Kishwar Ali

Somaclonal Variation in a Food Legume—Lathyrus sativus

Somaclones of Lathyrus sativus cv P-24 were obtained from leaf, internode and root derived callus cultures. These showed significantly decreased neurotoxin (ODAP) content up to 0.03% compared to that of parent cultivar P-24 (0.3%). The somaclones also showed higher seed yield than parent P24. In addition. somaclone Bio 1–22 showed significantly decreased time for flowering. Other heritable morphological variant features were leaf length, leaf breadth, internode length, flower colour, seed colour, 100 seed weight, neurotoxin content of leaves and red markings on the pods.

Role of phytosterols in drought stress tolerance in rice

Phytosterols are integral components of the membrane lipid bilayer in plants. They regulate membrane fluidity to influence its properties, functions and structure. An increase in accumulation of phytosterols namely campesterol, stigmasterol and β-sitosterol was observed in rice as seedlings matured. The levels of the major phytosterol, β-sitosterol in N22 (drought tolerant) rice seedlings was found to increase proportionately with severity of drought stress. Its levels were 145, 216, 345 and 364 μg/g FW after subjecting to water stress for 3, 6, 9 and 12 days respectively, while for IR64 (drought susceptible), levels were 137, 198, 227 and 287 μg/g FW at the same stages. Phytosterols were also found to increase with maturity as observed at 30, 50 and 75 days after planting. The activity of HMG-CoA reductase (EC 1.1.1.34) which is considered to be a key limiting enzyme in the biosynthesis of phytosterols was 0.55, 0.56, 0.78 and 0.85 μmol/min/L at 3, 6, 9 and 12 days of water stress in N22 and 0.31, 0.50, 0.54 and 0.65 μmol/min/L in case of IR64 respectively. The elevation in the levels of phytosterols as well as the activity of HMG-CoA reductase during drought stress indicates the role of phytosterols in providing tolerance to stress.

Functional validation of a drought-responsive AP2/ERF family transcription factor-encoding gene from rice in Arabidopsis

A water-deficit stress (WDS)-responsive transcription factor-encoding gene isolated from drought-tolerant Oryza sativa L. cv. N-22 corresponding to the AP2/ERF family and named AP2/ERF-N22, when overexpressed in Arabidopsis under a constitutive promoter, showed improved turgor and less wilting as compared to wild-type plants under WDS. However, the transgenic plants showed phenotypic aberrations such as stunted growth, smaller silique size, reduced number of seeds and delayed flowering. Physio-biochemical analysis of T2 plants revealed higher relative water content, membrane stability index, capacity to scavenge reactive oxygen species and increased levels of proline, carotenoids, wax and abscisic acid content under WDS. Higher abscisic acid content resulted in reduced stomatal conductance and transpiration rate. The reduced chlorophyll content also leads to reduced photosynthetic rate and efficiency. Thus, this gene may be exploited for enhanced drought tolerance in crop plants.

Physiological and Biochemical Analysis of Lathyrus sativus Somaclones Derived from Root and Internode Explants

Low ODAP somaclones from root and internode explants were evaluated for physiological and biochemical parameters. Considerable variation was observed in relation to leaf width, leaf length, internodal length and leaf area. All the somaclones during development had substantially lower ODAP content in leaves compared to parent P 24. Amongst somaclones, Bio R 202 had the maximum leaf area, biomass as well as photosynthetic rate. Harvest index and seed yield were highest in Biol 222 whereas ODAP content was the least in it.

Regulation of phytosterol biosynthetic pathway during drought stress in rice

Plants respond to drought stress in the form of various physio-biochemical and molecular changes at both cellular and molecular levels. Drought stress causes the destruction of cell membranes by disintegration of membrane lipids. One of the major groups of membrane lipids that plays important role in preserving the integrity of cell membranes is phytosterols. HMG-CoA reductase (HMGR) is the principal enzyme in the biosynthesis of plant sterols, synthesized via mevalonic acid pathway. Phospholipid: sterol acyltransferase (PSAT) is another important enzyme that plays an important role in turnover of phytosterols into steryl esters and helps maintain homeostasis of membrane lipids. In this study, the expression of both HMGR and PSAT genes in drought sensitive (IR64) and drought tolerant (N22) rice cultivars under applied drought conditions were found to be elevated. The increase in expression of these genes was proportional to the level of severity of applied drought stress. This is substantiated by the negative correlation of HMGR and PSAT expression to relative water content (RWC) and membrane stability index (MSI). Expression of PSAT was also found to be positively correlated to ABA content and HMGR expression.

Abscisic acid biosynthesis under water stress: anomalous behavior of the 9- cis -epoxycarotenoid dioxygenase1 ( NCED1 ) gene in rice

The gene NCED1 encodes 9-cis-epoxycarotenoid dioxygenase, which catalyzes oxidative cleavage of 9-cis-epoxycarotenoids neoxanthin and violaxanthin to xanthoxin, a key step in the biosynthesis of abscisic acid (ABA) in higher plants. In the present study, the complete NCED1 of 1 917 bp was cloned and characterized from rice (Oryza sativa L. cv. N22) as no earlier reports were available for its characterization from the indica cultivar. The NCED1 had no intron and encoded a protein of 639 amino acids with a predicted molecular mass of 68.62 kD and pI of 6.07. The aliphatic index and grand average of hydropathicity were found to be 77.04 and -0.148, respectively. Multiple alignment analysis revealed that the sequence shared a high identity with the Oryza sativa japonica group (100 %) followed by Triticum aestivum (90 %), Hordeum vulgare (90 %), and Zea mays (89 %). The enzyme had a RPE65 domain of 476 amino acid residues. The RPE65 domain requires Fe(II) as a cofactor coordinated with 4 histidine residues and 3 glutamic acid residues. The phylogenic tree shows that NCED1 of japonica rice and NCED1 of indica rice were in the same group. They might have been evolved from a common ancestor. Analysis with a PSORT III tool shows that NCED is a chloroplastic protein. The real-time quantitative PCR and RNA-sequencing studies show that the expression of NCED1 was progressively reduced with increasing water stress, and a negative correlation between expression of OsNCED1 and severity of stress was established. Further, NCED1 expression negatively correlated with ABA accumulation under water stress whereas in some other species, its expression increased along with ABA accumulation. This might be due to feedback inhibition of the ABA biosynthesis in rice.