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 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 Generation of a New Glutinous Photothermosensitive Genic-Male-Sterile (PTGMS) Line by CRISPR/Cas9-Directed Mutagenesis of Wx in Rice (Oryza sativa L.)

Agriculture ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1044
Author(s):  
Kaichong Teng ◽  
Xin Wang ◽  
Xinying Guo ◽  
Yaoguang Liu ◽  
Rongbai Li
Keyword(s):  
Mutation Rate ◽  
Hybrid Rice ◽  
Amylose Content ◽  
Rice Variety ◽  
High Yield ◽  
Male Sterile ◽  
High Quality ◽  
Glutinous Rice ◽  
Homozygous Mutant ◽  
Wx Gene

The Photothermosensitive Genic-Male-Sterile (PTGMS) line, Y58S, an indica rice variety, combines high-quality and high-light-efficiency use, disease and stress resistance, and excellent plant type and mating force. Y58S is widely used to assemble two-line hybrid rice varieties, especially super hybrids. The Wx gene is the main effector gene for controlling amylose synthesis, which determines the amylose content (AC) of rice grains. By editing this gene, a glutinous line with a low AC can be obtained. In this study, the CRISPR/Cas9 system was used to mediate the editing of the Wx gene, which caused ultra-low AC mutations that produced a PTGMS glutinous rice strain with excellent waxiness. The results showed that 18 positively transformed plants were obtained from the T0 generation, with a mutation rate of 64.29%, of which six were homozygous mutant plants, indicating that the gene-editing target had a higher targeting efficiency and a higher homozygosity mutation rate. Compared to the wild type, the AC of the mutants was significantly lower. Through molecular marker detection and screening of T1 and T2 generations, five homozygous T-DNA-free mutant strains were identified that were consistent with Y58S in fertility and other agronomic traits except for AC. Among these, the AC of the W-1-B-5 homozygous mutant, the glutinous PTGMS line wx-Y58S, was as low as 0.6%. Our research revealed that the Wx gene of excellent PTGMS rice can be edited to generate a new waxy PTGMS line using the CRISPR/Cas9 system. This study provided a simple and effective strategy for breeding high-yield, high-quality, and glutinous two-line hybrid rice, and provided excellent sterile lines for their large-scale application. Once put into use, waxy hybrid rice will greatly improve the yield of glutinous rice and increase social benefits.

scholarly journals Comparison on the Biochemical Characteristics between Alfalfa Sterile Line and Maintainer Line

2020 ◽  
Vol 203 ◽  
pp. 02002
Author(s):  
YAN Dong ◽  
XU Xiaobo ◽  
GUO Qiang ◽  
XU Bo
Keyword(s):  
Antioxidant Enzyme ◽  
Soluble Protein ◽  
Soluble Sugar ◽  
Pollen Grain ◽  
Abnormal Development ◽  
Free Proline ◽  
Male Sterile ◽  
Male Sterile Line ◽  
Maintainer Line ◽  
Cytoplasmic Male Sterile

To study on the morphological character of cytology and physiological and biochemical mechanism of alfalfa cytoplasmic male sterile line, this paper took sterile line MS-GN1A and maintainer line MS-GN1B as raw material, separately conducted microscopic observation to anther in different development stage and determined the content of soluble sugar, starch, soluble protein, free proline and malondialdehyde, and the activity of antioxidant enzyme (catalase, peroxidase and superoxide dismutase). The results showed that: 1. Compared with maintainer line, the pollen grain of alfalfa cytoplasmic male sterile line presented shriveled appearance, smaller size with deletion and few; during the tetrad period of pollen, tapetum cell abnormally grew; callose and epidermis didn’t disintegrate and couldn’t release pollen grain; pollen abortion of alfalfa cytoplasmic sterile line was related to the abnormal development of connective vascular bundle. 2. With the growth and development of bud and the development and maturity of pollen, the content of soluble sugar, starch, soluble protein, and free proline showed deficient phenomenon in each development period of bud of sterile line MS-GN1A; as peroxidation index of membranous substance, the activity of malondialdehyde and antioxidant enzyme was higher than it of homotype maintainer line MS-GN1B.

scholarly journals Expression profiling of immature florets of IR58025A, a wild-abortive cytoplasmic male sterile line of rice and its cognate, isonuclear maintainer line, IR58025B

3 Biotech ◽  
2019 ◽  
Vol 9 (7) ◽  
Author(s):  
K. Pranathi ◽  
M. B. Kalyani ◽  
B. C. Viraktamath ◽  
S. M. Balachandran ◽  
S. K. Hajira ◽  
...  
Keyword(s):  
Expression Profiling ◽  
Male Sterile ◽  
Male Sterile Line ◽  
Maintainer Line ◽  
Cytoplasmic Male Sterile

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.

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