Development of Thermo-Sensitive Male Sterile Rice with CRISPR/Cas9 Technology

WU Ming-ji; LIN Yan; LIU Hua-qing; CHEN Jian-min; FU Yan-ping; YANG Shao-hua; WANG Feng

doi:10.19303/j.issn.1008-0384.2018.10.001

Volume 33 Issue 10

Mar. 2019

Turn off MathJax

Article Contents

Article Navigation > Fujian Journal of Agricultural Sciences > 2018 > 33(10): 1011-1015

WU Ming-ji, LIN Yan, LIU Hua-qing, CHEN Jian-min, FU Yan-ping, YANG Shao-hua, WANG Feng. Development of Thermo-Sensitive Male Sterile Rice with CRISPR/Cas9 Technology[J]. Fujian Journal of Agricultural Sciences, 2018, 33(10): 1011-1015. doi: 10.19303/j.issn.1008-0384.2018.10.001

Citation:

WU Ming-ji, LIN Yan, LIU Hua-qing, CHEN Jian-min, FU Yan-ping, YANG Shao-hua, WANG Feng. Development of Thermo-Sensitive Male Sterile Rice with CRISPR/Cas9 Technology[J]. Fujian Journal of Agricultural Sciences, 2018, 33(10): 1011-1015. doi: 10.19303/j.issn.1008-0384.2018.10.001

Citation:

PDF( 1578 KB)

Development of Thermo-Sensitive Male Sterile Rice with CRISPR/Cas9 Technology

doi: 10.19303/j.issn.1008-0384.2018.10.001

Institute of Biotechnology, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350003, China

Received Date: 2018-08-16
Rev Recd Date: 2018-09-29
Publish Date: 2018-10-28

Abstract

Abstract

Breeding of photoperiod or thermo-sensitive male sterile line is the key for the development of two-line hybrid rice. The allele of tms5 is the major TGMS genetic resource for two-line hybrid rice breeding in China. Targeted editing on TMS5 of fertile rice using CRISPR/Cas-9 technology could effectively create thermo-sensitive genic male sterile rice. This study used the approach to edit the TMS5 gene successfully. The CRISPR/Cas-9 vector of TMS502 was constructed, then the T-DNA was transformed into GH89. Ten transgenic plants were thus regenerated, with 6 of them containing either the insertion or deletion mutations. The T₀ plant of tms5-l was a homozygous mutant, and tms 5-4 a bi-allelic mutant. Both of them displayed thermo-sensitive male sterile phenotype. There were 4 homozygous mutant genotypes in the T₁ generation of the non-transgenic plants. The mutants showed the same characteristics as shown in the T₀ generation without any new mutations. Male fertility of T₂ generation of tms5 mutants was tested in natural high temperature environment and in phytotrons at 3 temperature levels. The results indicated that the critical sterility-inducing temperature of the tms5 sterile lines was approximately 24 C, which satisfied the requirements for two-line male sterile line breeding. The successful conversion of GH89 line into a two-line male sterile rice using the gene editing technology suggested that CRISPR/Cas-9 could be applied to facilitate the development of two line hybrid rice breeding in the future.
- hybrid rice,
- two-line male sterile line,
- gene,
- CRISPR/Cas-9,
- tms5

FullText(HTML)

References(17)

References

[1]	朱英国.杂交水稻研究50年[J].科学通报, 2016, 61(35):3740-3747. http://www.cnki.com.cn/Article/CJFDTOTAL-KXTB201635007.htm
[2]	余四斌, 熊银, 肖景华, 等.杂交稻与绿色超级稻[J].科学通报, 2016, 61(35):3797-3803. http://www.cnki.com.cn/Article/CJFDTOTAL-KXTB201635013.htm
[3]	牟同敏.中国两系法杂交水稻研究进展和展望[J].科学通报, 2016, 61(35):3761-3769. http://www.cnki.com.cn/Article/CJFDTOTAL-KXTB201635009.htm
[4]	范优荣, 曹晓风, 张启发.光温敏雄性不育水稻的研究进展[J].科学通报, 2016, 61(35):3822-3832. http://d.old.wanfangdata.com.cn/Periodical/hunannykx201111002
[5]	FAN Y, YANG J, MATHIONI S M, et al. PMS1T, producing phased small-interfering RNAs, regulates photoperiod-sensitive male sterility in rice[J].Proceedings of the National Academy of Sciences, 2016, 113(52):15144-15149. doi: 10.1073/pnas.1619159114
[6]	DING J, LU Q, OUYANG Y, et al. A long noncoding RNA regulates photoperiod-sensitive male sterility, an essential component of hybrid rice[J]. Proceedings of the National Academy of Sciences, 2012, 109(7):2654-2659. doi: 10.1073/pnas.1121374109
[7]	ZHOU H, ZHOU M, YANG Y, et al. RNase Z^S1 processes UbL40 mRNAs and controls thermosensitive genic male sterility in rice[J]. Nature Communications, 2014(5):4884. http://www.nature.com/ncomms/2014/140911/ncomms5884/abs/ncomms5884.html
[8]	ZHANG H, XU C, He Y, et al. Mutation in CSA creates a new photoperiod-sensitive genic male sterile line applicable for hybrid rice seed production[J]. Proceedings of the National Academy of Sciences, 2013, 110(1):76-81. doi: 10.1073/pnas.1213041110
[9]	YU J, HAN J, KIM Y, et al. Two rice receptor-like kinases maintain male fertility under changing temperatures[J]. Proceedings of the National Academy of Sciences, 2017, 114(46):12327-12332. doi: 10.1073/pnas.1705189114
[10]	ZHANG J, ZHANG H, BOTELLA J R, et al. Generation of new glutinous rice by CRISPR/Cas9-targeted mutagenesis of the Waxy gene in elite rice varieties[J]. Journal of Integrative Plant Biology, 2018, 60:369-375. doi: 10.1111/jipb.v60.5
[11]	WANG F, WANG C, LIU P, et al. Enhanced rice blast resistance by CRISPR/Cas9-Targeted Mutagenesis of the ERF transcription factor gene OsERF922[J]. PLoS ONE, 2016(1):e0154027. http://www.ncbi.nlm.nih.gov/pubmed/27116122
[12]	TANG L, MAO B, LI Y, et al. Knockout of OsNramp5 using the CRISPR/Cas9 system produces low Cd-accumulating indica rice without compromising yield[J]. Scientific Reports, 2017(7):1-12. http://europepmc.org/abstract/MED/29089547
[13]	SUN Y, ZHANG X, WU C, et al. Engineering herbicide resistant rice plants through CRISPR/Cas9-Mediated homologous recombination of acetolactate synthase[J]. Molecular Plant, 2016(9):628-631. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=fzzw-e201604017
[14]	MACOVEI A, SEVILLA NR, CANTOS C, et al. Novel alleles of rice eIF4G generated by CRISPR/Cas9-targeted mutagenesis confer resistance to Rice tungro spherical virus[J]. Plant Biotechnology Journal, 2018, 1-10.doi: 10.1111/pbi.12927.
[15]	LI Q, ZHANG D, CHEN M, et al. Development of japonica photo-sensitive genic male sterile rice lines by editing carbon starved anther using CRISPR/Cas9[J]. Journal of Genettics Genomics, 2016, 43:415-419. doi: 10.1016/j.jgg.2016.04.011
[16]	ZHOU H, HE M, LI J, et al. Development of commercial thermo-sensitive genic male sterile rice accelerates hybrid rice breeding using the CRISPR/Cas9-mediated TMS5 editing system[J]. Scientific Reports, 2016(6):1-12. http://www.ncbi.nlm.nih.gov/pubmed/27874087
[17]	HIEI Y, KOMARI T, KUBO T. Transformation of rice mediated by Agrobacterium tumefaciens[J]. Plant Molecular Biology, 1997, 35:205-218. doi: 10.1023/A:1005847615493