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.

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 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 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.

scholarly journals Genetics of fertility restoration of the A4 cytoplasmic-nuclear male sterility system in pearl millet

2012 ◽  
Vol 48 (No. 2) ◽  
pp. 87-92 ◽  
Author(s):  
S.K. Gupta ◽  
K.N. Rai ◽  
M. Govindaraj ◽  
A.S. Rao
Keyword(s):  
Male Sterility ◽  
Pearl Millet ◽  
Fertility Restoration ◽  
Single Gene ◽  
Segregation Pattern ◽  
Gene Control ◽  
Male Sterile ◽  
Nuclear Male Sterility ◽  
Line Breeding ◽  
Breeding Efficiency

Inheritance of fertility restoration of the A<sub>4</sub> system of cytoplasmic-nuclear male sterility in pearl millet was investigated using six crosses between two diverse male sterile lines (A-lines) and three diverse restorers (R-lines). The segregation pattern of male sterile (S) and male fertile (F) plants observed in F<sub>2</sub>, and BC<sub>1 </sub>in two seasons at ICRISAT, Patancheru, indicated the dominant single-gene control of male fertility restoration. The segregation pattern in BC<sub>1</sub>F<sub>2</sub> progenies derived from the fertile BC<sub>1</sub> plants evaluated for one season provided further evidence for the single-gene control. The season did not have much effect on fertility restoration. The information on the single-gene control of fertility restoration will help in diversifying the restorer genetic base of the A<sub>4&nbsp;</sub>CMS system and enhance R-line breeding efficiency in pearl millet.

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 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.

scholarly journals The Dynamics of Gynodioecy in Plantago lanceolatu L. II. Mode of Action and Frequencies of Restorer Alleles

Genetics ◽  
1997 ◽  
Vol 147 (3) ◽  
pp. 1317-1328
Author(s):  
Anita A de Haan ◽  
Hans P Koelewijn ◽  
Maria P J Hundscheid ◽  
Jos M M Van Damme
Keyword(s):  
Cytoplasmic Male Sterility ◽  
Male Sterility ◽  
Allele Frequency ◽  
Mode Of Action ◽  
Male Fertility ◽  
Fertility Restoration ◽  
Plantago Lanceolata ◽  
Allele Frequencies ◽  
Male Sterile ◽  
Male Fertility Restoration

Male fertility in Plantago lanceolata is controlled by the interaction of cytoplasmic and nuclear genes. Different cytoplasmic male sterility (CMS) types can be either male sterile or hermaphrodite, depending on the presence of nuclear restorer alleles. In three CMS types of P. lanceolata (CMSI, CMSIIa, and CMSIIb) the number of loci involved in male fertility restoration was determined. In each CMS type, male fertility was restored by multiple genes with either dominant or recessive action and capable either of restoring male fertility independently or in interaction with each other (epistasis). Restorer allele frequencies for CMSI, CMSIIa and CMSIIb were determined by crossing hermaphrodites with “standard” male steriles. Segregation of male steriles vs. non-male steriles was used to estimate overall restorer allele frequency. The frequency of restorer alleles was different for the CMS types: restorer alleles for CMSI were less frequent than for CMSIIa and CMSIIb. On the basis of the frequencies of male steriles and the CMS types an “expected” restorer allele frequency could be calculated. The correlation between estimated and expected restorer allele frequency was significant.

scholarly journals INHERITANCE OF FERTILITY RESTORATION INVOLVING WILD ABORTIVE CYTOPLASMIC MALE STERILITY SYSTEM IN RICE (Oryza sativa L.)

2011 ◽  
Vol 24 (1) ◽  
pp. 33-40
Author(s):  
M. J. Hasan ◽  
M. U. Kulsum ◽  
A. Ansari ◽  
A. K. Paul ◽  
P. L. Biswas
Keyword(s):  
Oryza Sativa ◽  
Cytoplasmic Male Sterility ◽  
Male Sterility ◽  
Gene Action ◽  
Oryza Sativa L ◽  
Fertility Restoration ◽  
Dominant Gene ◽  
Recessive Gene ◽  
Male Sterile ◽  
Male Sterile Line

Inheritance of fertility restoration was studied in crosses involving ten elite restorer lines of rice viz. BR6839-41-5-1R, BR7013-62-1-1R, BR7011-37-1-2R, BR10R, BR11R, BR12R, BR13R, BR14R, BR15R and BR16R and one male sterile line Jin23A with WA sources of cytoplasmic male sterility. The segregation pattern for pollen fertility of F2 and BC1 populations of crosses involving Jin23A indicated the presence of two independent dominant fertility restoring genes. The mode of action of the two genes varied in different crosses revealing three types of interaction, i.e. epistasis with dominant gene action, epistasis with recessive gene action, and epistasis with incomplete dominance.DOI: http://dx.doi.org/10.3329/bjpbg.v24i1.16997

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