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A comparative study of morphological characteristics in diploid and tetraploid (auto and allotetraploids) Citrullus genotypes

Alim Aydin
Alim Aydin
 y 
Halit Yetişir
Halit Yetişir
   | 26 jun 2023
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Article Category: ORIGINAL ARTICLE
Publicado en línea: 26 jun 2023
Páginas: 33 - 48
Recibido: 28 jul 2022
Aceptado: 21 dic 2022
DOI: https://doi.org/10.2478/fhort-2023-0003
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© 2023 Alim Aydin et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

Stomal diameter and length in diploid (A) and tetraploid (B) plant leaves (magnified 400×).
Stomal diameter and length in diploid (A) and tetraploid (B) plant leaves (magnified 400×).

Figure 2

Ploidy level of diploid (A) and tetraploid watermelon (B).
Ploidy level of diploid (A) and tetraploid watermelon (B).

Figure 3

Seedling of diploid (A) CXN5 and tetraploid (B) M1CXN5-1 genotypes, leaf of diploid (C) CXN5 and tetraploid (D) M1CXN5-1 genotypes, male flower of diploid (E) N5 and tetraploid (F) M1N5-1 genotypes and seeds of diploid (G) CXN3 and tetraploid (H) M1CXN3-1 genotypes.
Seedling of diploid (A) CXN5 and tetraploid (B) M1CXN5-1 genotypes, leaf of diploid (C) CXN5 and tetraploid (D) M1CXN5-1 genotypes, male flower of diploid (E) N5 and tetraploid (F) M1N5-1 genotypes and seeds of diploid (G) CXN3 and tetraploid (H) M1CXN3-1 genotypes.

Figure 4

Diploid N7 (A) and tetraploid M1N7-1 (B) genotypes and diploid CXN3 (C) and tetraploid M1-CXN3-1 (D) genotype plants grown in hydroponic culture for 21 days after transplanting.
Diploid N7 (A) and tetraploid M1N7-1 (B) genotypes and diploid CXN3 (C) and tetraploid M1-CXN3-1 (D) genotype plants grown in hydroponic culture for 21 days after transplanting.

Figure 5

PCA of diploid (D) and tetraploid (T) watermelon genotypes based on hydroponic culture morphological features. PCA, principal component analysis; PL, plant length; PSD, plant stem diameter; RD, root diameter; RDW, root dry weight; RFW, root fresh weight; RL, root length; RV, root volume; SDW, shoot dry weight; SFW, shoot fresh weight.
PCA of diploid (D) and tetraploid (T) watermelon genotypes based on hydroponic culture morphological features. PCA, principal component analysis; PL, plant length; PSD, plant stem diameter; RD, root diameter; RDW, root dry weight; RFW, root fresh weight; RL, root length; RV, root volume; SDW, shoot dry weight; SFW, shoot fresh weight.

List, genotype codes and numbers of plant materials applied with colchicine.

Genotype Genotype code Plants used in colchicine application (number)
Calhoun Gray (Citrullus lanatus) C 180
W1482 (Citrullus lanatus var. citroides) N3 210
W1832 (Citrullus lanatus var. citroides) N5 210
W2001 (Citrullus lanatus var. citroides) N7 180
CxW1482 CxN3 210
CxW1832 CxN5 180
CxW2001 CxN7 210
Total 1,380

Number of plants applied, the number of surviving plants and their ratio (%).

Genotype Plants applied (number) Surviving plants (number) Surviving plants (%)
C 180 120 66.67
N3 210 119 56.67
N5 210 110 52.38
N7 180 158 87.78
CXN3 210 116 55.24
CXN5 180 119 66.11
CXN7 210 116 55.24
Total/mean 1,380 858 62.17

RFW, RDW, RL, RV and RD of the plants grown in hydroponic culture for 21 days after transplanting.

Genotype code Ploidy level RFW (g · plant-1) RDW (g · plant-1) RL (cm) RV (cm3 · plant-1) RD (mm)
M1Calhon Gray Tetraploid 9.81 mn 1.15 pr 1,319.16 m 3.31 l–n 0.40 g
M1N3 Tetraploid 10.50 l–n 1.22 o–r 1,702.86 m 1.82 o 0.37 i
M1N5-1 Tetraploid 22.13 fg 2.26 h–k 4,818.78 ef 3.52 k–m 0.44 f
M1N5-2 Tetraploid 22.94 d–f 2.46 f–i 3,570.95 g–k 3.85 kl 0.38 i
M1N5-3 Tetraploid 23.61 d–f 2.53 e–h 5,175.32 de 5.92 gh 0.53 c
M1N5-4 Tetraploid 26.78 c–e 2.73 d–g 3,470.66 h–l 5.25 h–j 0.44 f
M1N5-5 Tetraploid 20.26 f–h 2.20 h–k 3,514.55 h–k 4.32 l 0.40 g
M1N5-6 Tetraploid 23.48 d–f 2.43 f–i 3,660.05 g–j 4.48 i–k 0.58 a
M1N7-1 Tetraploid 13.97 k–m 1.52 n–p 4,046.80 f–i 4.44 i–k 0.37 i
M1N7-2 Tetraploid 14.51 j–l 1.63 m–o 2,764.20 kl 3.95 kl 0.55 b
M1N7-3 Tetraploid 20.65 f–h 2.19 h–k 3,722.38 g–j 5.38 h–j 0.44 f
M1N7-4 Tetraploid 21.48 fg 2.33 g–j 6,083.45 c 2.81 mn 0.37 i
M1CXN3-1 Tetraploid 15.73 i–k 1.68 l–n 2,658.33 l 4.35 j–l 0.46 e
M1CXN3-2 Tetraploid 28.37 c 2.93 de 4,223.48 f–h 6.53 fg 0.46 e
M1CXN3-3 Tetraploid 22.50 fg 2.43 f–i 3,333.78 i–l 6.67 fg 0.51 d
M1CXN3-4 Tetraploid 18.24 g–j 1.93 j–n 2,905.59 j–l 3.79 k–m 0.40 g
M1CXN5-1 Tetraploid 33.02 b 3.41 bc 7,678.59 b 11.09 b 0.44 f
M1CXN7-1 Tetraploid 27.09 cd 2.86 d–f 6,001.24 c 7.40 d–f 0.40 g
M1CXN7-2 Tetraploid 19.91 f–i 2.05 i–m 4,416.07 e–g 4.50 i–k 0.35 j
M1CXN7-3 Tetraploid 20.60 f–h 2.23 h–k 3,658.58 g–j 7.28 ef 0.51 d
M1CXN7-4 Tetraploid 28.94 c 3.04 c–e 4,104.75 f–i 8.21 de 0.51 d
M1CXN7-5 Tetraploid 22.82 d–f 2.45 f–i 5,825.66 cd 8.29 d 0.44 f
Calhoun Gray Diploid 3.16 o 0.48 s 344.85 n 0.78 p 0.44 f
CXN3 Diploid 1.70 o 0.22 s 173.10 n 0.27 p 0.46 e
CXN5 Diploid 19.29 f–i 2.09 h–l 3,645.02 g–j 4.22 l 0.40 g
CXN7 Diploid 16.70 h–k 1.83 k–n 4,428.47 e–g 5.51 hi 0.40 g
N3 Diploid 8.37 n 0.89 r 1,454.43 m 1.82 o 0.40 g
N5 Diploid 13.90 k–n 1.57 n–p 3,591.94 g–k 4.31 l 0.40 g
N7 Diploid 10.77 n 1.16 pr 1,719.21 m 2.44 no 0.44 f
C. tide Citrullus lanatus 1.19 o 0.24 s 148.66 n 0.32 p 0.37 i
RS841 C. maxima × C. moschata 38.17 a 4.00 a 11,098.84 a 13.04 a 0.39 h
Argentario Lagenaria siceraria 34.00 b 3.50 b 11,000.44 a 9.70 c 0.34 k

Mean stomatal diameter, stomatal length, stomatal density and chloroplast number in presumed polyploid and diploid plants.

Stomatal diameter (μm) Stomatal length (μm) Stomatal density (number · mm-2) Chloroplast number (number)
Ploidy level Diploid Tetraploid Diploid Tetraploid Diploid Tetraploid Diploid Tetraploid
Maximum 16.09 20.56 23.30 31.42 627.30 341.75 10.18 16.41
Minimum 18.90 23.87 26.04 34.01 927.77 470.34 12.03 19.37
Mean 17.50 22.22 24.67 32.72 777.54 406.05 11.11 17.89

The number and percentage of tetraploid plants obtained as a result of microscope and flow cytometry analyses.

Genotype Number of tetraploid plants Flow cytometry analysis percentage of tetraploid plants (%)
Stomatal examinations (number) Flow cytometry analysis
C 8 1 12.50
N3 15 1 6.67
N5 11 6 54.55
N7 12 4 33.33
CXN3 14 4 28.57
CXN5 11 1 9.09
CXN7 13 5 38.46
Total/mean 84 22 26.19

Pearson’s correlation coefficients (r values) between DNA content and stomatal measurement in 22 tetraploid and 62 diploid watermelon genotypes.

Stomatal diameter Stomatal length Stomatal density Chloroplast number
DNA content 0.96** 0.90** -0.79** 0.88**

List of genotypes used in hydroponic culture.

Genotype code Ploidy level Genotype code Ploidy level
M1Calhon Gray Tetraploid M1CXN5-1 Tetraploid
M1N3 Tetraploid M1CXN7-1 Tetraploid
M1N5-1 Tetraploid M1CXN7-2 Tetraploid
M1N5-2 Tetraploid M1CXN7-3 Tetraploid
M1N5-3 Tetraploid M1CXN7-4 Tetraploid
M1N5-4 Tetraploid M1CXN7-5 Tetraploid
M1N5-5 Tetraploid Calhoun Gray Diploid
M1N5-6 Tetraploid CXN3 Diploid
M1N7-1 Tetraploid CXN5 Diploid
M1N7-2 Tetraploid CXN7 Diploid
M1N7-3 Tetraploid N3 Diploid
M1N7-4 Tetraploid N5 Diploid
M1CXN3-1 Tetraploid N7 Diploid
M1CXN3-2 Tetraploid Crimson Tide Diploid
M1CXN3-3 Tetraploid RS841 Diploid
M1CXN3-4 Tetraploid Argentario Diploid

PSD, plant height, SFW and SDW of the plants grown in hydroponic culture.

Genotype code Ploidy level PSD (mm) PL (cm) SFW (g · plant-1) SDW (g · plant-1)
M1Calhon Gray Tetraploid 5.82 b–d 30.83 mn 12.58 no 1.42 op
M1N3 Tetraploid 5.61 b–d 35.53 k–m 21.37 k–m 2.30 l–n
M1N5-1 Tetraploid 5.53 b–d 67.40 c–f 49.14 de 4.96 d–f
M1N5-2 Tetraploid 5.57 b–d 67.67 c–f 37.50 f–i 3.92 g–j
M1N5-3 Tetraploid 5.17 b–d 73.67 cd 39.17 f–h 4.08 g–i
M1N5-4 Tetraploid 5.23 b–d 70.33 cd 39.17 f–h 3.97 g–j
M1N5-5 Tetraploid 5.26 b–d 54.00 e–j 30.33 h–j 3.21 i–k
M1N5-6 Tetraploid 4.94 b–d 51.12 f–k 30.83 h–j 3.16 jl
M1N7-1 Tetraploid 5.32 b–d 49.70 g–l 23.33 j–m 2.45 k–n
M1N7-2 Tetraploid 4.88 cd 65.05 c–g 25.00 j–m 2.68 k–m
M1N7-3 Tetraploid 5.19 b–d 64.25 c–h 40.83 e–g 4.21 f–h
M1N7-4 Tetraploid 5.38 b–d 75.65 c 43.33 e–g 4.51 e–h
M1CXN3-1 Tetraploid 6.05 b–d 46.67 i–m 30.83 h–j 3.19 jk
M1CXN3-2 Tetraploid 5.90 b–d 48.18 h–l 58.33 c 5.93 c
M1CXN3-3 Tetraploid 5.77 b–d 67.67 c–f 45.00 e–g 4.68 e–h
M1CXN3-4 Tetraploid 6.13 b–d 52.02 f–k 25.83 j–l 2.69 k–m
M1CXN5-1 Tetraploid 6.48 b–d 45.47 j–m 46.33 ef 4.75 e–g
M1CXN7-1 Tetraploid 6.12 b–d 64.00 c–h 44.67 e–g 4.62 e–h
M1CXN7-2 Tetraploid 5.93 b–d 34.55 l–n 40.17 fg 4.07 g–i
M1CXN7-3 Tetraploid 5.82 b–d 27.67 no 36.35 g–i 3.81 h–j
M1CXN7-4 Tetraploid 6.05 b–d 60.62 c–j 55.00 cd 5.65 cd
M1CXN7-5 Tetraploid 6.53 b–d 62.17 c–i 49.33 de 5.10 d–f
Calhoun Gray Diploid 6.07 b–d 17.83 o–r 5.08 op 0.67 pr
CXN3 Diploid 3.86 d 15.55 o–r 3.60 p 0.41 r
CXN5 Diploid 5.48 b–d 44.67 j–m 30.83 h–j 3.25 i–k
CXN7 Diploid 5.07 b–d 58.52 d–j 29.67 k 3.13 jl
N3 Diploid 4.76 d 26.22 n–p 16.52 mn 1.70 no
N5 Diploid 4.96 b-d 37.23 k–n 20.82 mn 2.26 mn
N7 Diploid 4.11 d 30.83 mn 17.33 l–n 1.81 no
C. tide Citrullus lanatus 2.79 d 11.00 p–r 2.72 p 0.39 r
RS841 C.maxima × C. moschata 9.73 a 95.83 b 84.00 a 8.58 a
Argentario Lagenaria siceraria 9.62 ab 115.83 a 73.33 b 7.43 b

Pearson’s correlation coefficients (r values) between plant growth parameters and direct (DNA content) and indirect (stomatal measurement) ploidy determination methods in 22 tetraploid and seven diploid watermelon genotypes.

PSD PL SFW SDW RFW RDW RL RV RD
DNA content 0.33* 0.48* 0.60** 0.60** 0.60** 0.61** 0.45* 0.47* -0.02ns
Stomatal diameter –0.15ns –0.49* –0.56** –0.56** -0.58** -0.58** -0.53** -0.44* 0.26ns
Stomatal length 0.26ns 0.50** 0.49* 0.49* 0.52* 0.53* 0.38* 0.32* -0.15ns
Stomatal density 0.30ns 0.50** 0.55** 0.55** 0.59** 0.59** 0.43* 0.40* -0.12ns
Chloroplast number –0.19ns 0.19ns –0.10 ns –0.10 ns 0.02ns 0.01ns -0.05ns -0.24ns -0.26ns

Thousand seed weights, germination rate, cotyledon width, cotyledon length, stem diameter and % change in genome size and 1,000 seed weights.

  Ploidy level % Change
Tetraploid Diploid
1,000 seed weight (g) 162.67 144.12 11.40
Germination rate (%) 65.8 94.8 -30.59
Cotyledon width (mm) 27.92 25.89 7.76
Cotyledon length (mm) 39.99 35.53 12.58
Hypocotyl diameter (mm) 3.14 2.57 20.62
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