MORPHOLOGICAL, MOLECULAR AND PATHOGENIC CHARACTERIZATION OF <i>Colletotrichum gloeosporioides</i> ISOLATED FROM MANGO

SILVA NETO, JORGE ALVES DA; AMBRÓSIO, MÁRCIA MICHELLE DE QUEIROZ; ARAÚJO, MARIA BRUNA MEDEIROS; SILVA, ROSECLEIDE MAIA DA; PINTO, PEDRO SIDARQUE LIMA; HOLANDA, IONÁ SANTOS ARAÚJO

doi:10.1590/1983-21252022v35n302rc

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Agronomy • Rev. Caatinga 35 (3) • Jul-Sep 2022 • https://doi.org/10.1590/1983-21252022v35n302rc copy

MORPHOLOGICAL, MOLECULAR AND PATHOGENIC CHARACTERIZATION OF Colletotrichum gloeosporioides ISOLATED FROM MANGO

CARACTERIZAÇÃO MORFOLÓGICA, MOLECULAR E PATOGÊNICA DE Colletotrichum gloeosporioides ISOLADO DE MANGA

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT

Mango (Mangifera indica L.) is a tropical fruit of great importance in the world. However, fruit postharvest losses are caused by anthracnose, a disease caused mainly by phytopathogenic fungi from the Colletotrichum genus. Thus, this work aimed to study morphology, genetic similarity, and pathogenicity of Colletotrichum spp. isolates from mango in Rio Grande do Norte state, Brazil. Morphological evaluation of 28 isolates, based on colony color, size of conidia and appressoria, and growth rate, showed that the isolates belong to the C. gloeosporioides species. It was later confirmed with species-specific oligonucleotides. Also, combined evaluation with RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter Simple Sequence Repeats) showed a genetic similarity of 0.23 between Isolates 12 and 21, and 0.71 between Isolates 2 and 4, an indication of high variability among isolates, with dissimilarity over 75%. Moreover, the dendrogram generated by the UPGMA method indicated that the isolates were arranged in eight groups, which did not correlate with the collection site. Based on pathogenicity, although most isolates showed low virulence for Tommy Atkins cultivar, some of them proved to be highly aggressive (Isolates 9, 15, 20, 21, 24, 26, and 27). Also, pathogenicity did not correlate with clusters. In conclusion, the results revealed high genetic and pathogenic variability in C. gloeosporioides isolated from mango cultivars in one of the main exporting regions in Brazil.

Keywords:
Anthracnose; Genetical diversity; Pathogenicity; Phytopathogens

Farm	Plant	Isolate	Plant tissue	Cultivar	City
Farm 1	Plant 1	1	Fruit	Palmer	Ipanguaçu
	Plant 2	2	Fruit	Palmer	Ipanguaçu
	Plant 3	3	Fruit	Palmer	Ipanguaçu
	Plant 4	4	Fruit	Palmer	Ipanguaçu
		5	Leaf	Palmer	Ipanguaçu
	Plant 5	6	Fruit	Palmer	Ipanguaçu
Farm 2	Plant 6	7	Fruit	Tommy Atkins	Ipanguaçu
	Plant 7	8	Fruit	Tommy Atkins	Ipanguaçu
Farm 3	Plant 8	9	Leaf	Palmer	Ipanguaçu
	Plant 9	10	Fruit	Palmer	Ipanguaçu
		11	Leaf	Palmer	Ipanguaçu
	Plant 10	12	Fruit	Palmer	Ipanguaçu
	Plant 11	13	Fruit	Palmer	Ipanguaçu
	Plant 12	14	Fruit	Palmer	Ipanguaçu
	Plant 13	15	Fruit	Palmer	Ipanguaçu
		16	Leaf	Palmer	Ipanguaçu
	Plant 14	17	Fruit	Palmer	Ipanguaçu
	Plant 15	18	Fruit	Palmer	Ipanguaçu
		19	Leaf	Palmer	Ipanguaçu
	Plant 16	20	Leaf	Palmer	Ipanguaçu
Farm 4	Plant 17	21	Leaf	Tommy Atkins	Pendências
	Plant 18	22	Leaf	Tommy Atkins	Pendências
	Plant 19	23	Leaf	Tommy Atkins	Pendências
	Plant 20	24	Fruit	Tommy Atkins	Pendências
		25	Leaf	Tommy Atkins	Pendências
	Plant 21	26	Fruit	Tommy Atkins	Pendências
	Plant 22	27	Leaf	Tommy Atkins	Pendências
Farm 5	Plant 23	28	Fruit	Palmer	Assú

Primer number	Primer code	Primer sequence (5’ → 3’)	Annealing temperature
ISSR primer
1	DiGA5’CR	CRGAGAGAGAGAGAGAGA	50
2	TriCAC’RC	CACCACCACCACCACRC	55
3	TriGTG	GTGGTGGTGGTGGTG	55
4	TriGTG3’yC	GTGGTGGTGGTGGTGyC	52
5	TriGTG5’Cy	CyGTGGTGGTGGTGGTG	55
6	TriTGT5’Cy	CyTGTTGTTGTTGTTGT	50
7	TriTCA3’RC	TCATCATCATCATCARC	50
8	TriTCC3’RC	TCCTCCTCCTCCTCCRC	50
9	TriCGA3’RC	CGACGACGACGACGARC	45
RAPD primer
1	OPD 02	GGACCCAACC	40
2	OPD 03	GTCGCCGTCA	40
3	OPD 07	TTGGCACGGG	40
4	OPD 08	GTGTGCCCCA	40
5	OPD 13	GGGGTGACGA	40
6	OPD 18	GAGAGCCAAC	40
7	OPD 20	ACCCGGTCAC	40
8	OPM 01	GTTGGTGGCT	40
9	OPM 02	ACAACGCCTC	40
10	OPM 12	GGGACGTTGG	40
11	OPM 15	GACCTACCAC	40
12	OPM 20	AGGTCTTGGG	40

Isolate	Presence of setae	Growth rate (mm day¹)	Conidium			Appressorium
Isolate	Presence of setae	Growth rate (mm day¹)	Width (μm)	Length (μm)	Most found shape	Width (μm)	Length (μm)	Most found shape
1	−	15.47±0.32 e	3.85±0.32 c	11.23±0.86 f	1	6.21±0.78 e	8.62±1.17 e	3
2	+	13.93±0.19 d	3.54±0.27 b	11.34±1.06 f	1	6.34±0.70 f	9.10±1.08 f	2
3	−	14.53±0.24 d	4.55±0.37 f	7.75±0.87 a	1	6.15±1.43 e	8.19±1.29 d	3
4	−	13.20±0.22 c	4.17±0.28 e	11.26±0.97 f	1	6.32±0.70 f	8.21±0.95 d	3
5	−	13.93±0.27 d	3.95±0.29 e	12.17±0.69 h	3	5.13±0.44 a	7.39±0.88 a	3
6	−	13.73±0.26 d	2.82±0.43 a	9.46±1.40 c	1	6.03±0.75 d	8.08±1.19 c	3
7	−	12.40±0.30 b	2.93±0.37 b	10.91±0.59 e	1	5.27±0.67 b	7.41±1.07 a	3
8	−	11.20±0.15 b	2.46±0.45 a	8.32±1.12 b	3	5.88±0.77 d	8.31±0.81 d	2
9	−	13.40±0.16 c	3.54±0.28 c	11.20±0.51 e	1	6.10±0.69 d	7.91±1.30 c	2
10	−	15.60±0.22 e	2.10±0.39 a	10.98±1.27 e	1	5.78±0.79 c	7.73±1.11 b	2
11	−	13.73±0.16 d	3.52±0.32 b	11.86±0.84 g	1	4.89±0.80 a	7.48±1.29 a	2
12	−	14.20±0.26 d	3.65±0.34 c	11.63±1.36 f	1	6.54±0.87 f	8.43±0.97 d	2
13	−	14.33±0.14 d	2.89±0.59 b	9.89±1.23 c	1	5.17±0.75 a	7.13±1.20 a	2
14	+	17.73±0.38 g	3.50±0.23 b	10.52±0.57 d	1	5.56±0.68 b	7.41±0.83 a	3
15	−	14.07±0.23 d	4.03±0.19 e	11.73±0.55 g	1	5.32±0.80 b	7.72±1.17 b	3
16	−	17.40±0.30 g	4.19±0.34 e	12.20±0.62 h	3	6.24±0.64 e	9.30±1.32 g	3
17	+	16.07±0.19 e	4.09±0.37 e	11.67±0.76 g	1	6.09±0.66 d	8.24±0.95 d	3
18	+	14.07±0.22 d	2.94±0.52 b	11.15±1.29 e	1	5.86±0.50 c	8.13±0.68 d	2
19	+	16.27±0.13 f	4.27±0.40 f	12.79±0.84 i	1	6.87±0.59 g	9.60±0.88 h	3
20	+	11.33±0.14 a	4.29±0.58 f	14.28±2.62 j	1	5.90±0.74 d	8.62±1.06 e	2
21	+	15.53±0.25 e	4.32±0.33 f	12.28±0.67 h	1	7.05±0.76 g	9.90±1.01 h	3
22	+	15.13±0.23 e	3.98±0.21 e	11.83±1.48 g	1	5.74±0.46 c	7.92±0.78 c	3
23	−	14.80±0.29 e	3.88±0.26 d	11.31±0.75 f	1	5.67±0.45 c	7.91±0.81 c	3
24	−	12.87±0.14 c	3.65±0.38 c	12.29±0.74 h	1	5.85±0.62 c	8.91±0.95 f	3
25	−	15.40±0.38 e	3.92±0.25 d	13.04±0.98 i	1	5.44±0.50 b	8.16±0.86 d	3
26	−	11.13±0.27 a	3.88±0.23 d	11.68±0.69 g	1	5.21±0.81 b	7.82±1.25 c	3
27	−	16.00±0.36 f	4.26±0.54 f	10.85±1.18 d	1	5.12±0.90 a	7.34±1.92 a	3
28	−	16.47±0.08 f	4.02±0.29 d	12.45±0.85 h	1	6.30±0.79 e	9.59±1.84 g	3

Primer	TL	ML	PL	%P	PIC
RAPD primers
OPD 02	10	1	9	90	0.387
OPD 03	8	2	6	75	0.356
OPD 07	9	1	8	88.9	0.440
OPD 08	7	2	5	71.4	0.328
OPD 13	12	7	5	41.7	0.363
OPD 18	17	3	14	82.4	0.290
OPD 20	20	2	18	90	0.328
OPM 01	22	2	20	90.9	0.406
OPM 02	17	3	14	82.4	0.265
OPM 12	15	8	7	46.7	0.414
OPM 15	12	3	9	75	0.378
OPM 20	18	5	13	72.2	0.408
Total	167	39	128	76.6	-
Mean	13.9	3.25	10.7	-	0.364

ISSR primers
DiGA5’CR	8	0	8	100	0.356
TriCAC’RC	7	1	6	85.7	0.215
TriGTG	10	0	10	100	0.353
TriGTG3’yC	8	2	6	75	0.460
TriGTG5’Cy	7	0	7	100	0.439
TriTGT5’Cy	5	0	5	100	0.364
TriTCA3’RC	6	0	6	100	0.401
TriTCC3’RC	6	0	6	100	0.446
triCGA3’RC	6	2	4	66.7	0.385
Total	63	5	58	92.1	-
Mean	7	0.5	6.4	-	0.380

Isolate	Lesion (cm)	Isolate	Lesion (cm)
Control	0.7 a	15	2.1 ef
1	0.82 ab	16	0.95 b
2	0.88 ab	17	0.85 ab
3	0.78 ab	18	0.77 ab
4	0.75 ab	19	0.72 ab
5	0.7 a	20	2.3 f
6	1.2 c	21	2.6 g
7	0.73 ab	22	0.73 ab
8	0.7 a	23	0.7 a
9	2.1 e	24	2.7 g
10	0.82 ab	25	0.8 ab
11	0.82 ab	26	1.65 d
12	0.75 ab	27	1.77 d
13	0.75 ab	28	0.82 ab
14	0.95 b

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E-mail: caatinga@ufersa.edu.br

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