Effect of different antimitotic agents on polyploid induction of anise hyssop (Agastache foeniculum L.)

Figures & data

Table 1. Influence of different concentrations of antimitotic agents on the survival rate and ploidy level of the apical meristems.

Apical meristem treatment
Treatment	Survival rate (%)	Diploid (%)	Mixoploid (%)	Tetraploid (%)
Control	96.00 a*	96.00 a	0.00 g	0.00 d
Colchicine	–	–	–	–
5000 μM	98.00 a	98.00 a	0.00 g	0.00 d
12500 μM	62.00 cd	46.00 c	16.00 ef	0.00 d
17500 μM	54.00 d	26.00 d	12.00 f	16.00 a
Oryzalin	–	–	–	–
10 μm	84.00 b	64.00 b	20.00 de	0.00 d
50 μm	68.00 c	20.00 d	34.00 a	14.00 ab
100 μm	56.00 d	20.00 d	26.00 bc	10.00 bc
Trifloralin	–	–	–	–
10 μm	82.00 b	64.00 b	18.00 e	0.00 d
50 μm	66.00 c	26.00 d	30.00 ab	12.00 abc
100 μm	54.00 d	22.00 d	24.00 cd	8.00 c

^* Means with the same letters in each column are not significantly different at p < 0.01%.

Table 2. Influence of different concentrations of antimitotic agents on the survival rate and ploidy level of the plant seeds and seedlings.

		Seed treatment				Seedling treatment
Treatment	Hour	Survival rate (%)	Diploid (%)	Mixoploid (%)	Tetraploid (%)	Survival rate (%)	Diploid (%)	Mixoploid (%)	Tetraploid (%)
Colchicine		–	–	–	–	–	–	–	–
Control	6	98.00 ab*	100.00 a	0.00 g	0.00 f	100.00 a	100.00 a	0.00 l	0.00 f
	12	96.00 bc	96.00 abc	0.00 g	0.00 f	98.00 a	98.00 ab	0.00 l	0.00 f
	24	96.00 bc	96.00 abc	0.00 g	0.00 f	94.00 b	94.00 bc	0.00 l	0.00 f
5000 μM	6	82.00 fg	82.00 de	0.00 g	0.00 f	94.00 b	90.00 cd	2.00 l	0.00 f
	12	80.00 g	80.00 def	0.00 g	0.00 f	86.00 de	82.00 f	4.00 kl	0.00 f
	24	84.00 ef	84.00 d	0.00 g	0.00 f	46.00 k	24.00 nop	14.00 fg	8.00 c
12500 μM	6	76.00 h	76.00 fg	0.00 g	0.00 f	78.00 g	68.00 i	10.00 hi	0.00 f
	12	74.00 hi	64.00 h	12.00 e	0.00 f	42.00 l	26.00 no	10.00 hi	6.00 cd
	24	72.00 i	58.00 ij	12.00 e	0.00 f	38.00 m	80.00 fg	16.00 ef	6.00 cd
17500 μM	6	80.00 g	80.00 def	0.00 g	0.00 f	76.00 g	60.00 J	10.00 hi	4.00 de
	12	74.00 hi	62.00 hi	16.00 cd	0.00 f	36.00 mn	12.00 r	16.00ef	8.00 c
	24	74.00 hi	52.00 k	18.00 bc	4.00 de	24.00 o	40.00 l	12.00 gh	4.00 de
Oryzalin		–	–	–	–	–	–	–	–
Control	6	92.00 d	92.00 c	0.00 g	0.00 f	100.00 a	100.00 a	0.00 l	0.00 f
	12	96.00 bc	96.00 abc	0.00 g	0.00 f	100.00 a	100.00 a	0.00 l	0.00 f
	24	100.00 a	98.00 ab	0.00 g	0.00 f	96.00 ab	96.00 ab	0.00 l	0.00 f
10 μm	6	80.00 g	80.00 def	0.00 g	0.00 f	100.00 a	100.00 a	0.00 l	0.00 f
	12	82.00 fg	82.00 de	0.00 g	0.00 f	82.00 f	74.00 h	10.00 hi	0.00 f
	24	84.00 ef	78.00 efg	6.00 f	0.00 f	64.00 i	52.00 k	8.00 ij	4.00 de
50 μm	6	84.00 ef	84.00 d	0.00 g	0.00 f	90.00 c	90.00 cd	0.00 l	0.00 f
	12	74.00 hi	54.00 jk	14.00 de	4.00 de	66.00 i	34.00 m	32.00 c	4.00 de
	24	56.00 m	30.00 mn	12.00 e	14.00 b	48.00 k	20.00 pq	28.00 d	12.00 b
100 μm	6	76.00 h	76.00 fg	0.00 g	0.00 f	82.00 f	76.00 gh	6.00 jk	0.00 f
	12	62.00 k	42.00 l	14.00 de	6.00 d	60.00 J	12.00 r	40.00 a	8.00 c
	24	56.00 m	32.00 m	4.00 f	20.00 a	34.00 n	22.00 op	16.00 ef	2.00 ef
Trifluralin			–	–	–				–
Control	6	94.00 cd	94.00 bc	0.00 g	0.00 f	98.00 a	98.00 ab	0.00 l	0.00 f
	12	96.00 bc	96.00 abc	0.00 g	0.00 f	98.00 a	98.00 ab	0.00 l	0.00 f
	24	98.00 ab	98.00 ab	0.00 g	0.00 f	98.00 a	98.00 ab	0.00 l	0.00 f
10 μm	6	82.00 fg	82.00 de	0.00 g	0.00 f	94.00 b	94.00 bc	0.00 l	0.00 f
	12	84.00 ef	80.00 def	4.00 f	0.00 f	86.00 de	82.00 f	4.00 kl	0.00 f
	24	86.00 e	82.00 de	4.00 f	0.00 f	60.00 J	48.00 k	12.00 gh	0.00 f
50 μm	6	80.00 g	80.00 def	0.00 g	0.00 f	88.00 cd	88.00 de	0.00 l	0.00 f
	12	74.00 hi	60.00 hi	12.00 e	2.00 ef	70.00 h	28.00 n	26.00 d	16.00 a
	24	60.00 kl	26.00 n	20.00 b	14.00 b	48.00 k	24.00 nop	18.00 e	8.00 c
100 μm	6	74.00 hi	74.66 g	0.00 g	0.00 f	84.00 ef	84.00 ef	6.00 jk	0.00 f
	12	66.00 J	50.00 k	12.00 e	4.00 e	60.00 J	16.00 qr	36.00 b	8.00 c
	24	58.00 lm	26.00 n	24.00 a	8.00 c	38.00 m	28.00 n	10.00 hi	0.00 f

^* Means with the same letters in each column are not significantly different at p < 0.01%.

Figure 1. Flow cytometric analysis histograms of (A) tetraploid, 2n=4x=36; (B) diploid 2n=2x=18; and (C) mixoploids (IS = internal standard, Pisum sativum).

Table 3. Morphophysiological characteristics of the diploid and tetraploid plants.

Characteristic	Diploid	Tetraploid
Stomatal density (number mm^–2)	35.50± 5.16 a*	11.66± 1.36 b
Chloroplast number	11.16± 0.98 b	20.16± 3.18 a
Chlorophyll index	46.23± 4.45 b	64.16± 1.39 a
Leaf length (mm)	45.14± 0.72 b	57.23± 0.43 a
Leaf width (mm)	40.45± 1.01 b	46.14± 0.90 a
Internode space (mm)	26.18± 0.76 a	11.99± 0.38 b
Leaf area (mm^2)	12.66± 0.81 b	22.16± 0.98 a
Plant height on 10 leaf (mm)	118.11± 0.36 a	85.98± 0.59 b
Net photosynthesis (μmol Co2 m^−2s^−1)	9.05±0.33 b	9.38±0.33 a
CAT activity (μmol min^−1 mg^−1 protein)	1.08± 0.10 b	2.32± 0.18 a
POD activity (μmol min^−1 mg^−1 protein)	0.67±0.07 b	1.17± 0.02 a
Protein content (mg g^−1 FW)	0.23± 0.03 b	0.39± 0.03 a
Stomatal length (μm)	24.05± 0.55 b	32.55± 0.42 a
Stomatal diameter (μm)	10.98± 0.84 b	16.95±0.34 a
Fresh weight (g)	7.37±0.24 b	12.28±0.39 a
Dry weight (g)	0.77±0.10 b	1.34±0.08 a
Spike length (cm)	5.19±0.22 b	16.82±0.25 a

^* Means with the same letters in row are not significantly different at p < 0.01%.

Figure 2. Difference in stomatal size between the tetraploid (A) and diploid (B) plants.

Figure 3. Difference in stomatal density between the tetraploid (A) and diploid (B) plants (scale bar = 20 μm).

Figure 4. Difference in chloroplast number between the tetraploid (A) and diploid (B) plants (scale bar = 20 μm).

Figure 5. Morphological changes between the diploid and tetraploid anise hyssop plants.

Figure 6. The chromosomes of diploid (2n=2x=18) and tetraploid (2n=4x=36) plants (scale bar = 10 μm).