Macrophomina phaseolina (Tassi) Goid remains the prevailing causal agent of charcoal rot disease that significantly suppresses the yield of a variety of oilseed crops. Its wide host range and ability to survive under arid conditions, coupled with the ineffective use of fungicides against it, have spurred scientific endeavours for alternative avenues to control this phytopathogen. Hence, the present study aimed to provide empirical evidence of the efficacy of three fungal isolates (T2, T10 and T12) of Trichoderma harzianum as biological control agents against charcoal rot in soybean (Glycine max L.). The results of the in vitro studies revealed that all three fungal isolates significantly inhibited the growth of M. phaseolina phytopathogen, with T12 showing considerably higher inhibition effect than T2 and T10 isolates. T12 inhibited the growth of M. phaseolina in the dual culture (72.31%) and volatile production (63.36%) assays, and the hyperparasitism test indicated cell lysis following the interactions with T12 mycelia. T12 isolate was mostly effective in field experiments, observable in the attained minimum plant disease indices both in the soil incorporation (11.98%) and seed inoculation (5.55%) treatments, in comparison to isolates T2 and T10. Moreover, the stem and root lengths, as well as the seed weight, were considerably increased, as compared to the control. Hence, the findings reported in the present study supported the applicability of T12 isolate as possible alternative to fungicides for the control of charcoal rot in soybean.

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Genomic DNA of the mutant lines of the three potato cultivars, Cardinal, Diamant and Desiree, with respect to controls were isolated and analyzed for polymorphisms by using random amplified polymorphic DNA (RAPD) markers. Four 10 bp random fragment primers, S-13, S-18, S-19 and R-17 were studied and all of them gave the amplification of genomic DNA. All of the mutant lines gave different banding pattern against different primers with respect to control plants of the three varieties, and bands are present at 50 bp to 1500 bp. All these primers with specific banding pattern were unique in their polymorphic behavior. Different banding pattern of total protein contents were also observed by PAGE analysis of all the mutant lines as compared with the control plants. It is therefore suggested that RAPD and protein analyses would be important tools to detect the polymorphism in mutated lines of potato.

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Bacteria blight is one of the most serious bacterial diseases of rice worldwide. The identification of genetic potential against bacterial blight in the existing rice resources is a prerequisite to develop multigenic resistance to combat the threat of climate change. This investigation was conducted to evaluate alleles variation in 38 Malaysian cultivars using thirteen Simple Sequences Repeats markers and one Sequence Tagged Sites (STS) marker which were reported to be linked with the resistance to bacterial blight. Based on molecular data, a dendrogram was constructed which classified the rice cultivars into seven major clusters at 0.0, 0.28 and 0.3 of similarity coefficient. Cluster 5 was the largest group comprised of ten rice cultivars where multiple genes were identified. However, xa13 could not be detected in the current rice germplasm, whereas xa2 was detected in 25 cultivars. Molecular analysis revealed that Malaysian rice cultivars possess multigenic resistance.

Due to the toxicity and inefficiency of chemical fungicides to control infestation of Macrophomina phaseolina (Tassi) Goid which causes charcoal rot in plants, a biotechnological approach using βglucosidase (EC.3.2.1) as the alternative bioactive ingredient in fungicide is hereby, proposed. The extracellular enzyme was isolated from a highly efficient fungal antagonist, Trichoderma harzianum T12. The highly similar molecular masses obtained using SDS-PAGE (96 kDa) and MALDI-TOF mass spectrometry (98.3 kDa) affirmed that the β-glucosidase was purified to homogeneity. Consequently, optimum catalytic parameters that rendered the highest enzyme activity were found to be: 45˚C, pH 7, inoculum size of 10 % (w/v), supplementation with metal ions Zn 2+ and Mn 2+ ions, and Tween 80. Addition of wheat bran and (NH 4 ) 2 SO 4 as carbon and nitrogen sources also improved enzyme activity. BLASTn showed the sequence of β-glucosidase T12 was highly identical to other β-glucosidases viz. T. harzianum strain IOC-3844 (99%), T. gamsii and T. virens bgl1 (86 %) as well as T. reesei strain SJVTR and T. viride strain AS 3.3711 (84 %). Kinetic assessment showed that β-glucosidase T12 catalyzes hydrolytic activity is characterized by a K m of 0.79 mM and V max of 8.45 mM min -1 mg -1 protein, with a corresponding k cat of 10.69 s -1 .

Rice production is decreasing by abiotic stresses like heavy metals. In such circumstances, producing food for growing human population is a challenge for plant breeders. Excess of Al3+ in soil has become threat for high yield of rice. Improvement of crop is one of potential solution for high production. The aim of this study was to develop the new method for optimization of Al3+ toxicity tolerance in indica rice at germination stag using two-way ANOVA and Duncan's multiple-range test (DMRT). Seeds of two indica rice cultivars (Pokkali and Pak Basmati) were exposed in different concentrations (control, 5 mM, 15 mM, and 20 mM) of Al3+ toxicity at pH 4 ±0.2 for two weeks. Germination traits such as final germination percentage (FG%), germination energy (GE), germination speed (GS), germination index (GI), mean time of germination (MGT), germination value (GV), germination velocity (GVe), peak value of germination (GPV), and germination capacity (GC) and growth traits such as root length (RL), shoot length (SL), total dry biomass (TDB), and germination vigour index (GVI) were measured. To obtain the maximum number of significance (≤ 0.01%) parameters in each concentration of Al3+ toxicity with control, two-way ANOVA was established and comparison of mean was done using DMRT. The results showed that 5 mM, 10 mM, and 15 mM have less significant effects on the above-mentioned parameters. However, 20 mM concentration of Al3+ produced significant effects (≤ 0.01%). Therefore, 20 mM of Al3+ is considered optimized limit for indica cultivars (Pokkali and Pak Basmati).