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A de novo floral transcriptome reveals clues into Phalaenopsis orchid flower development.

Huang JZ, Lin CP, Cheng TC, Chang BC, Cheng SY, Chen YW, Lee CY, Chin SW, Chen FC - PLoS ONE (2015)

Bottom Line: PhAGL6a, PhAGL6b and PhMADS4 were strongly expressed in the labellum and significantly upregulated in lip-like petals and lip-like sepals of peloric-mutant flowers.In addition, PhAGL6b was significantly downregulated in the labellum of the big lip mutant, with no change in expression of PhAGL6a.Our study provides new insights into how the orchid labellum differs and why the petal or sepal converts to a labellum in Phalaenopsis floral mutants.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Industry, National Pingtung University of Science and Technology, Pingtung 91201, Taiwan.

ABSTRACT
Phalaenopsis has a zygomorphic floral structure, including three outer tepals, two lateral inner tepals and a highly modified inner median tepal called labellum or lip; however, the regulation of its organ development remains unelucidated. We generated RNA-seq reads with the Illumina platform for floral organs of the Phalaenopsis wild-type and peloric mutant with a lip-like petal. A total of 43,552 contigs were obtained after de novo assembly. We used differentially expressed gene profiling to compare the transcriptional changes in floral organs for both the wild-type and peloric mutant. Pair-wise comparison of sepals, petals and labellum between peloric mutant and its wild-type revealed 1,838, 758 and 1,147 contigs, respectively, with significant differential expression. PhAGL6a (CUFF.17763), PhAGL6b (CUFF.17763.1), PhMADS1 (CUFF.36625.1), PhMADS4 (CUFF.25909) and PhMADS5 (CUFF.39479.1) were significantly upregulated in the lip-like petal of the peloric mutant. We used real-time PCR analysis of lip-like petals, lip-like sepals and the big lip of peloric mutants to confirm the five genes' expression patterns. PhAGL6a, PhAGL6b and PhMADS4 were strongly expressed in the labellum and significantly upregulated in lip-like petals and lip-like sepals of peloric-mutant flowers. In addition, PhAGL6b was significantly downregulated in the labellum of the big lip mutant, with no change in expression of PhAGL6a. We provide a comprehensive transcript profile and functional analysis of Phalaenopsis floral organs. PhAGL6a PhAGL6b, and PhMADS4 might play crucial roles in the development of the labellum in Phalaenopsis. Our study provides new insights into how the orchid labellum differs and why the petal or sepal converts to a labellum in Phalaenopsis floral mutants.

No MeSH data available.


Real-time PCR analysis of genes expressed in different floral mutants of Phalaenopsis orchid.Total RNAs isolated from the sepal, petal, lip and column of mature flowers from Fig 5. The wild-type ([A] Phalaenopsis Brother Spring Dancer ‘KHM190’; [B] Phalaenopsis aphrodite and [C] Phalaenopsis ‘NPU1458’) and (N), lip-like petal mutant (P), lip-like sepal mutant (M) and big lip mutant (BP) were used as templates to detect the expression of CUFF.17763(PhAGL6a), CUFF.17763.1 (PhAGL6b), CUFF.25909 (PhMADS4), CUFF.39479.1 (PhMADS5) and CUFF.36625.1(PhMADS1). S, sepal (include upper and lateral sepals); SU, upper sepal; SD, lateral sepal; P, petal; L, lip; C, column.
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pone.0123474.g006: Real-time PCR analysis of genes expressed in different floral mutants of Phalaenopsis orchid.Total RNAs isolated from the sepal, petal, lip and column of mature flowers from Fig 5. The wild-type ([A] Phalaenopsis Brother Spring Dancer ‘KHM190’; [B] Phalaenopsis aphrodite and [C] Phalaenopsis ‘NPU1458’) and (N), lip-like petal mutant (P), lip-like sepal mutant (M) and big lip mutant (BP) were used as templates to detect the expression of CUFF.17763(PhAGL6a), CUFF.17763.1 (PhAGL6b), CUFF.25909 (PhMADS4), CUFF.39479.1 (PhMADS5) and CUFF.36625.1(PhMADS1). S, sepal (include upper and lateral sepals); SU, upper sepal; SD, lateral sepal; P, petal; L, lip; C, column.

Mentions: To reveal putative roles in labellum development of Phalaenopsis flower, we evaluated the association of the expression of five TFs (PhMADS1, PhMADS4, PhMADS5, PhAGL6a and PhAGL6b) in the floral organs (sepal, petal, labellum and gynostemium) and different floral morphologic features of Phalaenopsis orchid mutants (lip-like petal, lip-like sepal, and big lip) from our collection (Fig 5; Fig E in S1 File). We estimated the association of the expression of the five TFs and labellum development by real-time PCR. The PhMADS1 transcript was highly expressed in the gynostemium (Fig 6). The expression of PhMADS5 was significantly reduced in lip-like petals of peloric-mutant flowers (Fig 6A) but not in the lip-like sepal of peloric-mutant flowers (Fig 6B). The expression of three TFs (PhMADS4, PhAGL6a and PhAGL6b) was increased in the labellum of the wild-type and mutant (Fig 6). In contrast, PhMADS4 and PhAGL6b transcripts were significantly reduced in the labellum of the big-lip mutant (Fig 6C). Furthermore, the transcript levels of PhAGL6a and PhAGL6b were significantly increased in the lip-like petal and lip-like sepal of peloric- mutant flowers (Fig 6A and 6B). Thus, PhMADS4, PhAGL6a and PhAGL6b may play different roles in the development of the labellum in Phalaenopsis, especially PhAGL6b in peloric-mutant flowers and labellum development.


A de novo floral transcriptome reveals clues into Phalaenopsis orchid flower development.

Huang JZ, Lin CP, Cheng TC, Chang BC, Cheng SY, Chen YW, Lee CY, Chin SW, Chen FC - PLoS ONE (2015)

Real-time PCR analysis of genes expressed in different floral mutants of Phalaenopsis orchid.Total RNAs isolated from the sepal, petal, lip and column of mature flowers from Fig 5. The wild-type ([A] Phalaenopsis Brother Spring Dancer ‘KHM190’; [B] Phalaenopsis aphrodite and [C] Phalaenopsis ‘NPU1458’) and (N), lip-like petal mutant (P), lip-like sepal mutant (M) and big lip mutant (BP) were used as templates to detect the expression of CUFF.17763(PhAGL6a), CUFF.17763.1 (PhAGL6b), CUFF.25909 (PhMADS4), CUFF.39479.1 (PhMADS5) and CUFF.36625.1(PhMADS1). S, sepal (include upper and lateral sepals); SU, upper sepal; SD, lateral sepal; P, petal; L, lip; C, column.
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Related In: Results  -  Collection

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pone.0123474.g006: Real-time PCR analysis of genes expressed in different floral mutants of Phalaenopsis orchid.Total RNAs isolated from the sepal, petal, lip and column of mature flowers from Fig 5. The wild-type ([A] Phalaenopsis Brother Spring Dancer ‘KHM190’; [B] Phalaenopsis aphrodite and [C] Phalaenopsis ‘NPU1458’) and (N), lip-like petal mutant (P), lip-like sepal mutant (M) and big lip mutant (BP) were used as templates to detect the expression of CUFF.17763(PhAGL6a), CUFF.17763.1 (PhAGL6b), CUFF.25909 (PhMADS4), CUFF.39479.1 (PhMADS5) and CUFF.36625.1(PhMADS1). S, sepal (include upper and lateral sepals); SU, upper sepal; SD, lateral sepal; P, petal; L, lip; C, column.
Mentions: To reveal putative roles in labellum development of Phalaenopsis flower, we evaluated the association of the expression of five TFs (PhMADS1, PhMADS4, PhMADS5, PhAGL6a and PhAGL6b) in the floral organs (sepal, petal, labellum and gynostemium) and different floral morphologic features of Phalaenopsis orchid mutants (lip-like petal, lip-like sepal, and big lip) from our collection (Fig 5; Fig E in S1 File). We estimated the association of the expression of the five TFs and labellum development by real-time PCR. The PhMADS1 transcript was highly expressed in the gynostemium (Fig 6). The expression of PhMADS5 was significantly reduced in lip-like petals of peloric-mutant flowers (Fig 6A) but not in the lip-like sepal of peloric-mutant flowers (Fig 6B). The expression of three TFs (PhMADS4, PhAGL6a and PhAGL6b) was increased in the labellum of the wild-type and mutant (Fig 6). In contrast, PhMADS4 and PhAGL6b transcripts were significantly reduced in the labellum of the big-lip mutant (Fig 6C). Furthermore, the transcript levels of PhAGL6a and PhAGL6b were significantly increased in the lip-like petal and lip-like sepal of peloric- mutant flowers (Fig 6A and 6B). Thus, PhMADS4, PhAGL6a and PhAGL6b may play different roles in the development of the labellum in Phalaenopsis, especially PhAGL6b in peloric-mutant flowers and labellum development.

Bottom Line: PhAGL6a, PhAGL6b and PhMADS4 were strongly expressed in the labellum and significantly upregulated in lip-like petals and lip-like sepals of peloric-mutant flowers.In addition, PhAGL6b was significantly downregulated in the labellum of the big lip mutant, with no change in expression of PhAGL6a.Our study provides new insights into how the orchid labellum differs and why the petal or sepal converts to a labellum in Phalaenopsis floral mutants.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Industry, National Pingtung University of Science and Technology, Pingtung 91201, Taiwan.

ABSTRACT
Phalaenopsis has a zygomorphic floral structure, including three outer tepals, two lateral inner tepals and a highly modified inner median tepal called labellum or lip; however, the regulation of its organ development remains unelucidated. We generated RNA-seq reads with the Illumina platform for floral organs of the Phalaenopsis wild-type and peloric mutant with a lip-like petal. A total of 43,552 contigs were obtained after de novo assembly. We used differentially expressed gene profiling to compare the transcriptional changes in floral organs for both the wild-type and peloric mutant. Pair-wise comparison of sepals, petals and labellum between peloric mutant and its wild-type revealed 1,838, 758 and 1,147 contigs, respectively, with significant differential expression. PhAGL6a (CUFF.17763), PhAGL6b (CUFF.17763.1), PhMADS1 (CUFF.36625.1), PhMADS4 (CUFF.25909) and PhMADS5 (CUFF.39479.1) were significantly upregulated in the lip-like petal of the peloric mutant. We used real-time PCR analysis of lip-like petals, lip-like sepals and the big lip of peloric mutants to confirm the five genes' expression patterns. PhAGL6a, PhAGL6b and PhMADS4 were strongly expressed in the labellum and significantly upregulated in lip-like petals and lip-like sepals of peloric-mutant flowers. In addition, PhAGL6b was significantly downregulated in the labellum of the big lip mutant, with no change in expression of PhAGL6a. We provide a comprehensive transcript profile and functional analysis of Phalaenopsis floral organs. PhAGL6a PhAGL6b, and PhMADS4 might play crucial roles in the development of the labellum in Phalaenopsis. Our study provides new insights into how the orchid labellum differs and why the petal or sepal converts to a labellum in Phalaenopsis floral mutants.

No MeSH data available.