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Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (2): 225-234.doi: 10.3724/SP.J.1006.2019.083033

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Identification of fertility restoration and molecular mapping of restorer genes in two maize restore lines of CMS-C

Bi-Tao MOU1,2,Zhuo-Fan ZHAO1,Ling YUE1,Chuan LI1,Jun ZHANG3,Zhang-Bo LI3,Han SHEN3,Mo-Ju CAO1,*()   

  1. 1 Maize Research Institute, Sichuan Agricultural University / Key Laboratory of Maize Biology and Genetics and Breeding of Southwest China, Ministry of Agriculture, Chengdu 611130, Sichuan, China
    2 Edible Fungus Sericulture Research Institute, Yibin Academy of Agricultural Sciences, Yibin 644000, Sichuan, China
    3 Inner Mongolia Zhenjin Seed S&T Co., Ltd, Ordos 014300, Inner Mongolia, China;
  • Received:2018-04-16 Accepted:2018-10-08 Online:2019-02-12 Published:2018-11-05
  • Contact: Mo-Ju CAO E-mail:caomj@sicau.edu.cn
  • Supported by:
    This study was supported by the National Thirteenth Five-Year National Research and Development Program(2016YFD0101206)

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Abstract:

The objective of the present study was to identify novel and powerful restorer lines for CMS-C. So maize inbred lines Z16 and 7250-14-1 were crossed with both isonuclear alloplasmic and isoplasmic allonuclear CMS-C, CMS-T, and CMS-S male sterile lines. Self-cross and back-cross were conducted for some of the fertility restored F1 for genetic analysis and restorer gene mapping. Male fertility expression was investigated for all the F1, F2 and backcross populations, showing that Z16 and 7250-14-1 could restore the fertility for C Huangzaosi, C478, C698-3, and CMo17 completely, and partly restore the fertility for C48-2. Z16 could not restore the fertility for G48-2, EC48-2, ES48-2, RB48-2, and Lei48-2, while 7250-14-1 could partly restore the fertility for G48-2, EC48-2, and ES48-2, and maintain the sterility for RB48-2 and Lei48-2. Both Z16 and 7250-14-1 couldn’t restore the fertility of CMS-T, and partly restore the fertility for CMS-S. Genetic analysis showed that the fertility restoration was controlled by a pair of dominant genes for Z16 when crossed with C478 or C Huangzaosi. But for 7250-14-1, the fertility restoration was controlled by a pair of dominant genes for C Huangzaosi, and two pairs of complementary dominant genes for C478. Both of the restorer genes for Z16 and 7250-14-1 were mapped on the short arm of chromosome 8 by molecular markers. For Z16, it was mapped within a physical distance of 494 kb from the marker B-1 to the end of the chromosome, and for 7250-14-1, it was located between B-1 and Chr8-86080, with physical distance of 249 kb. This study not only provides some information for the practical application of Z16 and 7250-14-1, but also lays a foundation for the cloning and functional analysis of restorer genes.

Key words: maize, cytoplasmic male sterility, restorer gene, molecular mapping

Table 1

Isonuclear alloplasmic and isoplasmic allonuclear sterile lines of maize"

C型不育系
CMS-C		 T型不育系
CMS-T		 S型不育系
CMS-S		 C型亚组不育系
CMS-C subgroup
CMo17 TMo17 SMo17 G48-2
C698-3 T698-3 S698-3 EC48-2
C黄早四
C Huangzaosi		 ES48-2
C478 RB48-2
C48-2 类48-2 Lei 48-2

Fig. 1

Fertility phenotype of some F1 combinations I: no emerged of anthers; II: from 0 to 25% anthers emerged; III: more than 25% to 50% anthers emerged; IV: more than 50% to 75% anthers emerged; V: more than 75% anthers emerged."

Table 2

Results of fertility identification of F1 hybrids crossed between CMS-C lines and Z16"

组合
Combination		 雄穗育性等级 Fertility grade of tassel 花粉可染率 Pollen staining rate
I II III IV V 可染
Fully staining (%)		 半染
Semi-staining (%)		 不染
Not staining (%)
CMo17×Z16 0 0 0 0 28 98.6 0 1.4
C698-3×Z16 0 0 0 0 25 96.7 0 3.3
C黄早四×Z16 C Huangzaosi×Z16 0 0 0 0 29 97.4 0 2.6
C478×Z16 0 0 0 0 28 97.8 0 2.2
C48-2×Z16 0 4 24 0 0 39.0 26.9 34.1
G48-2×Z16 28 0 0 0 0 0 0 100
EC48-2×Z16 30 0 0 0 0 0 0 100
ES48-2×Z16 29 0 0 0 0 - - -
RB48-2×Z16 28 0 0 0 0 - - -
类48-2×Z16 Lei48-2×Z16 28 0 0 0 0 - - -

Table 3

Results of fertility identification of F1 hybrids crossed between CMS-T, CMS-S lines, and Z16"

组合
Combination		 雄穗育性等级Fertility grade of tassel 花粉可染率Pollen staining rate
I II III IV V 可染
Fully staining (%)		 半染
Semi-staining (%)		 不染
Not staining (%)
TMo17×Z16 28 0 0 0 0 — — —
T698-3×Z16 29 0 0 0 0 — — —
SMo17×Z16 0 5 24 0 0 22.0 12.1 65.9
S698-3×Z16 30 0 0 0 0 29.2 33.3 37.5

Table 4

Results of fertility identification of F1 hybrids crossed between CMS-C lines and 7250-14-1"

组合
Combination		 雄穗育性等级Fertility grade of tassel 花粉可染率Pollen staining rate
I II III IV V 可染
Fully staining (%)		 半染
Semi-staining (%)		 不染
Not staining (%)
CMo17×7250-14-1 0 0 0 0 28 96.6 3.3 0
C698-3×7250-14-1 0 0 0 0 29 91.9 2.7 5.4
C黄早四×7250-14-1
C Huangzaosi×7250-14-1		 0
 0
 0
 0
 30
 98.8
 0
 1.2
C478×7250-14-1 0 0 0 0 29 97.8 0 2.2
C48-2×7250-14-1 0 0 29 0 0 83.3 11.1 5.6
G48-2×7250-14-1 0 0 28 0 0 94.1 0 5.9
EC48-2×7250-14-1 0 0 28 0 0 22.6 25.0 52.4
ES48-2×7250-14-1 0 2 26 0 0 96.6 0 3.4
RB48-2×7250-14-1 29 0 0 0 0 0 0 100
类48-2×7250-14-1
Lei48-2×7250-14-1		 28
 0
 0
 0
 0
 0
 0
 100

Table 5

Results of fertility identification of F1 hybrids crossed between CMS-T, CMS-S lines, and 7250-14-1"

组合
Combination		 雄穗育性等级Fertility grade of tassel 花粉可染率Pollen staining rate
I II III IV V 可染
Fully staining (%)		 半染
Semi-staining (%)		 不染
Not staining (%)
TMo17×7250-14-1 29 0 0 0 0 — — —
T698-3×7250-14-1 29 0 0 0 0 — — —
SMo17×7250-14-1 0 0 0 29 0 14.1 15.3 70.6
S698-3×7250-14-1 0 0 0 28 0 43.9 43.9 12.2

Table 6

Fertility results of F2 and BC1 populations crossed between CMS lines and Z16"

组合
Combination		 可育株
No. of fertile plants		 不育株
No. of sterile plants		 总株数
Total No. of
plants		 理论比例
Theoretical
ratio		 χ2 年份和地点
Year and place
(C黄早四×Z16)F2
(C Huangzaosi×Z16) F2		 287 115 402 3:1 1.33 2016景洪 Jinghong
299 106 405 3:1 0.15 2017温江 Wenjiang
1820 616 2436 3:1 0.05 2017崇州 Chongzhou
(C黄早四×Z16)×黄早四
(C Huangzaosi×Z16)×Huangzaosi		 33 36 69 1:1 0.06 2016温江 Wenjiang
98 107 205 1:1 0.20 2017温江 Wenjiang
(C478×Z16)F2 251 107 358 3:1 2.15 2017温江 Wenjiang
(C478×Z16)×478 134 135 269 1:1 0 2017温江 Wenjiang

Table 7

Fertility results of F2 and BC1 populations crossed between CMS lines and 7250-24-1"

组合
Combination		 可育株
No. of fertile plants		 不育株
No. of sterile plants		 总株数
Total No. of
plants		 期望比例
Theoretical
ratio		 χ2 年份、地点
Year and place
(C黄早四×7250-14-1)F2
(C Huangzaosi×7250-14-1) F2		 271 85 356 3:1 0.12 2016景洪 Jinghong
319 104 423 3:1 0.02 2017温江 Wenjiang
1386 475 1861 3:1 0.14 2017崇州 Chongzhou
(C黄早四×7250-14-1)×黄早四
(C Huangzaosi×7250-14-1) ×Huangzaosi		 23 22 45 1:1 0 2016温江 Wenjiang
124 141 265 1:1 0.55 2017温江 Wenjiang
(C478×7250-14-1)F2 163 110 273 9:7 1.19 2016温江 Wenjiang
243 159 392 9:7 1.44 2017温江 Wenjiang
(C478×7250-14-1)×478 59 191 247 1:3 0.37 2017温江 Wenjiang

Supplementary table 1

Polymorphic markers for restoring gene mapping in Z16"

引物
Primer		 引物类型
Primer type		 正向序列
Forward sequence (5'-3')		 反向序列
Reverse sequence (5'-3')
Chr8-86080 InDel CGTCGTTGAGGTGAGAAGAG CTCCGAACCTGATCCGAGTA
B-2 InDel ACGAATACGATACGTAGCCA GTGAATCTGCGGTGAACAAA
B-6-1 InDel GGATGGAATATATAAAGTTTGCT GGCTCATTACCTTGGTGTCA
B-1 InDel GATCGTTCCGGCCCAAGAAG TAGCCGTGGAGTTGGTAGCC
m-1 SSR CATTGACCGGGGTAGGAAGT CATTGACCGGGGTAGGAAGT
TIDP5557 InDel CATGAGATCAACGGGATGC AGTAGAGATCCGGGAGGTGG
Chr8-1330080 InDel CCAAGTTGGATACAACGACAGA AGAAGCAACGTCTGCAGGAT
IDP8573 InDel CGAGTCAGTTGCTTACGGG AATTGCCGAGTGGATACAGG

Fig. 2

Linkage map of the Rf on chromosome 8 in Z16"

Fig. 3

Linkage map of the Rf on chromosome 8 in 7250-14-1"

Supplementary table 2

Polymorphic markers for restoring gene mapping in 7250-14-1"

引物
Primer		 引物类型
Primer type		 正向序列
Forward sequence (5'-3')		 反向序列
Reverse sequence (5'-3')
m-10 SSR AGCGCTCGATTCCTGTAGTG GGGTGTCGTTGGTTGGGATT
Chr8-86080 InDel CGTCGTTGAGGTGAGAAGAG CTCCGAACCTGATCCGAGTA
B-2 InDel ACGAATACGATACGTAGCCA GTGAATCTGCGGTGAACAAA
B-6-2 InDel CCAATGTTTTGATGGAAGTCCT AATTGCCATGTTCTTACCTGT
B-1 InDel GATCGTTCCGGCCCAAGAAG TAGCCGTGGAGTTGGTAGCC
Chr8-398180 InDel GCCAGTTCGGAGACAGGAT ACCGCCATCCAATTAACAAG
IDP8319 InDel TTGACCCTCCTGTTACGTGC GAGCATGGACCACATGACC
IDP500 InDel CACTGCCGTAGAGTAGTGCG GGCTTCAAGATCAGTCCG
IDP7866 InDel GGACGAAGCGATCGAGTACC AGATGAGGGAAGTGAGCAGC
Chr8-1330080 InDel CCAAGTTGGATACAACGACAGA AGAAGCAACGTCTGCAGGAT
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