scholarly journals Development of High Yielding Glutinous Cytoplasmic Male Sterile Rice (Oryza sativa L.) Lines through CRISPR/Cas9 Based Mutagenesis of Wx and TGW6 and Proteomic Analysis of Anther

Agronomy ◽  
2018 ◽  
Vol 8 (12) ◽  
pp. 290 ◽  
Author(s):  
Yue Han ◽  
Dengjie Luo ◽  
Babar Usman ◽  
Gul Nawaz ◽  
Neng Zhao ◽  
...  
Keyword(s):  
Target Genes ◽  
Agronomic Traits ◽  
Amylose Content ◽  
Oryza Sativa L ◽  
Protein Biosynthesis ◽  
Starch Content ◽  
Male Sterile ◽  
Maintainer Line ◽  
Cytoplasmic Male Sterile ◽  
Cms Line

Development of high yielding and more palatable glutinous rice is an important goal in breeding and long-standing cultural interaction in Asia. In this study, the TGW6 and Wx, major genes conferring 1000 grain weight (GW) and amylose content (AC), were edited in a maintainer line by CRISPR/Cas9 technology. Four targets were assembled in pYLCRISPR/Cas9Pubi-H vector and T0 mutant plants were obtained through Agrobacterium mediated transformation with 90% mutation frequency having 28% homozygous mutations without off-target effects in three most likely sites of each target and expression level of target genes in mutant lines was significantly decreased (P < 0.01), the GW and gel consistency (GC) were increased, and the AC and gelatinization temperature (GT) were decreased significantly and grain appearance was opaque, while there was no change in starch content (SC) and other agronomic traits. Mutations were inheritable and some T1 plants were re-edited but T2 generation was completely stable. The pollen fertility status was randomly distributed, and the mutant maintainer lines were hybridized with Cytoplasmic Male Sterile (CMS) line 209A and after subsequent backcrossing the two glutinous CMS lines were obtained in BC2F1. The identified proteins from anthers of CMS and maintainer line were closely associated with transcription, metabolism, signal transduction, and protein biosynthesis. Putative mitochondrial NAD+-dependent malic enzyme was absent in CMS line which caused the pollen sterility because of insufficient energy, while upregulation of putative acetyl-CoA synthetase and Isoamylase in both lines might have strong relationship with CMS and amylose content. High yielding glutinous CMS lines will facilitate hybrid rice breeding and investigations of proteins linked to male sterility will provide the insights to complicated metabolic network in anther development.

scholarly journals Breeding of Indica glutinous cytoplasmic male sterile line WX209A via CRISPR/Cas9 mediated genomic editing

2019 ◽  
Vol 55 (No. 3) ◽  
pp. 93-100 ◽  
Author(s):  
Xin Wang ◽  
Yue Han ◽  
Xuan Feng ◽  
Yun-Zhen Li ◽  
Bao-Xiang Qin ◽  
...  
Keyword(s):  
Amylose Content ◽  
Rice Variety ◽  
High Yield ◽  
Male Sterile ◽  
Male Sterile Line ◽  
Maintainer Line ◽  
Cytoplasmic Male Sterile ◽  
Glutinous Rice ◽  
Short Period ◽  
Cms Line

Glutinous cytoplasmic male sterile (CMS) line is necessary to select hybrid glutinous rice combination with high yield and quality. To develop glutinous CMS with low amylose content, in this study, we firstly knocked out the granule-bound starch synthase OsWaxy in 209B using CRISPR/Cas9 mediated genome editing technology and successfully obtained a glutinous maintainer line WX209B. Comparing with maintainer line 209B, WX209B showed decreased amylose contents and similar agronomic characters. And then, through one generation of hybridization and two generations of backcrossing with WX209B as the male parent and 209A as the female parent, the glutinous CMS line WX209A was successfully achieved. Our study provides a strategy to efficiently breed for the glutinous cytoplasmic male sterile line by combining CRISPR/Cas9-mediated gene editing technology with conventional backcross breeding method in a short period, which prepares the ground for further breeding of hybrid glutinous rice variety.  

scholarly journals Development of cytoplasmic male sterile lines and restorer lines of various elite Indica Group rice cultivars using CW-CMS/Rf17 system

Rice ◽  
2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Kinya Toriyama ◽  
Tomohiko Kazama ◽  
Tadashi Sato ◽  
Yoshimichi Fukuta ◽  
Masaaki Oka
Keyword(s):  
Nuclear Gene ◽  
Oryza Rufipogon ◽  
Rice Cultivars ◽  
Male Sterile ◽  
Cytoplasmic Male Sterile ◽  
Cms Line

Abstract Background A cytoplasm of CW-type cytoplasmic male sterile (CMS) line is derived from Oryza rufipogon strain W1 and fertility is restored by a single nuclear gene, Rf17. We have previously reported that CW-CMS were effective for breeding CMS lines of Indica Group rice cultivars, IR 24 and IR 64. The applicability of this CW-CMS/Rf17 system to produce other elite Indica Group rice cultivars with CMS was explored. Findings Out of seven elite Indica Group rice cultivars, complete CMS lines were obtained for six cultivars: NSIC Rc 160, NSIC Rc 240, Ciherang, BRRI dhan 29, NERICA-L-19, and Pusa Basmati. The fertility of these six lines was restored when Rf17 was present. A CMS line was not obtained for the cultivar Samba Mahsuri. Conclusions The CW-CMS/Rf17 system will be useful to produce CMS lines and restorer lines of various elite Indica Group rice cultivars.

Study on the expression of specific chimeric-orfs of C type cytoplasmic male sterile in maize

2013 ◽  
Vol 61 (1-4) ◽  
pp. 25-36 ◽  
Author(s):  
Jiyue Wang ◽  
Lihua Zeng ◽  
Shengqing Wang ◽  
Jing Wang ◽  
Yanli Lu ◽  
...  
Keyword(s):  
Base Substitution ◽  
Pcr Analysis ◽  
Male Sterile ◽  
Maintainer Line ◽  
Cytoplasmic Male Sterile ◽  
Real Time Quantitative Pcr ◽  
Substitution Mutations ◽  
Membrane Spanning ◽  
Insertion Mutations ◽  
Online Software

In the present study, a mutated chimericorf364, namedorf366-c, was identified in a maize C type cytoplasmic male sterile line (CMS-C) by PCR and RT-PCR. Four base pair adjacent base substitution mutations (CAAA to TTTT) and eight bp insertion mutations were found inorf366-ccompared withorf364. ORF366-C was predicted to contain one membrane-spanning domain using TMHMM online software. Real-time quantitative PCR analysis showed thatorf366-cwas upregulated in the CMS-C line in comparison to its maintainer line at the uninucleate stage. The protokaryotic protein expression oforf366-cinEscherichia colishowed that ORF366-C could be a cytotoxic protein. All the results indicated thatorf366-cmay be associated with maize CMS-C.

Production of Cytoplasmic Male Sterile Rice (Oryza Sativa L.) by Cell Fusion

1989 ◽  
Vol 7 (11) ◽  
pp. 1171-1174
Author(s):  
Junko Kyozuka ◽  
Takeo Kaneda ◽  
Ko Shimamoto
Keyword(s):  
Oryza Sativa ◽  
Cell Fusion ◽  
Oryza Sativa L ◽  
Male Sterile ◽  
Cytoplasmic Male Sterile

scholarly journals Identification of Suitable Restorers for WA-CMS Lines of Rice (Oryza sativa L.) through Conventional and Molecular Breeding Methods

2020 ◽  
Author(s):  
Anirban Nath ◽  
Disharee Nath ◽  
Chand Kumar Santra ◽  
Tapash Dasgupta
Keyword(s):  
Genetic Background ◽  
Molecular Breeding ◽  
Oryza Sativa L ◽  
Fertility Restoration ◽  
Male Sterile ◽  
Maintainer Line ◽  
Fertility Restorer ◽  
Restorer Genes ◽  
Restoration Potential ◽  
Fertility Restorer Genes

The current study extensively evaluates 51 genotypes for their fertility restoration potential using test crosses with five WA (wild abortive) Cytoplasmic Male Sterile lines namely IR58025A, IR6897A, IR79156A, IR80559A and APMS6A. Also the genotypes were screened using SSR markers RM6100 and RM10313, tightly linked with the fertility restorer genes Rf4 and Rf3 respectively. The two way approach helped in identifying potential restorers for five WA-CMS lines and also detected the presence of dominant Rf genes in their genetic background. The R-lines identified can be safely presumed to be strong restorers for consecutive A-lines they were crossed with. The study also identified a potential maintainer CN1039-9 for the A-line IR58025A. The maintainer line identified can be later exploited for developing new CMS lines.

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