Contrary to chemical herbicides, the use of plant pathogenic fungi have been suggested as one of the best possible mean for controlling parthenium weed (Parthenium hysterophorus L.). Mass Production of mycoherbicides by solid substrate fermentation using four different strains of Alternaria macrospora Zimm. was standardized. Selection of suitable inexpensive and easily available agro-waste substrate for the mass production of mycoherbicidal agents was the main aim of the present study.  Mycelial growth was recorded on seven of the inoculated solid substrates. The outcome of the study indicates that chopped parthenium leaves were the most suitable solid substrate for the mass production of A. macrospora MKP1 and A. macrospora MKP3 while wheat bran for A. macrospora MKP2 and A. macrospora MKP4. The highest sporulation was observed with wheat bran. Contrary to these, wood shavings failed to support the growth and sporulation of the pathogens. Parthenium leaves (chopped and powdered) and wheat bran colonized inocula incited severe infection and were responsible for significant mortality of parthenium plant.