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ARTICLE

Genetic Analysis on Short Culm and the Rice Blast Resistance of Namil(SA)-bl5, a Japonica Rice Mutant Line


Published online: August 31, 2014

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*Corresponding Author (E-mail: jrnj@korea.kr Tel: +82-31-290-6728, Fax: +82-31-290-6730)
• Received: July 29, 2014   • Revised: September 12, 2014   • Accepted: September 16, 2014

© 2014 by the Korean Society of Breeding Science

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    Kuan-Lin Lo, Yi-Nian Chen, Min-Yu Chiang, Mei-Chun Chen, Jerome P. Panibe, Chung-Chun Chiu, Lu-Wei Liu, Liang-Jwu Chen, Chun-Wei Chen, Wen-Hsiung Li, Chang-Sheng Wang
    Rice.2022;[Epub]     CrossRef

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Genetic Analysis on Short Culm and the Rice Blast Resistance of Namil(SA)-bl5, a Japonica Rice Mutant Line
Korean. J. Breed. Sci.. ;46(3):238-249.   Published online September 30, 2014
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Genetic Analysis on Short Culm and the Rice Blast Resistance of Namil(SA)-bl5, a Japonica Rice Mutant Line
Korean. J. Breed. Sci.. ;46(3):238-249.   Published online September 30, 2014
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Genetic Analysis on Short Culm and the Rice Blast Resistance of Namil(SA)-bl5, a Japonica Rice Mutant Line
Image Image Image
Fig. 1. Agronomic performances of the wild type, Namil (left) and the mutant line of Namil, Namil(SA)-bl5 (right) (A) and reaction patterns against virulent rice blast isolate, 94-254 of Namil, Milyang23, and Namil(SA)-bl5, sequentially from left to right (B).
Fig. 2. Graphical comparisons of F2 mapping populations derived from two crosses between Namil (wild type) or Namil(SA)-bl5 and Milyang23 on the agronomic trait, culm length (A) and virulent rice blast isolate, 94-254 (B). To measure the agronomic trait, culm length of F2 progeny lines, harvested seeds from each F2 plant (F2:3 seeds) were planted in a raw. The descriptive statistics on the agronomic trait, culm length of 103 F2:3 lines of Namil x Milyang23, over-all mean, standard deviation, skewness, and kurtosis are 84.7, 7.4, 0.0, and -0.7, respectively. In 94 F2:3 lines of Namil(SA)-bl5 x Milyang23 on the agronomic trait, culm length, over-all mean, standard deviation, skewness, and kurtosis are 72.5, 6.7, 0.0, and -0.5, respectively. To estimate the resistant levels of F2 progeny lines, harvested seeds from each F2 plant (F2:3 seeds) were planted in a raw. Fourteen days after inoculation of rice blast isolate (94-254), individuals were evaluated. On virulent rice blast, 94-254, the descriptive statistics of 103 F2:3 lines of Namil x Milyang23, over-all mean, standard deviation, skewness, and kurtosis are 4.0, 0.6, -0.8, and 0.6, respectively. In 94 F2:3 lines of Namil(SA)-bl5 x Milyang23 on virulent rice blast isolate, 94-254, over-all mean, standard deviation, skewness, and kurtosis are 2.3, 1.5, 0.2, and -1.2, respectively.
Fig. 3. Linkage map skeleton of the agronomic trait, culm length and rice blast resistance on rice chromosomes. In Namil x Milyang23, wild type, preliminary gene locus concerned with culm length is on rice chromosome 1, and putative gene loci of rice blast resistance are on rice chromosome 5 and 11 (A). In Namil(SA)-bl5, a mutant line, preliminary gene loci concerned with culm length are on rice chromosome 6 and 7, and putative gene loci of rice blast resistance are on the same rice chromosome 6 and 12 (B). SSR markers were applied based on F-statistics from single-locus ANOVA results to narrow done the putative location responsible for rice blast resistance and brown Planthopper resistance. The genetic intervals are shown in centimorgan (cM).
Genetic Analysis on Short Culm and the Rice Blast Resistance of Namil(SA)-bl5, a Japonica Rice Mutant Line

Reactions of the mutant line, Namil(SA)-bl5 and cultivars against selected Korean virulent blast isolates.

Lines Rice blast isolate (race)z
90-002 (KI215) 90-008 (KI1113) 93-093 (KI197) 94-254 (ND)y 97-260 (KJ103) 02-319 (KJ105) 02-039 (KJ201)
Namil 4 4 5 4 4 5 5
Milyang23 4 3 4 4 2 4 2
Junam 5 5 5 5 5 5 3
Onnuri 4 4 5 5 4 1 4
Chilbo 2 5 5 5 5 4 4
Namil(SA)-bl5 0 1 1 0 1 1 1

Rice blast isolates were collected from Korean famers’ fields. Resistance degrees of virulent rice blast were classified five levels from 1 (resistance) to 5 (susceptibility). Korean rice cultivars, Junam, Onuri, and Chilbo were included as check varieties

Not determined

Comparisons between Namil, the wild type and Namil(SA)-bl5, a mutant line in terms of important agronomic traits.z

Lines HD (month.day) CL (cm) PL (cm) TN (No.) SN (No.) RGP (%) TWD (g) YDB (Kg/10a)

Namil 7.29 ± 0.0 77 ± 2.0 24 ± 0.6 11 ± 1.0 147 ± 7.8 74.4 ± 7.1 24.8 ± 0.0 599 ± 24.0
Namil(SA)-bl5 7.25 ± 0.6 65 ± 1.7 19 ± 0.0 17 ± 2.1 95 ± 0.6 74.2 ± 7.5 20.5 ± 0.2 578 ± 21.4

Heading date (HD), Culm length (CL), Panicle length (PL), Tiller number (TN), Spikelet number (SN), Ripened grain percentage (RGP), 1,000-grain weight of hulled grain (TWD), and Yield of brown rice (YDB). Note that the agronomic traits were evaluated by harvesting 100 plant for each replication plot (n=3), which were planted as three seedlings per plant during transplanting. All numerical values were mean ± standard deviation.

List of SSR primer sets, their e-Landing mediated physical positions on rice Pseudomolecule 6, corresponding BAC/PAC clones with their determined cM positions, and the segregating (χ2 test) on the F2 progenies derived from the cross between Namil(SA)-bl5 and Milyang23.

Markerz Physical informationy
Mirror mapx
Segregation Testw
Ch Start Stop e-PCR e % BAC/PAC cM% cM A H B M χ2

RM3252 1 299,681 299,852 172 0.7 AP002818 0.2 0.3 19 53 22 0 1.4
RM0600 1 9,461,346 9,461,566 221 21.7 AP001081 27.3 49.6 21 49 24 0 0.2
RM0449 1 15,305,619 15,305,758 140 35.1 AP008247 40.0 72.8 29 44 21 0 1.4
RM1349 1 25,398,082 25,398,300 219 58.3 AP002744 56.7 103.1 24 46 24 0 0.0
RM1003 1 33,803,800 33,803,927 128 77.5 AP003345 75.1 136.6 28 42 24 0 1.1
RM6321 1 43,251,019 43,251,186 168 99.2 AP003277 100.0 181.8 26 42 26 0 0.8

RM0154 2 1,083,895 1,084,077 183 3.0 AP005851 3.0 4.7 16 62 16 0 8.6 *
RM5699 2 8,981,409 8,981,575 167 25.0 AP005803 26.7 42.1 19 53 22 0 1.4
RM1211 2 18,450,427 18,450,588 162 51.4 AP005299 37.7 59.5 19 55 20 0 2.3
RM3220 2 28,458,108 28,458,248 141 79.2 AP004118 74.8 118.1 13 52 29 0 5.8
RM3850 2 35,425,798 35,425,907 110 98.6 AP003989 100.0 157.9 13 55 26 0 5.6

RM6301 3 2,631,471 2,631,613 143 7.2 AC090485 8.1 13.3 25 48 21 0 0.2
RM0007 3 9,808,540 9,808,710 171 27.0 AC134232 27.0 44.4 15 47 31 1 4.8
RM1164 3 14,840,558 14,840,757 200 40.8 AC084766 41.8 68.7 21 44 29 0 1.4
RM6266 3 23,778,063 23,778,222 160 65.4 AC091246 57.7 94.9 24 45 25 0 0.1
RM1350 3 28,632,514 28,632,682 169 78.8 AC087181 77.1 126.8 28 44 22 0 0.9
RM3585 3 36,080,616 36,080,784 169 99.3 AC128647 98.4 161.7 27 42 25 0 0.9

RM0551 4 168.620 168,811 192 0.5 AL606442 2.4 3.1 22 48 24 0 0.0
RM5633 4 13,059,370 13,059,580 211 37.1 AL731595 15.4 19.9 13 54 27 0 5.5
RM1155 4 20,328,759 20,328,906 148 57.7 AL606453 45.4 58.9 24 35 35 0 8.0 *
RM3839 4 23,870,755 23,870,972 218 67.7 AL606614 56.2 72.8 19 38 37 0 9.4 **
RM3217 4 30,083,469 30,083,662 194 85.4 AL606683 77.7 100.7 23 47 18 6 0.7
RM0559 4 35,117,645 35,117,804 160 99.6 AL606637 100.0 129.6 22 56 16 0 3.7

RM5693 5 441,872 442,071 200 1.5 AC129716 3.8 4.6 26 51 17 0 2.0
RM5844 5 9,128,802 9,128,996 195 30.6 AC120989 43.7 53.5 27 46 18 3 1.4
RM5558 5 21,168,727 21,168,899 173 70.9 AC105769 70.3 86.0 22 48 24 0 0.0
RM1054 5 29,144,035 29,144,184 150 97.6 AC98598 99.8 122.0 25 44 25 0 0.2

RM3353 6 435,582 435,697 116 1.4 AP001129 1.1 1.4 18 50 26 0 1.4
RM3370 6 6,434,836 6,435,012 177 20.6 AP005545 27.6 34.3 18 49 27 0 1.5
RM0539 6 8,170,600 NA 26.1 AP005619 41.0 51.3 15 49 26 4 2.9
RM0527 6 9,863,290 9,863,522 233 31.6 AP005695 45.3 56.3 16 50 26 2 2.4
RM6818 6 16,581,413 16,581,542 130 53.1 AP004012 52.9 65.8 18 51 25 0 1.4
RM3628 6 23,737,032 23,737,157 126 76.0 AP003612 68.6 85.4 16 51 27 0 2.7
RM6274 6 26,319,462 26,319,530 69 84.2 AP003568 80.6 100.3 23 40 29 2 2.0
RM5604 6 29,048,075 29,048,152 78 93.0 AP003772 92.0 113.4 22 46 26 0 0.2
RM5753 6 30,966,850 30,967,050 201 99.1 AP004685 100.0 124.4 18 52 23 1 1.5

RM1093 7 668,161 668,310 150 2.3 AP003746 2.1 2.5 22 53 19 0 1.4
RM0481 7 2,876,165 2,876,333 169 9.7 AP005632 14.8 11.0 20 53 20 1 1.4
RM1243 7 3,555,641 3,555,797 157 12.0 AP004267 21.4 25.4 21 55 17 1 2.9
RM1377 7 12,782,829 12,783,009 181 43.1 AP004305 41.9 49.7 33 40 21 0 4.5
RM3743 7 19,342,334 19,342,513 180 65.2 AP003815 56.5 67.0 30 44 20 0 2.0
RM5623 7 23,110,498 23,110,681 184 77.8 AP005908 68.4 81.1 30 46 17 1 3.1
RM3555 7 27,891,580 27,891,733 154 93.9 AP004378 89.1 105.7 25 45 21 3 0.2
RM0172 7 29,560,592 29,560,750 159 99.6 AP005199 100.0 118.6 23 45 26 0 0.2

RM0408 8 119,935 120,063 129 0.4 AP005406 0.4 0.5 24 50 20 0 0.5
RM0547 8 5,586,058 5,586,291 234 19.7 AP004746 33.2 40.2 20 51 23 0 0.6
RM3262 8 22,248,334 22,248,500 167 78.6 AP005483 71.5 86.7 24 51 19 0 0.9
RM5545 8 28,141,927 28,142,083 157 99.4 AP004623 100 121.2 19 48 27 0 1.1

RM23654 9 151,453 151,639 187 0.7 AP006059 0.0 0.0 28 55 11 0 8.0 *
RM0316 9 1,074,933 1,075,126 194 4.7 AP005860 0.9 0.8 28 55 11 0 8.0 *
RM0219 9 7,887,585 NA 34.3 AP005912 22.1 20.7 27 53 14 0 4.5
RM0566 9 14,704,798 14,705,036 239 63.9 AP005397 54.2 50.7 24 48 22 0 0.0
RM0205 9 22,720,646 22,720,801 156 98.7 AP005546 100.0 93.5 20 44 30 0 2.0

RM7492 10 39,037 39,181 145 0.2 AC166065 0.0 0.0 21 55 18 0 2.4
RM0311 10 9,487,243 9,487,406 164 41.5 AC090482 20.0 16.8 22 50 22 0 0.2
RM1375 10 16,386,764 16,386,943 180 71.6 AC025905 51.0 42.7 27 44 23 0 0.5
RM0590 10 22,784,993 22,785,130 138 99.6 AC018727 100.0 83.8 16 56 22 0 3.7

RM0286 11 383,839 383,960 122 1.3 BX000497 1.2 1.4 15 54 24 1 3.6
RM6901 11 1,494,196 NA 5.2 AC123516 5.5 6.5 14 47 27 6 3.6
RM3428 11 13,445,211 13,445,367 157 47.2 AC128642 47.7 56.2 15 46 33 0 6.2 *
RM5349 11 19,148,807 19,148,929 123 67.3 AC134925 67.1 79.1 21 42 30 1 2.2
RM0144 11 28,246,930 28,247,154 225 99.2 AC134045 98.6 116.2 20 48 26 0 0.6

RM8215 12 1,585,781 1,586,001 221 5.8 BX000498 8.9 9.7 10 37 47 0 31.6 ***
RM3472 12 3,520,117 3,520,331 215 12.8 AL713902 24.8 27.1 15 31 48 0 32.3 ***
RM1337 12 11,935,984 11,936,165 182 43.4 BX000556 47.2 51.5 17 34 42 1 18.8 ***
RM0277 12 18,290,458 NA 66.5 AL831799 57.0 62.2 14 39 41 0 16.9 ***
RM1300 12 25,998,975 25,999,139 165 94.4 AL713940 93.4 100.9 8 42 44 0 27.0 ***
RM0017 12 26,954,668 26,954,835 168 98.0 AC027133 98.4 107.4 7 44 43 0 26.4 ***

The primer sequences were used as the query to localize on Rice Pseudomolecule 6

When both primer sequences successfully recognized their physical locations to be annealed, the expected PCR product size (e-PCR:‘Stop’-‘Start’+1bp, NA=not available) was used to judge PCR products. If any primer failed for e-Landing, the other primer's result was adopted to estimate the corresponding point. The determined physical regions followed by e-Landing were used to match corresponding BAC/PAC clones depend on the ‘Rice Pseudomolcule Information and Gene Search’ (http://rice.plantbiology.msu.edu/)

The determined cM positions for each BAC/PAC clones was directly adopted from the ‘Rice Pseudomolcule Information and Gene Search’ which were estimated using the marker sequences obtained from the Cornell Rice Genes Database and the Japanese Rice Genome Program. To take some convenient in comparing the cM positions from various mapping populations having different total lengths, the percentage expressed relative genetic positions, ‘cM%’, was also calculated

A and B are homozygous for the Namil(SA)-bl5 and Milyang23 allele types, H is for heterozygous progenies at the tested locus, and M indicates the number of missing. Segregation distorted loci are indicated with asterisk; χ2 values for the significant levels are less than 0.05

significant at P<0.0001

significant at P < 0.01

significant at P < 0.05.

Summary of association analysis between DNA marker genotypes and phenotype performances of F2 progenies derived from the cross between Namil(SA)-bl5 and Milyang23z for culm length (CL) and rice blast resistance (BL).

Trait Evaluated Mirror Mapy
No. of Genotypesx
Genotype meanw
Single-locus ANOVAv
Genetic Effectu
Locus Ch cM cM% A H B A H B SSM SSE F-value R2 Add Dom DeD

CL (cm) N=94 Mean=72.6 RM1003 1 136.6 77.5 28 42 24 73.2 74.5 68.4 576.9 3640.4 7.21 ** 0.137 -2.38 3.68 -1.55

RM0154 2 4.7 2.2 16 62 16 77.8 71.2 72.4 554.7 3662.6 6.89 ** 0.132 -2.69 -3.92 1.46

RM0007 3 44.4 27.0 15 47 31 74.3 73.6 70.1 279.4 3935.5 3.19 * 0.066 -2.07 1.42 -0.69
RM1350 3 126.8 78.8 28 44 22 70.9 72.0 75.8 327.5 3889.9 3.83 * 0.078 2.46 -1.40 -0.57

RM0551 4 3.1 0.5 22 48 24 75.1 72.5 70.2 278.8 3938.5 3.22 * 0.066 -2.46 -0.15 0.06
RM5633 4 19.9 37.1 13 54 27 73.5 73.7 69.8 289.8 3927.5 3.36 * 0.069 -1.88 2.03 -1.08

RM3628 6 85.4 76.0 16 51 27 68.1 73.1 74.1 392.4 3824.9 4.67 * 0.093 2.97 2.02 0.68
RM6274 6 100.3 84.2 23 40 29 68.6 72.6 75.8 663.7 3544.6 8.33 *** 0.158 3.60 0.44 0.12

RM1093 7 2.5 2.3 22 53 19 69.2 73.5 73.7 325.1 3892.2 3.80 * 0.077 2.28 2.05 0.90
RM5623 7 81.1 77.8 30 46 17 69.5 73.9 74.4 426.9 3788.0 5.07 ** 0.101 2.47 1.95 0.79
RM0172 7 118.6 99.6 23 45 26 71.3 71.1 76.2 489.5 3727.9 5.97 ** 0.116 2.48 -2.68 -1.08
RM3555 7 105.7 93.9 25 45 21 71.1 70.8 78.1 841.6 3366.3 11.00 *** 0.200 3.53 -3.77 -1.07

BL (Degree: 0-5) N=86 Mean=2.3 RM1054 5 122.0 97.6 22 42 22 2.8 1.9 2.5 14.1 172.2 3.40 * 0.076 -0.12 -0.79 6.71

RM3370 6 34.3 21.2 18 41 27 1.5 2.3 2.7 17.7 168.6 4.35 * 0.095 0.64 0.21 0.33
RM0539 6 51.0 26.2 15 42 25 1.3 2.2 3.0 25.2 152.1 6.55 ** 0.142 0.82 0.09 0.12
RM6818 6 65.8 .53.1 18 45 23 1.4 2.2 3.1 27.3 159.0 7.13 ** 0.147 0.82 -0.03 -0.03
RM0527 6 56.3 31.6 16 44 24 1.4 2.2 3.1 31.3 149.2 8.50 *** 0.173 0.89 -0.06 -0.06

RM8215 12 9.7 5.8 9 34 43 1.3 2.1 2.6 13.9 172.3 3.36 * 0.075 0.64 0.14 0.21
RM3472 12 27.1 12.8 14 28 44 1.1 2.1 2.8 29.8 156.5 7.91 *** 0.160 0.81 0.11 0.13
RM0277 12 62.2 66.5 13 36 37 1.1 2.1 2.9 31.2 155.1 8.35 *** 0.168 0.87 0.09 0.11
RM1337 12 51.5 43.4 15 32 38 0.5 2.0 3.2 85.3 97.8 35.74 *** 0.466 1.37 0.12 0.09

Only significant loci to each trait tested are presented. 58 DNA markers were tested in 94 F2 progenies derived from the cross between Namil(SA)-bl5 and Milyang23.

Both cM positions and the percentage expressed relative genetic positions (cM%) are presented.

F2 progenies were split into three subgroups based on the revealed genotype by SSR markers. A and B are homozygous for the Namil(SA)-bl5 and Milyang23 allele types and H indicates heterozygous progenies at the tested locus.

The means of each subpopulation for each locus are calculated.

For the F-test, SSR markers having less than significant levels (p) were declared as significant empirically. Explainable phenotypic variation portion at the tested locus (R2) was also calculated. The significant levels are less than 0.05

significant at P<0.001

significant at P < 0.01

significant at P < 0.05.

Additive effect (Add), domiant effect (Do) and degree of dominance (DeD) were then estimated at the declared loci: Add=(Bmean-Amean)/2, Do= Hmean-(Bmean+Amean)/2, and DeD=Do/Add. where A and B are homozygous F2 individuals for Namil(SA)-bl5 and Milyang23, H is heterozygous individuals at the tested locus.

Table 2. Reactions of the mutant line, Namil(SA)-bl5 and cultivars against selected Korean virulent blast isolates.

Rice blast isolates were collected from Korean famers’ fields. Resistance degrees of virulent rice blast were classified five levels from 1 (resistance) to 5 (susceptibility). Korean rice cultivars, Junam, Onuri, and Chilbo were included as check varieties

Not determined

Table 1. Comparisons between Namil, the wild type and Namil(SA)-bl5, a mutant line in terms of important agronomic traits.z

Heading date (HD), Culm length (CL), Panicle length (PL), Tiller number (TN), Spikelet number (SN), Ripened grain percentage (RGP), 1,000-grain weight of hulled grain (TWD), and Yield of brown rice (YDB). Note that the agronomic traits were evaluated by harvesting 100 plant for each replication plot (n=3), which were planted as three seedlings per plant during transplanting. All numerical values were mean ± standard deviation.

Table 3. List of SSR primer sets, their e-Landing mediated physical positions on rice Pseudomolecule 6, corresponding BAC/PAC clones with their determined cM positions, and the segregating (χ2 test) on the F2 progenies derived from the cross between Namil(SA)-bl5 and Milyang23.

The primer sequences were used as the query to localize on Rice Pseudomolecule 6

When both primer sequences successfully recognized their physical locations to be annealed, the expected PCR product size (e-PCR:‘Stop’-‘Start’+1bp, NA=not available) was used to judge PCR products. If any primer failed for e-Landing, the other primer's result was adopted to estimate the corresponding point. The determined physical regions followed by e-Landing were used to match corresponding BAC/PAC clones depend on the ‘Rice Pseudomolcule Information and Gene Search’ (http://rice.plantbiology.msu.edu/)

The determined cM positions for each BAC/PAC clones was directly adopted from the ‘Rice Pseudomolcule Information and Gene Search’ which were estimated using the marker sequences obtained from the Cornell Rice Genes Database and the Japanese Rice Genome Program. To take some convenient in comparing the cM positions from various mapping populations having different total lengths, the percentage expressed relative genetic positions, ‘cM%’, was also calculated

A and B are homozygous for the Namil(SA)-bl5 and Milyang23 allele types, H is for heterozygous progenies at the tested locus, and M indicates the number of missing. Segregation distorted loci are indicated with asterisk; χ2 values for the significant levels are less than 0.05

significant at P<0.0001

significant at P < 0.01

significant at P < 0.05.

Table 4. Summary of association analysis between DNA marker genotypes and phenotype performances of F2 progenies derived from the cross between Namil(SA)-bl5 and Milyang23z for culm length (CL) and rice blast resistance (BL).

Only significant loci to each trait tested are presented. 58 DNA markers were tested in 94 F2 progenies derived from the cross between Namil(SA)-bl5 and Milyang23.

Both cM positions and the percentage expressed relative genetic positions (cM%) are presented.

F2 progenies were split into three subgroups based on the revealed genotype by SSR markers. A and B are homozygous for the Namil(SA)-bl5 and Milyang23 allele types and H indicates heterozygous progenies at the tested locus.

The means of each subpopulation for each locus are calculated.

For the F-test, SSR markers having less than significant levels (p) were declared as significant empirically. Explainable phenotypic variation portion at the tested locus (R2) was also calculated. The significant levels are less than 0.05

significant at P<0.001

significant at P < 0.01

significant at P < 0.05.

Additive effect (Add), domiant effect (Do) and degree of dominance (DeD) were then estimated at the declared loci: Add=(Bmean-Amean)/2, Do= Hmean-(Bmean+Amean)/2, and DeD=Do/Add. where A and B are homozygous F2 individuals for Namil(SA)-bl5 and Milyang23, H is heterozygous individuals at the tested locus.