A) US
3 Xanthomonas campestris pv. campestris Race 3 (5212) UK
4 Xanthomonas campestris pv. campestris Race 4 (CFBP 5817) UK
5 Xanthomonas campestris pv. campestris Race 5 (3880) Australia
6 Xanthomonas campestris pv. campestris Race 6 (6181) Portugal
7 Xanthomonas campestris pv. campestris Race 7 (8450A) UK

List of markers with their primer sequence used in the molecular screening for black rot resistance in cabbage.

Sl. No. Markers Chromosome Primer sequence (5′……3′) Tm (°C) Size (bp) Marker type Reference
1 BoESSR291 C03 F: AAGCTGGGGATGGAGAAGAT
R: GCACCTAATCGAACCCCTTA		 52 114 SSR Izzah et al. 2014
2 BoESSR216 C01 F: GGTTTCCGCTATGTCCAGAA
R: CGGAAGAAGACGTTGAGGAG		 52 317 SSR Izzah et al. 2014
3 BoESSR089 C01 F: ATGATCAGCGAAACCACTCC
R: TGATACATCCCGTTTGCTCA		 55 259 SSR Izzah et al. 2014
4 BoESSR145 C01 F: GGGCGAGGATGGTTACTACA
R: TCATACCCCAAGGCTATTTT		 55 240 SSR Izzah et al. 2014
5 BoESSR726 C01 F: CAATGGGTTACGCATGGTTT
R: CGTTTGTGAAACAGCCATTG		 55 228 SSR Izzah et al. 2014
6 SSR739 C03 F: TAGGGTGAAAGGGAAGCTCA
R: CGCTAATAATGGCGCTAAGG		 52 182 SSR Saha et al. 2014
7 BnGMS301 C01 F: AATATGCAGCATTCTAGACAAA
R: ATCATTCTCGTGATGACACA		 55 250 SSR Cheng et al. 2009
8 BoGMS0971 C08 F: TAATCCGAACAACACGAA
R: CACCCAATAAGCGATGAG		 52 344 SSR Li et al. 2011
9 OI10G06 C06 F: GACAAGTTCCCTTGTAATGGC
R:TGTAATCATCACACATTTTGGG		 52 109 SSR Li et al. 2011
10 Iso0857_371bc C09 F: AAGGAATTCCCCAGGATGTC
R: TTGGCAACCCTAATGCTTTT		 51 477 InDel Lee et al. 2015

Comparison of bioassay and molecular screening results for black rot resistance in cabbage.

Sl. No. Lines Race 1 OI10G06 BnGMS301 BoESSR726 BoESSR291 Race 2 BoESSR291 OI10G06 BoESSR726 BoGMS0971
Pz) Gy) Pz) Gy)
1 SCNU-C-106 HSu) + + + + HS + + + −
2 SCNU-C-107 n/at) − + + − HS − − + +
3 SCNU-C-3122 Sv) + − + + HS + + + +
4 SCNU-C-3270 S + − + + S + + + +
5 SCNU-C-3273 S + + + − HRx) − + + −
6 SCNU-C-3305 HS + + + + S + + + +
7 SCNU-C-3328 S + − − + HS + + − +
8 SCNU-C-3412 S + + + + HS + + + −
9 SCNU-C-3414 S − + + − n/a − − + −
10 SCNU-C-3449 HS + + + + HS + + + +
11 SCNU-C-3470 n/a + + − + n/a + + − +
12 SCNU-C-3631 S − + + − HR − − + −
13 SCNU-C-3637 S + + + − HR − + + −
14 SCNU-C-3639 S + + − + HR + + − +
15 SCNU-C-4059 S + + + + S + + + −
16 SCNU-C-4066 S + + + − S − + + −
17 SCNU-C-4071 S − + + + S + − + −
18 SCNU-C-4072 n/a − + + + HR + − + −
19 SCNU-C-4073 S + + + + HR + + + −
20 SCNU-C-4074 Rw) − − − + n/a + − − −
21 SCNU-C-4085 S + + + + S + + + −
22 SCNU-C-4105 S + + − + S + + − +
23 SCNU-C-4118 S + − − − S − + − +
24 SCNU-C-4161 HS + + + + S + + + −
25 SCNU-C-4168 S − + + − S − − + −
26 SCNU-C-4302 HS + + − + HS + + − +
27 SCNU-C-4320 HS + + + + HS + + + −
Matched 20.0 19.0 19.0 16.0 17.0 17.0 15.0 14.0
Not matched 4.0 5.0 5.0 9.0 6.0 6.0 8.0 9.0
% Adaptability 83.3 79.2 79.2 66.7 73.9 73.9 65.2 60.9
Sl. No. Lines Race 3 BnGMS301 BoESSR291 BoESSR726 OI10G06 Race 4 BnGMS301 BoESSR726 OI10G06 BoESSR291
Pz) Gy) Pz) Gy)
1 SCNU-C-106 HS + + + + n/a + + + +
2 SCNU-C-107 HS + − + − S + + − −
3 SCNU-C-3122 HS − + + + HS − + + +
4 SCNU-C-3270 HS − + + + S − + + +
5 SCNU-C-3273 R + − + + S + + + −
6 SCNU-C-3305 S + + + + HS + + + +
7 SCNU-C-3328 HS − + − + HS − − + +
8 SCNU-C-3412 HS + + + + S + + + +
9 SCNU-C-3414 S + − + − S + + − −
10 SCNU-C-3449 HS + + + + HS + + + +
11 SCNU-C-3470 S + + − + S + − + +
12 SCNU-C-3631 S + − + − S + + − −
13 SCNU-C-3637 S + − + + S + + + −
14 SCNU-C-3639 S + + − + S + − + +
15 SCNU-C-4059 S + + + + S + + + +
16 SCNU-C-4066 HS + − + + HS + + + −
17 SCNU-C-4071 HS + + + − S + + − +
18 SCNU-C-4072 HS + + + − n/a + + − +
19 SCNU-C-4073 HS + + + + S + + + +
20 SCNU-C-4074 HS − + − − HS − − − +
21 SCNU-C-4085 HS + + + + HS + + + +
22 SCNU-C-4105 HS + + − + S + − + +
23 SCNU-C-4118 R − − − + S − − + −
24 SCNU-C-4161 HS + + + + n/a + + + +
25 SCNU-C-4168 HS + − + − S + + − −
26 SCNU-C-4302 HS + + − + S + − + +
27 SCNU-C-4320 HS + + + + HS + + + +
Matched 22.0 21.0 20.0 19.0 19.0 18.0 17.0 16.0
Not matched 5.0 6.0 7.0 8.0 5.0 6.0 7.0 8.0
% Adaptability 81.5 77.8 74.1 70.1 79.2 75.0 70.8 66.7
Sl. No. Lines Race 5 BnGMS301 BoESSR726 BoESSR291 OI10G06 Race 6 BnGMS301 BoESSR726 BoESSR291 OI10G06
Pz) Gy) Pz) Gy)
1 SCNU-C-106 HS + + + + HS + + + +
2 SCNU-C-107 HS + + − − HS + + − −
3 SCNU-C-3122 S − + + + HS − + + +
4 SCNU-C-3270 R − + + + S − + + +
5 SCNU-C-3273 n/a + + − + HS + + − +
6 SCNU-C-3305 S + + + + HS + + + +
7 SCNU-C-3328 HS − − + + HS − − + +
8 SCNU-C-3412 S + + + + S + + + +
9 SCNU-C-3414 n/a + + − − HS + + − −
10 SCNU-C-3449 HS + + + + S + + + +
11 SCNU-C-3470 n/a + − + + R + − + +
12 SCNU-C-3631 S + + − − HS + + − −
13 SCNU-C-3637 HS + + − + HS + + − +
14 SCNU-C-3639 HS + − + + S + − + +
15 SCNU-C-4059 S + + + + S + + + +
16 SCNU-C-4066 HS + + − + HS + + − +
17 SCNU-C-4071 HS + + + − HS + + + −
18 SCNU-C-4072 HS + + + − HS + + + −
19 SCNU-C-4073 HS + + + + HS + + + +
20 SCNU-C-4074 n/a − − + − S − − + −
21 SCNU-C-4085 S + + + + HS + + + +
22 SCNU-C-4105 HS + − + + S + − + +
23 SCNU-C-4118 R − − − + R − − − +
24 SCNU-C-4161 S + + + + HS + + + +
25 SCNU-C-4168 HS + + − − HS + + − −
26 SCNU-C-4302 HS + − + + S + − + +
27 SCNU-C-4320 HS + + + + HS + + + +
Matched 21.0 18.0 17.0 16.0 22.0 22.0 19.0 18.0
Not matched 2.0 5.0 6.0 7.0 5.0 5.0 8.0 9.0
% Adaptability 91.3 78.3 73.9 69.6 81.5 81.5 70.4 66.7
Sl. No. Lines Race 7 BnGMS301 BoESSR291 OI10G06 BoESSR726
Pz) Gy)
1 SCNU-C-106 HS + + + +
2 SCNU-C-107 R + − − +
3 SCNU-C-3122 HS − + + +
4 SCNU-C-3270 HR − + + +
5 SCNU-C-3273 n/a + − + +
6 SCNU-C-3305 S + + + +
7 SCNU-C-3328 HS − + + −
8 SCNU-C-3412 HS + + + +
9 SCNU-C-3414 n/a + − − +
10 SCNU-C-3449 S + + + +
11 SCNU-C-3470 HR + + + −
12 SCNU-C-3631 HS + − − +
13 SCNU-C-3637 HS + − + +
14 SCNU-C-3639 S + + + −
15 SCNU-C-4059 HR + + + +
16 SCNU-C-4066 S + − + +
17 SCNU-C-4071 HS + + − +
18 SCNU-C-4072 HS + + − +
19 SCNU-C-4073 S + + + +
20 SCNU-C-4074 HS − + − −
21 SCNU-C-4085 HS + + + +
22 SCNU-C-4105 HS + + + −
23 SCNU-C-4118 HS − − + −
24 SCNU-C-4161 HS + + + +
25 SCNU-C-4168 HS + − − +
26 SCNU-C-4302 S + + + −
27 SCNU-C-4320 HS + + + +
Matched 18.0 17.0 17.0 16.0
Not matched 7.0 8.0 8.0 9.0
% Adaptability 72.0 68.0 68.0 64.0

z)P, phenotype based on disease reaction;

y)G, genotype on molecular marker screening;

x)HR, high resistant;

w)R, resistant;

v)S, susceptible;

u)HS, high susceptible;

t)n/a, bioassay data not available.

The tested molecular markers and their location on Brassica oleracea genome.

Maker name Locationz) Description
BoGMS0971 C8:31754531..31754753 Intergenic region (Bo8g092870 and Bo8g092880)
BnGMS301 C1:15386406..15386643 Intergenic region (Bo1g053300 and Bo1g053310)
BoESSR291 C3:21846613..21846644 Promoter region of a gene cystathionine gamma-synthase (Bo3g055310)
BoESSR726 C1:15967972..15968174 Intergenic region (Bo1g054960 and Bo1g054970)
OI10G06 C6:29898028..29898121 Intergenic region (Bo6g095520 and Bo6g095530)

z)The chromosomal locations are shown according to the ENSEMBL-based Brassica oleracea genome database.

Genes around the tested polymorphic molecular markers in Brassica oleracea genome.

Makers Gene ID Description
NCBIz) TAIRy) Bolbasex)
BoGMS0971 Bo8g092870 V-type proton ATPase subunit D-like Vacuolar ATP synthase subunit D V-type proton ATPase subunit D-like
Bo8g092880 Citrate synthase 2, peroxisomal-like Citrate synthase 1 Citrate synthase-like, large alpha subdomain
BnGMS301 Bo1g053300 (E)-beta-ocimene synthase, chloroplastic Terpene synthase 02 Terpene synthase-like
Bo1g053310 (E)-beta-ocimene synthase, chloroplastic Terpene synthase 02 Uncharacterized protein
BoESSR291 Bo3g055310 Cystathionine gamma-synthase Cystathionine gamma-synthase Cystathionine gamma-synthase (CGS)
BoESSR726 Bo1g054960 Uncharacterized ABC transporter family protein ABC transporter D family member 2, chloroplastic
Bo1g054970 Uncharacterized Mitochondrial transcription termination factor family protein Mitochodrial transcription termination factor-related
OI10G06 Bo6g095520 Uncharacterized Unknown protein Putative uncharacterized protein
Bo6g095530 Protein argonaute 7-like Argonaute family protein Protein argonaute

z)The National Center for Biotechnology Information (NCBI);

y)The Arabidopsis Information Resource (TAIR);

x)The Brassica oleracea Genome Database (Bolbase).

Table 1 Xanthomonas campestris pv. campestris (Xcc) races used for bioassay-based screening of cabbage lines.
Table 2 List of markers with their primer sequence used in the molecular screening for black rot resistance in cabbage.
Table 3 Comparison of bioassay and molecular screening results for black rot resistance in cabbage.

P, phenotype based on disease reaction;

G, genotype on molecular marker screening;

HR, high resistant;

R, resistant;

S, susceptible;

HS, high susceptible;

n/a, bioassay data not available.

Table 4 The tested molecular markers and their location on Brassica oleracea genome.

The chromosomal locations are shown according to the ENSEMBL-based Brassica oleracea genome database.

Table 5 Genes around the tested polymorphic molecular markers in Brassica oleracea genome.

The National Center for Biotechnology Information (NCBI);

The Arabidopsis Information Resource (TAIR);

The Brassica oleracea Genome Database (Bolbase).

Table 1

Table 2

Table 3

Table 4

Table 5

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