SEED TREATMENT AND FOLIAR SPRAY WITH FUNGICIDES TO CONTROL SEEDLING DISEASES AND CERCOSPORA LEAF SPOT OF MUNGBEAN
An investigation was undertaken to find out the influence of integrated use of seed treatment and foliar spray with fungicide on incidence and severity of Cercospora leaf spot and production of healthy seeds of mungbean. On the basis of preliminary experiments, three seed treating fungicides namely Provax-200, Metataf and Cupravit another three spray fungicides, Dithane M-45, Rovral 50 WP and Bavistin 50 WP were selected for the investigation. These were used in different combinations to have nine treatments. Highest seedlings emergence (55.64%) was recorded in seed treatment with Provax-200. On the other hand, the lowest seedlings infection was recorded in Provax-200 treated seed plots. Cupravit and Metataf also found effective to increase the emergence and to reduce diseases. All treatments with fungicides significantly reduced incidence and severity (PDI) of Cercospora leaf spot over control. Cercospora leaf spot incidence and PDI were observed lower in treatment, seed treatment with Provax + spaying of Bavistin. Treatment, seed treatment with Metataf + spaying of Bavistin and seed treatment with Cupravit + spaying of Bavistin were also promising in reducing Cercospora leaf spot incidence and PDI. The highest seed yield was recorded from the treatment when seed were treated with Provax and Bavistin was spayed. Harvested seeds under this treatment yielded the lowest prevalence of seed-borne of Aspergillus spp. and Fusarium oxysporum. Seeds treated with Provax and three sprays of seed crop with Bavistin 50 WP are recommended to control diseases and produce healthy seeds of mungbean.
Mungbean [Vigna radiata (L.) Wilczek] is one of the important food legumes in Bangladesh. It is extensively cultivated in the country during Kharif season in the gap period between Boro and Aus rice. Yield of legumes in farmer’s fields is usually less than 1 t ha-1 against the potential yield of 2 to 4 t ha-1 (Ramkrishna et al. 2000). Such yield gap of legumes indicates a great opportunity to increase the productivity of mungbean at farm level. Diseases are known to play a vital role in decreasing the yield and quality of the crop seeds. The crop suffers from at least 20 different diseases in Bangladesh. Of which 12 are caused by fungi (Bakr 1994). Cercospora leaf spot caused by C. cruenta is one of the major pulse diseases considering high incidence, wide distribution and detrimental effect on yield (Talukdar 1974). It can reduce yield up to 58% (Lal et al. 2001). Any variety with acceptable level of resistance against the disease is not available (Saifullah et al. 2005).
Seed is the vital input in agriculture. From the pathological point of view, quality of seed is very important to get disease free seedling and plant, and finally to achieve a satisfactory yield (Fakir et al. 2002). Seed treatment is an important measure for the control of seed transmitted diseases. It is unquestionable that proper seed treatment can substantially improve the quality of seed and seedling with satisfactory increase in the yield through increase of plant stand. Among the recommended practices, seed treatment with fungicides is the cheapest method of plant disease control. Treatments of seeds with fungicides have been proved to be quite effective in reducing seed-borne infection (Saraswathi et al. 1989, Dubey and Singh 2005, Barua et al. 2007).
In Bangladesh, use of seed treating fungicides like Vitavax-200 and Bavistin 50 WP may be useful to control seed-borne fungal pathogens. Disease management in the field is also an important prerequisite for the production of quality and healthy seeds. Foliar diseases of mungbean may be effectively controlled by spraying appropriate fungicides. According to Huq et al. (1991), Haque et al. (1994), Iqbal et al. (1995), Mian et al. (2000), Khunti et al. (2005), Rathore (2006), Saxena and Tripathi 2006) spraying of fungicides is the best way to control Cercospora leaf spot.
The present paper reports the results of an investigation conducted to evaluate the efficacy of three seed treating fungicides and another three foliar fungicides to control Cercospora leaf spot (C. cruenta) of mungbean with a view to select effective ones to control the disease.
MATERIALS AND METHODS
Necessary experiments were conducted in the Research farm of Bangabandhu Sheikh Mujibur Rahman Agricultural University (BSMRAU), Gazipur during 2011-2013 to screen six fungicides against Cercospora leaf spot (C. cruenta) of mungbean with a view to select effective ones to control the disease. Three of the fungicides namely Provax-200 (Oxathiin), Metataf (Metalaxyl) and Cupravit (Copperoxichloride) were tested as pre-sowing seed treating chemicals @ 2.5% (w/w). Other three namely Dithane M-45 (Mancozeb) @ 2.5 (w/v), Rovral 50 WP (Iprodione) @ 2.0% (w/v) and Bavistin 50 WP (Carbendazim) @ 2.0% (w/v) were tested as foliar spray applied at 15, 30 and 45 days after planting (DAS).
Only seed treatment with Provax-200, Metataf or Cupravit as well as only foliar spray with Dithane M-45, Rovral 50 WP or Bavistin 50 WP and seed treatment with a fungicide followed by foliar with one fungicide were tested in 15 treatments viz. Provax-200, Metataf, Cupravit, Dithane M-45, Rovral 50 WP, Bavistin 50 WP, Provax-200+ Dithane M-45, Provax-200+ Rovral 50 WP, Provax-200+ Bavistin 50 WP, Metataf+ Dithane M-45, Metataf+ Rovral 50 WP, Metataf+ Bavistin 50 WP, Cupravit+ Dithane M-45, Cupravit+ Rovral 50 WP and Cupravit + Bavistin 50 WP. A control trématent was maintained where seeds were not treated and no spray was applied.
Mungbean (Vigna radiata) variety BU mung-2 was grown following standard methods. The experiment was laid out in a randomized complete block design with 3 replications. The unit plot size was 1.5 m X 2.0 m and distance between plots was 1.0 m. Treated seeds were planted at a depth of 5 cm, maintaining 10 cm seed to seed and 30 cm row to row distances. Seed rate was 50 kg ha-1. Weeding, irrigation and other intercultural operations were done as and when needed.
Data on seedling emergence were recorded after 7 DAS and expressed as percentage based on total number of seeds sown. Seedling mortality caused by fungal pathogens was also recorded after two weeks of sowing and expressed in percentage. The incidence of Cercospora leaf spot (CLS) was recorded three times starting from 30 days after sowing (DAS) at 15 days interval. The incidence of CLS was recorded based on the basis visible symptoms as suggested by Bakr (1991) and expressed in percentage. The severity of CLS was indexed on a 0-9 scale (Singh et al. 1982)), where 0=no visible symptoms on leaves, plant growth, pod formation normal, 1=0.1- 5.0% foliage is affected by small pinhead sized brown lesions, 3=5.1- 10% foliage is covered with round brown spots, 5=10.1- 25% foliage is covered with bigger leaf spots, 7=25.1-50% foliage is covered with enlarging, coalescing leaf spots and 9=50.1-100% foliage is covered with very large, coalesced spots. The severity was expressed in percent disease index (PDI) which was computed using a standard formula (Mian et al. 2000).
The mungbean was harvested at full ripening stage. The harvested crop was sun dried and threshed manually. Seeds were sun dried, cleaned and stored at 4C for further use. Before harvesting 10 plants were selected in each plot for collection of seed yield data. Data on pod number plant-1, pod length (cm), seed number pod-1, 1000-seed weight (g) and seed yield (kg ha-1) were recorded.
Seed health of the harvested mungbean seeds was tested following blotter method to detect the seed-borne fungal pathogens following the International Rules for Seed Testing (Anon. 1999). Four hundred seeds from each sample were taken randomly and placed on moist filter paper in sixteen replicated Petri dishes at the rate of 25 seeds per plate. After seven days of incubation, the seeds were examined under steriobinocular microscope for the presence of seed-borne fungi and identified by observing their growth characters. In case of confusion, temporary mounts were prepared and examined under a compound microscope and identified the fungi using appropriate keys (Ellis 1993, Mathur and Kongsdal 2003). The prevalence of the pathogen was expressed in percentage based on total number of seeds plated.
Data were analyzed for Analysis of Variance using MSTAT-C software and before analysis data were transformed following arcsin transformation method. Mean separation was done following Duncan’s Multiple Range Test using the same computer program.
RESULTS AND DISCUSSION
Seedling emergence and seedling disease
At 7 days after sowing (DAS), the lowest seedling emergence of 41.12% was observed under control. Preplant seed treatment with Provax-200, Metataf and Cupravit caused significant increase in seedling emergence within the range of 50.64-55.64% over control. The highest emergence was achieved with Provax-200 followed by Cupravit and Metataf. Efficacy three fungicides to increase seedling emergence was statistically similar (Table 1).
At 14 DAS, seedling infection caused by soil borne fugal pathogens, Sclerotium rolfsii, Rhizoctonia solani and Fusarium oxysporum in a complex form were recorded. Maximum infection of 4.5% was recorded from control plot. It was reduced to 1.00-2.58% due to pre-sowing seed treatment with three fungicides. The reduction was significant over control. The lowest infection was recorded in Provax-200 treated seed plot followed by Cupravit and Metataf (Table 1).
Results of the experiment indicate that pre-sowing seed treatment with fungicides reduced seedling infection effectively. The findings are corroborated with the findings of other investigators (Welty et al.1988, Saraswathi et al. 1989, Barua et al. 2007). Saraswathi et al. (1989) and Mortuza et al. (1995) also reported that field emergence of seedling increased due to seed treatment with fungicides compared to control. Seed treating fungicides might destroy the seed-borne fungi and enhanced germination. Barua et al. (2007) reported that the seed-borne fungal pathogens reduced seed germination but chemical seed treatment reduced seed infection.
Table 1. Effect of pre-sowing seed treatment with fungicide on seedling emergence and seedling infection of mungbean
|Dose of seed treating fungicide (2.5% w/w)||Seedling emergence after 7 days sowing
|Seedling infection at 14 days after sowing
|Provax-200||55.64 a||1.00 c|
|Metataf||50.44 a||2.58 b|
|Cupravit||52.67 a||1.94 b|
|Control||41.12 c||4.50 a|
Values are average of three observations and those within the same column with a common letter(s) do not differ significantly (P = 0.05)
Incidence of Cercospora leaf spot
Incidence of Cercospora leaf spot was 1.20, 4.70 and 18.62% under control at 30, 45 and 60 DAS, respectively. Combination of pre-sowing seed treatment and foliar spray with fungicides in different combinations reduced the disease incidence over control within the range of 0.23-0.78% at 30 DAS, 1.27-3.84% at 45 DAS and 6.41-14.68% at 60 DAS. The reduction was significant at all stages of data collection compared to control (Table 2). At 30 DAS, the lowest disease incidence was achieved with combination of seed treatment with Metataf and foliar spray with Bavistin 50 WP, which was statistically similar to foliar spray of Bavistin alone, Provax + Dithane M-45, Provax + Bavistin, Cupravit + Rovral and Cupravit + Bavistin. The highest incidence was recorded from Metataf followed by Cupravit, Metataf + Dithane M-45 and Rovral alone. Their differences were not significant. At 45 DAS, the lowest incidence of Cercospora leaf spot was observed in treatment Provax + Bavistin, which was statistically similar to the treatments, seed treatment with Provax-200 + foliar spray with Dithane M-45, Seed treatment with Metataf + foliar spray with Bavistin 50 WP, Seed treatment with Cupravit + foliar spray with Rovral-200 and Seed treatment with Cupravit + foliar spray with Bavistin 50 WP (Table 2).
The results of the experiment indicate that effect of seed treatment and foliar spray with fungicides on the reduction of incidence of Cercospora leaf spot of mungbean is appreciable. The combination of seed treatment with Provax and foliar spray of Bavistin is superior to other treatments in reducing disease incidence. Effectiveness of Bavistin 50 WP in controlling CLS has been reported by other workers. Mian et al. (2000) found efficacy of Bavistin 50 WP and Indofil 80 WP to control CLS. Bakr (1993) reported that foliar application of Bavistin 50 WP at 0.1% successfully controlled CLS of mungbean.
Severity (PDI) of CLS
At 30 DAS, the lowest PDI of CLS was recorded from the in treatment combination of seed treatment with Provax and foliar spray with Bavistin 50 WP, which was statistically similar to the treatments, seed treatment with Cupravit + foliar spray with Dithane M-45) and foliar spray with Bavistin. The highest PDI of CLS was recorded from the treatment seed treatment with Cupravit, which was statistically similar to the treatments Seed treatment with Provax-200 + foliar spray with Dithane M-45 and Seed treatment with Metataf+ foliar spray with Rovral 50 WP. All other treatments also showed significant reduction in PDI compared to control. (Table 2).
Table 2. Effect of pre-sowing seed treatment and foliar spray of seed crop with fungicide on incidence and severity of Cercospora leaf spot
|Treatment||Incidence (%) at different days after sowing||Severity (%) at different days after sowing|
|Seed treatment with Provax-200 alone||0.51d*
|Seed treatment with Metataf alone||0.78 b
|Seed treatment with alone Cupravit||0.70 bc
|Foliar spray with Dithane M-45 alone||0.61 cd
|Foliar spray with Rovral||0.67 bc
|Foliar spray with Bavistin alone||0.36 e
|Seed treatment with Provax-200 + foliar spray with Dithane M-45||0.27 e
|Seed treatment with Provax-200 + foliar spray with Rovral 50 WP||0.56 cd
|Seed treatment with Provax-200 + foliar spry with Bavistin 50 WP||0.25e
|Seed treatment with Metataf + foliar spray with Dithane M-45T2+ T4||0.68 bc
|Seed treatment with Metataf+ foliar spray with Rovral 50 WP||0.55 cd
|Seed treatment with Metataf+ foliar spray with Bavistin 50 WP||0.23 e
|Seed treatment with Cupravit + foliar spray with Dithane M-45||0.57 cd
|Seed treatment with Cupravit + foliar spray with Rovral-200||0.29 e
|Seed treatment with Cupravit + foliar spray with Bavistin 50 WP||0.27e
*Values are average of three observations and those within the same column with a common letter(s) do not differ significantly (P = 0.05).
**Figures within parentheses are arcsin transformed value.
At 45 DAS, the lowest PDI of 10.73 was recorded in Provax + Bavistin 50 WP, which was statistically similar to the treatment Metataf + Bavistin, Cupravit + Bavistin and Provax + Rovral. The highest PDI of 12.20 was recorded which was significantly reduced over control by all treatments with fungicides applied as treatants as well as foliar sprays and their combinations. The second highest PDI of 10.86 was recorded from treatment seed treatment with Metataf followed by seed treatment with Cupravit, seed treatment with Provax-200 and foliar spray with Rovral (Table 2).
At 60 DAS, the lowest PDI of 10.14% was recorded from treatment Provax + Bavistin which was statistically similar to treatments Metataf + Bavistin, Cupravit + Bavistin, Provax + Rovral, Cupravit + Rovral, Cupravit + Dithane M-45, Provax-200 + Dithane M-45 and foliar spray of Bavistin. The maximum PDI of 26.96 was recorded from control. All treatments with fungicides caused significant reduction in the severity of CLS over control. (Table 2).
All the treatments controlled the pathogen effectively and reduced disease incidence and severity. Among the treatments, combination of seed treatment with Provax + foliar spray of Bavistin 50 WP performed better than other treatments to reduce incidence and severity of the CLS. Dubey and Singh (2005) reported that dry seed treatment and foliar sprays with Carbendazim gave maximum grain yield along with minimum PDI of Cercospora leaf spots. Khunti et al. (2005) found Carbendazim as a highly effective fungicide for the management of Cercospora leaf spot and powdery mildew. Singh et al. (1994) reported that Cercospora leaf spots could be effectively controlled with Carbendazim because of longer residual toxicity as compared to Mancozeb. The findings of the present study support the findings of (Khunti et al. 2005, Singh et al. 1994).
Yield and yield contributes
Pod length, seed number per pod and 1000-seed weight varied 7.85-9.00 cm, 8.85-9.64 and 8.49-64 g under different treatments including control with means of 8.25 cm, 9.08 and 45.64 g and standard deviation of 0.29 cm, 0.37 and 0.92 g, respectively. However, their variations were not significant and data are not shown in tabular form.
The number of pods plant-1 ranged from 37.23 to 53.33 under different fungicidal treatments including control. Effect of only seed treatment with Provax, Metataf and Cupravit, only foliar spray with Dithane M-45, Rovral, and Bavistin and combination of seed treatment with Metataf and foliar spray with Rovral on number of pods per plant was not significant compared to control. Other treatments with seed treatants as well as foliar spray with fungicides significantly increased pod number per plant compared to control. The highest number of pods plant-1 was recorded from the treatment, Provax + Bavistin 50 WP which was statistically similar to the treatments where fungicides were applied as foliar spray (Table 3).
Grain yield under different treatments including control ranged from 1465.13 to 1862.42 kg ha-1. The highest yield was recorded from the treatment Provax + Bavistin 50 WP, which was statistically similar to other treatments where foliar spray alone and in combination with seed treatments were used. The yield in these treatments was significantly higher over control. The maximum yield increase over control was recorded from the treatment Provax + Bavistin 50 WP followed by the treatment Metataf + Bavistin, Provax + Rovral, Cupravit + Bavistin (Table 3).
Mian et al. (2000) reported that increase in the number of pods plant-1, pod length, seeds pod-1 and 1000-seed weight might have resulted in maximum yield of mungbean. Haque et al. (1994) reported that three or four sprays with Bavistin was effective to control CLS and to increase yield of mungbean. Ayub et al. (1993) also reported that Bavistin, Bavistin + Dithane M-45, Calixin, Copac, Rovral and Tilt were effective against CLS and increased the seed yield of mungbean. Results of the present study reveal that seed treatment with Provax and thrice foliar spray with Bavistin 50 WP reduces the incidence of Cercospora leaf spot (CLS) and increases seed yield appreciably.
Seed health of mungbean
All the fungicidal treatments significantly reduced the prevalence of fungi associated with harvested seeds compared to control. Seed-borne fungal pathogens were Aspergillus spp. (A. flavus and A. niger) and Fusarium oxysporum. The lowest prevalence of Aspergillus spp. was recorded in treatment Provax + Bavistin, which was statistically similar to all other treatment combinations of seed treatment and foliar spray except Metataf + Dithane. The highest Aspergillus spp. (30.00 %) was recorded from control (Table 3).
The lowest prevalence of F. oxysporum was obtained with the treatment Provax + Bavistin, which was statistically similar to the treatment Cupravit + Bavistin. Significantly the highest prevalence of F. oxysporum was record under control. Effects of foliar spray with Dithane M-45, Rovral and Bavistin 50WP were statistically similar but significantly higher than seed treatment with Provax, Metataf and Cupravit alone (Table 3).
Table 3. Effect of pre-sowing seed treatment and foliar spray of seed crop with fungicide on yield and health of mungbean seeds
|Treatment||Pods number plant-1||Yield
|% seed-borne infection|
|Seed treatment with Provax||39.73 bcd||1612.08 bc||18.25 b||8.00 b|
|Seed treatment with Metataf||38.37 cd||1612.36 bc||18.25 b||8.00 b|
|Seed treatment with Cupravit||38.53 cd||1617.10 bc||18.00 bc||7.58 b|
|Foliar spray with Dithane M-45||45.45 abcd||1655.9 abc||14.75 def||5.42 c|
|Foliar spray with Rovral||43.47 abcd||1647.9 abc||16.75 bcd||5.83 c|
|Foliar spray with Bavistin||47.68 abcd||1766.86 ab||15.92 bed||4.67 cd|
|Seed treatment with Provax + Foliar spray with Dithene M-45||51.23 a||1752.37 ab||11.42 fg||4.50 cd|
|Seed treatment with Provax + Foliar spray with Rovral||50.98 ab||1794.59 ab||11.92 efg||4.58 cd|
|Seed treatment with Provax + Foliar spray with Bavistin||53.33 a||1862.42 a||10.00 g||3.17 e|
|Seed treatment with Metataf + Foliar spray with Dithene M-45||50.25 ab||1777.36 ab||14.83 cde||4.67 cd|
|T11 = Seed treatment with Metataf + Foliar spray with Rovral||48.03 abcd||1712.55 ab||11.75 efg||4.58 cd|
|Seed treatment with Metataf + Foliar spray with Bavistin||51.13 ab||1837.13 ab||11.08 g||4.58 cd|
|Seed treatment with Cupravit + Seed treatment with Cupravit||49.10 abc||1729.30 ab||11.67e fg||4.83 cd|
|Seed treatment with Cupravit + Foliar spray with Rovral||51.02 ab||1747.25 ab||12.67e fg||4.58 cd|
|Seed treatment with Cupravit + Foliar spray with Bavistin||51.80 a||1792.18 ab||11.50 efg||3.92 de|
|Control||37.23 d||1465.13 c||30.00 a||12.67 a|
* Values are average of four observations and those within the same column with the same letter(s) do not differ statistically (P = 0.05).
* Aspergillus spp. includes A. flavus and A. niger.
The significantly highest prevalence of Aspergillus spp. and F. oxysporum was recorded in control treatment than any other treatments indicating significant effect of fungicidal seed treatment and foliar spray on the control of Aspergillus spp. and F. oxysporum. The results of this study indicate that there was a significant effect of fungicidal seed treatment and foliar spray independently on the reduction of seed infection but combination of seed treatment + foliar spray performed better. Effectiveness of Vitavax-200 in controlling seed-borne fungi has also been reported by Barua (2004), who reported that seed treatment + foliar spray with vitavax-200 successfully controlled 85.5 to 90.1% seed-borne fungi of M. phaseolina and Fusarium sp. Seed-borne infection could be reduced if infected seeds are treated with Oxathiin @ 2.5 g kg-1 of seed (Agarwal and Verma,1975). Results of the present study reveal that seed treated with Provax + foliar spray with Bavistin 50 WP reduced the prevalence of seed-borne fungi in harvested mungbean seeds and thus improved seed health.
The findings of the present investigation reveal that seed treatment with Provax and three foliar spray of seed crop with Bavistin 50 WP is the best treatment to reduce the seedling infection, incidence and severity of Cercospora leaf spot (CLS) and to increase yield. The treatment also reduces the prevalence of seed-borne fungi in harvested seeds and improved seed health.
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