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Genetic intraspecific diversity of Meloidogyne javanica parasitizing vegetables in southern Iran

Reza Ghaderi
Reza Ghaderi
, 
Ali Asghar Dehghan
Ali Asghar Dehghan
, 
Abbas Mokaram Hesar
Abbas Mokaram Hesar
 oraz 
Akbar Karegar
Akbar Karegar
   | 23 kwi 2020
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Data publikacji: 23 kwi 2020
Zakres stron: 1 - 13
Otrzymano: 30 paź 2019
DOI: https://doi.org/10.21307/jofnem-2020-035
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© 2020 Reza Ghaderi et al., published by Sciendo.
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Figure 1:

Species-specific 670 bp band in the populations of Meloidogyne javanica recovered from southern Iran. (For population codes, see Table 1).
Species-specific 670 bp band in the populations of Meloidogyne javanica recovered from southern Iran. (For population codes, see Table 1).

Figure 2:

Species-specific 1200 bp band in the populations of Meloidogyne incognita recovered from southern Iran. (For population codes, see Table 1).
Species-specific 1200 bp band in the populations of Meloidogyne incognita recovered from southern Iran. (For population codes, see Table 1).

Figure 3:

RAPD (left) and ISSR (right) patterns for 19 populations of Meloidogyne javanica generated with the primer (CCA)5 and OPAD10. DNA Ladder = 200 bp. (For population codes, see Table 1).
RAPD (left) and ISSR (right) patterns for 19 populations of Meloidogyne javanica generated with the primer (CCA)5 and OPAD10. DNA Ladder = 200 bp. (For population codes, see Table 1).

Figure 4:

UPGMA cluster diagram based on Dice coefficient estimated from 180 RAPD markers for 37 populations of Meloidogyne javanica (For population codes, see Table 1).
UPGMA cluster diagram based on Dice coefficient estimated from 180 RAPD markers for 37 populations of Meloidogyne javanica (For population codes, see Table 1).

Figure 5:

UPGMA cluster diagram based on Dice coefficient estimated from 164 ISSR markers for 37 populations of the Meloidogyne javanica (For population codes, see Table 1).
UPGMA cluster diagram based on Dice coefficient estimated from 164 ISSR markers for 37 populations of the Meloidogyne javanica (For population codes, see Table 1).

Figure 6:

A Mantel test showing correlation between RAPD and ISSR datasets of the populations of Meloidogyne javanica recovered from southern Iran.
A Mantel test showing correlation between RAPD and ISSR datasets of the populations of Meloidogyne javanica recovered from southern Iran.

Average of the morphometrics of females, J2s and males of five populations of Meloidogyne javanica parasitizing vegetables in southern Iran.

Females Males Second-stage juveniles (J2s)†
Diagnostic characters 15 CV 10 CV MJ1 MJ4 MJ8 MJ14 MJ15 50 CV
L (including neck) 686±127 (506-1,010) 18 1482±155 (1189-1,694) 10 463 457 449 482 469 464±19 (429-497) 4
Body width (W) 420±99 (274-568) 23 33±2.8 (29-38) 9 14 14 15 15 16 15±1.5 (12-18) 9
Neck length 259±87 (153-540) 33 − − − − − − − − −
Stylet 15±2 (13-18) 14 18±1.2 (16-19) 6 9.7 10.4 10.5 10.1 10.2 10±0.5 (9-12) 5
DGO 3.1±0.6 (2.0-3.9) 20 3.3±0.2 (3.1-3.5) 5 2.7 2.1 2.2 1.9 2.0 2.2±0.5 (1.5-3.7) 22
Excretory pore 190±30 (145-270) 16 − − − − − − − − −
Vulval slit length 21±3 (15-27) 15 − − − − − − − − −
Vulva-anus distance 16±4 (12-28) 25 − − − − − − − − −
Inter-phasmidial distance 78 ± 16 (58-108) 20 − − − − − − − − −
Tail length − − 14±2.1 (10-17) 15 51 55 55 58 57 55±5 (40-66) 9
Hyaline length − − − − 11 12 11 16 16 13±4 (7-20) 27
Pharynx (glands end) − − 473±50 (390-552) 11 164 202 197 201 201 193±23 (90-216) 12
Body length to width (a) 1.7±0.4 (1.2-2.6) 22 45±4.5 (37-51) 10 38 32 29 33 30 31±3 (25-39) 10
b − − 6.6±1.3 (4.8-8.1) 20 6.4 4.9 4.7 5.0 5.2 5.2±1.4 (4.3-12.4) 27
b' − − 3.1±0.6 (2.4-3.9) 18 3.0 2.3 2.3 2.4 2.3 2.5±0.5 (2.0-5.2) 22
c − − 113±26 (87-169) 23 9.4 8.4 8.2 8.3 8.2 8.5±0.8 (7.2-11.3) 10
c' − − 0.5±0.1 (0.3-1.0) 19 4.6 5.3 5.5 5.9 5.2 5.7±0.5 (4.7-7.2) 10
Spicules − − 23±5.8 (15-29) 24 − − − − − − −
Gubernaculum − − 13±2.8 (8-17) 20 − − − − − − −

Nei’s (1978) unbiased genetic distance between pairs of Meloidogyne javanica populations detected by RAPD and ISSR analyses.

Population† Abdan Deir Bardkhon Jiroft Beiza Khafr Kazeron
Abdan – 0.032 0.070 0.074 0.091 0.084 0.088
Deir 0.082 – 0.054 0.076 0.068 0.088 0.091
Bardkhon 0.090 0.076 – 0.046 0.045 0.039 0.059
Jiroft 0.069 0.127 0.119 – 0.071 0.040 0.025
Beiza 0.043 0.080 0.119 0.108 – 0.073 0.071
Khafr 0.080 0.055 0.143 0.134 0.075 – 0.051
Kazeron 0.120 0.071 0.050 0.156 0.132 0.162 –

Analysis of molecular variance (AMOVA) of Meloidogyne javanica populations based on RAPD and ISSR data.

Marker Source of variation Degree of freedom Sum of squares Variance components Percentage of variation Fixation index (F ST )† Number of migrants (Nm)
Between populations 6 148.88 2.63* 19
RAPD Within populations 30 340.85 11.36* 81 0.188 1.079
Total 36 489.73 13.99
Between populations 6 90.68 1.34* 14
ISSR Within populations 30 247.80 8.26* 86 0.140 1.535
Total 36 338.49 9.60

Genetic diversity estimates of Meloidogyne javanica populations recovered from southern Iran by RAPD and ISSR analyses.

ISSR RAPD
Population %P† H I %P H I
Abdan (10)†† 25.61 0.110 0.158 36.11 0.150 0.218
Deir (5) 20.73 0.092 0.130 30.00 0.132 0.189
Bardkhon (7) 24.39 0.097 0.141 30.56 0.118 0.173
Jiroft (3) 10.37 0.039 0.058 17.78 0.079 0.113
Beiza (5) 21.34 0.088 0.128 22.78 0.095 0.137
Khafr (4) 18.29 0.076 0.110 16.11 0.064 0.093
Kazeron (3) 12.80 0.057 0.081 18.89 0.079 0.114
Mean 19.08 0.080 0.115 24.60 0.103 0.148

Populations of Meloidogyne species collected in this study.

Number Sampling area N E Sample Host plant Molecular code
1 Boshehr province-Abdan 28°11´00.38˝ 51°74´07.94˝ Soil and root Tomato MJ1
2 Boshehr province-Abdan 28°10´99.86˝ 51°74´08.46˝ Soil and root Tomato MJ2
3 Boshehr province-Abdan 28°11´00.58˝ 51°74´07.97˝ Soil and root Tomato MJ3
4 Boshehr province-Abdan 28°11´00.29˝ 51°74´08.27˝ Soil and root Tomato MJ4
5 Boshehr province-Abdan 28°11´00.31˝ 51°74´08.28˝ Soil and root Tomato MJ5
6 Boshehr province-Abdan 28°10´95.09˝ 51°74´04.66˝ Soil and root Tomato MJ6
7 Boshehr province-Abdan 28°10´94.93˝ 51°74´04.44˝ Soil and root Tomato MJ7
8 Boshehr province-Abdan 28°10´94.82˝ 51°74´04.43˝ Soil and root Tomato MJ8
9 Boshehr province-Abdan 28°10´94.56˝ 51°74´03.85˝ Soil and root Tomato MJ9
10 Boshehr province-Abdan 28°10´94.47˝ 51°74´03.81˝ Soil and root Tomato MJ10
11 Boshehr province-Deir Port 27°91´15.43˝ 51°98´02.27˝ Soil and root Tomato MJ11
12 Boshehr province-Deir Port 27°91´15.16˝ 51°98´02.71˝ Soil and root Tomato MJ12
13 Boshehr province-Deir Port 27°91´17.39˝ 51°98´03.58˝ Soil and root Tomato MJ13
14 Boshehr province-Deir Port 27°91´17.33˝ 51°98´03.47˝ Soil and root Tomato MJ14
15 Boshehr province-Deir Port 27°91´44.84˝ 51°98´43.36˝ Soil and root Tomato MJ15
16 Boshehr province-Bardkhon 28°07´54.24˝ 51°48´89.26˝ Soil and root Tomato MJ16
17 Boshehr province-Bardkhon 28°07´51.78˝ 51°48´90.21˝ Soil and root Tomato MJ17
18 Boshehr province-Bardkhon 28°36´13.93˝ 51°48´90.22˝ Soil and root Tomato MJ18
19 Boshehr province-Bardkhon 28°07´51.25˝ 51°48´84.44˝ Soil and root Tomato MJ19
20 Boshehr province-Bardkhon 28°07´51.92˝ 51°48´82.48˝ Soil and root Tomato MJ20
21 Boshehr province-Bardkhon 28°07´52.13˝ 51°48´82.42˝ Soil and root Tomato MJ21
22 Boshehr province-Bardkhon 28°07´52.15˝ 51°48´82.19˝ Soil and root Tomato MJ22
23 Kerman province-Jiroft 28°41´09.00˝ 57°40´13.81˝ Root Mallow MJ23
24 Kerman province-Jiroft 28°41´35.08˝ 57°39´25.18˝ Root Goosefoot MJ24
25 Kerman province-Jiroft 28°42´22.57˝ 57°43´05.86˝ Root Eggplant MJ25
26 Fars province-Beyza 29°55´30.53˝ 52°26´28.41˝ Soil and root Cucumber MJ26
27 Fars province-Beyza 29°55´51.85˝ 52°26´44.88˝ Soil and root Cucumber MJ27
28 Fars province-Beyza 29°55´59.49˝ 52°26´51.00˝ Soil and root Cucumber MJ28
29 Fars province-Beyza 29°55´16.00˝ 52°27´20.15˝ Soil and root Cucumber MJ29
30 Fars province-Beyza 29°55´11.72˝ 52°27´27.93˝ Soil and root Cucumber MJ30
31 Fars province-khafr 28°57´44.57˝ 53°11´58.46˝ Soil and root Okra MJ31
32 Fars province-khafr 28°57´35.80˝ 53°12´32.31˝ Soil and root Okra MJ32
33 Fars province-khafr 28°57´34.97˝ 53°12´28.38˝ Soil and root Okra MJ33
34 Fars province-khafr 28°58´56.08˝ 53°11´47.87˝ Soil and root Okra MJ34
35 Fars province-Kazeron 29°33´23.23˝ 51°45´56.27˝ Soil and root Tomato MJ35
36 Fars province-Kazeron 29°33´25.22˝ 51°45´42.76˝ Soil and root Tomato MJ36
37 Fars province-Kazeron 29°33´42.68˝ 51°45´08.64˝ Soil and root Tomato MJ37
38 Fars province-Fasa 29°00´46.56˝ 53°37´42.27˝ Soil and root Cucumber MI1
39 Fars province-Fasa 29°00´46.56˝ 53°37´42.27˝ Soil and root Cucumber MI2
40 Fars province-Fasa 29°00´46.56˝ 53°37´42.27˝ Soil and root Cucumber MI3
41 Fars province-Fasa 29°00´46.56˝ 53°37´42.27˝ Soil and root Cucumber MI4

All primers used for ISSR, RAPD and species-specific identification of Meloidogyne species during the present study.

ISSR RAPD
(CCA)5 5´-CCACCACCACCACCA-3' OPA8 5’-GTGACGTAGG-3'
(GTG)6 5'-GTGGTGGTGGTGGTGGTG-3' OPA10 5'-GTGATCGCAG-3'
(ATG)6 5'-ATGATGATGATGATGATG-3' OPA13 5'-CAGCACCCAC-3'
(GAG)4GC 5'-GAGGAGGAGGAGGC-3' OPAD10 5'-AAGAGGCCAG-3'
(GACA)4 5'-GACAGACAGACAGACA-3' OPE18 5'-GGACTGCAGA-3'
(GA)8C 5'-GAGAGAGAGAGAGAGAC-3' OPP17 5'-TGACCCGCCT-3'
(AC)8T 5'-ACACACACACACACACT-3' OPB11 5'-GTAGACCCGT-3'
(CA)8G 5'-CACACACACACACACAG-3' OPB17 5'-AGGGAACGAG-3'
(CTC)4GC 5'-CTCCTCCTCCTCGC-3' OPD8 5'-GTGTGCCCCA-3'
(GAGA)4GG 5'-GAGAGAGAGAGAGAGAGG-3' OPC8 5'-TGGACCGGTG-3'
Species-specific primers
M. javanica Fjav (5'-GGTGCGCGATTGAACTGAGC-3') M. incognita Finc (5'-CTCTGCCCAATGAGCTGTCC-3')
Rjav (5'-CAGGCCCTTCAGTGGAACTATAC-3') Rinc (5'-CTCTGCCCTCACATTAAG-3')
M. arenaria Far (5'-TCGGCGATAGAGGTAAATGAC-3') M. hapla F (5'- TGACGGCGGTGAGTGCGA-3')
Rar (5'-TCGGCGATAGACACTACAAACT-3') R (5'-TGACGGCGGTACCTCATAG-3')
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