Institutional Publications
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Item Effects of MULTIFOLIATE-PINNA, AFILA, TENDRIL-LESS and UNIFOLIATA genes on leafblade architecture in Pisum sativum(Springer, 2009) Mishra, Raghvendra Kumar; Chaudhary, Swati; Kumar, Anil; Kumar, SushilIn order to dissect the genetic regulation of leafblade morphogenesis, 16 genotypes of pea, constructed by combining the wild-type and mutant alleles of MFP, AF, TL and UNI genes, were quantitatively phenotyped. The morphological features of the three domains of leafblades of four genotypes, unknown earlier, were described. All the genotypes were found to differ in leafblade morphology. It was evident that MFP and TL functions acted as repressor of pinna ramification, in the distal domain. These functions, with and without interaction with UNI, also repressed the ramification of proximal pinnae in the absence of AF function. The expression of MFP and TL required UNI function. AF function was found to control leafblade architecture multifariously. The earlier identified role of AF as a repressor of UNI in the proximal domain was confirmed. Negative control of AF on the UNI-dependent pinna ramification in the distal domain was revealed. It was found that AF establishes a boundary between proximal and distal domains and activates formation of leaflet pinnae in the proximal domain.Item Mapping of the multifoliate pinna (mfp) leaf-blade morphology mutation in grain pea Pisum sativum(Indian Academy of Sciences, 2009) Mishra, Raghvendra Kumar; Kumar, Anil; Chaudhary, Swati; Kumar, SushilThe multifoliate pinna (mfp) mutation alters the leaf-blade architecture of pea, such that simple tendril pinnae of distal domain are replaced by compound pinna blades of tendrilled leaflets in mfp homozygotes. The MFP locus was mapped with reference to DNA markers using F2 and F2:5 RIL as mapping populations. Among 205 RAPD, 27 ISSR and 35 SSR markers that demonstrated polymorphism between the parents of mapping populations, three RAPD markers were found linked to the MFP locus by bulk segregant analyses on mfp/mfp and MFP/MFP bulks assembled from the F2:5 population. The segregational analysis of mfp and 267 DNA markers on 96 F2 plants allowed placement of 26 DNA markers with reference to MFP on a linkage group. The existence of common markers on reference genetic maps and MFP linkage group developed here showed that MFP is located on linkage group IV of the consensus genetic map of pea.Item 2-Hydroxyacetophenone, the main component of the essential oil of the roots of Carissa opaca Stapf ex Haines(Taylor & Francis Group, 2009) Mallavarupu, Gopal R.; Mishra, Raghvendra K.; Chaudhary, Swati; Pandey, Richa; Gupta, Sarika; Kumar, SushilThe volatile oil obtained by hydrodistillation of the roots of Carissa opaca was analyzed by GC and GC/MS to study its composition. Thirty-five compounds representing 98.3% of the oil were identified. Si-gel chromatography of the oil yielded 2-hydroxyacetophenone, which was characterized by spectral methods (1H-, 13C-NMR and mass), as the main component, amounting to 89.5% of the oil.Item Regulation of stipule development by COCHLEATA and STIPULE-REDUCED genes in pea Pisum sativum(Springer, 2009) Kumar, Sushil; Mishra, Raghvendra Kumar; Kumar, Anil; Srivastava, Suchi; Chaudhary, SwatiPisum sativum L., the garden pea crop plant, is serving as the unique model for genetic analyses of morphogenetic development of stipule, the lateral organ formed on either side of the junction of leafblade petiole and stem at nodes. The stipule reduced (st) and cochleata (coch) stipule mutations and afila (af), tendril-less (tl), multifoliate-pinna (mfp) and unifoliata-tendrilled acacia (uni-tac) leafblade mutations were variously combined and the recombinant genotypes were quantitatively phenotyped for stipule morphology at both vegetative and reproductive nodes. The observations suggest a role of master regulator to COCH in stipule development. COCH is essential for initiation, growth and development of stipule, represses the UNI-TAC, AF, TL and MFP led leafblade-like morphogenetic pathway for compound stipule and together with ST mediates the developmental pathway for peltate-shaped simple wild-type stipule. It is also shown that stipule is an autonomous lateral organ, like a leafblade and secondary inflorescence.Item Predominance of the serpentine route in monoterpenoid indole alkaloid pathway of Catharanthus roseus(Indian National Science Academy, 2008) Singh, Digvijay; Rai, Sanjay Kumar; Pandey-Rai, Shashi; Srivastava, Suchi; Mishra, Raghvendra Kumar; Sharma, Srikant; Kumar, SushilSingle runs of RP-HPLC procedure were used for simultaneous and organ-wise quantification of the terpenoid indole alkaloids (TIAs) serpentine, ajmalicine, vindoline, catharanthine, vincristine and vinblastine in whole adult plants of Catharanthus roseus, sampled from 81 accessions and F2 generation of a cross. The average total TIA content was 159 mg distributed among stem, root, leaf and inflorescence organs in 13:8:3:1 proportion. The root and stem barks, leaf-petiole and corolla petals were several fold TIA-richer than root and stem woods, leaf–lamina and rest of flower organs, respectively. TIAs were undetectable in seed but present in the seedling. Quantitatively, serpentine (+ ajmalicine), serpentine and catharanthine, vindoline, serpentine, catharanthine and vinblastine (+ vincristine) were the principal TIAs detected in stem, root, leaf and inflorescence organs, respectively. Significant amounts of vinblastine, vindoline and catharanthine were found present in the non-chlorophyllous corolla petals. Correlation studies showed that contents of catharanthine and serpentine in roots, catharanthine, vindoline and vinblastine in leaves, serpentine in roots and stems and stems and leaves were interdependent. Serpentine could be involved in suppressing the accumulation of other TIAs in leaves. The observed inter- and intra- organ contents of end-products from serpentine, catharanthine and vindoline TIA branch pathways and correlations between them showed predominance of the serpentine branch.Item Selection of leaf blight resistant Pelargonium graveolens plants regenerated from callus resistant to culture filterate of Alternaria alternata(Elsevier B.V., 2008) Saxena, Gauri; Verma, Praveen Chandra; Rahman, Laiq-ur; Banerjee, Suchitra; Shukla, R.S.; Kumar, SushilA protocol has been established for the induction of disease resistance in callus cultures of rose-scented geranium, Pelargonium graveolens cv. Hemanti against leaf blight disease caused by the fungal pathogen Alternaria alternata. The callus cultures were raised and maintained on semi-solid Murashige and Skoog's (MS) medium supplemented with 10 mg/l Kn and 1.0 mg/l NAA. The calli were subjected to various concentrations of culture filtrate (0%, 4%, 8%, 12%, 16% and 20%) obtained from A. alternata. Resistant calli were selected and placed on regeneration medium (MS supplemented with 0.5 mg/l BAP and 0.1 mg/l NAA). The regenerants were confirmed for A. alternata resistance by exposing their leaves to the same concentrations of culture filtrate as used previously. While the parental wild type demonstrated typical susceptibility, the leaves of putative resistant clones remained green and viable in the presence of toxin and regenerated shoots directly on the toxin-free regeneration medium (MS supplemented with 5 mg/l Kn and 1 mg/l NAA). The above experiment demonstrated the induction of disease resistance in rose-scented geranium plants at the cellular level. This approach could be successfully exploited in raising new disease-resistant cultivars in geranium against various fungal pathogens.Item Field performance of somaclones of rose scented geranium (Pelargonium graveolens L’ Her Ex Ait.) for evaluation of their essential oil yield and composition(Elsevier B.V., 2008) Saxena, Gauri; Rahman, Laiq-ur; Verma, Praveen Chandra; Banerjee, Suchitra; Kumar, SushilSeveral randomly selected glasshouse grown somaclones of rose scented geranium, Pelargonium graveolens L’Her Ex Ait. cv. Hemanti were successfully transferred to the field in Northern India for evaluation. Two distinct morphotypes were described on the basis of leaf dentation-one resembling the parental cultivar in having highly dentated leaves (HDL) and the other with less dentated leaves (LDL). After repeated field-testing for 3 consecutive years, the HDL clones closely resembled the parental cultivar with respect to the different quantity and quality determining traits, while the LDL group was clearly different. The field established LDL somaclones possessed higher herb yield, number of branches and other oil yield attributing traits as compared to the HDL clones and the parent cultivar. The chemical investigations of the essential oil revealed significant differences between the LDL clones, the HDL clones and the control. Selection of such somaclones, which are superior to the parental in most of the quantitative and qualitative traits and show better adaptability to different areas of cultivation, will help towards commercialization of geranium in India.Item SCAR markers for correct identification of Phyllanthus amarus, P. fraternus, P. debilis and P. urinaria used in scientific investigations and dry leaf bulk herb trade(Thieme E-Journals, 2008) Jain, Neeraj; Shasany, Ajit Kumar; Singh, Shefali; Khanuja, Suman Preet Singh; Kumar, SushilThe trade in Phyllanthus material as bulk herb is rampant and mainly involves herbaceous species such as Phyllanthus amarus, P. fraternus, P . debilis and P. urinaria. These species are very important in herbal medicines and have varied activities. In India these species grow sympatrically and there are chances of deliberate or ignorant adulteration of crude drugs, lowering the efficiency of the medication for its intended purpose. Secondly, incorrect identification may also lead to erroneous reports on activities/molecules. To overcome this problem in crude drug (dry leaf powder) and compliment morphological identification in live plant, we have developed SCAR markers for all four species. In each species, we selected one fragment as being monomorphic between accessions but differing in size between species. These species-specific fragments were selected, cloned and sequenced. Based on the sequences, primer pairs were designed and amplification conditions standardized. SCAR markers were isolated from population DNA amplification profiles and validated by sequencing. The species-specific SCAR primers could retrieve the same size and sequence of fragments as in the RAPD profile. These fragments are 1150 bp, 317 bp, 980 bp and 550 bp in size for P. amarus, P. fraternus, P. debilis and P. urinaria, respectively. Additional fragments in P. debilis and P. urinaria indicate different alleles. The retrieval of same size and sequence of species-specific unique SCAR markers from the respective accessions (mixed DNA sample of same accessions) indicates the usefulness to study natural hybridization between the species in addition to adulteration.Item Volatile metabolite compositions of the essential oil from aerial parts of ornamental and artemisinin rich cultivars of Artemisia annua(Taylor & Francis Group, 2008) Goel, Divya; Mallavarupu, Gopal Rao; Kumar, Sushil; Singh, Vijender; Ali, MohammedThe essential oils of the aerial parts (flowers, leaves and stems) of the artemisinin rich cultivar ‘Jeevanraksha’ and artemisinin poor ornamental type cultivar Suraksha of Artemisia annua, grown at New Delhi, were characterized for their metabolite composition by GC and GC/MS analyses. Ninety-seven compounds comprising 91.3% of the total oil of ‘Jeevanraksha’ were identified. Forty-three monoterpenes (56.6%), 32 sesquiterpenes (31.1%), and 2 diterpenes (0.2%) comprised bulk of the oil (87.9%). The oil was devoid of artemisia ketone and contained camphor (13.5%), 1,8-cineole (9.4%), trans-sabinol (7.1%), p-mentha-1(7), 5-dien-2-ol (6.3%), myrcene (4.7%), germacrene D (4.4%), (E)-β-farnesene (3.9%), β-caryophyllene (3.7%), dihydroartemisinic lactone (3.0%) and p-cymene (2.0%) as the major constituents. Eighty-six compounds representing 93.3% of the composition were identified in the Suraksha oil. This oil contained artemisia ketone (47%), 1,8-cineole (8.4%), camphor (5.9%) and α-pinene (5.2%) as the major components. Many compounds were detected in the A. annua oil for the first time. Consistent with hybrid origin of the cultivar ‘Jeevanraksha’, its oil possessed characteristics reported earlier for the corresponding essential oils of the A. annua accessions of Vietnamese origin and North-American accessions.Item Isolation and characterization of microsatellite markers for analysis of molecular variation in the medicinal plant Madagascar periwinkle (Catharanthus roseus (L.) G. Don.)(Elsevier B.V., 2007) Shokeen, Bhumika; Sethy, Niroj Kumar; Kumar, Sushil; Bhatia, SabhyataCatharanthus roseus (L.) G. Don, commonly known as Madagascar periwinkle, is a plant of great medicinal value. In this study microsatellite markers were developed and utilized for analysis of genetic diversity in C. roseus. In order to isolate microsatellite sequences, two small insert genomic libraries of C. roseus cv. Nirmal were constructed and screened with CA, CT, GC and GCG oligonucleotide repeats. Sixty-five microsatellite motifs were identified, from which 38 functional STMS primer pairs were designed and validated. Out of these, 24 STMS markers were used to evaluate the genetic polymorphism in 37 genotypes, which comprised of 32 accessions of C. roseus, a single accession each of two related species (C. trichophyllus and C. pusillus) and one accession each of three related genera (Vinca minor, Thevetia peruviana and Nerium indicum). The 24 STMS markers detected 26 loci with two markers amplifying more than one locus. A total of 124 alleles were amplified in the 37 genotypes ranging from 2 to 10 alleles with an average of 4.76 alleles per locus. The high average expected heterozygosity (H) value of 0.56 and observed heterozygosity (H) value of 0.52 established the efficiency of the STMS markers for discriminating the C. roseus genotypes. Nei and Li’s similarity coefficients were calculated and a UPGMA-based dendrogram was constructed which clearly distinguished all genotypes except two pairs. Sequence analysis of the length variant alleles at three STMS loci revealed that the variation in the copy number of repeat motifs was the major source of length polymorphism within C. roseus. However, isolated point mutations and indels in the microsatellite flanking regions (MFRs) of homologus loci from other species also contributed to size homoplasy and allelic size variation. This study is the first report of microsatellite development and utilization in C. roseus, providing significant insights into its genome structure and organization.