Use of Botanical Products for Breakdown the Parasitize of New Recorded Penicillium funiculosum of Aonla

The present investigation on was undertaken in the Department of Plant Pathology, B.A. College of Agriculture, A.A.U., Anand. The Penicillium funiculosum rot of aonla was found a new record in Gujarat and it was identified by Division of Mycology and Plant Pathology, Indian Agricultural Research Institute, New Delhi (ITCC No.: 7046.08). Bio-efficacy of nine phytoextracts at 10% were tested against the mycelial growth and sporulation of Penicillium funiculosum in vitro, among them significantly lowest mycelial growth of P. funiculosum was recorded in neem leaf extract (7.75 mm) showing 89.49% growth inhibition. Further it was also proved most effective in reducing the Penicillium rot severity both in pre- (20.44%) and post-inoculation (20.87%) treatments at 7 day after inoculation, respectively. The next best treatment in tested phytoextracts tulsi and garlic were found both in in vitro and in vivo condition.

INTRODUCTION

The Aonla (Emblica officinalis Gaertn., Syn. Phyllanthus emblica L.) is an important arid fruit crop in India. The area under aonla cultivation in India is about 50,000 hectares with an annual production 2,00,000 metric tonnes (Goyal et al., 2008). In Gujarat state has 12,000 hectares area under aonla cultivation with an annual production of one lakh MT (Shah et al., 2006). In India, nearly 20-25% of perishables are lost due to post harvest diseases (Sharma and Mashkoor Alam, 1998). It is reported that around 35% of produce is lost in storage and transits (Rawal and Saxena, 2005). In crop protection studies, various natural plant products have been identified and employed to control post harvest diseases of fruits.

MATERIALS AND METHODS

In vitro: Efficacy of different phytoextracts of plant species having medicinal value were tested in vitro by Poisoned Food Technique Method against aonla fruit rot pathogen (Nene and Thapliyal, 1979). All the phytoextracts were tested at 10% conc. Fresh and healthy 100 g plant parts of each species were thoroughly washed with tap water and then with sterile distilled water. They were macerated separately in grinder mixture by adding 100 mL ethanol.

The mixture was filtered through two fold sterilized muslin cloth and the filtrate was centrifuged at 5000 rpm for 10 min. and the clear supernatant extract was collected in sterilized conical flasks. After evaporating the ethanol from extract, the clear extract was collected and diluted with 100 mL distilled sterile water to make volume 1:1 (w/v). This was considered as 100 per cent concentration for the study to test the efficacy of plant extracts (Sinha and Saxena, 1989). Each phytoextracts (10%) were mixed thoroughly in sterilized 100 mL PDA medium filled in 250 mL flask under aseptic condition. The medium was supplemented with streptomycin sulphate @ 50 ppm to prevent bacterial contamination.

In vivo: The phytoextracts studied in vitro conditions were tested at 10% concentration by following pre- and post-inoculation methods.

Pre-inoculation method: The healthy, semi-ripe, uniform size aonla fruits of Gujarat Aonla-1 cultivar were surface sterilized by dipping in 0.1% HgCl₂ solution for one minute followed by three washings with distilled sterile water and inoculated separately by the stem end injury method. The spores from 7 day old culture of fruit rot pathogen were scrapped with the sterile needle and mixed in 200 mL distilled sterile water. The fruits were first dipped in phytoextracts (10%) separately and then inoculated with the pathogen by keeping 12 h interval. The fruits were injured with sterilized cork borer at stem-end and dipped in spore suspension (10⁶ spores mL) for 2 min and air dried for 15-20 min. The inoculated and uninoculated (dipped in distilled sterile water) fruits were placed in sterilized polythene bags. One fruit was accommodated in one bag. A piece of sterilized moist absorbent cotton swab was placed inside the bag and mouth of the bag was loosely tied. The bagged fruits were incubated at 27±1°C for 7 day.

Post-inoculation method: In post-inoculation treatment, the fruits were first inoculated with the pathogen and then treated with the phytoextracts. Further procedure was followed as mentioned above.

RESULTS AND DISCUSSION

In vitro: All the phytoextracts screened were found significantly superior in inhibiting the mycelial growth and sporulation of P. funiculosum over control. The results presented in Table 1 revealed that significantly highest mycelial growth inhibition was recorded in neem leaf extract (89.49%) followed by tulsi leaf extract (87.79%) and garlic cloves extract (84.74%). The next best treatment in order of merit were lemon grass, datura leaf, bittergourd fruit, alovera leaf and ginger rhizome extract giving 77.28, 69.49, 63.72, 61.69 and 59.66% growth inhibition, respectively. While, ardusi leaf extract found least effective in inhibiting the mycelial growth (39.66%). None of the phytoextracts supported spore production of P. funiculosum.

The present findings are supported by the work carried out by Godara and Pathak (1995). They screened five plant extracts in vitro and reported that Ocimum sanctum leaf extract proved highly effective against P. italicum and Botrydiplodia theobromae causing pre and post-harvest rots of sweet oranges. Khilare and Gangawane (1997) observed that leaf extract of Azadirachta indica (100 PGI) found most effective against Penicillium rot of mosambi (72.4 PGI). Raju and Naik (2006) reported that nimbicidin (2.5 and 5.0%) completely inhibited the mycelial growth of blue mould in onion (Penicillium digitatum) followed by garlic extract (64.19 and 67.46%).

In vivo: All the phytoextracts (10%) were found significantly superior in reducing the Penicillium rot severity over control at 4 and 7th d after inoculation both in pre - and post - inoculation methods (Table 2).

Pre-inoculation: The results presented in Table 2 revealed that the significantly lowest Penicillium rot severity was noted in fruits treated with neem leaf extract (15.00 and 20.44%) at 4 and 7th day of inoculation, respectively but it was at par with tulsi leaf extract (16.84%) at 4th d after inoculation. The next best treatment in order of merit was tulsi leaf extract (24.14%) at 7th day of inoculation followed by garlic cloves extract (17.15 and 25.08%), lemon grass leaf extract (20.23 and 25.26%), ginger rhizome extract (21.75 and 29.48%), datura leaf extract (23.37 and 28.98%), bittergourd fruit extract (24.52 and 30.47%) and ardusi leaf extract (24.90 and 32.07%) at 4 and 7th day after inoculation, respectively. Alovera leaf extract proved least effective in reducing the Penicillium rot (26.00 and 31.44%).

Post-inoculation: Significantly lowest Penicillium rot severity was recorded in fruits treated with neem leaf extract (14.71 and 20.87%) at 4 and 7th day of inoculation, respectively. The next best treatment in order of merit was tulsi leaf extract (17.61 and 23.75%) followed by garlic cloves extract (18.84 and 25.60%), lemon grass leaf extract (22.14 and 26.36%), ginger rhizome extract (22.14 and 30.96%), datura leaf extract (25.08 and 30.12%), bittergourd fruit extract (25.27 and 31.44%) and ardusi leaf extract (25.81 and 33.96%) at 4 and 7th day of inoculation, respectively.

Table 1:	Bio-efficacy of phytoextracts against Penicillium funiculosum in in vitro
PGI: Percent growth inhibition

Table 2:	Bio-efficacy of phytoextracts on the severity of Penicillium rot of aonla
*Values parenthesis are original value, while outside are the arc sine transformed values

Alovera leaf extract found least effective in controlling the Penicillium rot (26.72 and 32.02%).

The results of present investigation are in consonance with the results obtained by Kaur and Verma (2004). They noted that neem dry leaf extracts (5x10⁴ μg mL^-1) found most effective in pre- (94.44%) and post-inoculation (50.00%) in controling green mould rot (Penicillium digitatum) of kinnow fruits. Five% extract of Azadirachta indica found better than Curcuma longa, Zingeber officinale, Cartharanthus roseus, Ocimum basilicum and Allium sativum in controlling the Penicillium rot of aonla both in pre and post-inoculation treatments (Meena, 2006). Singh and Sumbali (2003) noted that leaf extract of Azadirachta indica was most effective in controlling the Penicillium expansum infecting apple fruits.

CONCLUSION

In in vitro all nine phytoextracts screened (10%) were found significantly superior in inhibiting the mycelial growth of P. funiculosum over control. Lowest mycelial growth was recorded in neem leaf extracts (7.75 mm) showing 89.49 per cent growth inhibition over control. The next best treatment in order of merit was tulsi leaf extract (87.79 PGI). While, ardusi leaf extract found least effective in inhibiting the mycelial growth (39.66%).

Neem leaf extract (10%) proved highly effective in reducing the Penicillium rot severity (20.44 and 20.87%) followed by tulsi leaf extract (24.14 and 23.75%) both in pre- and post-inoculation at 7th day of inoculation, respectively. Alovera leaf extract found least effective in controlling the Penicillium rot.

ACKNOWLEDGMENT

Authors are grateful to the Professor and Head, Department of Plant Pathology and Principal, B.A. College of Agriculture, Anand Agricultural University, Anand (Gujarat) for providing necessary facilities.