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Phenotype and transcriptome analysis reveals chloroplast development and pigment biosynthesis together influenced the leaf color formation in mutants of Anthurium andraeanum 'Sonate'.

Yang Y, Chen X, Xu B, Li Y, Ma Y, Wang G - Front Plant Sci (2015)

Bottom Line: Knowledge of mechanisms in anthuriums to produce leaves with different shades of green would help to effectively select desirable traits.From the 27,539 (67.1%) unigenes with annotated functions, 858 significantly differently expressed genes (DEGs) were identified, consisting of 446 up-regulated genes and 412 down-regulated genes.And the possible formation pathway of leaf color mutant of A. andraeanum 'Sonate' is deduced based on our results.

View Article: PubMed Central - PubMed

Affiliation: Department of Horticulture, Nanjing Agricultural University Nanjing, China.

ABSTRACT
Leaf color is one of the well-sought traits in breeding program for Anthurium andraeanum Lind. Knowledge of mechanisms in anthuriums to produce leaves with different shades of green would help to effectively select desirable traits. In this study, the micro- and ultra-structural and physiological features of leaves on wild type and leaf color mutants (dark green, rubescent, etiolated, albino) in A. andraeanum 'Sonate' were analyzed. Results show that chloroplasts of leaf color mutants exhibited abnormal morphology and distribution. Using next generation sequencing technology followed by de novo assembly, leaf transcriptomes comprising of 41,017 unigenes with an average sequence length of 768 bp were produced from wild type and rubescent mutant. From the 27,539 (67.1%) unigenes with annotated functions, 858 significantly differently expressed genes (DEGs) were identified, consisting of 446 up-regulated genes and 412 down-regulated genes. Genes that affect chloroplasts development and division, and chlorophyll biosynthesis were included in the down-regulated DEGs. Quantitative real-time PCR (qRT-PCR) analysis validated that the expression level of those genes was significantly lower in the rubescent, etiolated, and albino mutant compared to wild type plants, which concurs with the differences in micro- and ultra-structures and physiological features between these two types of plants. Conclusively, the leaf color formation is greatly affected by the activity of chloroplast development and pigment biosynthesis. And the possible formation pathway of leaf color mutant of A. andraeanum 'Sonate' is deduced based on our results.

No MeSH data available.


Related in: MedlinePlus

qRT-PCR expression analysis of genes involved in chloroplasts development, division and pigment biosynthesis in different tissues in the wild type Anthurium and 4 leaf color mutants. In these pictures, WT, Wild type; DG, dark green; RB, rubescent; ET, etiolated; AB, albino. (A) the relative expression level of AaGLK;(B)AaARC5;(C)AaMinE;(D)AaAPRR2;(E)AaDFR;(F)AaCHS;(G)AaF3H;(H)AaHEMC;(I)AaCRD1;(J)AaPORC. Different small letter with in lines indicate significant difference by LSD's multiple range test at 5% level in the leaves and spathes, respectively.
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Figure 9: qRT-PCR expression analysis of genes involved in chloroplasts development, division and pigment biosynthesis in different tissues in the wild type Anthurium and 4 leaf color mutants. In these pictures, WT, Wild type; DG, dark green; RB, rubescent; ET, etiolated; AB, albino. (A) the relative expression level of AaGLK;(B)AaARC5;(C)AaMinE;(D)AaAPRR2;(E)AaDFR;(F)AaCHS;(G)AaF3H;(H)AaHEMC;(I)AaCRD1;(J)AaPORC. Different small letter with in lines indicate significant difference by LSD's multiple range test at 5% level in the leaves and spathes, respectively.

Mentions: qRT-PCR was performed to validate the differential expression level of target genes in mutant and wild type plants. Results indicate that the transcript levels of AaGLK, AaARC5, and AaAPRR2 in wild type were lower than that of dark green in the leaf, and higher than that of rubescent, etiolated, and albino in leaf (Figures 9A–D). Consistent with leaf color differences, the expression of AaDFR (anthocyanin biosynthesis) in wild type leaf was lower than that in dark green, rubescent, and etiolated mutants. And in spathe of dark green the expression of AaDFR was higher than that in the wild type (Figure 9E). Interestingly, AaCHS (chalcone and stilbene biosynthesis) showed a higher expression in tissues of wild type than that of dark green (leaf and spathe), rubescent, etiolated, and albino mutants (leaf) (Figure 9F). Similar to AaCHS, AaF3H (flavanone and anthocyanin biosynthesis) expression was down-regulated in the leaf of 4 leaf color mutants but up-regulated in the spathe mutant (of dark green) (Figure 9G). AaHEMC, AaCRD1 (Dicarboxylate diiron protein) and AaPORC (Protochlorophyllide oxidoreductase C) (chlorophyll biosynthesis) expressed a higher quantity in the leaf from the wild and dark green type than that in rubescent, etiolated and albino mutants (Figures 9H–J). Thus, the expression pattern of AaCHS and AaF3H reflects the complex process of color formation.


Phenotype and transcriptome analysis reveals chloroplast development and pigment biosynthesis together influenced the leaf color formation in mutants of Anthurium andraeanum 'Sonate'.

Yang Y, Chen X, Xu B, Li Y, Ma Y, Wang G - Front Plant Sci (2015)

qRT-PCR expression analysis of genes involved in chloroplasts development, division and pigment biosynthesis in different tissues in the wild type Anthurium and 4 leaf color mutants. In these pictures, WT, Wild type; DG, dark green; RB, rubescent; ET, etiolated; AB, albino. (A) the relative expression level of AaGLK;(B)AaARC5;(C)AaMinE;(D)AaAPRR2;(E)AaDFR;(F)AaCHS;(G)AaF3H;(H)AaHEMC;(I)AaCRD1;(J)AaPORC. Different small letter with in lines indicate significant difference by LSD's multiple range test at 5% level in the leaves and spathes, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356079&req=5

Figure 9: qRT-PCR expression analysis of genes involved in chloroplasts development, division and pigment biosynthesis in different tissues in the wild type Anthurium and 4 leaf color mutants. In these pictures, WT, Wild type; DG, dark green; RB, rubescent; ET, etiolated; AB, albino. (A) the relative expression level of AaGLK;(B)AaARC5;(C)AaMinE;(D)AaAPRR2;(E)AaDFR;(F)AaCHS;(G)AaF3H;(H)AaHEMC;(I)AaCRD1;(J)AaPORC. Different small letter with in lines indicate significant difference by LSD's multiple range test at 5% level in the leaves and spathes, respectively.
Mentions: qRT-PCR was performed to validate the differential expression level of target genes in mutant and wild type plants. Results indicate that the transcript levels of AaGLK, AaARC5, and AaAPRR2 in wild type were lower than that of dark green in the leaf, and higher than that of rubescent, etiolated, and albino in leaf (Figures 9A–D). Consistent with leaf color differences, the expression of AaDFR (anthocyanin biosynthesis) in wild type leaf was lower than that in dark green, rubescent, and etiolated mutants. And in spathe of dark green the expression of AaDFR was higher than that in the wild type (Figure 9E). Interestingly, AaCHS (chalcone and stilbene biosynthesis) showed a higher expression in tissues of wild type than that of dark green (leaf and spathe), rubescent, etiolated, and albino mutants (leaf) (Figure 9F). Similar to AaCHS, AaF3H (flavanone and anthocyanin biosynthesis) expression was down-regulated in the leaf of 4 leaf color mutants but up-regulated in the spathe mutant (of dark green) (Figure 9G). AaHEMC, AaCRD1 (Dicarboxylate diiron protein) and AaPORC (Protochlorophyllide oxidoreductase C) (chlorophyll biosynthesis) expressed a higher quantity in the leaf from the wild and dark green type than that in rubescent, etiolated and albino mutants (Figures 9H–J). Thus, the expression pattern of AaCHS and AaF3H reflects the complex process of color formation.

Bottom Line: Knowledge of mechanisms in anthuriums to produce leaves with different shades of green would help to effectively select desirable traits.From the 27,539 (67.1%) unigenes with annotated functions, 858 significantly differently expressed genes (DEGs) were identified, consisting of 446 up-regulated genes and 412 down-regulated genes.And the possible formation pathway of leaf color mutant of A. andraeanum 'Sonate' is deduced based on our results.

View Article: PubMed Central - PubMed

Affiliation: Department of Horticulture, Nanjing Agricultural University Nanjing, China.

ABSTRACT
Leaf color is one of the well-sought traits in breeding program for Anthurium andraeanum Lind. Knowledge of mechanisms in anthuriums to produce leaves with different shades of green would help to effectively select desirable traits. In this study, the micro- and ultra-structural and physiological features of leaves on wild type and leaf color mutants (dark green, rubescent, etiolated, albino) in A. andraeanum 'Sonate' were analyzed. Results show that chloroplasts of leaf color mutants exhibited abnormal morphology and distribution. Using next generation sequencing technology followed by de novo assembly, leaf transcriptomes comprising of 41,017 unigenes with an average sequence length of 768 bp were produced from wild type and rubescent mutant. From the 27,539 (67.1%) unigenes with annotated functions, 858 significantly differently expressed genes (DEGs) were identified, consisting of 446 up-regulated genes and 412 down-regulated genes. Genes that affect chloroplasts development and division, and chlorophyll biosynthesis were included in the down-regulated DEGs. Quantitative real-time PCR (qRT-PCR) analysis validated that the expression level of those genes was significantly lower in the rubescent, etiolated, and albino mutant compared to wild type plants, which concurs with the differences in micro- and ultra-structures and physiological features between these two types of plants. Conclusively, the leaf color formation is greatly affected by the activity of chloroplast development and pigment biosynthesis. And the possible formation pathway of leaf color mutant of A. andraeanum 'Sonate' is deduced based on our results.

No MeSH data available.


Related in: MedlinePlus