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Research Article

Breeding Hybrid Rice with Genes Resistant to Diseases and Insects Using Marker-Assisted Selection and Evaluation of Biological Assay

Me-Sun Kim1, Sothea Ouk1, Kuk-Hyun Jung2, Yoohan Song1, Le Van Trang1, Ju-Young Yang1, Yong-Gu Cho1,*
Plant Breeding and Biotechnology 2019;7(3):272-286.
Published online: September 1, 2019

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*Yong-Gu Cho, ygcho@cbnu.ac.kr, Tel: +82-43-261-2514, Fax: +82-43-273-1598
• Received: August 14, 2019   • Revised: August 21, 2019   • Accepted: August 21, 2019

Copyright © 2019 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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Breeding Hybrid Rice with Genes Resistant to Diseases and Insects Using Marker-Assisted Selection and Evaluation of Biological Assay
Plant Breed. Biotech.. 2019;7(3):272-286.   Published online September 1, 2019
DOI: https://doi.org/10.9787/PBB.2019.7.3.272
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Breeding Hybrid Rice with Genes Resistant to Diseases and Insects Using Marker-Assisted Selection and Evaluation of Biological Assay
Plant Breed. Biotech.. 2019;7(3):272-286.   Published online September 1, 2019
DOI: https://doi.org/10.9787/PBB.2019.7.3.272
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Breeding Hybrid Rice with Genes Resistant to Diseases and Insects Using Marker-Assisted Selection and Evaluation of Biological Assay
Image Image Image Image
Fig. 1 PCR products for genotyping with each marker linked to disease and insect resistance in breeding lines. S5, Rf4, Pita, Pib, Pii, and Pi5 show band patterns on agarose gel electrophoresis; Rf3 indicates size differentiation by fragment analyzer gel electrophoresis. L, R, S indicates ladder marker, resistant, and susceptible control variety, respectively.
Fig. 2 PCR products for genotyping with each marker linked to hybried related genes and blast resistance genes in breeding lines. Xa3, Xa7, xa13, and Xa21 show band patterns on gel electrophoresis; xa4, xa5, and tsv1 indicate size differentiation by fragment analyzer gel electrophoresis. Bph18(t) shows HRM curve profiles of 240 genetic resources and F1 hybrid combinations. L, R, S indicates ladder marker, resistant, and susceptible control variety, respectively.
Fig. 3 (A) Frequency distribution of disease and insect resistant genes among 240 genetic resources and F1 hybrid combinations, and (B) Frequency of rice lines having different numbers of resistant genes.
Fig. 4 Panicle type and growth performance of F1 hybrid combinations, KR0695H, and KR0696H. (A) Panicle length and morphologies of KR0695H and KR0696H; (B) Growth performance of KR0695H and KR0696H at Hai Hau field in Vietnam.

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Breeding Hybrid Rice with Genes Resistant to Diseases and Insects Using Marker-Assisted Selection and Evaluation of Biological Assay

List of 240 genetic resources and hybrid rice combinations used in this study.

Entry No. Designation Cross/Origin Remark Source
1 9019 China Restorer 811001
2 Basmati 370 India Aromatic rice 811005
3 Chiherang Cambodia Local variety 811007
4 Cigeulis Cambodia Local variety 811009
5 Ciliwung Cambodia Local variety 811011
6 FFZ1 China Local variety 811017
7 Giza178 Egypt Local variety 811019
8 HHZ12-SAL2-Y3-Y1 China Breeding line 811025
9 HHZ12-SAL8-Y1-Y2 China Breeding line 811027
10 IR05N412 IR72875-94-3-3-2/IR73707-45-3-2-3 Breeding line, IRRI 811029
11 IR05N412 IR72875-94-3-3-2/IR73707-45-3-2-3 Breeding line, IRRI 811030
12 IR06A145 IR02A127/JANAKI Breeding line, IRRI 811031
13 IR09N538 IRRI 132/PR 30138-35-2//IR04N114 Breeding line, IRRI 811033
14 IR10A267 IR02A483/IRBB 60-1 Breeding line, IRRI 811035
15 IR10N305-1 IRRI Breeding line, IRRI 811037
16 IR78581-12-3-2-2-1 IRRI Breeding line, IRRI 811041
17 IR98070-kB13-1-2-2-1-1-1 IR72903-131-1-2-3R/IR72998-78-1-3-2 R Restorer 811047
18 IR98070-kB14-1-2-3-1-4-1 IR72903-131-1-2-3R/IR72998-78-1-3-2 R Restorer 811051
19 IR98102-kB9-1-1-1-1-1-1 IR65622-151-2-2-2R/IR73885-1-4-3-2-1-10R Restorer 811055
20 IR98102-kB9-1-1-3-1-1-B2-1 IR65622-151-2-2-2R/IR73885-1-4-3-2-1-10R Restorer 811059
21 IR98229-9-2-1-k1-1-6-1-1 IR08A138/IR72998-93-3-3-2R Restorer 811060
22 IR98229-24-1-1-k1-1-1-1-1 IR08A138/IR72998-93-3-3-2R Restorer 811062
23 IR98241-24-2-1-k1-1-1-1 IR06N172/IR86612-38-2-2-1-1-1-1-1 Breeding line, IRRI 811068
24 IR101872-46-1-K1-1-2-1 MingHui63/IR86590-22-2-2-1-3-1-1-1 Breeding line, IRRI 811078
25 L24 China Local variety 811084
26 NSIC 238 Philippines Local variety 811094
27 KR3R MY3R(SSLR-12, Myanma Col. 2012) Local variety 811096
28 OM52 Vietnam Local variety 811098
29 TLR353 Vietnam Local variety 811102
30 TLR363 Vietnam Local variety 811104
31 Vietnam collection 1 Vietnam Local variety 811106
32 WEED TOLERANT RICE 1-1 IRRI Breeding line, IRRI 811108
33 Zhong419 China Local variety 811110
34 6527 Unknown Local variety 811113
35 Com. Collection 3 Com Col. (Cambodia Col. 2015) Local variety 811114
36 HHZ1-Y4-Y1 HUANG-HUA-ZHAN*2/YUE-XIANG-ZHAN Breeding line, China 811120
37 HUA564 China Breeding line 811122
38 IR02A127 IR00A107/IR62243-41-1-3-3 Breeding line, IRRI 811124
39 IR05N359 IR72158-11-5-2-3/Ir72903-121-2-1-2 Breeding line, IRRI 811126
40 IR06A181 IR71718-59-1-2-3/IR72 Breeding line, IRRI 811128
41 IR08N136 IR72967-12-2-3/PR 31090-33-2-1 Breeding line, IRRI 811130
42 IR10K153 HR 24580-15-1/IR03K105 Breeding line, IRRI 811132
43 IR11A303 IR04A427/IR72875-94-3-3-2 Breeding line, IRRI 811134
44 IR11A334 IR04A427/IRRI 115 Breeding line, IRRI 811136
45 Japonica 1 Philippines Local variety 811138
46 SACG4 IRRI Local variety 811150
47 SAGC-02 IRRI Local variety 811152
48 ZH1 China Local variety 811156
49 HHZ11-Y10-DT3-Y3 China Breeding line 811158
50 HHZ5-DT1-DT1 China Breeding line 811160
51 HHZ5-SAL12-DT3-Y2 China Breeding line 811162
52 HHZ5-SAL8-DT2-SAL1 China Breeding line 811164
53 HHZ8-SAL14-SAL1-SUB1 China Breeding line 811166
54 HHZ8-SAL6-SAL3-SAL1 China Breeding line 811168
55 HHZ8-SAL6-SAL3-Y1 China Breeding line 811170
56 HHZ8-SAL6-SAL3-Y2 HUANG-HUA-ZHAN*2/PHALGUNA Breeding line 811172
57 IR06M150 MEM BERANO/PADI ABANG GOGO Breeding line, IRRI 811174
58 IR72 (IR72) IR19661-9-2-3-3/IR15795-199-3-3//IR9129-209-2-2-2-1 Advanced variety, IRRI 811178
59 Teqing China Local variety 811180
60 TME80518 TME 80518 Local variety 811182
61 KR2R MR2R(Myanma Col. 2012) Restorer 811184
62 A 69-1 IRRI Breeding line 811186
63 BR 28-SalTol Bangladesh Breeding line 811188
64 Chulsa Cambodia Local variety 811190
65 IR04A395 IRRI Breeding line 811192
66 IR07A234 NSIC RC 138/IRRI 123 Breeding line 811194
67 IR10A 227 IR01A154/IR72870-19-2-2-3//Irri 123 Breeding line 811196
68 IR65482-4-136 IRRI Breeding line 811198
69 AN 424627 IRRI Local variety 811200
70 BR 26 Bangladesh Local variety 811202
71 Daerip H-R11-2-1-1-1 78/Daelipbyeo F1 Breeding line 811204
72 IR64 Sub1 IRRI Breeding line 811206
73 IR68897H-B24-B-1-1-2 IRRI Breeding line 811208
74 IR09A228 PR29232-B-17-2-1-1/IR 64 Breeding line, 811210
75 J.P.5-IR946-2-2-2/IR1635-1F IRRI Breeding line 811212
76 IR97727-82-1-2-2 IRRI Breeding line 811214
77 IR98073-3-1-1-K1-1 IR72903-131-1-2-3R/IR85485-106-B-B-1-1-1-1 Breeding line 811216
78 IR98107-kB3-1-1-2-1-1 IR71604-4-1-4-4-4-2-2-2R/IR65622-151-2-2-2R Breeding line 811218
79 IR98108-kB13-1-2-3-1-2-1 IRRI Breeding line 811220
80 IR98161-2-1-1-k2-2-2 IR86409-3-1-1-1-1-1/IRBB66 Breeding line 811222
81 IR98194-9-2-1-k1-1-1 IRRI Breeding line 811224
82 IR101861-7-1-K1-1-1 MingHui63/IR03A550 Breeding line 811226
83 IRRI 102 IR4215-301-2-2-6/BG90-2//IR19661-131-1-2 Advanced variety, IRRI 811234
84 Jasponica Bulk Aroma4-1 Philippines Breeding line 811236
85 Jasponica Bulk Aroma5-1 Philippines Breeding line 811238
86 KCD1 IRRI Local variety 811240
87 MY1H-R23-3-1-1-1-1 MY 1 A/R Breeding line 811242
88 MY1H-R23-3-2-1-1-1 MY 1 A/R Breeding line 811244
89 NSIC 222 Philippines Local variety 811246
90 OM100411 Vietnam Local variety 811248
91 OM10375 Vietnam Local variety 811250
92 OM4900 Vietnam Local variety 811252
93 OM7347 Vietnam Local variety 811254
94 OM8108 Vietnam Local variety 811256
95 OMCS 2012 Vietnam Local variety 811258
96 Pearl riceH-R28-3-2-1-1 IRRI Breeding line 811260
97 Pearl riceH-R52-2-1-1-1 IRRI Breeding line 811262
98 PHB73H-R9-2-1-1-1 IRRI Breeding line 811264
99 Phka Romeat Cambodia Local variety 811266
100 Phka Rumchang Cambodia Local variety 811268
101 Phka Rumchek Cambodia Local variety 811270
102 Phka Rumdeng Cambodia Local variety 811272
103 Phka Rumduol Cambodia Local variety 811274
104 Popoul Cambodia Local variety 811276
105 Rumpe Cambodia Local variety 811278
106 S430 China Local variety 811280
107 San pidao Cambodia Local variety 811283
108 TH82H-R2-1-1-1-1-1 Vietnam Breeding line 811284
109 TLR405 Vietnam - 811286
110 TLR407 Vietnam - 811288
111 WC467-2-1-1-1-2-2-1-1 (Milyang154/Norin PL9//Milyang154)/Milyang154 Breeding line 811290
112 WC467-2-3-2-1-2-1-1-1 (Milyang154/Norin PL9//Milyang154)/Milyang154 Breeding line 811292
113 WC468-2-1-3-1-2-3-1-1 (Milyang154/Norin PL9//Milyang154)/Milyang154 Breeding line 811294
114 WC488-6-1-1-2-1-1-1-1 (Milyang23//Norin PL9/Dular///Milyang23)/Milyang23 Breeding line 811296
115 WC488-6-1-1-2-1-3-1-1 (Milyang23//Norin PL9/Dular///Milyang23)/Milyang23 Breeding line 811298
116 WC495-1-1-1-1-2-3-1-1 (Milyang160//Norin PL9/Dular///Areumbyeo)/Areumbyeo Breeding line 811300
117 WC509-4-1-2-1-2-3-1-1 (Jangsungbyeo/Dular//Jangsungbyeo)/Jangsungbyeo Breeding line 811302
118 WC540-2-1-3-1-1-1-1-1 (Milyang160/CPSLO 17//Areumbyeo)/Areumbyeo Breeding line 811304
119 WC540-2-1-3-1-2-2-1-1 (Milyang160/CPSLO 17//Areumbyeo)/Areumbyeo Breeding line 811308
120 WC540-2-3-3-1-2-3-1-1 (Milyang160/CPSLO 17//Areumbyeo)/Areumbyeo Breeding line 811312
121 WC549-1-1-2-1-1-1-1-1 (Yongmunbyeo/CPSLO 17//Yongmunbyeo)/Yongmunbyeo Breeding line 811314
122 WC570-2-1-3-1-1-1-1-1 (Samgangbyeo//Dular/Samgangbyeo///Samgangbyeo)/Samganbyeo Breeding line 811316
123 WC634-1-1-2-1-2-1-1-1 (Yongjubyeo//N22/Yongjubyeo)/Yongjubyeo Breeding line 811320
124 WC647-1-1-1-1-1-2-1-1 Ilpumbyeo/IR65600-96-1-2-2//Ilpumbyeo)/Ilpumbyeo Breeding line 811324
125 WC962-1-2-1-1-1-1 02428-97-2/Areumbyeo//3*Yeonghaebyeo Breeding line 811328
126 WC964-1-1-2-1-3-1-1-1 Sambaekbyeo/02428-97-1//3*Sambaekbyeo Breeding line 811332
127 WC972-3-3-1-1-1-1 02428/3*Yongmunbyeo Breeding line 811334
128 WC972-4-2-1-3-2-1 02428/3*Yongmunbyeo Breeding line 811340
129 Corn rice China Local variety 811375
130 Restorer 1-1 Unknown Breeding line 811377
131 Restorer 2-1 Unknown Breeding line 811379
132 Restorer 3-1 Unknown Breeding line 811381
133 Indonesia col. 1(2016) Indonesia Col. (Cambodia Col. 2016) Local variety 811383
134 Indonesia col. 2(2016) Indonesia Col. (Cambodia Col. 2016) Local variety 811385
135 HYT 116H-3-1-3-2-1-1 HYT 116 H Breeding line 811391
136 HYT 116H-17-2-1-1-2-1 HYT 116 H Breeding line 811395
137 HYT 116H-31-2-3-1-1-1 HYT 116 H Breeding line 811397
138 HYT 116H-46-1-1-1-1-1 HYT 116 H Breeding line 811401
139 HYT 116H-50-1-1-2-1-1 HYT 116 H Breeding line 811405
140 HYT 119H-18-2-2-2-1-1 HYT 119 H Breeding line 811407
141 HYT 119H-18-3-2-2-2-2 HYT 119 H Breeding line 811411
142 HYT 119H-21-1-3-2-2-1 HYT 119 H Breeding line 811414
143 HYT 123H-13-3-3-1-2-1 HYT 123H Breeding line 811416
144 HYT 124H-3-3-1-1-2-1 HYT 124H Breeding line 811420
145 HYT 128H-8-2-2-2-2-1 HYT 128H Breeding line 811426
146 HYT 128H-11-2-1-1-2-1 HYT 128H Breeding line 811428
147 HYT 130H-3-2-3-1-1-1 HYT 130H Breeding line 811432
148 CASH 1H-1-2-3-1-1-1 CASH 1H Breeding line 811434
149 CASH 1H-4-3-2-1-1-1 CASH 1H Breeding line 811440
150 CASH 1H-34-2-2-2-1-1 CASH 1H Breeding line 811444
151 Hipa Jatim 1H-5-2-2-3-2-1 Hipa Jatim 1 H Breeding line 811446
152 IR68897H-94 B-1-3-3-1-1 IR68897H Breeding line 811448
153 IR101922-BK-KB-2-2-1 Maybelle/PSBRC80 Breeding line 811452
154 IR101922-BK-KB-6-1-1 Maybelle/PSBRC80 Breeding line 811456
155 IR101923-BK-KB-1-2-1 IR86427-29-6-1-3-1-1-1-1/IR86508-4-2-3-2-1-1-2-1 Breeding line 811458
156 IR101923-BK-KB-3-2-1 IR86427-29-6-1-3-1-1-1-1/IR86508-4-2-3-2-1-1-2-1 Breeding line 811460
157 IR101923-BK-KB-4-2-1 IR86427-29-6-1-3-1-1-1-1/IR86508-4-2-3-2-1-1-2-1 Breeding line 811462
158 IR101923-BK-KB-4-3-1 IR86427-29-6-1-3-1-1-1-1/IR86508-4-2-3-2-1-1-2-1 Breeding line 811464
159 IR101923-BK-KB-7-1-1 IR86427-29-6-1-3-1-1-1-1/IR86508-4-2-3-2-1-1-2-1 Breeding line 811466
160 IR101924-BK-KB-2-3-1 IR86427-29-6-1-3-1-1-1-1/IR86519-12-1-3-1-1-1-1-1 Breeding line 811468
161 IR101924-BK-KB-7-2-1 IR86427-29-6-1-3-1-1-1-1/IR86519-12-1-3-1-1-1-1-1 Breeding line 811470
162 IR101924-BK-KB-10-2-1 IR86427-29-6-1-3-1-1-1-1/IR86519-12-1-3-1-1-1-1-1 Breeding line 811472
163 IR101924-BK-KB-11-3-1 IR86427-29-6-1-3-1-1-1-1/IR86519-12-1-3-1-1-1-1-1 Breeding line 811474
164 IR101924-BK-KB-14-1-1 IR86427-29-6-1-3-1-1-1-1/IR86519-12-1-3-1-1-1-1-1 Breeding line 811476
165 IR101924-BK-KB-14-2-1 IR86427-29-6-1-3-1-1-1-1/IR86519-12-1-3-1-1-1-1-1 Breeding line 811478
166 IR101933-BK-KB-2-1-1 IR86505-6-4-3-1-1-1-1-1/MingHui63 Breeding line 811480
167 IR101937-BK-KB-3-2-1 IR86505-6-4-3-1-1-1-1-1/IR86612-26-1-1-1-1-2-1-1 Breeding line 811482
168 IR101937-BK-KB-9-3-1 IR86505-6-4-3-1-1-1-1-1/IR86612-26-1-1-1-1-2-1-1 Breeding line 811484
169 KR0301-B-12-1-1-1 OM 052/NSIC RC 238 Breeding line 811485
170 KR0302-B-5-2-2-1 OM 052/Minghui 63 Breeding line 811490
171 KR0302-B-5-3-1-1 OM 052/Minghui 63 Breeding line 811492
172 KR0302-B-14-2-1-1 OM 052/Minghui 63 Breeding line 811496
173 KR0302-B-17-1-3-1 OM 052/Minghui 63 Breeding line 811498
174 KR0302-B-10-2-1-1 OM 052/Minghui 63 Breeding line 811500
175 KR0302-B-10-2-2-1 OM 052/Minghui 63 Breeding line 811502
176 KR0302-B-13-1-1-1 OM 052/Minghui 63 Breeding line 811506
177 GR19-1-8-2-2-1 2A/3m164 Breeding line 811510
178 GR20-1-3-1-1-1 2A/3m170 Breeding line 811512
179 PHB 73H-1-2-3-1-2-1 PHB 73 Breeding line 811514
180 PHB 73H-18-2-2-3-1-1 PHB 73 Breeding line 811522
181 Matibay H-5-3-1-1-1-1 Matibay Breeding line 811526
182 ABp H-20-2-1-1-1-1 Arize Bigante plusH Breeding line 811532
183 ABp H-29-2-1-1-2-1 Arize Bigante plusH Breeding line 811538
184 ABp H-33-2-3-2-1-1 Arize Bigante plusH Breeding line 811540
185 Hipa Jatim 2H-1-3-3-1-2-1 Hipa Jatim 2 H Breeding line 811542
186 HYT 106H-17-2-1-2-1-1 HYT 106H Breeding line 811544
187 HYT 106H-17-2-2-3-2-1 HYT 106H Breeding line 811548
188 HYT 106H-17-2-3-1-2-1 HYT 106H Breeding line 811551
189 HYT 106H-17-3-1-1-1-1 HYT 106H Breeding line 811559
190 HYT 106H-17-3-2-3-2-1 HYT 106H Breeding line 811563
191 HYT 108H-10-2-3-1-3-1 HYT 108H Breeding line 811565
192 Jasponica-4-1-1-2-2-1 SH 9 Breeding line 811569
193 Jasponica-12-1-1-2-3-1 SH 9 Breeding line 811575
194 Jasponica-14-3-1-1-1-1 SH 9 Breeding line 811577
195 Jasponica-15-1-1-1-1-1 SH 9 Breeding line 811583
196 Jasponica-26-1-1-1-3-1 SH 9 Breeding line 811585
197 Jasponica-29-3-1-1-2-1 SH 9 Breeding line 811587
198 Jasponica-40-3-1-1-2-1 SH 9 Breeding line 811599
199 IR24 IRRI Advanced variety 811601
200 IRBB1 IRRI Bacterial Blight 811603
201 IRBB3 IRRI Bacterial Blight 811605
202 IRBB7 IRRI Bacterial Blight 811607
203 IRBB13 IRRI Bacterial Blight 811609
204 IRBB55 IRRI Bacterial Blight 811611
205 IRBB59 IRRI Bacterial Blight 811613
206 Dasan Korea Local variety 810801
207 Hanareum Korea Local variety 810802
208 KR0203H Korea Hybrid rice F1 810803
209 KR0695H Korea Hybrid rice F1 810804
210 KR0696H Korea Hybrid rice F1 810805
211 KR1454H Korea Hybrid rice F1 810806
212 KR1455H Korea Hybrid rice F1 810807
213 KR1354H Korea Hybrid rice F1 810808
214 KR1497H Korea Hybrid rice F1 810809
215 KR1444H Korea Hybrid rice F1 810810
216 KR1487H Korea Hybrid rice F1 810811
217 KR1994H Korea Hybrid rice F1 810812
218 KR2116H Korea Hybrid rice F1 810813
219 KR2117H Korea Hybrid rice F1 810814
220 KR 1B Korea Maintainer 812501
221 KR 1A Korea CGMS 812503
222 KR 2B Korea Maintainer 812526
223 KR 2A Korea CGMS 812528
224 KR 211B Korea Maintainer 812551
225 KR 211A Korea CGMS 812553
226 MingHui 63 China Local variety, Restorer 812571
227 IR75589-31-27-8-33S-5-1 IRRI Breeding line, TGMS 812608
228 IR102100-KB5S2-1-4-1-1-1-1 IRRI Breeding line, TGMS 812618
229 OM052 Vietnam Local variety, Restorer 812636
230 IR102100-KB14-2S2-1-22-2-1-2 IRRI Breeding line, TGMS 812718
231 IR98070-kB14-1-2-3-1-1 IRRI Breeding line 812736
232 IR98229-2-2-1-K1-1-1 IRRI Breeding line 812756
233 IR98229-9-2-1-K1-1-1 IRRI Breeding line 812771
234 IR98229-9-2-1-K1-1-3 IRRI Breeding line 812776
235 IR101861-7-1-K1-1-1 IRRI Breeding line 812786
236 IR98241-24-2-1-k1-1-1 IRRI Breeding line 812806
237 IR98102-kB9-1-1-3-1-1-1 IRRI Breeding line 812821
238 IR102100-KB12S1-1-25-1-1-1-1 IRRI Breeding line, TGMS 812823
239 IR98102-kB21-1-3-1-3-1 IRRI Breeding line 812836
240 IR102452-KB3-1-2-2-1-1 IRRI Breeding line 812851

Gene-specific PCR primers and their primer sequences used for analysis of related genes.

Target Gene Marker name Type Primer sequence (5′→3′) Exspected size (bp) Reference
Genes related to hybrid S5 S5-Indel Fw CCTACGTTTGACTGCCTGCCTG 281/417 Sundaram et al. 2010
Rv CTACACGCGGCTTCGGGAAAGC
Indica-specific SNP Fw GACAGCAGCATCAACGACTTCC 527
Rv TCGTCAGTGGGCAAGCAGTAGCTG
Japonica-specific SNP Fw ACCCTGATATTCTGAGTTACAAGGCATTA 325
Rv GCTCTTGATGTCCGGTGATACC
Rf3 DRRM-RF3-5 Fw GATGGCACAGCTTCAGAACA 120/134 Suresh et al. 2012
Rv CTAATTCTGGGCGAGCAAAG
DRRM-RF3-10 Fw TCACCTCTTCCTGCTTCGAC 180/195
Rv CTCCACCAGTGCAGGTTTTT
Rf4 DRCG-RF4-14 Fw GCAATGCTTGTATTCAGCAAA 845/885 Tang et al. 2014
Rv TCCAGCTGTAAATCCGTCAA
M19280 Fw ATCTGTCCAGACACCATTTTC 147/141
Rv TCCACTGATGAGTGCATTG
Blast Pita YL155/87 Fw AGCAGGTTATAAGCTAGGCC 1042 Jia et al. 2002, 2004
Rv CTACCAACAAGTTCATCAAA
YL183/87 Fw AGCAGGTTATAAGCTAGCTAT
Rv CTACCAACAAGTTCATCAAA
Pib NSb Fw ATCAACTCTGCCACAAAATCC 629 Kwon et al. 2008
Rv CCCATATCACCACTTGTTCCCC
Pi5 JJ817 Fw GATATGGTTGAAAAGCTAATCTCA 1450 Kwon et al. 2008
Rv ATCATTGTCCTTCATATTCAGAGT
Pii JJ113 Fw GGATGATGTGATCTGCAGAG 484 Jeon 2002
Rv CTCTTGGTGATCTTTGTTAC
Bacterial blight (BB) Xa3 BB3-Sus Fw CGGAGCGACACAGCTATCAT 743 Hu et al. 2013
Rv CGTGAGGTTCCCTATGGCGATT
BB3-Re Fw CCACAATGCCATGTCAGGTGGCATCCCTGCA 255 Hu et al. 2013
Rv AGGTGTTGGAGGATTGGCAT
Xa4 RM224 Fw ATCGATCGATCTTCACGAGG 150/120 McCouch et al. 2002
Rv TGCTATAAAAGGCATTCGGG
Xa5 RM122 Fw GCACTGCAACCATCAATGAATC 236/232 Chen et al. 1997
Rv CCTAGGAGAAACTAGCCGTCCA
Xa7 M5 Fw CGATCTTACTGGCTCTGCAACTCTGT 294/1170 Porter et al. 2013
Rv GCATGTCTGTGTCGATTCGTCCGTACGA
Xa13 Xa13prom Fw GGCCATGGCTCAGTGTTTAT 1000/520 Zhang et al. 1996; Singh et al. 2011
Rv GAGCTCCAGCTCTCCAAATG
Xa21 pTA248 Fw AGACGCGGAAGGGTGGTTCCCGGA 1000/750 Huang et al. 1997
Rv AGACGCGGTAATCGAAAGATGAAA
BPH Bph18(t) Bph18(t) SNP23 CGATGGATTACCCTATCACCT CAA HRM Developed in this study
SNP24 AACCCTCTGCACACCATCGG
Tungro virus tsv1 RM6152 Fw GAATTCACCGCTCTCCAGTC 206 Lee et al. 2010
Rv AGGAGGATCTCCTCCAGGAG

List of control varieties used for identification of resistance genes.

Gene Resistant controls Susceptible controls
Xa3 IRBB3 IR24
Xa4 IRBB4 IR24
xa5 IRBB5 IR24
Xa7 IRBB7 IR24
xa13 IRBB13 IR24
Xa21 IRBB21 IR24
Pib IRBL-b LTH
Pita IRBL-ta(K1), IRBL-ta(CT2), IRBL-ta2(Pi) LTH
Pi5, Pii Pi3, Pi5(t), Pii LTH
Bph18(t) Anmi, Anda IR24, Ilpum
tsv1 Utri merah, N22 Nipponbare, TN1

List of eleven hybrid rice breeding lines and their cross combinations used for evaluating agronomic traits and screening disease/insect resistance in the Red River Delta in Vietnam.

F1 entry Cross combination Remark
KR0203H KR2A/Minghui 63 CGMS
KR0695H KR1A/OM052 CGMS
KR0696H KR2A/OM052 CGMS
KR1454H KR1A/MY2R (= KR2R) CGMS
KR1455H KR2A/MY2R (= KR2R) CGMS
KR1354H KR2A/IR98070-KB14-1-2-3-1-1 CGMS
KR1497H IR75589-31-27-8-33S/IR98229-9-2-1-k1-1 TGMS
KR1444H IR75589-31-27-8-33S/IR102452-KB3-1-2-2-1-1 TGMS
KR1487H HYT 108 S8-1-25-2/IR98102-kB9-1-1-3-1-1 TGMS
KR1994H IR75589-31-27-8-33S/Minghui 63 TGMS
KR2116H IR102100-KB12S2-1-19-1/IR98102-kB21-1-3-1-3-1 TGMS

Identification patterns of disease and insect resistant genes in eleven hybrid rice combinations.

Entry Bacterial Blight No. of R genes Blast No. of R genes Bph 18(t) tsv1 Total of R genes
Xa3S Xa3R Xa4 xa5 Xa7 xa13 Xa21 Pi-ta Pib Pii Pi5
KR0203H + − − +/− − − − 0 + − + + + 3 − +/− 3
KR0695H − − − − − − − 0 + − + + + 3 − +/− 3
KR0696H + − − − − − − 0 + + + + + 4 − +/− 4
KR1454H + − − − + − − 1 + − + + + 3 − − 4
KR1455H + − − − + − − 1 + − + + + 3 − − 4
KR1354H + + − +/− − − − 1 + − + + + 3 − +/− 4
KR1497H + + − − − − − 1 + + + + + 4 − − 5
KR1444H + − − − − − − 0 + + + + + 4 − − 4
KR1487H + − +/− − − − − 0 + − + − + 2 − − 2
KR1994H − − +/− − − − − 0 − + + + + 4 − − 4
KR2116H + + +/− +/− − − − 1 + + + + + 4 − + 6

To get the whole information of 240 genetic resources and hybrid rice combinations, please see Supplementary Table S1.

Relative levels of resistance on hybrid rice combinations and control varieties upon inoculation separately by disease and insect in the Red River Delta region in Vietnam.

Entryz) Brown Planthopper Blast Bacterial Blight
6 DARy) 8 DAR 28 DAI 35 DAIy) 42 DAI
Score Reaction Score Reaction Score Reaction Score Reaction Score Reaction
KR0203H 6.0 MSx) 8.0 S 0.0 HR 1.0 HR 5.0 MS
KR0695H 7.0 S 9.0 HS 0.0 HR 0.0 HR 1.0 HR
KR0696H 6.0 MS 9.0 HS 0.0 HR 0.0 HR 3.0 R
KR1454H 5.0 MS 8.0 S 0.0 HR 0.0 HR 3.0 R
KR1455H 6.0 MS 8.0 S 0.0 HR 0.0 HR 3.0 R
KR1354H 7.0 S 8.0 S 1.0 HR 1.0 HR 3.0 R
KR1497H 4.0 MR 8.0 S 0.0 HR 0.0 HR 1.0 HR
KR1444H 7.0 S 9.0 HS 0.0 HR 0.0 HR 3.0 R
KR1487H 4.0 MR 9.0 HS 1.5 HR 2.0 HR 3.0 R
KR1994H 3.0 R 6.0 MS 4.0 MR 5.0 MS 0.0 HR
KR2116H 5.0 MS 9.0 HS 0.0 HR 1.0 HR 1.0 HR
BTP 33 1.0 R 1.0 R - - - - - -
TN 1 9.0 HS 9.0 HS - - - - - -
OM 1490 - - - - 9.0 HS 9.0 HS 5.0 MS

z)All of hybrid rice breeding lines, KR0203H to KR2116H, were in generation of F1.

y)DAR: Days after release, DAI: Day after inoculation.

x)HR: Highly resistance, R: Resistance, MR: Medium resistance, MS: Medium susceptible, S: Susceptible, HS: Highly susceptible.

Agronomic traits and yields of F1 hybrid combinations with disease/insect resistantance genes introduced by MAS and control varieties.

Entry HD (mm.dd) GP (day) PH (cm) PL (cm) PET (%) PRG (%) GW (g) Yield (tonne/ha)
KR0203H 5.18 121 114.5 29.4 64.8 78.1 27.0 10.1
KR0695H 5.15 119 122.6 27.5 74.0 65.2 26.5 11.8
KR0696H 5.15 119 122.9 27.9 75.3 71.8 27.0 11.1
KR1454H 5.19 123 118.2 27.4 71.7 68.4 26.0 9.7
KR1455H 5.18 122 114.4 27.4 72.6 73.8 26.4 9.6
KR1354H 5.19 126 113.6 31.1 71.5 69.9 26.4 10.7
KR1497H 5.15 119 111.6 28.3 76.2 88.5 26.2 10.0
KR2116H 5.15 122 113.8 29.0 83.6 68.5 26.1 10.3
IIA838 5.12 113 118.3 28.1 79.3 90.4 30.3 14.6
BC15 5.16 115 112.8 28.1 69.8 74.3 22.1 9.1

HD: Heading date, GP: Growing period, PH: Plant height, PL: Panicle length, PET: Percent effective tillers, PRG: Percent ripened grain, GW: 1000-grain weight.

Table 1 List of 240 genetic resources and hybrid rice combinations used in this study.
Table 2 Gene-specific PCR primers and their primer sequences used for analysis of related genes.
Table 3 List of control varieties used for identification of resistance genes.
Table 4 List of eleven hybrid rice breeding lines and their cross combinations used for evaluating agronomic traits and screening disease/insect resistance in the Red River Delta in Vietnam.
Table 5 Identification patterns of disease and insect resistant genes in eleven hybrid rice combinations.

To get the whole information of 240 genetic resources and hybrid rice combinations, please see Supplementary Table S1.

Table 6 Relative levels of resistance on hybrid rice combinations and control varieties upon inoculation separately by disease and insect in the Red River Delta region in Vietnam.

All of hybrid rice breeding lines, KR0203H to KR2116H, were in generation of F1.

DAR: Days after release, DAI: Day after inoculation.

HR: Highly resistance, R: Resistance, MR: Medium resistance, MS: Medium susceptible, S: Susceptible, HS: Highly susceptible.

Table 7 Agronomic traits and yields of F1 hybrid combinations with disease/insect resistantance genes introduced by MAS and control varieties.

HD: Heading date, GP: Growing period, PH: Plant height, PL: Panicle length, PET: Percent effective tillers, PRG: Percent ripened grain, GW: 1000-grain weight.

Table 1

Table 2

Table 3

Table 4

Table 5

Table 6

Table 7

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