Publications of NIPGR Scientists

Permanent URI for this communityhttps://ndkr-library.nipgr.ac.in/handle/123456789/1

Browse

Author: search.filters.author.Kumar, SushilSubject: search.filters.subject.Catharanthus roseus

Search Results

Now showing 1 - 6 of 6
  • Thumbnail Image
    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, Sushil
    Single 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.
  • Thumbnail Image
    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, Sabhyata
    Catharanthus 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.
  • Thumbnail Image
    Item
    Construction of genetic linkage map of the medicinal and ornamental plant Catharanthus roseus
    (Indian Academy of Sciences, 2007) Gupta, Sarika; Pandey-Rai, Sashi; Srivastava, Suchi; Naithani, Subhas Chandra; Prasad, Manoj; Kumar, Sushil
    An integrated genetic linkage map of the medicinal and ornamental plant Catharanthus roseus, based on different types of molecular and morphological markers was constructed, using a F2 population of 144 plants. The map defines 14 linkage groups (LGs) and consists of 131 marker loci, including 125 molecular DNA markers (76 RAPD, 3 RAPD combinations; 7 ISSR; 2 EST-SSR from Medicago truncatula and 37 other PCR based DNA markers), selected from a total of 472 primers or primer pairs, and six morphological markers (stem pigmentation, leaf lamina pigmentation and shape, leaf petiole and pod size, and petal colour). The total map length is 1131.9 cM (centiMorgans), giving an average map length and distance between two markers equal to 80.9 cM and 8.6 cM, respectively. The morphological markers/genes were found linked with nearest molecular or morphological markers at distances varying from 0.7 to 11.4 cM. Linkage was observed between the morphological markers concerned with lamina shape and petiole size of leaf on LG1 and leaf, stem and petiole pigmentation and pod size on LG8. This is the first genetic linkage map of C. roseus.
  • Thumbnail Image
    Item
    Transcript profiling of terpenoid indole alkaloid pathway genes and regulators reveals strong expression of repressors in Catharanthus roseus cell cultures
    (Springer, 2007) Dutta, Ajaswrata; Singh, Digvijay; Kumar, Sushil; Sen, Jayanti
    The understanding of the complexities and molecular events regulating genes and the activators involved in terpenoid indole alkaloid (TIA) metabolism is known to a certain extent in cell cultures of an important TIA yielding plant, Catharanthus roseus, though it is not yet complete. Recently, the repressors of early TIA pathway genes have also been identified. However, their roles in the regulation of TIA pathway in C. roseus cell cultures remains yet unknown. We have made a comparative profiling of genes catalyzing the important steps of 2-C methyl-D-erythritol-4- phosphate (MEP), shikimate and TIA biosynthetic pathways, their activator and repressors using macro- array, semiquantitative RT-PCR and northern analyses in a rotation culture system of C. roseus comprising differentiated and proliferated cells. Our results demonstrate that TIA biosynthetic pathway genes and their activators show variable expression pattern, which was correlated with the changes in the cellular conditions in these systems. Under similar conditions, TIA pathway repressors show strong and consistent expression. The role of repressors in the complex regulation of the TIA pathway in C. roseus cell cultures is discussed. The results were supported by HPLC data, which demonstrated that the molecular program of cellular differentiation is intimately linked with TIA pathway gene expression and TIA production in C. roseus cell cultures.
  • Thumbnail Image
    Item
    Volatile compounds of leaves and flowers of periwinkle Catharanthus roseus (L.) G. Don from New Delhi
    (Wiley-Blackwell, 2006) Pandey-Rai, Shashi; Mallavarapu, Gopal Rao; Naqvi, A. A.; Yadav, Anju; Rai, Sanjay Kumar; Srivastava, Suchi; Singh, Digvijay; Mishra, Raghvendra; Kumar, Sushil
    The hydrodistilled waxy oils of the leaves and flowers of Catharanthus roseus (L.) G. Don cv. Nirmal plants grown at New Delhi were analysed by GC and GC–MS. A total of 52 compounds were identified, accounting for the 91.3% of the leaf oil.Among these the main constituents were as follows: (E,E )-2,4-hexadienal (7.7%), citronellol (7.9%), geraniol (7.9%), p-cresol (4.7%), (Z,E )-pentadecanal (6.6%), hexadecanoic acid (4.9%), palmitic acid (4.9%) and phytol (6.4%). A total of 41 compounds were identified, accounting for 99.4% of the flower oil, whose main constituents were heneicosane (20.8%), tricosane (37.9%), tetracosane (6.1%) and docosane (4.3%). A total of 24 compounds occurred in both the leaf and flower oils.
  • Thumbnail Image
    Item
    Growth and terpenoid indole alkaloid production in Catharanthus roseus hairy root clones in relation to left- and right-termini-linked Ri T-DNA gene integration
    (Springer, 2004) Batra, Jyoti; Dutta, Ajaswrata; Singh, Digvijay; Kumar, Sushil; Sen, Jayanti
    Hairy root cultures of Catharanthus roseus var. Prabal were established by infecting the leaves with Agrobacterium rhizogenes agropine-type A4 strain. Two hundred and fifty independent root clones were evaluated for growth, morphology, number of integration of Ri T-DNA genes and alkaloid contents. On the basis of growth pat- tern, type of branching and number of lateral roots we were able to separate the hairy root clones into four categories. However based on the integration of the Ri TL-DNA and TR-DNA genes, there were only three different categories of independent hairy root clones–C1 (rolA&B+/ags+), C2 (rolA&B-/ags+) and C3 (rolA&B+/ags ). Southern hybridization analysis revealed both single and multiple copies of T-DNA integration in the root clones. The accumulation of considerable amounts of the root-specific alkaloids ajmalicine and serpentine was observed in the presence of both the TL-DNA and TR-DNA genes (C1) and the TL-DNA gene (C3) alone. Two rolA&B but ags+ clones (C2) accumulated much less or only very negligible amounts of ajmalicine. The possible role of the TL-DNA and TR-DNA genes on growth and alkaloid accumulation in these root clones is discussed