Genetic control of flower petal number in Rosa x Damascena Mill f. trigintipetala

Figures & data

Table 1. Primers used for analysis of expression of flower homeotic genes in early stages of flower development in Rosa x damascena by qPCR.

Gene	Primer designation	Primer sequence
R. hybrida AGAMOUS [19]	RhAG-s1	5′-AAAAGGGAACTTGACTTGCACAAC-3′
	RhAG-as1	5′-TCAAACTAATTGAAGGGAAATCTGG-3′
R. x damascena euAPETALA3 (euAP3 line) (Rusanov et al., unpublished data)	AP3QPCRse	5′-TGGCGAGAGGTAAGATCCAG-3′
	AP3QPCRas	5′-TGCTTTGTCGTGGTGGAG-3′
R. x damascena APETALA1/FUL (Rusanov et al., unpublished data)	AP1QPCRas	5′-TCGACACTACTAATGTGAGAAGACG-3′
	AP1QPCRse	5′-GAAAATAAGGAATTACACGGGAGA-3′
R. hybrida TCTP [19]	RhTCTPs	5′-GATGCTGATGAGGGTGTTGA-3′
	RhTCTPas	5′-CTTGGTTGCTCCCTCAATGT-3′
R. hybrida EF1-alpha [19]	RhEF1-QS1	5′-GGGTAAGGAGAAGGTTCACATC-3′
	RhEF1-QAS1	5′-CAGCCTCCTTCTCAAACCTCT-3′

Table 2. Primers used for analysis of microsatellite loci.

Locus	Primer pair sequence*	т annealing (°C)
Rh58	Fw 5′-AAACGACGGCCAGTACAATTTAGTGCGGATAGAACAAC-3′ Rev 5′-GGAAAGCCCGAAAGCGTAAGC-3′	59
Rh50	Fw 5′-AAACGACGGCCAGTTGATGAAATCATCCGAGTGTCAG-3′ Rev 5′-TCACTTTCATTGGAATGCCAGAAT-3′	59
Rh65	Fw 5′-AAACGACGGCCAGTAGTACGCCGACGCAGATCCAGTGA-3′ Rev 5′-ACGGCGTTGTAGGTCGTCATTCTC-3′	67
RhI402	Fw 5′-AAACGACGGCCAGTTCCCATCTTGCTAAGTGCCTT-3′ Rev 5′-GTTTCTTCAGGGTAACTGAGCCGATT-3′	62
Rw11E5	Fw 5′-AAACGACGGCCAGTGATACCGCGAAGGTGTAGT-3′ Rev 5′-GAGTGAAAACTCTGCAATCA-3′	55

* Note: The M13 tail in each forward primer is shown in bold.

Table 3. Flower biometric analysis* of plants obtained after self-pollination of Rosa x damascena f. trigintipetala.

Flower type	Value	Flower diameter, mm	Number of petals	Number of stamens
Simple flower	Aver.	62 ± 6	6 ± 1	110 ± 12
	Min.	50	5	82
	Max.	75	7	132
Semi-double flower	Aver.	61 ± 8	16 ± 5	76 ± 24
	Min.	46	8	36
	Max.	74	20	107
Double flower	Aver.	60 ± 10	43 ± 20	70 ± 28
	Min.	41	21	0
	Max.	112	103	134

* Note: Mean values ± standard deviation. Simple flower defined as 6 ± 1 petals, semi-double flower as >7 ≤ 20 petals, double flower as >20 petals.

Figure 1. Variation in flower morphology in plants obtained after self-pollination of Rosa x damascena Mill. f. trigintipetala. Simple flower (a), semi-double flower (b), double flower (c and d).

Figure 2. Box-and-whiskers plot showing the distribution of data for number of petals among plants with different allele phenotype of the microsatellite marker Rh58 in plants from a segregating population obtained after self-pollination of Rosa x damascena f. trigintipetala. A, B, C and D designate the scored allele sizes (A = 222 bp, B = 228 bp, C = 248 bp and D = 260 bp).

Figure 3. Expression level of Rosa x damascena AGAMOUS (RdAG), R. x damascena APETALA1/FUL-like (RdAP1/FUL-like) and R. x damascena euAPETALA3 (RdAP3). RdD, early stage flower buds of R. x damascena f. trigintipetala with double flowers. RdS, early stage flower buds of a simple flower plant (6 ± 1 petals) obtained by self-pollination of R. x damascena f. trigintipetala.

Figure 4. Box-and-whiskers plot showing the distribution of data for number of petals among plants with different allele phenotypes of Rosa x damascena AGAMOUS in a segregating population of 143 plants obtained by self-pollination of R. x damascena f. trigintipetala. A, B, C and D designate the 4 different alleles of R. x damascena AGAMOUS scored by using the AGM2 primer pair (A = 147 bp, B = 153 bp, C = 155 bp and D = 163 bp).

Dubois A, Raymond O, Maene M, et al. Tinkering with the C-function: a molecular frame for the selection of double flowers in cultivated roses. PLoS One. 2010;5:e9288.

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