Suitable Integrated Approach for Management of Fusarium Wilt of Tomato caused by Fusarium oxysporum f. sp. lycopersici (Sacc.)

Among the different integrated approaches for management of Fusarium wilt and their effect on growth and yield parameters of tomato revealed that soil application of FYM @ 100gm/ pot + Neem cake @ 100gm/pot + seedling treatment with bio-formulation of Azotobater @ 5% + foliar spray of Carbendazim @ 0.1% was showing minimum disease incidence with 6.23, 10.11 and 15.03 per cent at 7, 14 and 21 days after inoculation, respectively. The observations on plant height of tomato was found in T3 treatment representing the value 17.00, 18.85, 20.66, 22.10, 24.10 and 27.30 cm at 5, 10, 15, 20, 25 and 30 days age of seedling, respectively against the minimum plant height i.e. 10.42, 10.92, 11.56, 11.76, and 12.55 in case of control (T10). The effect of integrated approach on branching of shoot in tomato was estimated at 85 days age of plant which revealed the maximum number of branch with 5.00 was found in case of soil application of FYM @ 100gm/pot + Neem cake@ 100gm/pot + seedling treatment with bio-formulation of Azotobater @ 5% + foliar spray of Carbendazim @ 0.1% whereas, in case of control it was only 2.33. The morphological character of roots was examined and recorded developed robust root system in T3 treatment while the less fibrous, weakly developed roots in control. The maximum yield was recorded per plant in T3 treatment (soil application of FYM @ 100gm/pot + Neem cake @ 100gm/pot + seedling treatment with bio-formulation of Azotobator @ 5% +foliar spray of Carbendazim @0.1%.) represented the value 490.30g per plant. Similarly, the maximum large size tuber with 4 in number was recorded in treatment T3 (soil application of FYM@ 100gm/pot+ Neem cake@ 100gm/pot + seedling treatment with bioformulation of Azotobator@5% +foliar spray of Carbendazim @ 0.1%) followed by treatment T9 (Soil application of FYM @ 100 gm/pot + Neem cake @ 100 gm +seedling treatment with bio-formulation of T. viride + foliar spray Carbendazim @0.1%) as 03.

Tomato (Lycopersicon esculentum Mill) belongs to the family Solanaceae and is one of the most remunerable and widely grown vegetable in the world. Among vegetable crops, tomato ranks second important vegetable in the world next to potato and first among the processing crops. The worldwide production of tomato is about 130 million tonnes in the year 2014. China is the largest producer in the world with a production of 48.577 million tonnes (FAOSTAT 2013-14). In India, production of tomato during 2014-15 was 182.86 lakh tones and total area under tomato cultivation was 777.0 ha. This constitutes 9.8 per cent of total vegetable area and 11.2 per cent of total vegetable production. The productivity of tomato in India is 20.7 MT/ha during 2014-15 which is very low as compare to other country of the world like USA, 81.0 tones /ha. One of main reasons of low productivity of tomato in India is disease which is caused by fungi, bacteria, virus, nematode and other biotic factors (Blanchard, 1992). Among them, Fusarium wilt incited by Fusarium oxysporum f sp. lycopersici (Sacc) is world's most destructive disease which causes 50 to 86 per cent loss in fruit yield in different parts of country. (Mathur and Shekhawat, 1986 etc. But all these methods have some limitations and cannot solely control the disease. Therefore, there is need to integrate all these methods in a suitable and sustainable manner for management of plant diseases in near future. Morajdhwaj et al. (2015) reported that soil application of mustard cake along with tuber treatment and foliar spray with Trichoderma viride was found best in reducing disease severity of late blight, stimulating germination and increased plant height of potato. Yogesh et al. (2015) also reported that among the different integrated approaches, soil application of FYM + seedling treatment with bio-formulation of Trichoderma harzianum + foliar spray of mancozeb reduced the disease severity of early blight of tomato and increased the growth parameters of plant. Bio-fertilizers viz., Azotobacter chroococum, PGPR, Trichoderma harzianum, Trichoderma viride, PSB, Rhizobium, Carbandazim, have also found effective in management of spot blotch of wheat and increase growth parameters and yield of wheat . Keeping all the point in view, the study was under taken in the present investigations as "Suitable Integrated Approach for Management of Fusarium Wilt of Tomato caused by Fusarium oxysporum f. sp. lycopersici (Sacc.)"

MATERIAL AND METHODS
The experiment was conducted in the Wire House Complex Department of Plant Pathology, C.S.A. University of Agriculture & Technology, Kanpur during Rabi season 2014-16.

Collection of disease sample
The disease samples were collected from Vegetable Research Farm, C.S.A. University of Agriculture and Technology, Kalayanpur (Kanpur). Infected plants apparently showing wilt like symptoms were collected and bring to the laboratory for initial examination. The specimen diseased sample were pressed in between the fold of sterilized blotting paper and preserved at 4-6°C in refrigerator for further study. The entire specimen were collected and examined in the laboratory for the presence of the causal organism and virulence study.

Isolation of pathogen
Infected plant showing typical symptoms was selected for isolation of pathogen. Initially the diseased stem of tomato was washed thoroughly with distilled water to remove dust particles. Then diseased portion of stem were cut in to small pieces by a sterilized knife and each piece is having small bits of disease and healthy tissues. These pieces were dipped in 0.1% mercuric chloride solution for 30 seconds and then thoroughly washed thrice in distilled water to remove the traces amount of mercuric chloride solution. Excess moisture was removed by putting these pieces between the folds of sterilized blotting paper under aseptic conditions. The two pieces were then transferred to sterilized Petri plates containing 2% potato dextrose Agar (PDA) medium in inoculation chamber with the help of sterilized forceps. The Petri plates were then kept at room temperature. The Petri plates were observed daily to notice the presence of mycelia growth around the bits.
The pathogen was then purified by the transfer of mycelium from the tip of the colony to another Petri plate which was previously poured with sterilized PDA in aseptic condition. The purified culture was then characterised based on morphological and cultural behaviour as per described by Sacc. Synder and Hansen (1940).
Effect of integrated approach on disease incidence, growth parameters and yield of tomato.
The experiment was conducted at Glass house complex, Department of Plant Pathology, C. S. A. University Agriculture & Technology Kanpur. The Tomato variety 'Azad T-6' was sown in the glass house in 30 cm earthen pot, which was previously filled with a mixture of sandy loam and Farm Yard Manure in the ratios of 2:1. The treatments were given as follows- In each pot one treated seedling of tomato were sown and irrigated regularly. The experiment design was laid out in simple CRD. Three replications per treatment were kept and three pots were sown with untreated seedling served as control. The foliar spray scheduled was given at 3 days after artificial inoculation. Observation pertaining to effect of different treatment was recorded as per following parameters and days.

Seedling treatment
Tomato seedlings of a variety were treated by root dip method. The seedlings were dipped in Azotobactor solution @ 20% conc. for a period of four hrs. The packets of Azotobactor containing 200 gm inoculums were obtained from Department of Soil Science, C. S. A. University of Agriculture & Technology Kanpur.
On the other hand, seedling was also treated with bio-formulation of Trichoderma viride @ 5% formulation.

Effect of integrated approach on growth parameters of tomato and development of disease Growth parameter Shoot length
The shoot length of tomato was measured after 10 days of transplanting at every 5 days interval up to 30 days age of tomato plant with help of meter scale. This is well to know that same height of seedling was choice for transplanting.

Root length
Prior to measure the root lengths of tomato plants, pots were irrigated and the seedlings were up rooted carefully. The roots of seedling were separated from the shoots and washed with water to remove soil particles and then root length (cm) was measured with the help of meter scale.

Disease development Inoculation of pathogen
At 45 days after transplanting, plants were inoculated with spore suspension of Fusarium f. sp. lycopersici at the concentration of conidia @ 10 6 conidia / ml. The spore suspension was prepared from seven days old culture of pathogen. The homogenized, spore suspensions were inoculated at the base of plant @ 1 ml / plant. The plants were then kept on the bench of wire house. Two controls were kept. In first case, plants were inoculated with pathogen and served as control 1 while in other case; plants were sprayed with distilled water and served as control-2. Three replications were kept for each treatment. Observation on disease severity were recorded at 7, 14, 21 days after final spraying.

Measurement of disease severity
Observation measuring on the disease severity was taken after 10 days of pathogen inoculation. The disease severity was recorded using 0-4 scale (Weitang et al. 2004) where zero representing no infection and four denoting completely infected plant. 2 representing moderate infection and 3 denoting extensive infection.
The 0-4 scale of the disease incidence was classified as follows: • Complete infection, the whole plant leaves become yellow, 100% of leaves become wilted, and the plants die. The percentage of disease incidence was determined using the formulas as given by Weitang, et al., (2004)

Yield/plant
The edible fruits were harvested twice a week from each treated plant and weighted with the help of physical balance. The total weight of all picking was recorded after adding weight of fruits at each picking. The yield of the crop of each treatment was calculated separately.

Effect of integrated approach on growth parameters of tomato Plant height
The effects of integrated approach on plant height of tomato were studied under Wire House Complex in pot culture experiment, Department of Plant Pathology, Chandra Shekhar Azad University of Agriculture and Technology, Kanpur. The observations of plant height were taken after 10 days of transplanting at every 5 days of interval up to 30 days age of tomato plant. The data presented in Table-1 showed that the plant height of tomato was increase in all the treatments over control. The maximum plant height was found in T 3 treatment (soil application of FYM@ 100gm/ pot+ Neem cake@ 100gm/pot + seedling treatment with bio-formulation of Azotobater @ 5% +foliar spray of Carbendazim @ 0.1%) representing the value 17.00, 18 reported that shoot height and shoot diameter, fresh and dry weight of shoot in tomato seedlings were interestingly increased when sown in T. harzianum T969 fortified soil. Datnoff et al. (1995) found that Trichoderma sp. enhanced the growth of tomato plants.

Branching of shoots in tomato
The effect of various treatments on branching of shoot in tomato was estimated at 85 days age of plant and the data presented in table-1 revealed that all the treatments significantly enhance branching of shoot. Among the treatments maximum number of branch with 5.00 was recorded in T 3 treatment (soil application of FYM@ 100gm/pot+ Neem cake@ 100gm/pot + seedling treatment with bio-formulation of Azotobator@5% +foliar spray of Carbendazim0.05%). whereas in case of control it was only 2.33. The T 9 treatment was showing average 4.48 branches indicating second highest among the treatments followed by the T 5 and T 7 with the value of 4.33, and 4.00, respectively. Yogesh et al. (2015) also reported that among the different integrated approaches, soil application of FYM + seedling treatment with bio-formulation of Trichoderma harzianum + foliar spray of mancozeb reduced the disease severity of early blight of tomato and increased the growth parameters and branching pattern of plant. Tippannavar et al. (2005) observed that the Azotobacter significantly increased the tillering, dry matter accumulation and growth parameter in wheat.

Root length
Eighty five days after transplanting, the tomato plant was uprooted and the root length was measured separately by using scale. The data presented in Table-4 showed that all the treatments were able to significantly increased root length over control at 85 days age of tomato (Fig. 1). Among the treatments, the maximum root length was recorded in treatment T 3 (soil application of FYM@ 100gm/pot+ Neem cake@ 100gm/pot + seedling treatment with bio-formulation of Azotobator@5% +foliar spray of Carbendazim@0.1%) representing 15.00cm followed by treatment T 9 (Soil application of FYM@ 100 gm/pot + Neem cake @100 gm + seedling treatment with bio-formulation of T. viride + foliar spray of Carbendazim @0.1%) and treatment T 5 (Soil application of FYM @ 100 gm/ pot +Mustard cake @100 + seedling treatment with T. viride + Foliar spray Carbendazim@0.1%) with the value of 14.00cm and 10.00.00cm, respectively. On the other, maximum width of root zone was also recorded in T 3 treatment, representing 16.00 cm, followed by T 9 treatment (15.32 cm). From the table, it is cleared that the all the treatments were able to increase root length and width over control. Ravindra et al. (2015) found that root length was highest in case of seed treatment with T. viride + soil application of Neem cake powder + foliar spray of Carbendazim at 90 days age of tomato crop. Gopinathan and Prakesh (2014) found that vermicompost enriched with bio-fertilizer increased plant height, root length, number of branches, number of leaves and the productivity of tomato.

Morphological character of roots
Integrated approach has also found that it changes the morphology of root. The data presented in the table-1 showed that well developed robust root system is found in T 3 treatment (soil application of F Y M @ 1 0 0 g m / p o t + N e e m cake@ 100gm/pot + seedling treatment with bio-formulation of Azotobactor @5% +foliar spray of CarbendazimHYPERLINK "mailto:Carbendazim@0.1%25" HYPERLINK "mailto:Carbendazim@0.1%25"@0.1%) whereas, in case of control, poorly developed, less branching and less fibrous root system are found which is indicated that integrated approach have ability to change the morphology of root.

Effect of different treatments on development of disease
The data presented in Table-3 found that all treatments were able to reduce the disease incidence over control. The minimum disease severity was recorded in the treatment T 3 (soil application of FYM@ 100gm/pot+ Neem cake@ 100gm/pot + seedling treatment with bioformulation of Azotobator @5% +foliar spray of Carbendazim @0.1%) representing 6.23, 10.11 and 15.03 per cent at 7, 14 and 21 days after inoculation. The treatment T 9 (Soil application of FYM@ 100 gm/pot + Neem cake @100 gm + seedling treatment with bio-formulation of T. viride + foliar spray Carbendazim @0.1% ) was showing 8.13, 14.22 and 17.50 per cent disease severity, representing the second lowest among the treatments. Among the treatments, the maximum disease severity was recorded in treatment T 2 (Soil application of FYM @ 200 gm/pot + seedling treatment with Trichoderma viride) i.e. 14.35, 17.75, 20.12 per cent at 7, 14 and 21 days, respectively. Varma et al. (2008) reported that the foliar spray of Trichoderma viride (10 7 CFU s ml -1 ) 24 hrs before challenge inoculation with the test fungus was found effective in reducing the disease severity under screen house condition.