Secondary metabolite induced tolerance to Fusarium oxysporum f.sp. cubense TR4 in banana cv. Grand Naine through in vitro bio-immunization: a prospective research translation from induction to field tolerance.

An innovative tissue culture mediated incorporation of metabolite-based biomolecule (Bio-immune) at in vitro stage itself in banana cv. Grand Naine was developed and validated for the production of Fusarium oxysporum f.sp. cubense TR4 tolerant plantlets. The novel bio-immune formulation developed by us, exhibited a significant antifungal potency against Foc TR4 with a high percent inhibition (100%) at a 2.5% concentration of bio-immune on the 5th, 7th, and 9th DAI. Bio-immune integrated during in vitro shoot proliferation stage in banana cv. Grand Naine recorded significant enhancement in the growth of roots and shoots. Bio-immune (0.5%) fortified media produced 12.67 shoots per clump whereas control registered only 9.67 shoots per clump. Similarly, maximum root numbers (7.67) were observed in bio-immune plants which were significantly higher over control (5.0). The bio-immunized banana transplants recorded a higher survival rate (97.57%) during acclimatization as compared to the control (94.53%). Furthermore, evaluation of the bio-immunized plants in pot experiments revealed that unimmunized plants treated with FocTR4 (TF) exhibited mortality between 60 and 90 days. On the 90th day after planting, a high mean disease severity index (DSI) of 3.45 was observed with unimmunized plantlets while the bio-immunized plants (TFBI) and ICAR-FUSICONT treated plants (TFTR) showed substantially reduced DSI (0.20 and 1.00) compared to FocTR4 treated control (TF). Significant increases in polyphenol oxidase (PPO), peroxidase (POD), ß-1,3-glucanase, phenylalanine ammonia-lyase (PAL), chitinase activities, and enhanced phenol contents were recorded in bio-immunized plants compared to unimmunized plants. Field experiments at two different locations in Bihar, India revealed that bunch weight, no. of hands/bunch, and no. of fingers/hand of bio-immune treated plants were significantly higher compared to the control.

Papaya Leaf Curl Virus (PaLCuV) Infection on Papaya (Carica papaya L.) Plants Alters Anatomical and Physiological Properties and Reduces Bioactive Components.

Papaya leaves are used frequently for curing scores of ailments. The medicinal properties of papaya leaves are due to presence of certain bioactive/pharmacological compounds. However, the papaya leaf curl virus (PaLCuV), a geminivirus, is a major threat to papaya cultivation globally. During the present investigation, we observed that PaLCuV infection significantly altered the anatomy, physiology, and bioactive properties of papaya leaves. As compared to healthy leaves, the PaLCuV-infected leaves were found to have reduced stomatal density (76.83%), stomatal conductance (78.34%), photosynthesis rate (74.87%), water use efficiency (82.51%), chlorophyll (72.88%), carotenoid (46.63%), osmolality (48.55%), and soluble sugars (70.37%). We also found lower enzymatic activity (superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT)-56.88%, 85.27%, and 74.49%, respectively). It was found that the size of guard cells (50%), transpiration rate (45.05%), intercellular CO2 concentration (47.81%), anthocyanin (27.47%), proline content (74.17%), malondialdehyde (MDA) (106.65%), and electrolyte leakage (75.38%) was elevated in PaLCuV-infected leaves. The chlorophyll fluorescence analysis showed that the infected plant leaves had a significantly lower value of maximal quantum yield of photosystem II (PSII (Fv/Fm), photochemical quantum yield of photosystem I (PSI (Y(I)), and effective quantum yield of PSII (Y(II)). However, in non-photochemical quenching mechanisms, the proportion of energy dissipated in heat form (Y(NPQ)) was found to be significantly higher. We also tested the bioactivity of infected and healthy papaya leaf extracts on a Caenorhabditis elegans (C. elegans) model system. It was found that the crude extract of papaya leaves significantly enhanced the life span of C. elegans (29.7%) in comparison to virus-infected leaves (18.4%) on application of 100 µg/mL dose of the crude extract. Our research indicates that the PaLCuV-infected leaves not only had anatomical and physiological losses, but that pharmacological potential was also significantly decreased.

Studies on bio-chemical profiling of Indian gooseberry (Emblica officinalis) for genetic diversity.

Biochemical profiling of physiologically mature fruits of 51 diverse Indian gooseberry (Emblica officinalis Gaertn) germplasm accessions was collected from Vindhyan hill region of Madhya Pradesh, with a view to select nutraceutically rich genotypes based on important biochemical traits. The mean ascorbic acid and total phenol (tannin) content amongst different accessions was recorded as 496.47 mg 100 g⁻¹ and 4.88% with highest value found in CISH A-12 (654.50 mg 100 g⁻¹) and CISH A-30 (7.18%), respectively. Apart from the above, wide range of variability in the composition of other important biochemical attributes viz., total soluble solids (8.60-17.70°Brix), acidity (1.61-2.94%), total sugar (4.15-9.17), reducing sugar (2.19-4.45%) and TSS/acid ratio (3.89-8.33) was also recorded. Highest significant and positive correlation was observed between total sugar and TSS (0.895) followed by reducing sugar and TSS (0.882). Significant positive correlation between ascorbic acid and tannins (0.551) was an indication to be associated with binding capacity of ascorbic acid over a longer period of storage.

In vitro clonal propagation of bael (Aegle marmelos Corr.) CV. CISH-B1 through enhanced axillary branching.

Rapid clonal micropropagation protocol of Aegle marmelos (L.) Corr. cv. CISH-B1 was achieved by nodal stem segment of mature bearing tree. Three centimeter long shoots having one axillary bud excised from 10-15th nodal region of shoots during September gave quick in vitro bud burst (5.33 days) when cultured on MS medium supplemented with BAP, 8.84 µM + IAA 5.7 µM. The maximum number of proliferated shoots (9.0/explant) were obtained on same medium supplemented with BAP 8.84 µM + IAA 5.7 µM. The micro shoots were rooted (100 %) on + IAA 5.7 µM. In vitro rooted plants were acclimatized on autoclaved coconut husk containing plant salt mixture and under shade net house (50 % shade 70-80 % RH). The plants were established in the field after acclimatization. The micropropagated plants were tested for its genetic fidelity using 13 RAPD, 3 ISSR and 2 DAMD primers. Profile obtained by all the three Single Primer Amplification Reaction (SPAR) technique from mother tree and micropropagated plants revealed genetic integrity of micropropagated plants with that of mother tree.