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Etiology of Trichosporium vesiculosum Butl. Causing Blister Bark Disease in Casuarina equisetifolia Forst

A. Karthikeyan, T. Muthukumar and I. Sureka

Blister bark caused by Trichosporium vesiculosum Butl. is a lethal fungal disease of Casuarina equisetifolia Forst. In plantations, blister bark is more prevalent in trees that are more than two and a half years old. So in this present study, the influence of climatic and edaphic factors on blister bark disease development in C. equisetifolia under controlled conditions were investigated. The pathogen isolated from the disease plant tissues was maintained on Potato Dextrose Agar medium. Conidial suspension of the pathogen was inoculated onto 3 to 6 months old. C. equisetifolia seedlings raised on different soil types (Alfisol, Vertisol and Ultisol). The pathogenecity study indicated that T. vesiculosum attacked C. equisetifolia seedlings as early as 5 months old. A temperature range of 25 to 30°C and a relative humidity (%) range of 45 to 60 favoured the conidial development and fungal spread in the seedlings. Similarly soil pH of 5.5 to 6 favoured the pathogen and disease development whereas soil pH<7.5 deters disease development. It was also found that regular watering prevented disease development. Soil type seems to play a major role in disease development as disease incidence and severity was more in seedlings raised in Vertisol than in other soil types. From this experiment it was understood that the favourable temperature, soil factors and soil type for this pathogen to be taken care at the time of planting of C. equisetifolia seedlings so as to avoid the blister bark disease.

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A. Karthikeyan, T. Muthukumar and I. Sureka, 2011. Etiology of Trichosporium vesiculosum Butl. Causing Blister Bark Disease in Casuarina equisetifolia Forst. Plant Pathology Journal, 10: 85-88.

DOI: 10.3923/ppj.2011.85.88

Received: April 19, 2011; Accepted: May 25, 2011; Published: July 20, 2011


Casuarina equisetifolia Forst. is an important multipurpose tree species mainly grown for its fuel wood. It plays a major role as windbreak and shelterbelt along coasts in several tropical countries including India (Nicodemus, 2009). It is also a nitrogen fixing tree and grows up to 50 m in height and 50 cm dbh. It is widely planted in Andhra Pradesh, Orissa, Tamilnadu and Pondicherry with an area of 50,000 ha (Murthy et al., 2006). The average yield of C. equisetifolia in India is about 50 to 60 tonnes per ha, under rainfed and 90 to 100 tonnes under irrigated conditions (Karthikeyan et al., 2009). It is also used as agroforestry crop and the uses are scaffolding for building construction, ornamental and soil improvement (Nicodemus, 2009). After the devastation of coastal areas by a Tsunami in the year 2004, C. equisetifolia gained much importance as the matured trees up to 20 m tall successfully reduced the force of the Tsunami and saved the coastal areas in few parts of Tamilnadu, India (Karthikeyan et al., 2009). Though this tree is very strong it is highly susceptible to a destructive fungal pathogen Trichosporium vesiculosum Butl. This disease was first reported in India during 1905 and in recent years this disease has been known to cause large-scale mortalities of C. equisetifolia in India, China, Kenya and Vietnam (Narayanan et al., 1996). This fungus is able to multiply rapidly and spread over the main stem of the tree that causing death of the tree. This disease because of its rapid spread affects many places in Tamil Nadu and Pondicherry. Though the researchers on management of this have carried out considerable research, no concrete control measure is available for want of information on etiology of the disease. Therefore, in this study the mode and spread of infection of this destructive pathogen on the specific host of C. equisetifolia were studied under controlled conditions through etiology.


This study was conducted in Institute of Forest Genetics and Tree Breeding IFGTB), Coimbatore, India from April to October 2008.

Isolation of pathogen: Diseased plantations were identified in Panampally (Kerala) situated at 11° 7'N and 77° 7' E has typical warm-humid climate with average annual rainfall of 3,020 mm and mean monthly temperature ranging from 17.5 to 29.5°C and the mean monthly relative humidity range from 75 to 92%. The total area of the affected C. equisetifolia is 1 ha (Narayanan et al., 1996).

Culture of pathogen: Culture of T. vesiculosum was developed by plating the spores collected from the disease trees on to Potato Dextrose Agar (PDA) Medium (potato extract 200 g, Dextrose 20 g and Agar 20 g in 1000 mL of sterile distilled water). Fungal colonies developed after seven days under normal room temperature conditions (21 to 24°C). To identify the influence of temperature on the mycelial growth of T. vesiculosum the pathogen was placed under seven temperature conditions (10, 15, 20, 25, 30, 35 and 40°C) in the laboratory using growth gradient chamber. Each treatment was replicated at five times. The colony development was measured after 72 h. The growth incidence was calculated according to the methods of Karthikeyan et al. (2003).

Inoculation of pathogen in C. equisetifolia seedlings under different environmental conditions: Five millilitre of T. vesiculosum spores (106 spores mL-1) was inoculated to three months old C. equisetifolia according to Davison (1994) and kept under different temperatures (10-40°C) in an environmental chamber (NK system, Nippon chemicals and Laboratory Co. Ltd. Japan). Seedlings inoculated with T. vesiculosum were regularly observed for the presence of disease symptoms. Similarly, a second set of inoculated seedlings were kept under different humidity levels (30 to 60°C) in a growth chamber. These seedlings were also periodically observed for the development of blister bark disease.

Influence of irrigation frequency: Seedlings inoculated with T. vesiculosum were watered at different intervals (Daily, once in two days, once in 4 days and once in a week) and monitored up to 6 months. The influence of water in development of disease symptoms were also noticed and recorded.

Influence of soil type, moisture and pH: C. equisetifolia seedlings were planted and inoculated with T. vesiculosum in three different soil types. Vertisol soil was collected from Panampally (Kerala), Alfisol was collected from Sadivayal (Coimbatore, T.N) and Ultisol was collected from Coimbatore (T.N). The plants were monitored for 6 months and the soil moisture and soil pH of each soil type was calculated according to Jackson (1973).

Statistical analysis: All the data were statistically analysed by using the software SPSS (ver.10). The data were presented as means and their standard errors. The data were subjected to Critical difference and Pearson’s coefficient correlation was used to determine the degree of association between soil characteristics, growth and disease incidence of T. vesiculosum (Zar, 1984).


Growth incidence (%): The growth incidence (%) was higher under temperature of ranged from 25 to 40°C. A significant positive correlation (p<0.01) was also occurred between Temperature and growth of T. vesiculosum (Table 1).

Disease symptoms under different environmental conditions: Under temperature above 25°C the conidial formation was occurred in the seedlings. Similarly the relative humidity between 40 to 60% was influenced the disease as the diseases symptoms occurred (Table 2). Influence of water was also noticed that caused disease in T. vesiculosum inoculated C. equisetifolia seedlings daily watered seedlings did not exhibit any disease symptoms. Significant positive (p<0.01) correlations were established among Temperature, RH (%), percentage of infection and disease incidence (Table 3).

Table 1: Growth of T. vesiculosum under different temperature conditions (mean of 5 replicates)
*Values significant at 5% of C.D; ** Correlations are significant at p = 0.01

Table 2: Influence of temperature and humidity on T. vesiculosum inoculated seedlings of C. equisetifolia (mean of 5 replicates)
DSI = 0: No symptoms; 1: Yellowing of needles; 2: Wilting and withering of needles, 3: Drying and dead of seedlings, *±SE of the mean

Table 3: Correlation co-efficient (r) for Temperature, RH (%), Percentage of infection and disease incidence (n = 5)
Correlations are significant at p = 0.01

Fig. 1: Influence of soil pH and soil moisture of different soil types on disease severity of T. vesiculosum inoculated seeding of C. equisetifolia

Influence of soil moisture and pH: Soil moisture and pH are greatly influence the disease as showed disease symptoms in seedlings of C. equisetifolia. Soil pH ranges between 5.5 to 6 highly favours the disease and soil moisture <40% showed the symptoms (Fig. 1).


The disease blister bark affects C. equisetifolia trees of different ages causing large scale mortalities as high as 90% (Sharma, 1994). Few chemical control measures were developed to control the disease (Narayanan et al., 1996; Narayanan, 2000). However; these were not successful in curing the disease in the field (Narayanan and Sharma, 1996). So, to control this disease in field it is essential to understand the edapho climatic factors which may help to prevent the disease in field. Generally the symptoms of the disease include yellowing of the needles (cladophylls) and wilting of the needles. In advanced stage of the disease, small blister appear on the stem and roots. The black spores are exposed when the bark split bursting open blisters. The symptoms of yellowing and wilting of needles were evident at the temperature range of 25°C and above which indicates that the optimum temperature for the pathogen is 25°C and above. It also reflects the water stress which showed chlorotic appearance (Dell and Malajczuk, 1989). Jabnoun-Khiareddine et al. (2006) found similar results with Verticillium wilt of tomato under 30 °C temperature conditions. Pensuk et al. (2010) supported these results from their experiment stating that peanut bud necrosis virus inoculated pea nut plant to low temperature did not increase the infection. Further, in an earlier study it was showed that the incubation condition of 25°C induced the optimal growth of Fusarium solani, F. oxysporum, Verticillium thalia and Rhizoctonia solani (Fayzalla et al., 2008). These all fungi are soil pathogens and belong to fungi imperfecti category. T. vesiculosum also falls under the fungi imperfecti category hence the similar results were found in this study. The positive correlation between edapho climatic factors and disease severity showed that favorable environmental conditions activate the growth and disease severity of pathogen on seedlings of C. equisetifolia. Similar results were found by Chang et al. (2006) in root rot severity at 5-15°C in Cone flowers. Hiber et al. (2006) also stated that optimum temperature was more favorable to disease development by fungal pathogens. However, seedlings watered daily failed to exhibit any symptoms in spite of T. vesiculosum inoculation. Routine watering may help to prevent the disease development through control of plant temperature. This effect was also observed in plantations of Tamilnadu and Pondicherry where there is a good irrigation. Less soil moisture of vertisol soils are also indicated the results of severe yellowing, drying of needles and formation of black conidial spores. It showed the impact of environment and soil influence the disease (Shearer and Tippett, 1989). The acidic pH causes the disease whereas the pH<7.5 did not show any symptoms. These results supported by Fayzalla et al. (2008) as F. solani, F. oxysporum, V. thalia and R. solani found grown well under pH 9. Similar results also found in earlier study on impact of soil pH on Phytophthora cinnamomi in Cedrus deodara (Karthikeyan et al., 2000). In this present study, vertisol type soils favours suitable for the pathogen to proliferate and survive. Similarly, Nerey et al. (2010) found calcisol soil type is the most suitable soil for Rhizoctonia solani causing hypocotyl rot in Phaseolous vulgaris which proved that certain soil types are more favourable to particular pathogen than any other soil types. The Temperature, humidity, soil factors and soil types definitely influences the pathogen development which may also cause sever damage in field conditions according to Aigbe and Remison (2010). Hence, it was understood that to prevent this blister bark disease in C. equiestifolia seedlings necessary steps have to be undertaken while planting viz., (1) adjusting soil pH to an alkaline range, (2) providing good irrigation system and (3) avoidance of planting sites at vertisol type soils.


T. vesiculosum is a severe pathogen which causing blister bark disease in C. equisetifolia trees. To control this disease understanding the etiology of the pathogen is very much essential. From this experiment it was understood that the favourable climatic conditions soil factors, soil type for this pathogen to be avoided while planting so as to control the blister bark disease in C. equisetifolia trees.


The author (A.K) thanked Indian Council of Forestry Research and Education, Govt. of India for financial support in form of a research project.

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