Chinese Journal OF Rice Science ›› 2016, Vol. 30 ›› Issue (2): 143-151.DOI: 10.16819/j.1001-7216.2016.5154
• Orginal Article • Previous Articles Next Articles
Zheng-fu YANG1, Ying-xin ZHANG1, Lian-ping SUN1,2, Pei-pei ZHANG1, Dan-dan XUAN1, Ling LIU1,3, Xia HU1, Zi-he LI1, Xiao-deng ZHAN1, Wei-xun WU1, Li-yong CAO1,*(), Shi-hua CHENG1,*()
Received:
2015-10-19
Revised:
2015-12-11
Online:
2016-03-10
Published:
2016-03-10
Contact:
Li-yong CAO, Shi-hua CHENG
About author:
# These authors contributed equally to this work;
杨正福1, 张迎信1, 孙廉平1,2, 张沛沛1, 轩丹丹1, 刘嶺1,3, 胡霞1, 李紫荷1, 占小登1, 吴玮勋1, 曹立勇1,*(), 程式华1,*()
通讯作者:
曹立勇,程式华
作者简介:
# 共同第一作者;
基金资助:
CLC Number:
Zheng-fu YANG, Ying-xin ZHANG, Lian-ping SUN, Pei-pei ZHANG, Dan-dan XUAN, Ling LIU, Xia HU, Zi-he LI, Xiao-deng ZHAN, Wei-xun WU, Li-yong CAO, Shi-hua CHENG. Identification and Gene Mapping of Male Sterile Mutant gamyb5 in Rice[J]. Chinese Journal OF Rice Science, 2016, 30(2): 143-151.
杨正福, 张迎信, 孙廉平, 张沛沛, 轩丹丹, 刘嶺, 胡霞, 李紫荷, 占小登, 吴玮勋, 曹立勇, 程式华. 水稻雄性不育突变体gamyb5的鉴定与基因定位[J]. 中国水稻科学, 2016, 30(2): 143-151.
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URL: http://www.ricesci.cn/EN/10.16819/j.1001-7216.2016.5154
引物名称 Primer name | 上游引物 Forward primer (5'-3') | 下游引物 Reverse primer (5'-3') | 实验目的 Purpose |
---|---|---|---|
RD0114 | GCTTGTGGCAATTGGG | CGCCTGATGGATGTCG | 精细定位Fine mapping |
ZF-26 | GATCATACCATCATCAGAAGCAGT | CAAGCAGCCCACATCACAGC | 精细定位Fine mapping |
ZF-28 | TCTCGCTCCTCTTCGCCCAAAC | ACCCGCACCAGTACGTGACCC | 精细定位Fine mapping |
ZF-29 | GGCAGCAGCAATCACAACAC | TCGGCACTTTCTTTACTCAACT | 精细定位Fine mapping |
ZF-31 | CAGCCAGGCACCGCAGCAAA | AAGAAAGGCAGAGCAAGAAGG | 精细定位Fine mapping |
ZF-32 | GCACGCTTCTCGCTACGCTAT | GCTCCAGGAACACCACCAACTC | 精细定位Fine mapping |
ZF-38 | TGGGTGCGAGGTTTCTGTGA | AATGAATGCGTGTTTCCTGT | 精细定位Fine mapping |
InDel15 | CAACCCCTCCAAATACCTGA | ACCGTGTTCATGCCTTTCAC | 精细定位Fine mapping |
GAMYB5 | ACTGGAGCACTGCTGATCTTC | GCACATCTTCTCAGTTGCCAG | gamyb5位点检测gamyb5 deletion site detection |
cDNA | CCCCTGCGAGTCCAATCTAC | ACCGGCTTATCTCCATGCAC | cDNA测序cDNA sequencing |
Table 1 Makers used for fine mapping and sequencing.
引物名称 Primer name | 上游引物 Forward primer (5'-3') | 下游引物 Reverse primer (5'-3') | 实验目的 Purpose |
---|---|---|---|
RD0114 | GCTTGTGGCAATTGGG | CGCCTGATGGATGTCG | 精细定位Fine mapping |
ZF-26 | GATCATACCATCATCAGAAGCAGT | CAAGCAGCCCACATCACAGC | 精细定位Fine mapping |
ZF-28 | TCTCGCTCCTCTTCGCCCAAAC | ACCCGCACCAGTACGTGACCC | 精细定位Fine mapping |
ZF-29 | GGCAGCAGCAATCACAACAC | TCGGCACTTTCTTTACTCAACT | 精细定位Fine mapping |
ZF-31 | CAGCCAGGCACCGCAGCAAA | AAGAAAGGCAGAGCAAGAAGG | 精细定位Fine mapping |
ZF-32 | GCACGCTTCTCGCTACGCTAT | GCTCCAGGAACACCACCAACTC | 精细定位Fine mapping |
ZF-38 | TGGGTGCGAGGTTTCTGTGA | AATGAATGCGTGTTTCCTGT | 精细定位Fine mapping |
InDel15 | CAACCCCTCCAAATACCTGA | ACCGTGTTCATGCCTTTCAC | 精细定位Fine mapping |
GAMYB5 | ACTGGAGCACTGCTGATCTTC | GCACATCTTCTCAGTTGCCAG | gamyb5位点检测gamyb5 deletion site detection |
cDNA | CCCCTGCGAGTCCAATCTAC | ACCGGCTTATCTCCATGCAC | cDNA测序cDNA sequencing |
Fig. 1. Phenotypic comparison of wild type Zhonghui 8015(WT)and gamyb5 . A, Plant of Zhonghui 8015 and the gamyb5 mutant at the heading stage; B, Panicle of Zhonghui 8015 and the gamyb5 mutant; C, Spikelet of the wild type Zhonghui 8015 and gamyb5 ; D, Anther of Zhonghui 8015 and gamyb5 mutant; E, Pollen fertility of Zhonghui 8015 and gamyb5.
组合 Cross | Seed-setting rate of F1 /% | F2 | |||
---|---|---|---|---|---|
野生型植株数 Number of wild type plants | 突变体植株数 Number of mutant plants | ||||
gamyb5/02428 | 86.8 | 1895 | 642 | 0.14 | 3.84 |
gamyb5/中恢8015 gamyb5/Zhonghui 8015 | 87.2 | 234 | 70 | 0.63 | |
中恢8015/02428 Zhonghui 8015/02428 | 87.6 | 960 | 0 | - |
Table 2 Genetic analysis of the gamyb5 mutant.
组合 Cross | Seed-setting rate of F1 /% | F2 | |||
---|---|---|---|---|---|
野生型植株数 Number of wild type plants | 突变体植株数 Number of mutant plants | ||||
gamyb5/02428 | 86.8 | 1895 | 642 | 0.14 | 3.84 |
gamyb5/中恢8015 gamyb5/Zhonghui 8015 | 87.2 | 234 | 70 | 0.63 | |
中恢8015/02428 Zhonghui 8015/02428 | 87.6 | 960 | 0 | - |
Fig.2. Positional cloning of the gamyb5 mutated gene. A, Preliminary mapping of gamyb5; B, Fine mapping of gamyb5; C, Prediction of the candidate gene within the fine-mapped region; D, The structure of candidate gene LOC_Os01g0812000 and the mutation site in gamyb5 mutant (8 bases deleted).
Fig. 3. Transverse section analyses of the wild type and gamyb5 anthers in various of developmental stages. A to E, Wild type; G to L, The gamyb5 mutant. A and G, Cross-section of anthers at the stage 7; B and H, Cross-section of anthers at the stage 8; C and I, Cross-section of anthers at the stage 9; D and J, Cross-section of anthers at the stage 10; E and K, Cross-section of anthers at the stage 11; F and L, Cross-section of anthers at the stage 12. Ep, Epidermis; En, Endothecium; ML, Middle layer; T, Tapetum; Ms, Microsporocyte; Dy, Dyad cell; Msp, Microspore; BP, Biceullar pollen; MP, Mature pollen. Bars = 20 μm.
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