Micropropagation and Cellular Behaviour Changes During <i>in vitro</i> Flowering of <i>Impatiens balsamina</i>

MOHAMED, N.; TAHA, R.M.; RAZAK, U.N.A.A.

doi:10.1590/S0100-83582019370100037

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Article • Planta daninha 37 • 2019 • https://doi.org/10.1590/S0100-83582019370100037 copy

Micropropagation and Cellular Behaviour Changes During in vitro Flowering of Impatiens balsamina

Micropropagação e Mudanças de Comportamento das Células Durante o Florescimento in vitro de Impatiens balsamina

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT:

In this study, the micropropagation of Impatiens balsamina was established from stem and shoot explants. The effects of GA3 and glutathione on the morphogenesis of this species were also investigated, in order to induce in vitro flowering. It was found that the optimum in vitro plant regeneration was achieved on MS medium supplemented with 1.0 mg L-1 GA3 and in vitro flowering was also obtained from the same medium after 4 weeks of culture. To understand cellular behavior during in vitro flowering, Mitotic Index (MI), chromosome counts, measurement of mean cell and nuclear areas, DNA measurements and ploidy levels were analyzed from in vivo plants, in vitro grown plants and plantlets that flowered in vitro. The chromosome count was the same for all, 2x=2n=14 or n=7. However, it was observed that in vitro flowering plants of Impatiens balsamina had the highest percentage of polyploid cells (30.7%), based on a histogram plotted by the AxioVision 4.7 software. It was found that plant growth regulators, especially GA3, increased the polyploidy level of the meristematic root cells.

Keywords:
regeneration; gibberellic acid; glutathione; flowering response; cellular behavior and acclimatization

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[1] * Corresponding author: <normadiha@yahoo.com>

MS + PGR (mg L^-1)	Mean no. of shoots per explant	Mean height of plantlet (cm)	Mean no. of flowers per explant
Control	0.70±0.16 d	1.50±0.26 d	0.90±0.12 d
0.5GA₃	5.00±0.05 b	2.80±0.03 bc	1.60±0.10 bc
1.0GA₃	9.00±0.06 a	4.00±0.02 a	3.50±0.03 a
1.5GA₃	6.20±0.03 b	2.50±0.04 c	2.00±0.04 b
2.0GA₃	6.00±0.03 b	3.30±0.08 b	2.50±0.13 b
0.5Glu	1.10±0.22 c	3.00±0.13 b	n/a
1.0Glu	1.00±0.19 c	2.50±0.17 c	n/a
1.5Glu	1.20±0.31 c	2.40±0.12 c	n/a
2.0Glu	0.80±0.24 d	1.80±0.22 d	n/a

MS + Hormone (mg L^-1)	Mean no. of shoots per explant	Mean height of plantlet (cm)	Mean no. of flowers per explant
Control	0.63±0.12 d	0.55±0.21 d	n/a
0.5GA₃	3.80±0.15 b	2.50±0.01 ab	n/a
1.0GA₃	4.40±0.10 a	2.70±0.04 a	n/a
1.5GA₃	4.20±0.02 a	3.00±0.05 a	n/a
2.0GA₃	3.20±0.04 b	2.00±0.06 b	n/a
0.5Glu	1.20±0.25 c	2.80±0.19 ab	n/a
1.0Glu	0.70±0.19 d	1.80±0.11 c	n/a
1.5Glu	1.30±0.23 c	2.40±0.20 ab	n/a
2.0Glu	1.10±0.24 c	2.20±0.28 b	n/a

Type of growth media	Survival rate (%) of plants	Observation
Black soil+red soil (1:1)	75.0±0.6 a	Plantlets survived and showed healthy growth
Red soil	10.3±0.2 c	Plantlets survived and showed slow growth
Black soil	48.0±0.3 b	Plantlets survived and showed healthy growth

Characteristic	In vivo plant (control)	In vitro flowering plants
Plant height (cm)	48 ± 0.1	18 ± 0.4
Leaf
Arrangement	Alternate	Alternate
Type	Simple	Simple
Margin	Serrate	Serrate
Shape	Lanceolate	Linear to Lanceolate
Venation	Pinnate	Pinnate
Length (cm)	5 ± 0.62	3 ± 0.44
Color	Green	Green
No. of leaves per plant	42 ± 0.52	33 ± 0.35
Flower
No. of flowers per plant	18 ± 0.42	12 ± 0.25
Symmetry	Actinomorphic	Actinomorphic
Color	Red	Red
Flower size (cm)	3 ± 0.57	3 ± 0.24

Growth media	Mitotic Index, MI (%)	Mean no. of Chromosomes
MS basal (in vitro)	43.0 ± 0.78	14.33 ± 0.68
MS + 1.0 mg L^-1 GA₃ (in vitro flowering)	53.8 ± 1.34	14.07 ± 0.34
Soil (in vivo)	36.33 ± 1.75	14.27 ± 0.55.

Impatiens balsamina source	Mean areas (μm²)
Impatiens balsamina source	Nucleus (N)	Cell (C)	Ratio (N/C)
MS basal (in vitro)	49.31 ± 0.66	225.45 ± 0.45	0.22 ± 0.12
MS + 1.0 mg L^-1 GA₃ (in vitro flowering)	65.18 ± 0.49	276.35 ± 0.34	0.24 ± 0.18
In vivo	47.94 ± 0.54	248.63 ± 0.67	0.19 ± 0.04

Impatiens balsamina source	Cell cycle phase (%)			Polyploidy (%)
Impatiens balsamina source	G1	S	G2
MS basal (in vitro)	25	40.7	20.3	14
MS + 1.0 mg L^-1 GA₃ (in vitro flowering)	14	21.3	34.0	30.7
In vivo	74	20.6	5.4	0