scholarly journals Unraveling the Genetic Basis of Fertility Restoration for Cytoplasmic Male Sterile Line WNJ01A Originated From Brassica juncea in Brassica napus

2021 ◽  
Vol 12 ◽  
Author(s):  
Qian Yang ◽  
Xiaoyi Nong ◽  
Jize Xu ◽  
Fan Huang ◽  
Fang Wang ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Bulked Segregant Analysis ◽  
Fertility Restoration ◽  
26S Proteasome ◽  
Morphological Observation ◽  
Male Sterile ◽  
Bac Clone ◽  
Cell Stage ◽  
Cms Line ◽  
Candidate Interval

Crosses that lead to heterosis have been widely used in the rapeseed (Brassica napus L.) industry. Cytoplasmic male sterility (CMS)/restorer-of-fertility (Rf) systems represent one of the most useful tools for rapeseed production. Several CMS types and their restorer lines have been identified in rapeseed, but there are few studies on the mechanisms underlying fertility restoration. Here, we performed morphological observation, map-based cloning, and transcriptomic analysis of the F2 population developed by crossing the CMS line WNJ01A with its restorer line Hui01. Paraffin-embedded sections showed that the sporogenous cell stage was the critical pollen degeneration period, with major sporogenous cells displaying loose and irregular arrangement in sterile anthers. Most mitochondrial electron transport chain (mtETC) complex genes were upregulated in fertile compared to sterile buds. Using bulked segregant analysis (BSA)-seq to analyze mixed DNA pools from sterile and fertile F2 buds, respectively, we identified a 6.25 Mb candidate interval where Rfw is located. Using map-based cloning experiments combined with bacterial artificial chromosome (BAC) clone sequencing, the candidate interval was reduced to 99.75 kb and two pentatricopeptide repeat (PPR) genes were found among 28 predicted genes in this interval. Transcriptome sequencing showed that there were 1679 DEGs (1023 upregulated and 656 downregulated) in fertile compared to sterile F2 buds. The upregulated differentially expressed genes (DEGs) were enriched in the Kyoto Encyclopedia of Genes and Genomes (KEGG) lysine degradation pathway and phenylalanine metabolism, and the downregulated DEGs were enriched in cutin, suberine, and wax biosynthesis. Furthermore, 44 DEGs were involved in pollen and anther development, such as tapetum, microspores, and pollen wall development. All of them were upregulated except a few such as POE1 genes (which encode Pollen Ole e I allergen and extensin family proteins). There were 261 specifically expressed DEGs (9 and 252 in sterile and fertile buds, respectively). Regarding the fertile bud-specific upregulated DEGs, the ubiquitin–proteasome pathway was enriched. The top four hub genes in the protein–protein interaction network (BnaA09g56400D, BnaA10g18210D, BnaA10g18220D, and BnaC09g41740D) encode RAD23d proteins, which deliver ubiquitinated substrates to the 26S proteasome. These findings provide evidence on the pathways regulated by Rfw and improve our understanding of fertility restoration.

Alloplasmic male sterile Brassica napus with Enarthrocarpus lyratus cytoplasm: introgression and molecular mapping of an E. lyratus chromosome segment carrying a fertility restoring gene

Genome ◽  
2003 ◽  
Vol 46 (5) ◽  
pp. 792-797 ◽  
Author(s):  
H S Janeja ◽  
S K Banga ◽  
P B Bhaskar ◽  
S S Banga
Keyword(s):  
Brassica Napus ◽  
Cytoplasmic Male Sterility ◽  
Male Sterility ◽  
Molecular Mapping ◽  
Bulked Segregant Analysis ◽  
Fertility Restoration ◽  
Hybrid Seed Production ◽  
Male Sterile ◽  
Fertility Restorer ◽  
Donor Species

A cytoplasmic male sterility (CMS) system for Brassica napus (2n = 38; AACC) was developed by backcross substitution of its nucleus into the cytoplasm of a wild crucifer, Enarthrocarpus lyratus. Male sterility was complete, stable, and expressed in small flowers with rudimentary anthers. Since the B. napus germplasm lines were complete or partial maintainers of male sterility, the required fertility restorer gene (Rfl) was introgressed from the cytoplasm donor species. Inheritance studies carried out on F1 and F2 populations derived from hybridizing cytoplasmic male sterile and male fertile near-isogenic (PNILs) lines of B. napus 'Westar', revealed a monogenic dominant control for fertility restoration. Bulked segregant analysis with 215 RAPD primers helped in the identification of putative primers associated with fertility restoration. Co-segregation analysis of eight such primers with Rfl gene revealed two markers, OPK 15700 and OPZ 061300, which flank the Rfl locus on either side at a distance of 8.2 and 2.5 cM, respectively. These DNA markers will be useful in marker-assisted selection for improving the commercial potential of this newly developed CMS-fertility-restorer system for hybrid seed production programs in rapeseed.Key words: oilseed rape, hybrids, cytoplasmic male sterility, fertility restoration, RAPD mapping.

scholarly journals Molecular Analysis Uncovers the Mechanism of Fertility Restoration in Temperature-Sensitive Polima Cytoplasmic Male-Sterile Brassica napus

2021 ◽  
Vol 22 (22) ◽  
pp. 12450
Author(s):  
Qing Xiao ◽  
Huadong Wang ◽  
Hui Chen ◽  
Xiaohan Chen ◽  
Jing Wen ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Male Sterility ◽  
Rna Sequencing ◽  
Single Molecule ◽  
Bulked Segregant Analysis ◽  
Fertility Restoration ◽  
Differentially Expressed ◽  
Temperature Sensitive ◽  
Male Sterile ◽  
Line Breeding

Temperature-sensitive male sterility is a heritable agronomic trait affected by genotype-environment interactions. In rapeseed (Brassica napus), Polima (pol) temperature-sensitive cytoplasmic male sterility (TCMS) is commonly used for two-line breeding, as the fertility of pol TCMS lines can be partially restored at certain temperatures. However, little is known about the underlying molecular mechanism that controls fertility restoration. Therefore, we aimed to investigate the fertility conversion mechanism of the pol TCMS line at two different ambient temperatures (16 °C and 25 °C). Our results showed that the anthers developed and produced vigorous pollen at 16 °C but not at 25 °C. In addition, we identified a novel co-transcript of orf224-atp6 in the mitochondria that might lead to fertility conversion of the pol TCMS line. RNA-seq analysis showed that 1637 genes were significantly differentially expressed in the fertile flowers of 596-L when compared to the sterile flower of 1318 and 596-H. Detailed analysis revealed that differentially expressed genes were involved in temperature response, ROS accumulation, anther development, and mitochondrial function. Single-molecule long-read isoform sequencing combined with RNA sequencing revealed numerous genes produce alternative splicing transcripts at high temperatures. Here, we also found that alternative oxidase, type II NAD(P)H dehydrogenases, and transcription factor Hsfs might play a crucial role in male fertility under the low-temperature condition. RNA sequencing and bulked segregant analysis coupled with whole-genome sequencing identified the candidate genes involved in the post-transcriptional modification of orf224. Overall, our study described a putative mechanism of fertility restoration in a pol TCMS line controlled by ambient temperature that might help utilise TCMS in the two-line breeding of Brassica crops.

Molecular tagging of the male fertility restorer gene for the 501-8S cytoplasmic male sterility in rapeseed (Brassica napus L.)

2007 ◽  
Vol 58 (8) ◽  
pp. 753
Author(s):  
Haohua He ◽  
Liang Xu ◽  
Xiaosong Peng ◽  
Guangsheng Yang ◽  
Changlan Zhu ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Male Fertility ◽  
Mapping Population ◽  
Bulked Segregant Analysis ◽  
Fertility Restoration ◽  
Nuclear Gene ◽  
F1 Hybrids ◽  
Male Sterile ◽  
Brassica Napus L ◽  
Rf Gene

The cytoplasmic male sterile (CMS) system has been successfully used to explore heterosis in rapeseed (Brassica napus L.). A newly developed male sterile line (501-8S) was characterised for its male fertility response to temperature and photoperiod, and the inheritance of fertility restoration. Segregation analysis using F1 and F2, BC1, and F3 populations of the crosses between the 501-8S and fertile lines of B. napus revealed that fertility restoration was conferred by a dominant nuclear gene (Rf). The F2 population of the cross 501-8S × Yuyou1 was used as a mapping population to map the Rf gene. A combination of bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) methodology was used to identify putative markers linked to the Rf gene. Twenty-nine of the 1280 primer combinations tested revealed polymorphism between the 2 extreme bulks. Further testing of these primer combinations in individual plants identified 5 AFLP markers tightly linked to the Rf gene with a map distance of less than 5.0 cM. All 5 markers were on one side of the restoration gene in the coupling phase. The closest marker, EA02MG03-260, is only 0.4 cM from the Rf gene. The EA02MG03-260 marker was converted to a dominant sequence characterised amplified region (SCAR) marker (SCARE2M3-214). Amplification using this locus-specific primer generated specific bands with male fertile plants when tested using the mapping population. Specific amplification of SCARE2M3-214 was also detected in all 3 male sterile plants and their F1 hybrids, with 5 restorer lines used for verification. Thus, SCARE2M3-214 will be very useful for the development of new restorer lines by the transfer of the Rf gene into other breeding lines. It can also be used for isolating the Rf gene by means of map-based cloning.

scholarly journals Identification of Ms2, a Novel Locus Controlling Male-fertility Restoration of Cytoplasmic Male-sterility in Onion (Allium Cepa L.), and Development of Tightly Linked Molecular Markers

2021 ◽  
Author(s):  
Nari Yu ◽  
Sunggil Kim
Keyword(s):  
Cytoplasmic Male Sterility ◽  
Male Sterility ◽  
Allium Cepa ◽  
Male Fertility ◽  
Bulked Segregant Analysis ◽  
Fertility Restoration ◽  
Chromosome 2 ◽  
Rna Seq ◽  
Male Sterile ◽  
Male Fertility Restoration

Abstract Cytoplasmic male-sterility (CMS) has been exclusively used to produce F1 hybrid seeds of onion (Allium cepa L.). A single nuclear locus, Ms, is known to restore male-fertility of CMS in onions. Unstable male-sterile onions producing a small amount of pollen grains have been identified in a previous study. When such unstable male-sterile onions were crossed with stable male-sterile onions containing CMS-T cytoplasm, male-fertility was completely restored, although genotypes of the Ms locus were homozygous recessive. Inheritance patterns indicated that male-fertility restoration was controlled by a single locus designated as Ms2. A combined approach of bulked segregant analysis and RNA-seq was used to identify candidate genes for the Ms2 locus. High resolution melting (HRM) markers were developed based on single nucleotide polymorphisms (SNPs) detected by RNA-Seq. Comparative mapping of the Ms2 locus showed that Ms2 was positioned at the end of chromosome 2 with a distance of approximately 70 cM away from the Ms locus. Although 38 contigs containing reliable SNPs were analyzed using recombinants selected from 1,344 individuals, no contig showed perfect linkage to Ms2. Interestingly, transcription levels of orf725, a CMS-associated gene in onions, were significantly reduced in male-fertile individuals of segregating populations. However, no significant change in its transcription level was observed in individuals of a segregating population with male-fertility phenotypes determined by the Ms locus, suggesting that male-fertility restoration mechanism of Ms2 might be different from that of the Ms locus.

scholarly journals Resynthesis of new R lines in Brassica napus L.

2016 ◽  
Vol 41 (3) ◽  
pp. 529-540
Author(s):  
MA Miah ◽  
MG Rasul ◽  
MAK Mian
Keyword(s):  
Brassica Napus ◽  
Fertility Restoration ◽  
Winter Season ◽  
Nuclear Gene ◽  
Restorer Line ◽  
Male Sterile ◽  
Brassica Napus L ◽  
F1 Hybrid ◽  
First Case ◽  
F2 Generation

Identification of male fertility restorer genotypes for rapeseed CMS lines towards hybrid development in spring habit rapeseed (Brassica napus L.) adapted for short day winter season was studied. The experiment was conducted at the experimental farm and laboratory of Bangabandhu Sheikh Mujibur Rahman Agricultural University, Salna, Gazipur during October, 2008 to March, 2011. An exotic CMS-based F1 hybrid of rapeseed was selfed to get F2 generation with a view to resynthesizing restorer line. As a result a restorer line for Nap248A Z1 and Nap248A Z2 cytoplasmic male sterile lines was identified in the F3 generation of the exotic F1 rapeseed hybrid which appears as the first case so far reported as achievement in Bangladesh in this regard. Genetic analysis further revealed fertility restoration for Nap248A Z1 and Nap248A Z2 cytoplasmic male sterility was controlled by a single dominant nuclear gene as a simple genetic phenomenon.Bangladesh J. Agril. Res. 41(3): 529-540, September 2016

Sterility and variation resulting from the transfer of polima cytoplasmic male sterility from Brassica napus into Brassica chinensis

1997 ◽  
Vol 128 (3) ◽  
pp. 299-301 ◽  
Author(s):  
B. HU ◽  
F. CHEN ◽  
Q. LI
Keyword(s):  
Control System ◽  
Brassica Napus ◽  
Interspecific Crosses ◽  
Hybrid Seed Production ◽  
Male Sterile ◽  
Male Sterile Line ◽  
Brassica Chinensis ◽  
Full Flowering ◽  
Cms Line ◽  
Pollination Control

Male sterile combinations made from interspecific crosses between the polima CMS line of Brassica napus and varieties of B. chinensis were backcrossed to BC3. Twenty-six selfing lines from B. chinensis were tested for their ability to either maintain complete sterility or to restore fertility in crosses with the polima male sterile line. Results show that four of these hybrids were completely male sterile and two were fertile. The sterility of the B. chinensis with polima cytoplasm was much more stable than male sterile lines with B. campestris and B. chinensis cytoplasm, which were sterile before full flowering but progressively became fertile as flowering proceeded. The results suggest that polima cytoplasm could be a suitable male sterile-inducing allocytoplasm for B. chinensis, as both maintainers and restorers are available, and could supply a reliable pollination control system for hybrid seed production in this species.

scholarly journals Transcriptome and Hormone Comparison of Three Cytoplasmic Male Sterile Systems in Brassica napus

2018 ◽  
Vol 19 (12) ◽  
pp. 4022 ◽  
Author(s):  
Bingli Ding ◽  
Mengyu Hao ◽  
Desheng Mei ◽  
Qamar Zaman ◽  
Shifei Sang ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Stress Responses ◽  
Developmental Stages ◽  
Plant Mitochondria ◽  
Research Work ◽  
Male Sterile ◽  
Pollen Abortion ◽  
Tetrad Stage ◽  
Cms Line ◽  
Aba Content

The interaction between plant mitochondria and the nucleus markedly influences stress responses and morphological features, including growth and development. An important example of this interaction is cytoplasmic male sterility (CMS), which results in plants producing non-functional pollen. In current research work, we compared the phenotypic differences in floral buds of different Brassica napus CMS (Polima, Ogura, Nsa) lines with their corresponding maintainer lines. By comparing anther developmental stages between CMS and maintainer lines, we identified that in the Nsa CMS line abnormality occurred at the tetrad stage of pollen development. Phytohormone assays demonstrated that IAA content decreased in sterile lines as compared to maintainer lines, while the total hormone content was increased two-fold in the S2 stage compared with the S1 stage. ABA content was higher in the S1 stage and exhibited a two-fold decreasing trend in S2 stage. Sterile lines however, had increased ABA content at both stages compared with the corresponding maintainer lines. Through transcriptome sequencing, we compared differentially expressed unigenes in sterile and maintainer lines at both (S1 and S2) developmental stages. We also explored the co-expressed genes of the three sterile lines in the two stages and classified these genes by gene function. By analyzing transcriptome data and validating by RT-PCR, it was shown that some transcription factors (TFs) and hormone-related genes were weakly or not expressed in the sterile lines. This research work provides preliminary identification of the pollen abortion stage in Nsa CMS line. Our focus on genes specifically expressed in sterile lines may be useful to understand the regulation of CMS.

scholarly journals Integrated cyto-physiological and proteomic analyses reveal new insight into CMS mechanism in a novel upland cotton CMS line LD6A

2022 ◽  
Author(s):  
Zheng Jie ◽  
Aziz Khan ◽  
Zhou Bujin ◽  
Zhou Qiong ◽  
Najeeb Ullah ◽  
...  
Keyword(s):  
Fertility Restoration ◽  
Hybrid Vigor ◽  
Male Sterile ◽  
Maintainer Line ◽  
Tetrad Stage ◽  
Significant Differential Expression ◽  
Mitochondrial Ribosome ◽  
Cms Line ◽  
Analytical Approaches ◽  
Insight Into

Cytoplasmic male sterile (CMS) system has extensively been exploited for hybrid vigor in plant breeding programs. However, its application in many crops is limited due to poor understanding of molecular mechanism of fertility restoration. Using advanced analytical approaches, we elucidated molecular pathways regulating CMS induction and fertility restoration in cotton. Reproductive structures of a novel CMS (LD6A) and its maintainer (LD6B) line were analyzed for physiological and proteomic changes during the development process. Significant differential expression of proteins, such as Abrin, malate dehydrogenase, malic enzyme, isocitrate dehydrogenase, histone acetyltransferase was observed in CMS and its maintainer line. Transmission electron micrographs of anther tapetum showed that inner ridge of CMS mitochondria was relatively indistinct than that of LD6B with narrower membranous space at tetrad stage. Further, relatively higher reactive oxygen species were accumulated in the anther of CMS than its maintainer line at pollen mother cell and tetrad stage. We suggest that abnormal sequence of mitochondrial ribosome gene rps4 and rpl10 and high expression of ribosome-inactivating protein gene Abrin in CMS line damaged mitochondrial membrane and consequently induced pollen sterility. These data provide new insight into CMS mechanism in cotton crops and a tool to develop new CMS germplasm resources.

Meiotic behaviour of extra chromosomes in fertility restored Polima CMS Brassica napus plants

Genome ◽  
1988 ◽  
Vol 30 (3) ◽  
pp. 438-442 ◽  
Author(s):  
William Tai ◽  
Peter B. E. McVetty
Keyword(s):  
Brassica Napus ◽  
Fertility Restoration ◽  
Extra Chromosome ◽  
Pollen Grains ◽  
Backcross Progeny ◽  
Male Sterile ◽  
High Frequencies ◽  
B Genome ◽  
Mother Cells ◽  
Very High Frequencies

Crosses were made between Brassica napus (2n = 38) with the 'Polima' cytoplasmic male sterile system and variety 'Zem' of B. juncea. Fertility was partially restored in backcross progeny with an extra chromosome (2n = 39) that was believed to be a member of the B genome of B. juncea. Among more than 40 self-pollinated offspring plants studied, fertility restoration was transmitted only through those plants with two extra chromosomes (2n = 40). Anthers of these plants were either full or shriveled with small swollen protrusions. Full and round pollen grains were found in full anthers and in the swollen portion of shriveled anthers. These pollen grains were stained darkly with I2-KI solution and full seed set was obtained from bagged flowers. Meiotic studies in plants with 2n = 40 showed 18 bivalents + 1 quadrivalent at diakinesis in most of the pollen mother cells analyzed. The chromosomes segregated equally at anaphase I and behaved normally at other meiotic stages. The presence of a single quadrivalent in the backcross progeny was highly consistent and occurred in very high frequencies. It is believed that the extra chromosomes belong to the B genome of B. juncea and take part in the quadrivalent formation.Key words: Brassica napus, cytoplasmic male sterility, aneuploidy, Polima, Brassica juncea, rapeseed cytogenetics.

scholarly journals A single nucleotide polymorphism in an R2R3 MYB transcription factor gene triggers the male sterility in soybean ms6 (Ames1)

2021 ◽  
Author(s):  
Junping Yu ◽  
Guolong Zhao ◽  
Wei Li ◽  
Ying Zhang ◽  
Peng Wang ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Glycine Max ◽  
Male Sterility ◽  
Bulked Segregant Analysis ◽  
Soybean Protein ◽  
Anther Development ◽  
Hybrid Breeding ◽  
Myb Transcription Factor ◽  
Male Sterile ◽  
R2r3 Myb

Abstract Key message Identification and functional analysis of the male sterile gene MS6 in Glycine max. Abstract Soybean (Glycine max (L.) Merr.) is an important crop providing vegetable oil and protein. The male sterility-based hybrid breeding is a promising method for improving soybean yield to meet the globally growing demand. In this research, we identified a soybean genic male sterile locus, MS6, by combining the bulked segregant analysis sequencing method and the map-based cloning technology. MS6, highly expressed in anther, encodes an R2R3 MYB transcription factor (GmTDF1-1) that is homologous to Tapetal Development and Function 1, a key factor for anther development in Arabidopsis and rice. In male sterile ms6 (Ames1), the mutant allele contains a missense mutation, leading to the 76th leucine substituted by histidine in the DNA binding domain of GmTDF1-1. The expression of soybean MS6 under the control of the AtTDF1 promoter could rescue the male sterility of attdf1 but ms6 could not. Additionally, ms6 overexpression in wild-type Arabidopsis did not affect anther development. These results evidence that GmTDF1-1 is a functional TDF1 homolog and L76H disrupts its function. Notably, GmTDF1-1 shows 92% sequence identity with another soybean protein termed as GmTDF1-2, whose active expression also restored the fertility of attdf1. However, GmTDF1-2 is constitutively expressed at a very low level in soybean, and therefore, not able to compensate for the MS6 deficiency. Analysis of the TDF1-involved anther development regulatory pathway showed that expressions of the genes downstream of TDF1 are significantly suppressed in ms6, unveiling that GmTDF1-1 is a core transcription factor regulating soybean anther development.

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