scholarly journals A Semi-Quantitative Bulked Segregant Analysis Preliminarily Localizes a Maize Male-Sterility Gene

2021 ◽  
Vol 25 (03) ◽  
pp. 659-666
Author(s):  
Mingbo Wu
Keyword(s):  
Male Sterility ◽  
Target Gene ◽  
Dna Analysis ◽  
High Efficiency ◽  
Bulked Segregant Analysis ◽  
Mutant Gene ◽  
Polymorphic Markers ◽  
Chromosome 5 ◽  
Mutant Genes ◽  
Male Sterility Gene

Bulked segregant analysis (BSA) assists in map-based cloning of mutant genes. However, a traditional BSA requires many high-density markers for successful linkage analysis which is labor-intensive and time-consuming. In this study, a semi-quantitative DNA analysis program was optimized and combined it with BSA, resulting in a semi-quantitative BSA (sq-BSA). The sq-BSA approach allowed evaluation of the proportions of marker-defined individuals (dominant or recessive marker types) in bulks. The sq-BSA method was used to map a male-sterility (ms) gene, ms2016, in maize. Forty polymorphic markers were screened from one-third of each chromosome (from the head or tail) for mapping. Among these markers, seven were identified as candidate gene-linked markers, of which four markers (bnlg1046, umc1563, umc1171 and umc1722) were located on chromosome 5. Using group validation, ms2016 was anchored on chromosome 5 and was most closely linked to bnlg1046. Furthermore, four new InDel markers located near bnlg1046 were screened to map the preliminary location of ms2016. The ms2016 gene was mapped to an 8.7 Mb interval flanked by the InDel polymorphic markers I5-3 (chr5:14588060) and I5-12 (chr5:23308445). Thus, this improved BSA method (sq-BSA) requires only a small number of molecular markers to quickly localize a target gene, representing a high-efficiency tool for mutant gene mapping. © 2021 Friends Science Publishers

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.

Molecular mapping of a thermosensitive genetic male sterility gene in rice using bulked segregant analysis

Genome ◽  
1997 ◽  
Vol 40 (2) ◽  
pp. 188-194 ◽  
Author(s):  
P. K. Subudhi ◽  
R. P. Borkakati ◽  
S. S. Virmani ◽  
N. Huang
Keyword(s):  
Male Sterility ◽  
Molecular Mapping ◽  
Bulked Segregant Analysis ◽  
Pcr Primers ◽  
Rflp Markers ◽  
Arbitrary Primers ◽  
Repulsion Phase ◽  
Genetic Male Sterility ◽  
True Breeding ◽  
Efficient Alternative

The thermosensitive genetic male sterility (TGMS) system is considered to be a more efficient alternative to the cytoplasmic male sterility (CMS) system for hybrid rice. An F2 population from a cross between a TGMS mutant line (IR32364TGMS) and IR68 was used to map the TGMS gene tms3(t). Fertile and sterile bulks were constructed following the classification of F2 plants into true breeding sterile, fertile, and segregating fertile plants based on F3 family studies. From the survey of 389 arbitrary primers in bulked segregant analysis, four RAPD markers were identified in which three, OPF182600, OPB19750, and OPAA7550, were linked to tms3(t) in repulsion phase and one, OPAC3640, was linked to tms3(t) in coupling phase. The tms3(t) gene was flanked by OPF182600 and OPAC3640 on one side and by OPAA7550 and OPB19750 on the other side. All four markers were low-copy sequences and two of them (OPF182600 and OPAC3640) detected polymorphism when the markers were used to probe the genomic blots. Subsequently, OPAC3640 was mapped to the short arm of chromosome 6 using a mapping population available at IRRI. However, no RFLP markers from this region showed linkage to tms3(t) owing to the lack of polymorphism between the parents. All RAPD fragments were cloned and partially sequenced from both ends. Thus, PCR primers can be designed to develop PCR markers for marker-assisted breeding to facilitate the transfer of tms3(t) from one genetic background to another.Key words: bulked segregant analysis, gene tagging, marker-assisted selection, RAPD, TGMS.

scholarly journals Quinolone-resistant mutants of escherichia coli DNA topoisomerase IV parC gene.

1996 ◽  
Vol 40 (3) ◽  
pp. 710-714 ◽  
Author(s):  
Y Kumagai ◽  
J I Kato ◽  
K Hoshino ◽  
T Akasaka ◽  
K Sato ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Dna Gyrase ◽  
Mutant Gene ◽  
Missense Mutations ◽  
Topoisomerase Iv ◽  
Dna Topoisomerase ◽  
E Coli ◽  
A Subunit ◽  
Resistant Strains ◽  
Mutant Genes

Escherichia coli quinolone-resistant strains with mutations of the parC gene, which codes for a subunit of topoisomerase IV, were isolated from a quinolone-resistant gyrA mutant of DNA gyrase. Quinolone-resistant parC mutants were also identified among the quinolone-resistant clinical strains. The parC mutants became susceptible to quinolones by introduction of a parC+ plasmid. Introduction of the multicopy plasmids carrying the quinolone-resistant parC mutant gene resulted in an increase in MICs of quinolones for the parC+ and quinolone-resistant gyrA strain. Nucleotide sequences of the quinolone-resistant parC mutant genes were determined, and missense mutations at position Gly-78, Ser-80, or Glu-84, corresponding to those in the quinolone-resistance-determining region of DNA gyrase, were identified. These results indicate that topoisomerase IV is a target of quinolones in E. coli and suggest that the susceptibility of E. coli cells to quinolones is determined by sensitivity of the targets, DNA gyrase and topoisomerase IV.

scholarly journals Inactivation of the Neurospora crassa gene encoding the mitochondrial protein import receptor MOM19 by the technique of "sheltered RIP".

Genetics ◽  
1994 ◽  
Vol 136 (1) ◽  
pp. 107-118 ◽  
Author(s):  
T A Harkness ◽  
R L Metzenberg ◽  
H Schneider ◽  
R Lill ◽  
W Neupert ◽  
...  
Keyword(s):  
Neurospora Crassa ◽  
Target Gene ◽  
Protein Import ◽  
Selectable Marker ◽  
Mitochondrial Protein ◽  
Mutant Gene ◽  
Mitochondrial Protein Import ◽  
Gene Encoding ◽  
Precursor Proteins ◽  
Import Receptor

Abstract We have used a technique referred to as "sheltered RIP" (repeat induced point mutation) to create mutants of the mom-19 gene of Neurospora crassa, which encodes an import receptor for nuclear encoded mitochondrial precursor proteins. Sheltered RIP permits the isolation of a mutant gene in one nucleus, even if that gene is essential for the survival of the organism, by sheltering the nucleus carrying the mutant gene in a heterokaryon with an unaffected nucleus. Furthermore, the nucleus harboring the RIPed gene contains a selectable marker so that it is possible to shift nuclear ratios in the heterokaryons to a state in which the nucleus containing the RIPed gene predominates in cultures grown under selective conditions. This results in a condition where the target gene product should be present at very suboptimal levels and allows the study of the mutant phenotype. One allele of mom-19 generated by this method contains 44 transitions resulting in 18 amino acid substitutions. When the heterokaryon containing this allele was grown under conditions favoring the RIPed nucleus, no MOM19 protein was detectable in the mitochondria of the strain. Homokaryotic strains containing the RIPed allele exhibit a complex and extremely slow growth phenotype suggesting that the product of the mom-19 gene is important in N. crassa.

Fine mapping and expression analysis of thermosensitive genic male sterility gene (tms) in rice (Oryza sativa L.)

Plant Gene ◽  
2019 ◽  
Vol 19 ◽  
pp. 100186
Author(s):  
Sai Rekha Kadirimangalam ◽  
Rahman Hifzur ◽  
Saraswathi R ◽  
Kumar M ◽  
Raveendran M ◽  
...  
Keyword(s):  
Oryza Sativa ◽  
Male Sterility ◽  
Expression Analysis ◽  
Fine Mapping ◽  
Oryza Sativa L ◽  
Genic Male Sterility ◽  
Sterility Gene ◽  
Male Sterility Gene

scholarly journals Genetic Analysis and Fine Mapping of a Spontaneously Mutated Male Sterility Gene in Brassica rapa ssp. chinensis

2020 ◽  
Vol 10 (4) ◽  
pp. 1309-1318
Author(s):  
Tzu-Kai Lin ◽  
Ya-Ping Lin ◽  
Shun-Fu Lin
Keyword(s):  
Male Sterility ◽  
Fine Mapping ◽  
Genotyping By Sequencing ◽  
Snp Markers ◽  
Hybrid Seed Production ◽  
Spontaneous Mutant ◽  
Male Sterile ◽  
Sterility Gene ◽  
Intron 4 ◽  
Male Sterility Gene

Male sterility has been widely used in hybrid seed production in Brassica, but not in B. rapa ssp. chinensis, and genetic models of male sterility for this subspecies are unclear. We discovered a spontaneous mutant in B. rapa ssp. chinensis. A series of progeny tests indicated that male sterility in B. rapa ssp. chinensis follows a three-allele model with BrMsa, BrMsb, and BrMsc. The male sterility locus has been mapped to chromosome A07 in BC1 and F2 populations through genotyping by sequencing. Fine mapping in a total of 1,590 F2 plants narrowed the male sterility gene BrMs to a 400 kb region, with two SNP markers only 0.3 cM from the gene. Comparative gene mapping shows that the Ms gene in B. rapa ssp. pekinensis is different from the BrMs gene of B. rapa ssp. chinensis, despite that both genes are located on chromosome A07. Interestingly, the DNA sequence orthologous to a male sterile gene in Brassica napus, BnRf, is within 400 kb of the BrMs locus. The BnRf orthologs of B. rapa ssp. chinensis were sequenced, and one KASP marker (BrMs_indel) was developed for genotyping based on a 14 bp indel at intron 4. Cosegregation of male sterility and BrMs_indel genotypes in the F2 population indicated that BnRf from B. napus and BrMs from B. rapa are likely to be orthologs. The BrMs_indel marker developed in this study will be useful in marker-assisted selection for the male sterility trait.

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