D 1 -pyrroline-5-carboxylate synthetase (P5CS) is a proline biosynthetic pathway enzyme and is known for conferring enhanced salt and drought stress in transgenics carrying this gene in a variety of plant species; however, the wild-type P5CS is subjected to feedback control. Therefore, in the present study, we used a mutagenized version of this osmoregulatory gene-P5CSF129A, which is not subjected to feedback control, for producing transgenic indica rice plants of cultivar Karjat-3 via Agrobacterium tumefaciens. We have used two types of explants for this purpose, namely mature embryo-derived callus and shoot apices. Various parameters for transformation were optimized including antibiotic concentration for selection, duration of cocultivation, addition of phenolic compound, and bacterial culture density. The resultant primary transgenic plants showed more enhanced proline accumulation than their non-transformed counterparts. This proline level was particularly enhanced in the transgenic plants of next generation (T 1 ) under 150 mM NaCl stress. The higher proline level shown by transgenic plants was associated with better biomass production and growth performance under salt stress and lower extent of lipid peroxidation, indicating that overproduction of proline may have a role in counteracting the negative effect of salt stress and higher maintenance of cellular integrity and basic physiological processes under stress.

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The effect of increasing sodium chloride (NaCl; 0 to 300 mM) stress was investigated on plant growth, mineral nutrients, and proline accumulation in two indica rice cultivars differing in salt tolerance. The shoots and roots of 'Karjat-3' (salt sensitive cv.) showed greater reduction in fresh weight, dry weight, and water content under increasing salinity stress when compared to 'Panvel-3' (salt tolerant cv.). The magnitude of chloride (Cl) accumulation with increasing salinity varied between cultivars, with lesser accumulation in 'Panvel-3', whereas, calcium (Ca) content was more in 'Panvel-3' than 'Karjat-3' at all salinity levels. Increase in shoot-sodium (Na) did not show notable variation between cultivars under salinity stress; however, Na accumulation was notably lower in roots of 'Panvel-3' than 'Karjat-3'. Under increasing salinity stress, 'Panvel-3' maintained significantly lower sodium/potassium, sodium/calcium and sodium/nitrogen ratios than 'Karjat-3', both in shoots and roots, with more differential response in roots. Regardless of treatment, proline concentration was considerably higher in 'Panvel-3' than in 'Karjat-3'. The salt tolerance of cv. 'Panvel-3' was positively correlated with better growth, higher proline accumulation and enhanced uptakes of beneficial minerals such as potassium (K) and Ca.

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Helicteres isora is a medicinal plant effective against asthma, diabetes, hypolipidemia, HIV, polio besides a good source of diosgenin. Seed dormancy and low natural fruit production rate make this plant a perfect candidate for developing an in vitro regeneration method. However, to date, no such work has been procured in this plant. An efficient method for plant regeneration via shoot organogenesis from callus cultures has been developed using nodal explants in H. isora. Murashige and Skoog (MS) media counting 2,4-Dichlorophenoxyacetic acid (2,4-D, 2.26 to 13.57 μM), Indole-3-acetic acid (IAA, 2.85 to 17.13 μM), Indole-3-butyric acid (IBA, 2.46 to 14.70 μM), 6-Benzylaminopurine (BA, 2.22 to 13.32 μM) and Kinetin (Kin, 2.32 to 13.92 μM) either singly or in the following combinations (IAA + BA; IAA + Kin, and BA + Kin) produced granular callus except BA + Kin which resulted in compact, hard, greenish-white (CHGW) callus. The optimum CHGW callus (2.62 g fresh weight/ explant) was produced on MS media with 13.32 μM BA + 2.32 μM Kin with over 93% callus induction frequency. Optimum shoot organogenesis (67% frequency) was achieved in CHGW callus with lower level of BA (2.22 μM) and Kin (2.32 μM) and produced 3.2 shoots/0.5 g callus within 35 d of culture. Microshoots were rooted successfully (62% frequency) after 35 d of culture on 1/2MS containing 4.90 μM IBA and hardened off. Antioxidant enzymes such as catalase, peroxidase, polyphenol oxidase, and biochemical parameters viz. hydrogen peroxide, reducing and nonreducing sugars, starch, proteins, phenols, and proline contents were studied in regenerating and nonregenerating CHGW calluses to establish a correlation between these parameters and shoot morphogenesis. All the enzyme activities and biochemical parameters were found more in regenerating callus than in nonregenerating except phenols. induction frequency. Optimal shoot organogenesis (67% frequency) was achieved in CHGW callus with lower levels of BA (2.22 μM) and Kin (2.32 μM) and produced 3.2 shoots/0.5 g callus within 35 d of culture. Microshoots were rooted and hardened successfully (62% frequency) after 35 d of culture on 1/2-MS containing 4.90 μM IBA. Antioxidant enzymes (catalase, peroxidase, polyphenol oxidase) and biochemical parameters viz. hydrogen peroxide, reducing and nonreducing sugars, starch, proteins, phenols, and proline contents were studied in regenerating and nonregenerating CHGW calluses to establish a correlation between these parameters and shoot morphogenesis. With the exception of phenols, all the enzyme activities and biochemical parameters were found more frequently in regenerating than in nonregenerating callus.