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Identification of Genes Potentially Associated with the Fertility Instability of S-Type Cytoplasmic Male Sterility in Maize via Bulked Segregant RNA-Seq

View Article: PubMed Central - PubMed

ABSTRACT

S-type cytoplasmic male sterility (CMS-S) is the largest group among the three major types of CMS in maize. CMS-S exhibits fertility instability as a partial fertility restoration in a specific nuclear genetic background, which impedes its commercial application in hybrid breeding programs. The fertility instability phenomenon of CMS-S is controlled by several minor quantitative trait locus (QTLs), but not the major nuclear fertility restorer (Rf3). However, the gene mapping of these minor QTLs and the molecular mechanism of the genetic modifications are still unclear. Using completely sterile and partially rescued plants of fertility instable line (FIL)-B, we performed bulk segregant RNA-Seq and identified six potential associated genes in minor effect QTLs contributing to fertility instability. Analyses demonstrate that these potential associated genes may be involved in biological processes, such as floral organ differentiation and development regulation, energy metabolism and carbohydrates biosynthesis, which results in a partial anther exsertion and pollen fertility restoration in the partially rescued plants. The single nucleotide polymorphisms (SNPs) identified in two potential associated genes were validated to be related to the fertility restoration phenotype by KASP marker assays. This novel knowledge contributes to the understanding of the molecular mechanism of the partial fertility restoration of CMS-S in maize and thus helps to guide the breeding programs.

No MeSH data available.


Related in: MedlinePlus

Comparison of the transcript levels of the six potential associated genes from the partially rescued (F) and sterile (S) plants of FIL-B, as detected by QPCR.Each bar represents the mean±SE of the biological replicates. The values are calculated using Actin as an internal control. The asterisks show the statistically significant difference compared to the partially rescued plants, as determined by the analysis of variance: * (P<0.05), **(P<0.01), ***(P<0.001).
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pone.0163489.g005: Comparison of the transcript levels of the six potential associated genes from the partially rescued (F) and sterile (S) plants of FIL-B, as detected by QPCR.Each bar represents the mean±SE of the biological replicates. The values are calculated using Actin as an internal control. The asterisks show the statistically significant difference compared to the partially rescued plants, as determined by the analysis of variance: * (P<0.05), **(P<0.01), ***(P<0.001).

Mentions: QPCR was performed for the six genes using individual plants to validate the RNA-Seq data and to obtain more quantitative transcript level measurements. The relative expression of all of the six potential associated genes showed a 4- to 8-fold up regulation in the partially rescued plants compared to the sterile plants (Fig 5). The trends in regulation exhibited a consistent pattern between methods and thus, confirming the qualitative values of the RNA-Seq.


Identification of Genes Potentially Associated with the Fertility Instability of S-Type Cytoplasmic Male Sterility in Maize via Bulked Segregant RNA-Seq
Comparison of the transcript levels of the six potential associated genes from the partially rescued (F) and sterile (S) plants of FIL-B, as detected by QPCR.Each bar represents the mean±SE of the biological replicates. The values are calculated using Actin as an internal control. The asterisks show the statistically significant difference compared to the partially rescued plants, as determined by the analysis of variance: * (P<0.05), **(P<0.01), ***(P<0.001).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5036866&req=5

pone.0163489.g005: Comparison of the transcript levels of the six potential associated genes from the partially rescued (F) and sterile (S) plants of FIL-B, as detected by QPCR.Each bar represents the mean±SE of the biological replicates. The values are calculated using Actin as an internal control. The asterisks show the statistically significant difference compared to the partially rescued plants, as determined by the analysis of variance: * (P<0.05), **(P<0.01), ***(P<0.001).
Mentions: QPCR was performed for the six genes using individual plants to validate the RNA-Seq data and to obtain more quantitative transcript level measurements. The relative expression of all of the six potential associated genes showed a 4- to 8-fold up regulation in the partially rescued plants compared to the sterile plants (Fig 5). The trends in regulation exhibited a consistent pattern between methods and thus, confirming the qualitative values of the RNA-Seq.

View Article: PubMed Central - PubMed

ABSTRACT

S-type cytoplasmic male sterility (CMS-S) is the largest group among the three major types of CMS in maize. CMS-S exhibits fertility instability as a partial fertility restoration in a specific nuclear genetic background, which impedes its commercial application in hybrid breeding programs. The fertility instability phenomenon of CMS-S is controlled by several minor quantitative trait locus (QTLs), but not the major nuclear fertility restorer (Rf3). However, the gene mapping of these minor QTLs and the molecular mechanism of the genetic modifications are still unclear. Using completely sterile and partially rescued plants of fertility instable line (FIL)-B, we performed bulk segregant RNA-Seq and identified six potential associated genes in minor effect QTLs contributing to fertility instability. Analyses demonstrate that these potential associated genes may be involved in biological processes, such as floral organ differentiation and development regulation, energy metabolism and carbohydrates biosynthesis, which results in a partial anther exsertion and pollen fertility restoration in the partially rescued plants. The single nucleotide polymorphisms (SNPs) identified in two potential associated genes were validated to be related to the fertility restoration phenotype by KASP marker assays. This novel knowledge contributes to the understanding of the molecular mechanism of the partial fertility restoration of CMS-S in maize and thus helps to guide the breeding programs.

No MeSH data available.


Related in: MedlinePlus