Limits...
Comparative transcriptomic analysis of male and female flowers of monoecious Quercus suber.

Rocheta M, Sobral R, Magalhães J, Amorim MI, Ribeiro T, Pinheiro M, Egas C, Morais-Cecílio L, Costa MM - Front Plant Sci (2014)

Bottom Line: The assembly of the reads resulted in 14,488 contigs for female libraries and 10,438 contigs for male libraries.Moreover, we found differentially expressed genes that have not yet been characterized and others that have not been previously shown to be implicated in flower development.This transcriptomic analysis constitutes a major step toward the characterization of the molecular mechanisms involved in flower development in a monoecious tree with a potential contribution toward the knowledge of conserved developmental mechanisms in other species.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Recursos Naturais Ambiente e Território, Instituto Superior de Agronomia, Universidade de Lisboa Lisboa, Portugal.

ABSTRACT
Monoecious species provide a comprehensive system to study the developmental programs underlying the establishment of female and male organs in unisexual flowers. However, molecular resources for most monoecious non-model species are limited, hampering our ability to study the molecular mechanisms involved in flower development of these species. The objective of this study was to identify differentially expressed genes during the development of male and female flowers of the monoecious species Quercus suber, an economically important Mediterranean tree. Total RNA was extracted from different developmental stages of Q. suber flowers. Non-normalized cDNA libraries of male and female flowers were generated using 454 pyrosequencing technology producing a total of 962,172 high-quality reads with an average length of 264 nucleotides. The assembly of the reads resulted in 14,488 contigs for female libraries and 10,438 contigs for male libraries. Comparative analysis of the transcriptomes revealed genes differentially expressed in early and late stages of development of female and male flowers, some of which have been shown to be involved in pollen development, in ovule formation and in flower development of other species with a monoecious, dioecious, or hermaphroditic sexual system. Moreover, we found differentially expressed genes that have not yet been characterized and others that have not been previously shown to be implicated in flower development. This transcriptomic analysis constitutes a major step toward the characterization of the molecular mechanisms involved in flower development in a monoecious tree with a potential contribution toward the knowledge of conserved developmental mechanisms in other species.

No MeSH data available.


Relative expression of differentially expressed male and female genes chosen to validate RNAseq results. QsAMS, QsLAP3, QsLAP5, QsLAP6 were selected as male candidate genes, whereas QsAt4g27290, QsCYP78A9, QsPG1, and QsSTIG1 as female candidate genes. Transcript abundance was determined using qPCR, and normalized to QsPP2AA3 using cDNA synthesized from distinct pools of RNA covering either early (1F and 1M) or late (2F and 2M) stages of male or female flower development. Reactions were performed in three biological and technical replicates. Error bars indicate standard deviation (SD).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4222140&req=5

Figure 3: Relative expression of differentially expressed male and female genes chosen to validate RNAseq results. QsAMS, QsLAP3, QsLAP5, QsLAP6 were selected as male candidate genes, whereas QsAt4g27290, QsCYP78A9, QsPG1, and QsSTIG1 as female candidate genes. Transcript abundance was determined using qPCR, and normalized to QsPP2AA3 using cDNA synthesized from distinct pools of RNA covering either early (1F and 1M) or late (2F and 2M) stages of male or female flower development. Reactions were performed in three biological and technical replicates. Error bars indicate standard deviation (SD).

Mentions: As expected, the Q. suber homologous genes presented differential expression ratios between male and female libraries (Table 2), and thus were considerate good candidates for qRT-PCR analysis. The qRT-PCR results confirmed that genes involved in pollen exine formation (LAP3, LAP5, and LAP6) and in the tapetum cell development (AMS) were more expressed in the early stages of male flower development (Figure 3), whereas genes involved in stigma-specific recognition (STIG1), in the recognition of pollen (At4g27290) and in fruit growth and development (CYP78A9, PG1) were more expressed in the female flowers. These results were in close agreement with the RNAseq data (Table 2) suggesting the reliability of the transcriptomic profiling data.


Comparative transcriptomic analysis of male and female flowers of monoecious Quercus suber.

Rocheta M, Sobral R, Magalhães J, Amorim MI, Ribeiro T, Pinheiro M, Egas C, Morais-Cecílio L, Costa MM - Front Plant Sci (2014)

Relative expression of differentially expressed male and female genes chosen to validate RNAseq results. QsAMS, QsLAP3, QsLAP5, QsLAP6 were selected as male candidate genes, whereas QsAt4g27290, QsCYP78A9, QsPG1, and QsSTIG1 as female candidate genes. Transcript abundance was determined using qPCR, and normalized to QsPP2AA3 using cDNA synthesized from distinct pools of RNA covering either early (1F and 1M) or late (2F and 2M) stages of male or female flower development. Reactions were performed in three biological and technical replicates. Error bars indicate standard deviation (SD).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4222140&req=5

Figure 3: Relative expression of differentially expressed male and female genes chosen to validate RNAseq results. QsAMS, QsLAP3, QsLAP5, QsLAP6 were selected as male candidate genes, whereas QsAt4g27290, QsCYP78A9, QsPG1, and QsSTIG1 as female candidate genes. Transcript abundance was determined using qPCR, and normalized to QsPP2AA3 using cDNA synthesized from distinct pools of RNA covering either early (1F and 1M) or late (2F and 2M) stages of male or female flower development. Reactions were performed in three biological and technical replicates. Error bars indicate standard deviation (SD).
Mentions: As expected, the Q. suber homologous genes presented differential expression ratios between male and female libraries (Table 2), and thus were considerate good candidates for qRT-PCR analysis. The qRT-PCR results confirmed that genes involved in pollen exine formation (LAP3, LAP5, and LAP6) and in the tapetum cell development (AMS) were more expressed in the early stages of male flower development (Figure 3), whereas genes involved in stigma-specific recognition (STIG1), in the recognition of pollen (At4g27290) and in fruit growth and development (CYP78A9, PG1) were more expressed in the female flowers. These results were in close agreement with the RNAseq data (Table 2) suggesting the reliability of the transcriptomic profiling data.

Bottom Line: The assembly of the reads resulted in 14,488 contigs for female libraries and 10,438 contigs for male libraries.Moreover, we found differentially expressed genes that have not yet been characterized and others that have not been previously shown to be implicated in flower development.This transcriptomic analysis constitutes a major step toward the characterization of the molecular mechanisms involved in flower development in a monoecious tree with a potential contribution toward the knowledge of conserved developmental mechanisms in other species.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Recursos Naturais Ambiente e Território, Instituto Superior de Agronomia, Universidade de Lisboa Lisboa, Portugal.

ABSTRACT
Monoecious species provide a comprehensive system to study the developmental programs underlying the establishment of female and male organs in unisexual flowers. However, molecular resources for most monoecious non-model species are limited, hampering our ability to study the molecular mechanisms involved in flower development of these species. The objective of this study was to identify differentially expressed genes during the development of male and female flowers of the monoecious species Quercus suber, an economically important Mediterranean tree. Total RNA was extracted from different developmental stages of Q. suber flowers. Non-normalized cDNA libraries of male and female flowers were generated using 454 pyrosequencing technology producing a total of 962,172 high-quality reads with an average length of 264 nucleotides. The assembly of the reads resulted in 14,488 contigs for female libraries and 10,438 contigs for male libraries. Comparative analysis of the transcriptomes revealed genes differentially expressed in early and late stages of development of female and male flowers, some of which have been shown to be involved in pollen development, in ovule formation and in flower development of other species with a monoecious, dioecious, or hermaphroditic sexual system. Moreover, we found differentially expressed genes that have not yet been characterized and others that have not been previously shown to be implicated in flower development. This transcriptomic analysis constitutes a major step toward the characterization of the molecular mechanisms involved in flower development in a monoecious tree with a potential contribution toward the knowledge of conserved developmental mechanisms in other species.

No MeSH data available.