Plant parts of Acacia nilotica (L.) Willd. ex Delile and Sapindus mukorossi (L.) were collected from Campus of University of Karachi and microbial antagonists including Trichoderma harzianum Rifai (Th-6), Paecilomyces variotii Bain (Pv-14), Rhizobium meliloti Frank (Rm-5) and Bacillus thuringiensis Berliner (Bt-9) were obtained from Karachi University Culture Collection (KUCC). Aqueous extracts of A. nilotica and S. mukorossi plant parts were prepared with different concentrations and screened against root infecting fungi (Rhizoctonia solani Kühn, Fusarium oxysporum Schlect., Macrophomina phaseolina (Tassi) Goid) using agar well diffusion and paper disc diffusion methods. Paper disc diffusion method showed pronounced effect against root infecting fungi by aqueous leaves extract of A. nilotica (100 % w/v). Interaction of microbial antagonists with pathogenic fungi showed that both fungal and bacterial antagonists were effective in the inhibition of root rot fungi by producing the zone of inhibition. In pot experiments, leguminous and non-leguminous seeds (peanut, chickpea, sunflower and okra) were primed with plant parts (stem, leaves and seeds) extracts of A. nilotica and S. mukorossi for 5, 10, 20, and 40 minutes where seed-priming with A. nilotica and S. mukorossi leaves extracts for 10 minutes time interval was found to be the most effective for the control of root rot fungi and growth of all tested crop plants. Effects of seed bio-priming with microbial antagonists at different time intervals (5, 10, 20, and 40 minutes) showed that M. phaseolina and Fusarium spp were suppressed when seeds were bio-primed with R. meliloti, T. harzianum and B. thuringiensis conidial/cell suspensions at 5, 10 and 20 minutes whereas, root weight, shoot length, root length and shoot weight were increased prominently after seed bio-priming with T. harzianum and R. meliloti for 10 minutes. Biopriming of seeds with different doses of T. harzianum, R. meliloti and different concentrations of A. nilotica and S. mukorossi leaves extracts were performed in pot experiment which showed that 100 % concentration (stock) of leaves extracts was found to be most effective and (188 x 107 conidia/mL) conidal suspension of T. harzianum was found to be most effective for enhancing the growth and suppression of root rot fungi on leguminous and non-leguminous crops. Combined effect of different fertilizers (urea and di-ammonium phosphate) @ 0.1 and 0.01 % and seed bio-priming with R. meliloti, B. thuringiensis P. variotii and T. harzianum, A. nilotica and S. mukorissi plant parts for 5, 10, 20 and 40 minutes recorded that seed bio-priming with A. nilotica, S. mukorossi, T. harzianum and R. meliloti for 10 minutes and soil amendment with urea @ 0.1 % significantly control root rot disease as well as increased the growth of crop plants. Another pot experiment was performed where soil amendment with oil seed cakes (mustard, cotton, almond and Nigella sativa cakes) @ 0.1 and 1% w/w and priming of seeds with leaves extracts of A. nilotica and S. mukorossi, Trichoderma harzianum and Rhizobium melilotii conidial/cell suspension showed that T. harzianum conidial suspension and amendment of soil with mustard cake @ 1% was found to be most effective for the growth of leguminous and nonleguminous crops and reducing the root infecting fungi followed by R. meliloti primed seeds and cotton, almond and N. sativa cakes amendment respectively as compared to the control (non treated seeds and soil). The effect of different fungicides like mancozeb and dithane in combination with priming of leguminous and non-leguminous seeds with plant extracts was studied in pots experiments in which combined application of seed priming with A. nilotica and S. mukorossi leaves extracts and soil drenching with dithane and mancozeb fungicides @ 0.1 % was found to be most effective for enhancing the growth and reduction of root rot fungi. A. nilotica and S. mukorossi pellets (pyrophyllite mixed with leaves powder @ 50:50 ratio),capsules (empty shells of capsules filled with leaves powder @ 0.5 g) alone or in combination with biopriming of leguminous and non-leguminous seeds with T. harzianum and R. meliloti were also studied in pot experiment which showed that A. nilotica, S. mukorossi pellets and capsules in combination with bio-priming of seeds with T. harzianum were most effective for growth promotion and suppression of root infecting fungi on peanut, chickpea, okra and sunflower. Pot experiment was performed in which the resistance of different cultivars of okra (OH- 152, Arka anamika and unknown variety purchased from local market), peanut (BARI-II, Golden, BARI-2000), chickpea (Bakhar-2009, Pb-2008, Noor) and sunflower (S-278, Hysun-39 and unknown variety purchased from local market) against root rot fungi were screened after bio-priming with A. nilotica, S. mukorossi leaves extracts and T. harzianum, R. meliloti conidial/cells suspension for 10 and 20 minutes. In this experiment, variety S-278 and variety Pb-2008 after bio-priming with A. nilotica leaves extract for 10 minutes and T. harzianum conidial suspension for 20 minutes was found to be most effective for plants growth and complete inhibition of M. phaseolina followed by R. solani and Fusarium spp. Variety OH-152 bio-primed with A. nilotica leaves extract, T. harzianum and R. meliloti cells/conidal suspension for 10 minutes was recorded to be most effective for the complete reduction of M. phaseolina and significant elevation of plant growth. However, in peanut cultivars, bio-priming of seeds with A. nilotica leaves extracts and T. harzianum conidial suspension for 10 minutes was most effective and variety BARI-II found to be most resistant against root infecting pathogens and growth promotion of crop plants. Long-term resistance of seed bio-priming (with plant parts and microbial antagonists) was evaluated under different storage conditions both at room temperature and low temperature for 0, 30, 60, 90, and 180 days. T. harzianum and R. meliloti primed seeds under storage showed that storage of T. harzianum primed seeds for 90 and 180 days at low temperature (4°C) greatly enhanced the growth parameters as well as significantly inhibit the incidence of M. phaseolina followed by R. solani and Fusarium spp on crop plants. In case of plant parts bio-primed seeds under storage, it was noticed that storage of seeds primed with A. nilotica leaves extracts gave maximum growth and root rot inhibition of R. solani and seeds were effective for 180 days whereas, maximum growth of plants were noticed at 60 days of seed storage and 4°C temperature was the best temperature for storage of primed seeds. Field application of bio-primed seeds with plant parts and microbial antagonists was found to be effective when seeds of crop plants were bio-primed with A. nilotica leaves extracts and T. harzianum conidial suspension which not only enhances the growth parameters but also efficiently reduced soil borne root rot fungi. Field application of okra, peanut, chickpea and sunflower cultivars also observed that okra cultivar OH-152, peanut cultivar BARI-II, chickpea cultivar Pb-2008 and sunflower cultivar S-278 were the most resistant varieties against root infecting fungal pathogens and treatments of Acacia nilotica and T. harzianum were noticed as the most useful treatments in the elevation of growth and vigor of leguminous and nonleguminous crops. In BARI-II and OH-152, Catalase (CAT) enzyme activity increased when seeds bio-primed with A. nilotica leaves extract and T. harzianum conidial suspension while, S. mukorossi and A. nilotica bio-priming enhanced the level of Guaiacol peroxidase (GPX) enzyme as compared to the untreated seeds (control). In Pb-2008 and S-278, seeds bio-primed with A. nilotica showed maximum activity of Catalase enzyme whereas, Guaiacol peroxidase enzyme activity elevated with A. nilotica and T. harzianum treatments.