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Transcriptomic analysis of developmental features of Bombyx mori wing disc during metamorphosis.

Ou J, Deng HM, Zheng SC, Huang LH, Feng QL, Liu L - BMC Genomics (2014)

Bottom Line: Seventeen transcription factors were significantly up-regulated.Genes responsible for the degradation and de novo synthesis of chitin were significantly up-regulated.These results implied that during the larva-to-pupa metamorphosis, pupal wing development and transition might be mainly controlled by 20E signaling in B. mori.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular and Developmental Entomology, Guangdong Provincial Key Lab of Biotechnology for Plant Development, School of Life Sciences, South China Normal University, Guangzhou 510631, China. qlfeng@scnu.edu.cn.

ABSTRACT

Background: Wing discs of B. mori are transformed to pupal wings during the larva-to-pupa metamorphosis with dramatic morphological and structural changes. To understand these changes at a transcriptional level, RNA-seq of the wing discs from 6-day-old fifth instar larvae (L5D6), prepupae (PP) and pupae (P0) was performed.

Results: In total, 12,254 transcripts were obtained from the wing disc, out of which 5,287 were identified to be differentially expressed from L5D6 to PP and from PP to P0. The results of comprehensive analysis of RNA-seq data showed that during larvae-to-pupae metamorphosis, many genes of 20E signaling pathway were up-regulated and those of JH signaling pathway were down-regulated. Seventeen transcription factors were significantly up-regulated. Cuticle protein genes (especially wing cuticle protein genes), were most abundant and significantly up-regulated at P0 stage. Genes responsible for the degradation and de novo synthesis of chitin were significantly up-regulated. There were A and B two types of chitin synthases in B. mori, whereas only chitin synthase A was up-regulated. Both trehalose and D-fructose, which are precursors of chitin synthesis, were detected in the hemolymph of L5D6, PP and P0, suggesting de novo synthesis of chitin. However, most of the genes that are related to early wing disc differentiation were down-regulated.

Conclusions: Extensive transcriptome and DGE profiling data of wing disc during metamorphosis of silkworm have been generated, which provided comprehensive gene expression information at the transcriptional level. These results implied that during the larva-to-pupa metamorphosis, pupal wing development and transition might be mainly controlled by 20E signaling in B. mori. The 17 up-regulated transcription factors might be involved in wing development. Chitin required for pupal wing development might be generated from both degradation of componential chitin and de novo synthesis. Chitin synthase A might be responsible for the chitin synthesis in the pupal wing, while both trehalose and D-fructose might contribute to the de novo synthesis of chitin during the formation of pupal wing.

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Expression analysis of cuticular or cuticle protein coding genes. The RPKM values of undetected genes are viewed as 0.001. Cluster 3.0 software was used to plot the heat map (Similarity Metric: Euclidean distance, Clustering method: Complete linkage), and Treeview software was used to generate the heat map. The colors in the map display the relative values at the given 3 developmental stages. Green indicates the lowest level of expression, black indicates the intermediate level of expression, and red indicates the highest level of expression. (A) Distribution of cuticular or cuticle protein coding genes at the three stages. (B) Heat map of hierarchical clustering of 148 differentially expressed cuticle protein genes.
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Fig5: Expression analysis of cuticular or cuticle protein coding genes. The RPKM values of undetected genes are viewed as 0.001. Cluster 3.0 software was used to plot the heat map (Similarity Metric: Euclidean distance, Clustering method: Complete linkage), and Treeview software was used to generate the heat map. The colors in the map display the relative values at the given 3 developmental stages. Green indicates the lowest level of expression, black indicates the intermediate level of expression, and red indicates the highest level of expression. (A) Distribution of cuticular or cuticle protein coding genes at the three stages. (B) Heat map of hierarchical clustering of 148 differentially expressed cuticle protein genes.

Mentions: Cuticle proteins are important component of wing disc in the post-larva development. To study the relationship between expression of cuticle proteins and wing development, cuticle protein genes were analyzed. Totally 205 cuticle protein transcripts were identified using Blast search. Ninety-seven transcripts were expressed at all the three stages, while 7, 9 and 58 transcripts were specifically expressed at L5D6, PP and P0 stage respectively (Figure 5A). One hundred forty eight out of 205 transcripts were differentially expressed, with most of them being up-regulated during the larva-to-pupa metamorphosis, particularly from PP to P0 stage (Figure 5B). This result was consistent with the results by microarray analysis[36]. By similarity analysis of sequences, 10 WCPs genes were identified in our RNA-Seq dataset. Expression of WCP1a, WCP1b, WCP3, WCP4, WCP5, WCP8 and WCP9 was slightly up-regulated at the PP stage and then greatly increased at the P0 stage. Expression of WCP10 was slightly up-regulated at the PP stage and then greatly decreased at the P0 stage, whereas the expression of WCP11 was decreased at the PP stage and then greatly increased at the P0 stage (Table 4).Figure 5


Transcriptomic analysis of developmental features of Bombyx mori wing disc during metamorphosis.

Ou J, Deng HM, Zheng SC, Huang LH, Feng QL, Liu L - BMC Genomics (2014)

Expression analysis of cuticular or cuticle protein coding genes. The RPKM values of undetected genes are viewed as 0.001. Cluster 3.0 software was used to plot the heat map (Similarity Metric: Euclidean distance, Clustering method: Complete linkage), and Treeview software was used to generate the heat map. The colors in the map display the relative values at the given 3 developmental stages. Green indicates the lowest level of expression, black indicates the intermediate level of expression, and red indicates the highest level of expression. (A) Distribution of cuticular or cuticle protein coding genes at the three stages. (B) Heat map of hierarchical clustering of 148 differentially expressed cuticle protein genes.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4196006&req=5

Fig5: Expression analysis of cuticular or cuticle protein coding genes. The RPKM values of undetected genes are viewed as 0.001. Cluster 3.0 software was used to plot the heat map (Similarity Metric: Euclidean distance, Clustering method: Complete linkage), and Treeview software was used to generate the heat map. The colors in the map display the relative values at the given 3 developmental stages. Green indicates the lowest level of expression, black indicates the intermediate level of expression, and red indicates the highest level of expression. (A) Distribution of cuticular or cuticle protein coding genes at the three stages. (B) Heat map of hierarchical clustering of 148 differentially expressed cuticle protein genes.
Mentions: Cuticle proteins are important component of wing disc in the post-larva development. To study the relationship between expression of cuticle proteins and wing development, cuticle protein genes were analyzed. Totally 205 cuticle protein transcripts were identified using Blast search. Ninety-seven transcripts were expressed at all the three stages, while 7, 9 and 58 transcripts were specifically expressed at L5D6, PP and P0 stage respectively (Figure 5A). One hundred forty eight out of 205 transcripts were differentially expressed, with most of them being up-regulated during the larva-to-pupa metamorphosis, particularly from PP to P0 stage (Figure 5B). This result was consistent with the results by microarray analysis[36]. By similarity analysis of sequences, 10 WCPs genes were identified in our RNA-Seq dataset. Expression of WCP1a, WCP1b, WCP3, WCP4, WCP5, WCP8 and WCP9 was slightly up-regulated at the PP stage and then greatly increased at the P0 stage. Expression of WCP10 was slightly up-regulated at the PP stage and then greatly decreased at the P0 stage, whereas the expression of WCP11 was decreased at the PP stage and then greatly increased at the P0 stage (Table 4).Figure 5

Bottom Line: Seventeen transcription factors were significantly up-regulated.Genes responsible for the degradation and de novo synthesis of chitin were significantly up-regulated.These results implied that during the larva-to-pupa metamorphosis, pupal wing development and transition might be mainly controlled by 20E signaling in B. mori.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular and Developmental Entomology, Guangdong Provincial Key Lab of Biotechnology for Plant Development, School of Life Sciences, South China Normal University, Guangzhou 510631, China. qlfeng@scnu.edu.cn.

ABSTRACT

Background: Wing discs of B. mori are transformed to pupal wings during the larva-to-pupa metamorphosis with dramatic morphological and structural changes. To understand these changes at a transcriptional level, RNA-seq of the wing discs from 6-day-old fifth instar larvae (L5D6), prepupae (PP) and pupae (P0) was performed.

Results: In total, 12,254 transcripts were obtained from the wing disc, out of which 5,287 were identified to be differentially expressed from L5D6 to PP and from PP to P0. The results of comprehensive analysis of RNA-seq data showed that during larvae-to-pupae metamorphosis, many genes of 20E signaling pathway were up-regulated and those of JH signaling pathway were down-regulated. Seventeen transcription factors were significantly up-regulated. Cuticle protein genes (especially wing cuticle protein genes), were most abundant and significantly up-regulated at P0 stage. Genes responsible for the degradation and de novo synthesis of chitin were significantly up-regulated. There were A and B two types of chitin synthases in B. mori, whereas only chitin synthase A was up-regulated. Both trehalose and D-fructose, which are precursors of chitin synthesis, were detected in the hemolymph of L5D6, PP and P0, suggesting de novo synthesis of chitin. However, most of the genes that are related to early wing disc differentiation were down-regulated.

Conclusions: Extensive transcriptome and DGE profiling data of wing disc during metamorphosis of silkworm have been generated, which provided comprehensive gene expression information at the transcriptional level. These results implied that during the larva-to-pupa metamorphosis, pupal wing development and transition might be mainly controlled by 20E signaling in B. mori. The 17 up-regulated transcription factors might be involved in wing development. Chitin required for pupal wing development might be generated from both degradation of componential chitin and de novo synthesis. Chitin synthase A might be responsible for the chitin synthesis in the pupal wing, while both trehalose and D-fructose might contribute to the de novo synthesis of chitin during the formation of pupal wing.

Show MeSH
Related in: MedlinePlus