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Morphological, cytological and molecular variations induced by gamma rays in Chrysanthemum morifolium ‘Donglinruixue’

Ling Wang
Ling Wang
, 
Jianhui Wu
Jianhui Wu
, 
Feng Lan
Feng Lan
 et 
Pengfei Gao
Pengfei Gao
   | 18 mai 2020
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Article Category: Research Article
Publié en ligne: 18 mai 2020
Pages: 87 - 96
Reçu: 13 déc. 2019
Accepté: 31 mars 2020
DOI: https://doi.org/10.2478/fhort-2020-0009
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© 2020 Ling Wang et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

Effect of gamma radiation on the death rate of chrysanthemum ‘Donglinruixue’ plants at different radiation doses. Mean values were calculated from three experiments.
Effect of gamma radiation on the death rate of chrysanthemum ‘Donglinruixue’ plants at different radiation doses. Mean values were calculated from three experiments.

Figure 2

Effect of gamma radiation on average plant height of chrysanthemum ‘Donglinruixue’ plants at 120 days of plant culture after treatment with different radiation doses. (A) Effect of 0, 15, 20, 25, 30 and 35 Gy radiation. (B) Effect of gamma radiation on average plant height of chrysanthemum ‘Donglinruixue’ plants at different radiation doses. Mean values and SEM were calculated from three experiments. Within each set of experiments, bars with different letters are significantly different at the 0.05 level.
Effect of gamma radiation on average plant height of chrysanthemum ‘Donglinruixue’ plants at 120 days of plant culture after treatment with different radiation doses. (A) Effect of 0, 15, 20, 25, 30 and 35 Gy radiation. (B) Effect of gamma radiation on average plant height of chrysanthemum ‘Donglinruixue’ plants at different radiation doses. Mean values and SEM were calculated from three experiments. Within each set of experiments, bars with different letters are significantly different at the 0.05 level.

Figure 3

Morphological variations induced by gamma-ray irradiation in chrysanthemum ‘Donglinruixue’. (A) Variations in leaves: (a) original leaves on the control plant; (b) increased leaf crack; (c) leaves missing green colour; (d) deformed leaves; (e) lanceolate leaves; (f) leathery leaves and (g) rolled leaves. (B) Variations in flowers: (a) original flowers of the control plant; (b) reduced number of tubular flowers; (c) the top of the petal split; (d) reduced number of petals; (e) petals widened; (f) edge of flower with tubular flowers and (g) twisted lingulate flowers.
Morphological variations induced by gamma-ray irradiation in chrysanthemum ‘Donglinruixue’. (A) Variations in leaves: (a) original leaves on the control plant; (b) increased leaf crack; (c) leaves missing green colour; (d) deformed leaves; (e) lanceolate leaves; (f) leathery leaves and (g) rolled leaves. (B) Variations in flowers: (a) original flowers of the control plant; (b) reduced number of tubular flowers; (c) the top of the petal split; (d) reduced number of petals; (e) petals widened; (f) edge of flower with tubular flowers and (g) twisted lingulate flowers.

Figure 4

Meiotic behaviour in chrysanthemum ‘Donglinruixue’ as affected by gamma-ray abnormalities. (A) Normal metaphase I; (B) normal anaphase I; (C) normal anaphase II; (D) stickiness of metaphase I; (E) univalents (indicated by arrow); (F) unipolarity at prophase I; (G) unipolarity at metaphase I; (H) anaphase II with laggard (indicated by arrow); (I) anaphase I showing bridge and fragments (indicated by arrow); (J) micronuclei at prophase I (indicated by arrow); (K) stickiness at metaphase with precocious movement (indicated by arrow) and (L) tetrad with micronuclei (indicated by arrow). Bar = 10 μm.
Meiotic behaviour in chrysanthemum ‘Donglinruixue’ as affected by gamma-ray abnormalities. (A) Normal metaphase I; (B) normal anaphase I; (C) normal anaphase II; (D) stickiness of metaphase I; (E) univalents (indicated by arrow); (F) unipolarity at prophase I; (G) unipolarity at metaphase I; (H) anaphase II with laggard (indicated by arrow); (I) anaphase I showing bridge and fragments (indicated by arrow); (J) micronuclei at prophase I (indicated by arrow); (K) stickiness at metaphase with precocious movement (indicated by arrow) and (L) tetrad with micronuclei (indicated by arrow). Bar = 10 μm.

Figure 5

ISSR banding patterns in chrysanthemum ‘Donglinruixue’ by primer ISSR 30.
ISSR banding patterns in chrysanthemum ‘Donglinruixue’ by primer ISSR 30.

Figure 6

Dendrogram representing morphological variations based on similarity coefficients of ISSR data.
Dendrogram representing morphological variations based on similarity coefficients of ISSR data.

The abnormal cells with meiosis aberrations of chrysanthemum ‘Donglinruixue’ plants due to the irradiation effect

Radiation dose (Gy)Total PMC countNumber of cells with aberrationsTotal abnormal cells (%)*
StickinessUnivalentLaggardsBridgesMicronucleiUnipolarity
01,00012048002.40 f
151,02330126243251217.98 e
201,09834217376221621.22 d
251,05452287659262122.70 c
301,00378318979292332.79 b
351,1761023811595393135.71 a

Proximity matrix based on Jaccard’s coefficients

Control15 Gy20 Gy-120 Gy-225 Gy-125 Gy-225 Gy-330 Gy35 Gy-135 Gy-2
Control1.000
15 Gy0.5791.000
20 Gy-10.2920.2931.000
20 Gy-20.6130.4710.3101.000
25 Gy-10.5420.4530.2220.5931.000
25 Gy-20.2780.2330.3060.2300.3001.000
25 Gy-30.3240.3100.2250.1850.2180.4121.000
30 Gy0.4000.2900.2930.2860.3150.2090.2561.000
35 Gy-10.4420.4320.2540.4860.4060.2380.1770.2001.000
35 Gy-20.2920.2500.2970.3330.3750.2700.1950.2620.3621.000

Effect of gamma radiation on morphological variations of chrysanthemum ‘Donglinruixue’ plants

Radiation dose (Gy)Total numberVariations in leaves after 120 days of cultureVariations in flowers after 120 days of culture
Number of variantsRate of variation (%)*Number of variantsRate of variation (%)*
08700.00 f00.00 f
158467.14 e33.57 e
207279.72 d56.25 d
25661319.96 c711.36 c
30601423.33 b812.50 b
35441227.27 a613.81 a

List of primers, number of amplified products, polymorphic bands and polymorphism percentage

Primer namePrimer sequence (5′–3′)No. of amplified bandsNo. of polymorphic bandsPercentage of polymorphic bands
UBC873GACAGACAGACAGACA4375
UBC836AGAGAGAGAGAGAGAGYA5360
UBC857ACACACACACACACACCG3266.66
UBC835AGAGAGAGAGAGAGAGCC66100
UBC855ACACACACACACACACYT9888.89
UBC834AGAGAGAGAGAGAGAGYT66100
ISSR13TCTCTCTCTCTCTCCC99100
ISSR 20ACACACACACACACACGG5480
ISSR 25AGAGAGAGAGAGAGAGGA44100
ISSR 26AGAGAGAGAGAGAGAGGC44100
ISSR 30AGAGAGAGAGAGAGAGCG77100
ISSR 32AGAGAGAGAGAGAGAGAT4375
ISSR 34ACTCACTCACTCACTC6583.33
Total72641,128.88
Mean5.544.9286.84

List of mutants in chrysanthemum ‘Donglinruixue’ used for diversity analysis

Irradiation dose (Gy)Type of mutation
0 GyNo variations
15 GyIncreased leaf crack
20 Gy-1Reduced number of petals
20 Gy-2Leaves missing green colour, increased leaf crack
25 Gy-1Rolled leaves
25 Gy-2Lanceolate leaves, increased tubular flowers
25 Gy-3Lanceolate leaves, decreased number of petals
30 GyIncreased leaf cracks and reduced number of petals
35 Gy-1Rolled leaves
35 Gy-2Plant dwarfing, twisted ligulate flowers
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