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Effects of Macrophomina Phaseolina on Seed Viability, Vigour and Storability in Blackgram

M.S. Rahman
Bangladesh Sugarcane Research InstituteIshurdi, Pabna

S. Vearasilp
Department of Agronomy, Chiang Mai University, Thailand

S. Srichuwong
Department of Plant Pathology, Chiang Mai University, Thailand

Abstract/ Executive Summary:

Macrophomina phaseolina, a seed-borne fungus is a causal organism of charcoal rot disease of blackgram. To know the actual detrimental effects of this fungus on seed quality of blackgram, an experiment was undertaken by comparing inoculated and uninoculated seeds. Artificially inoculated seeds of two blackgram varieties with M. phaseolina showed about ?rd number of normal seedlings in comparison to uninoculated seeds. The mean shoot length, root length and dry weight of seedlings, which are the parameters of seedling vigour, were also found significantly reduced in both inoculated blackgram varieties. Accelerated aging test (AA-test) showed nearly half of normal seedlings in inoculated seeds compared to control in both the varieties, which indicates that M. phaseolina can reduce the storability of blackgram seeds greatly. In AA-test, the proportion of dead and rotten seeds in the inoculated treatments appeared to be nearly two folds higher than that of control treatment.

Keywords: Macrophomina phaseolina, Blackgram, Seed viability, Vigour, Storability.

Location: Thailand

Start Date: 00-00-2008

End Date: 00-00-0000

Program Area: Seed Technology

Commodity: Black gram

Objectives:

To know the actual detrimental effects of this fungus on seed quality of blackgram

Methodology:

The experiment was conducted in the Seed Science & Technology laboratory, Faculty of Agriculture, Chiang Mai University, Thailand.Seed Samples: The two blackgram varieties Uthong 2 and Phitsanulok 2 were grown in Chiang Mai University experimental farm, Thailand and newly harvested seed samples were used in the experiment. Isolation and pure culture of M. phaseolina: Seed sample of blackgram named as Uthong 2, which was obtained from Chai Nat Field Crops Research Centre, Thailand, was carrying about 24.0 percent M. phaseolina according to Blotter Test. Fifty seeds were soaked with 10% sodium hypochlorite solution followed by washing with sterilized water three times. Then the seeds were placed equidistantly at the rate of 10 seeds per plate, in sterilized 9 cm-diameter petriplates, containing 3-layered Whatman no. 1 blotting paper. After 3 days, the seeds with well-noticed pycnidia and sclerotia of M. phaseolina on seed coat and radicle were transplanted in sterilized petriplate containing about 20ml solidified potato dextrose agar (PDA) Difco. Seed was placed in the middle of the plate @ 1 seed/plate. The plates were sealed by Nesco film to avoid contamination by other aerial microorganisms. After 3 days, when the mycelia of M phaseolina were spread around the seed, 5mm diameter of PDA media with mycelia were cut by sterilized cork borer and placed on the middle of another sterilized petriplate containing PDA. These plates were kept under 12-hour alternating light and darkness after sealed by Nesco film. After 3 days, the blakish mycelia and sclerotia of M. phaseolina grown profusely in the plates were used as pure culture. Inoculation of seed: Newly grown seeds of two varieties of blackgram were inoculated by the pure culture of M. phaseolina. About one thousand seeds of each variety were surface sterilized with 10% sodium hypochlorite solution for 2 minutes followed by rinsing with sterilized distilled water for 3 to 4 times. Half amount of seed of each variety was used for control or check and the remaining half was inoculated with previously prepared mycelia and sclerotia of M. phaseolina. The mycelia and sclerotia were separated from PDA media by scrapping with a sterilized knife. The scrapped fungal structures were suspended with sterilized water at 50ml water per petriplate. The seeds were soaked for one hour in this suspension. The control seeds were also soaked with sterilized water in the same way for one hour. All the inoculated and non-inoculated or control seeds were dried under fan for overnight. Viability test: Viability test of seed was employed by Paper Towel Method (ISTA, 1976). Fifty seeds were placed on two layered moist germinating papers, covered with another moist germinating paper, and then rolled. Four such rolls of each containing 50 seeds were placed as four replicates. All the rolls were kept in germinating chamber (Temperature 30⁰C and relative humidity 85%). The inoculated and non-inoculated seeds were placed separately in the germinating chamber. After 6 days, data were recorded on normal seedlings, abnormal seedlings, rotten or dead seeds, and hard seeds. Vigour test: The vigour of seedling was estimated by the mean shoot length, root length and dry weight of 7 days old seedlings from inoculated and non-inoculated seeds. The seedlings were raised in the germinator with Paper Towel Method (ISTA, 1976). The average of seedlings shoot and root length was estimated from 50 seedlings per replication. Dry weight of 7 days old seedlings was taken after drying of the seedlings in air-dry oven at 60⁰C for 3 days. In every case, four replications were maintained. Storability test: It was done by Accelerated Aging test (AA-test) outlined in the AOSA’s Vigour Testing Handbook (AOSA, 1983). Delouche and Baskin (1973) developed an accelerated aging (AA) test to estimate the relative life span of seed in warehouse storage. The seeds are exposed to high humidity and high temperature. The conditions promote the growth of fungi on seeds and caused deterioration. For blackgram, exposing of seeds at 41⁰C temperature and 99 to 100 percent relative humidity for 96 hours is almost equivalent to store the seed for one year (Ross and Minalo, 1972). About 10g of inoculated and non-inoculated seeds of each variety were distributed uniformly in two layers in 10 cmx10 cm trays. The trays were kept on the stand just above one cm from the surface of 50ml of water glass container. Caution was taken to avoid direct contact between water and seeds. The containers were sealed with their lids and placed in an accelerated aging chamber maintained at 41⁰C temperature and 99 to 100% relative humidity. After 96 hours, seeds were removed and dried on paper towel on the table for two hours at room temperature. The aged seeds were tested by Paper Towel Method (ISTA, 1976) following the same procedure as the viability test. After 7 days, data were recorded on normal seedlings, abnormal seedlings, dead seeds and hard seeds.

Source of Information: PhD thesis of Mr Md. Sharif Uddin, Roll No. & Reg. No.: 01,11321 (1982-83 Department of Soil Science,Bangladesh Agricultural University, Mymensingh Bangladesh J. Seed Sci. Technol. 13(1&2): 1–4 (2009), ISSN 1029-8800

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

it is obvious that M. phaseolina can decrease the seed quality of blackgram by reducing viability, vigour and storability. As M. phaseolina is a seedborne pathogen hence the infested seed is the main cause of spread disease in the field. So, to keep the quality of seed, appropriate and effective control measure like seed treatment should be done before storing and planting in the field.

Knowledge Management: Journal, Thesis