Oxidative stress has been recognized as a central feature of smoke induced chronic obstructive pulmonary disease (COPD). Imbalance between oxidant and antioxidant enzymes is also an established fact in these patients. But studies in regard to stable COPD patients and effect of vitamin E supplementation are lacking. Thirty patients with COPD were included in the study. Their baseline clinical examination, spirometry, plasma malondialdehyde (MDA), alpha-tocopherol and red blood cell superoxide dismutase (SOD) levels were mea sured. Twenty healthy non-smokers who were matched for age and sex served as controls. All the above parameters were repeated after 12 weeks of supplementation with 400 IU of vitamin E daily. The mean malondialdehyde levels in the patients at baseline were higher than controls (5.91 +/- 1.23 nmol/ml vs 4.55 +/- 1.51 nmol/ml, P = 0 001), so also was plasma alpha-tocopherol levels (P < 0 001), while SOD levels were lower in the patients compared to controls (1692 +/- 259 units g/Hb vs 2451 +/- 131 units g/Hb, P < 0 001). Exogenous vitamin E (400 IU per day) supplementation did not bring about any significant change in plasma alpha-tocopherol and SOD levels. The Pearson s co-efficient of correlation between the levels of MDA, vitamin E, SOD; and spirometric measurements were not significant either on day 1 or after 12 weeks of vitamin E supplementation. The present study shows that initially the plasma lipid peroxide (MDA) levels are high and antioxidants (alpha-tocopherol and SOD) are low in patients with COPD. Exogenous supplementation with vitamin E does not have any significant effect on the spirometric measurements though it brings down the levels of MDA showing attenuation of further damage. However, inclusion of larger number of patients and supple mentation with vitamin E for longer periods may throw more light on free radical injury and protective effects of antioxidants.

The development and commercialization of genetically modified (GM) crops with enhanced insect and herbicide resistance, abiotic stress tolerance, and improved nutritional quality has expanded dramatically. Notwithstanding the huge potential benefits of GM crops, the perceived environmental risks associated with these crops need to be addressed in proper perspective. One critical concern is the adventitious presence or unintentional mixing of GM seed in non-GM seed lots, which can seriously affect the global seed market. It would therefore be necessary though a challenging task to develop reliable, efficient, and economical assays for GM detection, identification, and quantification in non-GM seed lots. This can be systematically undertaken by preliminary screening for control elements and selectable or scorable (reporter) marker genes. In this study, simplex and multiplex polymerase chain reaction (PCR) assays individually as well as simultaneously amplifying the commonly used selectable marker genes, i.e., aadA, bar, hpt, nptII, pat encoding, respectively, for aminoglycoside-3'-adenyltransferase, Streptococcus viridochromogenes phosphinothricin-N-acetyltransferase, hygromycin phosphotransferase, neomycin phosphotransferase, Streptococcus hygroscopicus phosphinothricin-N-acetyltransferase, and a reporter gene uidA encoding beta-d-glucuronidase, were developed as a reliable tool for qualitative screening of GM crops. The efficiency of the assays was also standardized in the test samples prepared by artificial mixing of transgenic seed samples in different proportions. The developed multiplex PCR assays will be useful in verifying the GM status of a sample irrespective of the crop and GM trait.

Waxy corn possesses 95-100% amylopectin, compared to 70-75% in normal maize, owing to mutation in gene encoding a granule-bound starch synthase I. Amylopectin is used as an ingredient in textile, adhesive and paper industries. Further, waxy green cob is popular as breakfast item in South Asia and an important constituent of diet in north-eastern states of India as well. We developed a series of waxy inbreds from diverse exotic sources and through introgression breeding. To characterize and unravel the genetic relationships, 24 diverse waxy inbreds were analysed using 77 SSRs distributed throughout the genome. The study generated a total of 203 polymorphic alleles, with a mean of 2.69 alleles per locus. A total of nine unique and 20 rare alleles were detected. The polymorphism information content ranged from 0.08 to 0.68 with an average value of 0.40. Molecular profiling suggested sufficient attainment of homozygosity among the inbreds. Jaccard's dissimilarity coefficient between pairs of genotypes varied from 0.26 to 0.83 which revealed the diverse nature of the inbred lines. Cluster analyses grouped 24 genotypes into three major clusters. Principle coordinate analysis based on SSR also depicted the diverse origin of the genotypes as per the pedigree more reliably than agro-morphological traits. These inbreds were also promising for various cob and grain characteristics including grain yield. The study identified a set of potential cross-combinations that can be planned to develop highly heterotic waxy hybrid combinations. This is the first report of development and characterization of waxy inbreds in India.

Maize grains are the important source of food and energy, but possess very low proA (< 2.5 µg/g) compared to target level of 15 µg/g set by HarvestPlus to alleviate VAD. Favorable allele having variation in 5' untranslated region (UTR) of () gene enhances concentration of proA in maize. To identify the sequence variation in 5' UTR of , a set of diverse 13 inbreds of indigenous and exotic origin was characterized for allelic constitution of. Inbreds possessed wide variation in proA (1.62-23.12 µg/g) with a mean of 9.64 µg/g. The proA in CIMMYT-HarvestPlus genotypes having favorable allele of was very high (22.28 µg/g), whereas the Indian inbreds with the same allele possessed very low proA (2.48 µg/g). Eight genotypes viz., HKI161, HKI163, HKI161-PV, HKI163-PV, HKI193-1-PV, HKI193-2-PV, HP704-22 and HP704-23 revealed the presence of favorable allele, while VQL1, DMRIL47, MGU-PV-123/C6, HKI193-1 and HKI193-2 showed the presence of unfavorable allele of gene. Sequence comparison of favorable allele of Indian (HKI161 and HKI163) and exotic genotypes (HP704-22 and HP704-23) revealed seven having three transitions ( and : G to A,: C to T) and four transversions (: C to G, : T to G,: G to C and : G to T). Four (: position 446, : position 458,: position 459 and : position 483) discriminated the low- and high- proA lines having favorable allele of. These hold significance in enrichment of proA in maize for marker development and their use in marker-assisted selection.

The genetically modified (GM) Bt crops expressing delta-endotoxins from Bacillus thuringiensis provide protection against a wide range of lepidopteron insect pests throughout the growing season of the plant. Bt cotton is the only commercialized crop in India that is planted on an area of 7.6 million hectares. With the increase in development and commercialization of transgenic crops, it is necessary to develop appropriate qualitative and quantitative methods for detection of different transgenic events. The present study reports on the development of a decaplex polymerase chain reaction (PCR) assay for simultaneous detection of transgene sequences, specific transgene constructs, and endogenous stearoyl acyl desaturase (Sad1) gene in two events of Bt cotton, i.e., MON531 and MON15985. The decaplex PCR assay is an efficient tool to identify and discriminate the two major commercialized events of Bt cotton, i.e., MON531 and MON15985, in India. Real-time PCR assays were also developed for quantification of cry1Ac and cry2Ab genes being employed in these two events. The quantitative method was developed using seven serial dilutions containing different levels of Bt cotton DNA mixed with a non-Bt counterpart ranging from 0.01 to 100%. The results revealed that the biases from the true value and the relative standard deviations were all within the range of ±20%. The limit of quantification (LOQ) of the developed real-time PCR method has also been established up to 0.01%.

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The recessive shrunken2-Reference (sh2-Ref) allele is traditionally used in sweet corn cultivar development worldwide. However, non-availability of suitable gene-based marker(s) for sh2-Ref has considerably delayed its utilization in molecular breeding. Here, 7,320 bp sequence of Sh2 gene among five wild-and six sh2-Ref-based inbreds was analysed by employing 13 overlapping primers. SNP583 (A in wild and G in mutant) and SNP755 (T in wild and C in mutant) in 5'UTR, and SNP5112 (C in wild and T in mutant) in intron-12 clearly differentiated dominant (Sh2) and recessive (sh2-Ref) allele. These SNPs were used to develop four gene-based PCR markers. The newly developed markers were validated in six F 2 populations segregating for sh2-Ref allele, and 230 diverse inbreds of normal and sweet corn types. These markers were further used in genotyping of BC 1 F 1 populations leading to successful selection of sh2-Ref allele in the marker-assisted breeding. Globally, this assumes great significance in molecular breeding of sweet corn.