Journal of Plant Pathology

FIRST REPORT of Diaporthe foeniculina CAUSING FRUIT ROT of LEMON in

TURKEY
--Manuscript Draft--

Manuscript Number: JPPY-D-19-00199R1

Full Title: FIRST REPORT of Diaporthe foeniculina CAUSING FRUIT ROT of LEMON in
TURKEY

Article Type: Disease Note

Abstract: Diaporthe spp. are pathogens, endophytes or saprobes on ornamentals, forest trees
and economically important crops, causing different symptoms such as dieback,
canker, leaf-spot, blight, fruit rot or wilt (Marin-Felix et al. 2019). During November
2018, a post harvest fruit rot was observed on lemon fruit (Citrus limon) in Mersin
Province, south Turkey, with an incidence of 10% of the fruit. Brown and rough decay
were present on fruit. Fungal colonies were obtained from single-spore isolation on
potato dextrose agar (PDA). Colonies appeared first white, turning creamy to Brown,
with black, globose conidiomata. Beta conidia were smooth, hyaline, filiform, base
truncate, straight or curved. To confirm the identity, the internal transcribed spacer
(ITS) region of the rDNA, was amplified and sequenced. The representative isolate (ET
88, GenBank accession no MK656290) showed 100 % of similarity with Diaporthe
foeniculina CBS 111553 (NR145303).
Pathogenicity tests were performed on healthy lemon fruit which were washed under
tap water and surface sterilized with 70% ethanol, then inoculated at the center with a
5 mm plugs obtained from 5-day old mycelial culture of the strain ET 88 grown on PDA
at 26° C for 7 days. Control fruit were inoculated with PDA plugs. The fruit were kept in
plastic containers with wet filter paper and incubated at room temperature for 10 days.
After 5 days, the symptoms appeared on inoculated lemons. Control fruit remained
healthy. The fungus was re-isolated only from diseased fruit, fulfilling Koch’s
postulates. Diaporthe foeniculina has been detected in Europe, USA and New Zeland,
causing various symptoms on Citrus spp. (Guarnaccia and Crous 2017). To our
knowledge, this is the first report of D. foeniculina causing lemon fruit rot in Turkey.

Response to Reviewers: Dear Editor,

We checked our MS and submited our revised manuscript in your Journal System.
Thank you very much for your interest.

Sincerely yours,

Elif Tozlu

Atatürk University Faculty of Agriculture

Department of Plant Protection
25240 Erzurum/TURKEY
Tel: +90 442 231 20 70
Fax: +90 442 231 14 69
e-mail: elifalpertozlu@atauni.edu.tr

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Manuscript Click here to access/download;Manuscript;Tekiner et al.,
Diaporthe first report.docx
Click here to view linked References

FIRST REPORT of Diaporthe foeniculina CAUSING FRUIT ROT of LEMON

1 in TURKEY
2
3 Nasibe Tekiner1 Elif Tozlu1 Vladimiro Guarnaccia2
4
1
5 Department of Plant Pathology, Faculty of Agriculture, Atatürk University, 25240, Erzurum, Turkey,
6 2
7 DiSAFA, University of Torino, Largo Paolo Braccini, 2, 10095 Grugliasco, TO, Italy.
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10 CORRESPONDING AUTHOR
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12 Elif Tozlu
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14 E-mail: elifalpertozlu@atauni.edu.tr
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16 Nasibe Tekiner ORCID: http://orcid.org/0000-0003-2396-7786
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18 Elif Tozlu ORCID: http://orcid.org/0000-0002-0016-9696
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20 Vladimiro Guarnaccia ORCID: http://orcid.org/0000-0003-3188-7743
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 Abstract
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2
Diaporthe spp. are known as important pathogens, endophytes or saprobes on economical important crops,
3
4
ornementals and forest trees (Marin-Felix et al. 2019), causing different symptoms such as dieback, cankers,
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6
leaf-spots, blights, fruit rot or wilt (Gomes et al. 2013). During November 2018, a post harvest fruit rot was
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observed on lemon fruit (Citrus limon) in Mersin Province, south Turkey. The prevalence of the fruit rot in the
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11 fruits was estimated at 10% incidence. Brown and rough decay, and discolouration were present on lemon fruit
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13 surface. Several fragments taken from the margin of infected area were cultured on potato dextrose agar (PDA).
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15 Fungal colonies were purified and incubated on PDA, at 25 ± 1°C (14-h fluorescent light, 10-h dark). Colonies
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17 appeared first white and later turned creamy with brown pigmentation. Conidiomata globose, black and beta
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19 conidia smooth, hyaline, filiform, base truncate, straight or curved. To confirm the identity at the molecular
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21 level, the internal transcribed spacer (ITS) region of the rDNA, was amplified and sequenced using primer sets
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23 ITS1/ITS4 (White et al. 1990). The represantative isolate (ET 88, GenBank accession no MK656290) showed
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25 100 % of similarity with Diaporthe foeniculina CBS 111553 (NR145303).
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Pathogenicity tests were performed on healthy lemon fruit which were washed under tap water and then surface
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sterilized with 70% ethanol and inoculated at the center with a 5 mm plugs obtained from 5-day old mycelial
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32 culture of the strain “ET 88” grown on PDA at 26°C for 7 days. The inoculated lemons were kept in closed
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34 plastic containers with wet filter paper and incubated at room temperature for 10 days. Control fruit were
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36 inoculated with PDA plugs. After 5 days, the first symptoms started on inoculated lemons, while control fruit
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38 remained healthy. The fungal pathogen was re-isolated from the disease lesions on the inoculated fruit. This re-
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40 isolated pathogen exhibited the same morphological characteristics as those of the original isolates, so Kochs
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42 postulates were completed.
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Previously, D. foeniculina has been recently detected in Europe, USA and New Zeland, causing various
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symptoms on Citrus spp. (Guarnaccia and Crous 2017). To our knowledge, this is the first report of D.
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foeniculina causing lemon fruit rot in Turkey.
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51 Keywords Phomopsis, Citrus, Molecular identification, Pathogenicity
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References:
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56 Gomes RR, Glienke C, Videira SIR, Lombard L, Groenewald JZ,, Crous PW (2013) Diaporthe: a genus of
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58 endophytic, saprobic and plant pathogenic fungi. Persoonia 31:1–41
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 Guarnaccia V, Crous PW (2017) Emerging citrus diseases in Europe caused by species of Diaporthe. IMA
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2 Fungus 8:317–334
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5 Marin-Felix Y, Hernández-Restrepo M, Wingfield MJ, Akulov A, Carnegie AJ, Cheewangkoon R, Gramaje D,
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7 Groenewald JZ, Guarnaccia V, Halleen F, Lombard L, Luangsaard J, Marincowitz S, Moslemi A,
8
9 Mostert L, Quaedvlieg W, Schumacher RK, Spies CFJ, Thangavel R, Taylor PWJ, Wilson AM,
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11 Wingfield BD, Wood AR, Crous PW (2019) Genera of phytopathogenic fungi: GOPHY2. Studies in
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13 Mycology 92:47–133
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16 White TJ, Bruns T, Lee S, et al. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for
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18 phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, et al. (eds), PCR Protocols: a guide to methods
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20 and applications: 315–322. Academic Press, San Diego, California.
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revise Manuscript Click here to access/download;Manuscript;Tekiner et al.,
Diaporthe first report, revise manuscript.docx
Click here to view linked References

FIRST REPORT of Diaporthe foeniculina CAUSING FRUIT ROT of LEMON

1 in TURKEY
2
3 Nasibe Tekiner1 Elif Tozlu1 Vladimiro Guarnaccia2
4
1
5 Department of Plant Pathology, Faculty of Agriculture, Atatürk University, 25240, Erzurum, Turkey,
6 2
7 DiSAFA, University of Torino, Largo Paolo Braccini, 2, 10095 Grugliasco, TO, Italy.
8
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10 CORRESPONDING AUTHOR
11
12 Elif Tozlu
13
14 E-mail: elifalpertozlu@atauni.edu.tr
15
16 Nasibe Tekiner ORCID: http://orcid.org/0000-0003-2396-7786
17
18 Elif Tozlu ORCID: http://orcid.org/0000-0002-0016-9696
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20 Vladimiro Guarnaccia ORCID: http://orcid.org/0000-0003-3188-7743
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 Abstract
1
2 Diaporthe spp. are pathogens, endophytes or saprobes on ornamentals, forest trees and economically important
3
4 crops, causing different symptoms such as dieback, canker, leaf-spot, blight, fruit rot or wilt (Marin-Felix et al.
5
6 2019). During November 2018, a post harvest fruit rot was observed on lemon fruit (Citrus limon) in Mersin
7
8 Province, south Turkey, with an incidence of 10% of the fruit. Brown and rough decay were present on fruit.
9
10 Fungal colonies were obtained from single-spore isolation on potato dextrose agar (PDA). Colonies appeared
11
12 first white, turning creamy to Brown, with black, globose conidiomata. Beta conidia were smooth, hyaline,
13
14 filiform, base truncate, straight or curved. To confirm the identity, the internal transcribed spacer (ITS) region of
15
16 the rDNA, was amplified and sequenced. The representative isolate (ET 88, GenBank accession no MK656290)
17
18 showed 100 % of similarity with Diaporthe foeniculina CBS 111553 (NR145303).
19
20 Pathogenicity tests were performed on healthy lemon fruit which were washed under tap water and surface
21
22 sterilized with 70% ethanol, then inoculated at the center with a 5 mm plugs obtained from 5-day old mycelial
23
24 culture of the strain ET 88 grown on PDA at 26° C for 7 days. Control fruit were inoculated with PDA plugs.
25
26 The fruit were kept in plastic containers with wet filter paper and incubated at room temperature for 10 days.
27
28 After 5 days, the symptoms appeared on inoculated lemons. Control fruit remained healthy. The fungus was re-
29
30 isolated only from diseased fruit, fulfilling Koch’s postulates. Diaporthe foeniculina has been detected in
31
32 Europe, USA and New Zeland, causing various symptoms on Citrus spp. (Guarnaccia and Crous 2017). To our
33
34 knowledge, this is the first report of D. foeniculina causing lemon fruit rot in Turkey.
35
36 Keywords Phomopsis, Citrus, Molecular identification, Pathogenicity
37
38
References:
39
40
41 Guarnaccia V, Crous PW (2017) Emerging citrus diseases in Europe caused by species of Diaporthe. IMA
42
43 Fungus 8:317–334
44
45 Marin-Felix Y, Hernández-Restrepo M, Wingfield MJ, Akulov A, Carnegie AJ, Cheewangkoon R, Gramaje D,
46
47 Groenewald JZ, Guarnaccia V, Halleen F, Lombard L, Luangsaard J, Marincowitz S, Moslemi A,
48
49 Mostert L, Quaedvlieg W, Schumacher RK, Spies CFJ, Thangavel R, Taylor PWJ, Wilson AM,
50
51 Wingfield BD, Wood AR, Crous PW (2019) Genera of phytopathogenic fungi: GOPHY2. Studies in
52
53 Mycology 92:47–133
54
55
56
57
58
59
60
61
62
63
64
65