








































American Journal of Agricultural Science, Engineering and Technology 

STUDIES ON THE INCIDENCE OF ANTHRACNOSE DISEASE IN DIFFERENT 

VARIETIES OF MANGO (MANGIFERA INDICA L.) AND IT’S IN VITRO 

BIOCONTROL MEASURES 

S. K. Ghosh1, S. K. Ray2 and   M. A. Hasan3 

ABSTRACT 

Anthracnose of mango caused by Colletotrichum gloeosporioides) is a very serious disease. 

Leaves, panicles, immature and mature fruits of mango are infected and damaged by this 

disease.  The symptoms of this disease on infected parts were noted. The incidence of disease 

on leaves was studied on fourteen varieties of mango viz. Himsagar, Dashehari, Vanraj, 

Farnandin, Mulgoa, Bombai, Kishanbhog, Bangalora, Alphanso, Langra, Mallika, Zardalu, 

Chausa and Suvarnrekha. In all the varieties the incidence of anthracnose ranged from 1 - 

15 per cent. The disease incidence was maximum (15.00 per cent) in Kishanbhog followed by 

Bombai (12.00 per cent), Himsagar (9.00 per cent) and Bangalora (8.00 per cent).  In contrast, 

the disease incidence was recorded minimum in Alphanso (1.00 per cent). Trichoderma viride 

and Beauveria bassiana were applied against Colletotrichum gloeosporioides in vitro and their 

antagonistic activity were recorded. 

Keywords: Anthracnose, mango, variety, Trichoderma, Beauveria 

----------------------------------------------------------------------------------------------------------- 
1Mycopathology Lab.,P.G. Dept. of Botany, Ramakrishna Mission Vivekananda 

Centenary College, Rahara, Kolkata -700118, West Bengal, India 

E-mail: swapan.krghosh@yahoo.com
2Dept. of Plant Pathology, Faculty of Agriculture, Bidhan Chanda Krishi Viswavidyalaya,

Mohanpur, Nadia, West Bengal, India
3Dept. of Fruits and Orchard Management, Faculty of Horticulture, Bidhan Chanda Krishi

Viswavidyalaya, Mohanpur, Nadia, West Bengal, India

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American Journal of Agricultural Science, Engineering and Technology 

INTRODUCTION 

Mango (Mangifera indica L.) is universally considered one of the most important fruit crop in India. India 

produces near about 54.2% of World’s mangos and exports Rs. 170.71 Crore of mangoes in 2008—

09(APEDA, 2010).  

Anthracnose, the most important mango disease, is caused by the fungus Colletotrichum gloeosporioides. 

Flower blight, fruit rot, and leaf spots are among the symptoms of this disease (Arauz, 2000). Fruits infected 

at mature stage carry the fungus into storage and cause considerable loss during storage, transit and 

marketing (Haggag, 2010). Generally to combat this disease, fungicides (e.g. bavistin, blitox -50) are 

applied but fungicides are environmental pollutant. The non-chemical or eco-friendly methods are now 

popularized in developed countries (America, U.K. etc.). Cultivation of disease resistant variety and bio 

control method are eco-friendly strategies to manage diseases.  

Therefore, the main objectives of this work are to record mango varietal differences in susceptibility to 

anthracnose and antagonistic efficacy of biocontrol agents - Trichoderma viride, and Beauveria bassiana 

against Colletotrichum gloeosporioides, causal organism of anthracnose of mango in vitro. 

MATERIAL AND METHODS 

a) Study of symptoms of anthracnose: The infected leaves, petioles, twigs, panicles, flower and fruits were

collected separately in sterilized biodegradable polythene bags and carried in laboratory and the

symptoms caused by anthracnose were studied with the help of simple microscope.

b) Isolation and purification of pathogen from diseased parts : The infected leaves , petioles , twigs ,

panicles , flower and fruits were collected separately in sterilized biodegradable polythene sheets

,carried in laboratory ,and isolation of the pathogen was done in PDA  medium in Petri dishes  at 28o C

by the method presented by Dhingra & Sinclair(1994).

For purification of isolated pathogen, single hyphal tip method was taken.

c) Identification or Characterization of the pathogen: The identification of the pathogen was done by

cultural and microscopical characteristics with the help of published fungal Key and books (Nagamani

et al., 2006; Domsch et al.; Bailey and Jeger, 1992; Freeman et al, 1998). The identifications of  isolates

of T. viride were done by IARI, Delhi, India

d) Pathogenecity test of the pathogen: It was done  by koch’s postulates

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American Journal of Agricultural Science, Engineering and Technology 

e) Mango varietal susceptibility test : The anthracnose disease incidence on mango

leaves  was recorded during the period of flowering and before the fruit set in the years of 2009 & 2010 

in the Gayaspur Farm , under BCKV , Mohanpure , Nadia , West Bengal ,India Fourteen mango 

varieties ( Himsagar , Dasehari , Vanraj ,Farnandin, Mulgoa ,Mumbai, Krishnabhog, Bangalore, 

Alfanso , Langra, Zardalu, Chausa, Mallika and Suvarnrekha ) were screened in randomized block for 

the study of incidence of anthracnose. The scale proposed by Khalid & Alam (2002) was taken to 

observe the incidence of anthracnose of mango.  

Standard Scale for assessment of anthracnose of mango (Khalid &Alam 2002) 

Disease grading Types of symptoms 

1—5 % Affected leaves per twig per plant -Mild 

6—10% Affected leaves per twig per plant –High 

11—15% Affected leaves per twig per plant -Severe 

f) Isolation and characterization of antagonistic fungi: Isolation of fungi from different soils from

different geographical regions of west Bengal, were done in the laboratory by dilution Plate Method

followed by Dhingra & Sinclair (1994).

Fungal colonies were isolated and sub-cultured repeatedly for getting pure colonies and then preserved

in slant tubes for further identification. . The fungal strains were identified after staining them with

cotton blue, by following the keys of Domsch et.al 1980...Nagamani et al, (2006) and

www.mycobank.org

g) Antagonistic potentiality test or rating of mycoparasitism of isolated antagonistic fungi: Five mm

diameter of mycelial colony from the margin of actively growing colony of C. gloeosporioides and that

of antagonist were incubated simultaneously at opposite ends of a Petri dish containing 25 ml of

sterilized PDA medium. The plates containing the paired culture were incubated at 280 ± 10 C for 9

days in a B.O.D. incubator and were subsequently scored for degrees of antagonism on a 1-5 scale (Bell

et al., 1982).

An isolate of the mycoparasite was considered highly antagonistic to the pathogen when the mean score for 

a given comparison (when rounded to the nearest whole class number) was ≤2, but not highly antagonistic 

if the number was ≥ 3. 

Among the Class—I antagonists, a comparative analysis was done on the basis of ability of antagonist to 

grow faster than other over C. gloeosporioides. 

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The selected cultures from pairing of mycoparasitized pathogen were observed under 

microscope (Leitz Laborlux K, Germany) to study of hyphal interactions between the antagonist and the 

pathogen and photographed when required. 

RESULT AND DISCUSSION 

Symptoms of the disease:  

i) Leaves: On the leaves  the symptom starts  as circular or oval shaped irregular brownish to black color

spots variable in sized( 2-6 mm) .Under conducive condition ( Humidity >90 and temperature 300C ) the 

spot increases in size and disease tissues become rotted. During dry weather, the spot cannot enlarge and it 

becomes dried and drops off (Fig. 1). The disease spots are observed on the petiole also.  

ii) Twig: On the young twig black spot appears on the young leaves. During rainy season, the disease

rapidly covers the twig causing it to turn black and it becomes dry. 

iii) Inflorescence: Spots appears as black, they enlarged and coalesced on the inflorescence or panicle axis,

the infected axis becomes dry and withered before fruit set (Fig.2). 

Fig .1 Symptom on leaves         Fig.2 Symptoms on panicle Fig.3 Symptom on fruit          

iv) Fruit: The symptoms are found both young as well as mature fruits.  Black spots are formed on the

fruits and their skin becomes discolored. The pulp beneath the infected skin becomes hard (Fig3). 

Similar symptoms were recorded by many workers (Ploetz, 1994; Arauz, 2000; Singh 2005). 

Identification of the pathogen: The isolated pathogen was Colletotrichum gloeosporioides (Penz) (Fig.4-

7).The identification of the pathogen was done by cultural and microscopical characteristics with the help 

of published fungal Key and books (Nagamani et al., 2006; Domsch et al.; Bailey and Jeger, 1992; Freeman 

et al, 1998). 

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American Journal of Agricultural Science, Engineering and Technology 

Conidia→ 

seta→ 

  Fig. 4 Acervulae of C. gloeosporioides  Fig. 5 Conidia & conidiophore 

  Fig. 6: Conidia of C.gloeosporioides Fig. 7: (Left) Dual culture 

Mango varietal susceptibility test: The presented in the table-1 exhibited that all varieties of mango trees 

tested were susceptible to anthracnose disease. The maximum (15) per cent of disease incidence (PDI) 

occurred in the Krishanbog followed by Bombai (12 per cent), Himsagar (9.00 per cent), Dashehari (8.50) 

and Bangalora (8.00 per cent).  

Table 1 Study of mango varietal susceptible test against anthracnose 

S. No. Mango variety Per cent of disease incidence Type of disease 

 2009 2010 Average 

1 Himsagar 08.50* 09.50 09.00 High 

2 Dashehari 08.75 08.25 08.50 High 

3 Vanraj 06.00 05.00 05.50 High 

4 Farnandin 02.00 02.00 02.00 Mild 

5 Mulgoa 05.00 08.00 06.50 High 

6 Bombai 09.00 15.00 12.00 Severe 

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American Journal of Agricultural Science, Engineering and Technology 

7 Kishanbog 13.00 17.00 15.00 Severe 

8 Bangalora 08.00 08.00 08.00 High 

9 Alphanso 01.00 01.00 01.00 Mild 

10 Langra 06.00 09.00 07.50 High 

11 Mallika, 09.00 06.00 07.50 High 

12 Zardalu 05.50 06.50 06.00 High 

13 Chausa 07.00 09.00 08.00 High 

14 Suvarnrekha. 05.50 07.50 06.50 High 

S.Em± 0.0012 

C.D. (P≤0.05) 0.3130 
*Average of three replicated plants

In contrast, the disease incidence was recorded minimum in Alphanso (1.00 per cent). Out of fourteen

mango varieties, two (Farnandin and Alphanso) were categorized as mild susceptible while ten varieties

(Himsagar, Deshahari , Bangolora Chausa,Langra, Mallika, Mulgoa,Subarnkeka, Zardalu andVanraj were

under high. And only two (Krisanbag and Bombai) were severe susceptible. In the year 2010 per cent of

disease incidence was higher in many varieties in comparison to 2009. It is due to prevailing weather during

this period.

Varietal differences in susceptibility have been noted in Egypt. Maximum damage was observed on

Alphanso, whereas variety Tommi and Fagr Kelan were recorded to be resistant (Haggag, 2010).

Antagonistic potentiality test or rating of mycoparasites of isolated antagonistic fungi:  A total of

seven fungal isolates were screened against C. gloeosporioides

under in vitro condition by Bell’s test (Fig.8) for determining

mycoparasitic activity. The data presented in the Table -2 indicate

that all seven isolates were rated as class—I mycoparasites

/antagonists .Out of them, five isolates(T1.T2.T3,T10 &T12)

identified by IARI as Trichoderma viride .Two isolates( BB& B2)  

were two strains of Beauveria bassiana.

    Fig. 8 T10 x pathogen 

Conidia of T viride →

Shrinkage of protoplast→

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Table-2 Rating of mycoparasites in dual culture plate by Bell’s method. 

Serial No. Organism or isolates Mean scores I  II  III 

1 T1 1.12 (1.00) 

2 T2 1.50(1.00) 

3 T3 2.00(2.00) 

4 T10 1.06(1.00) 

5 T12 1.40(1.00) 

6 BB 1.80(1.00) 

7 B2 2.00(2.00) 

I=Mean of three replications. 

II =Data are recorded 9 days after incubation. 

III =Scale of classes described by Bell et al.  (1982) was followed. ; The figures in 

parenthesis indicate the whole class. 

Further, the results from Table –3 revealed that out of five strains of T. viride ,T10 showed maximum (3.10) 

mycoparasitism over C. gloeosporioides followed by T12(3.00),T1(1.40) ,T3(1.40) and T2 (1.00). Out of 

two strains of B. bassiana, BB was better than B2 

Table 3: Comparative mycoparasitic activity of different isolates of hyperparasites showing 

class –I activities against C. gloeosporioides 

Serial 

No. 

IARI Herb. 

No. 

Organism or 

isolate 

Scientific name of 

mycoparasite 

Radial growth(cm) of 

mycoparasites over C. 

gloeosporioides at 24 hr. 

interval 

 24 48 72 

1 108 T1 Trichoderma viride 0.70* 1.09 1.40 

2 109 T2 Trichoderma viride 0.45 0.86 1.00 

3 110 T3 Trichoderma viride 0.61 1.00 1.30 

4 112 T10 Trichoderma viride 1.00 2.25 3.10 

5 115 T12 Trichoderma viride 0.80 2.02 3.00 

6 -- BB Beauveria bassiana 0.20 0.42 1.00 

7 -- B2 Beauveria bassiana 0.15 0.32 0.60 

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S.Em± 0.093 

C.D. (P≤0.05) 0.452 

*Each insertion is average of three replications

Kefialew and Ayalew( 2008) reported that four isolates of bacteria, five yeasts and two filamentous fungi 

were evaluated in this study. Cell suspensions and culture filtrates of the isolates inhibited spore 

germination and hyphal growth of C. gloeosporioides in vitro. 

Mango plants treated with Trichoderma's pellet and spraying with spore suspension of Trichoderma 

harzianum PC01 and T. hamatum PC02 (404 x 1010) and some chemical fungicide treated ones. It was 

observed that the biological treatments gave better yield than the chemical fungicides (Carbendazim, Zinep, 

Manep and Copper oxychloride) treatment. (Noiaium and Soytong, 2010). 

Interaction between Trichoderma viride (T10) and C. gloeosporioides 

under microscope showed that spores of T10 adhered on the wall of C. 

gloeosporioides and shrinkage of protoplast of the latter were recorded 

(Fig. 9).The Fig. - 9 depicted that the hyphae of C. gloeosporioides       

was surrounded by T 10 and the shrinkage of the protoplast of the  

      Fig. 9: T10 xPathogen   pathogen. 

Similar phenomenons were reported in the interaction between mycoparasites (Trichoderma viride) and 

plant pathogens by other workers (Chat et al., 1981; Pan and Ghosh, 1997). 

In conclusion, all varieties of mangoes are susceptible to anthracnose from mild to severe. Trichoderma 

viride strains are good biocontrol agents as tested in vitro and this experiment encourages other to apply 

this biocontrol agent in the field of mango (in vivo). 

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