Fig 1.
Morphologic and Phylogenetic analysis of different-colored mutants of Cymbidium sinense 'Dharma.'
(A) The UPGMA cluster of ten leaf-color mutants of Cymbidium sinense 'Dharma' based on AFLP markers; (B) Morphologic characteristics of six leaf-color mutants of Cymbidium sinense 'Dharma' studied in this work.
Fig 2.
The variations in the contents of photosynthetic pigments between the wild type and leaf-color mutants.
(A) Chlorophyll content in wild-type and color-mutant leaves of Cymbidium sinense 'Dharma'(mg g-1 FW); (B) The relative contents of chlorophyll synthesis precursors PBG, Proto IX, Mg-Proto, and Pchlide, and chlorophyll degradation metabolite Pheide a (Take each intermediate product content in the wild type as 100%, comparing it with its mutant correspondent). Values are means ±standard deviation (n > 10). Mean-Whitney U-test significant at *P<0.05 between the mutants and the wild type control.
Fig 3.
The size distribution of assembled isotigs.
The x-axis represents the sequence length in base pairs. The y-axis represents the isotigs number.
Table 1.
Summary of sequencing and de novo assembling of the transcriptome in Cymbidium sinense 'Dharma'.
Table 2.
Summary of the blast hits against the known protein database.
Fig 4.
Characteristics of homology search of the isotigs.
(A) Similarity distribution of the best Blast hits in Nr database; (B) Similarity distribution of the best Blast hits in Uniprot database; (C) Similarity distribution of the best Blast hits in Swissport database.
Fig 5.
GO classification of unigenes of assembled isotigs.
Fig 6.
KOG function classification of assembled isotigs.
Table 3.
Functional categories of assembled isotigs in KEGG pathways.
Fig 7.
Assembled isotigs involved in the chlorophyll biosynthesis pathway pathway of Cymbidium sinense 'Dharma'.
Abbreviation: HEMC, hydroxymethylbilane synthase; HEMD, uroporphyrinogen-III synthase; HEME, uroporphyrinogen decarboxylase; CHLH, magnesium chelatase; CHLD, magnesium chelatase subunit ChlD; CHLI, magnesium chelatase subunit ChlI; CHLM, magnesium-protoporphyrin O-methyltransferase; PORA, protochlorophyllide oxidoreductase A; CHLG, Chlorophyll synthase; CAO, chlorophyllide a oxygenase.
Table 4.
Putative key enzyme-encoding genes in chlorophyll metabolism pathway.
Fig 8.
Assembled isotigs involved in the chlorophyll degradation pathway pathway of Cymbidium sinense 'Dharma'.
Abbreviation: NOL, chlorophyll b reductase; CLH, chlorophyllase; PAO, pheophorbide a oxygenase, RCCR, red Chl catabolite reductase.
Fig 9.
Northern blotting analysis of the expression of identified genes in different tissues of Cymbidium sinense 'Dharma'.
Each lane contained 15 g total RNA isolated from roots, pseudobulbs and six-month old leaves, rRNA served as a loading control (bottom of the panel).
Fig 10.
The qRT-PCR analysis of the expressions of two enzyme-encoding genes in chl degradation pathway (A) and five key enzyme-encoding genes in chl biosynthesis pathway (B). The y-axis indicates fold change in expression among the samples. The Lg(Relative Quantitation) of the genes in the wild-type leaves was calibrated as zero. RNA was extracted from six-month-old normal growth leaves. ACT gene served as the internal control. Error bars indicate the standard deviation of the mean (SD) (n = 3). Three biological replicates were analyzed, with similar results.