Browsing by Author "Srivastava, Suchi"

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Characterization of variation and quantitative trait loci related to terpenoid indole alkaloid yield in a recombinant inbred line mapping population of Catharanthus roseus
(Indian Academy of Sciences, 2012) Sharma, Vishakha; Chaudhary, Swati; Srivastava, Suchi; Pandey, Richa; Kumar, Sushil
Improved Catharanthus roseus cultivars are required for high yields of vinblastine, vindoline and catharanthine and/or serpen- tine and ajmalicine, the pharmaceutical terpenoid indole alkaloids. An approach to derive them is to map QTL for terpenoid indole alkaloids yields, identify DNA markers tightly linked to the QTL and apply marker assisted selection. Towards the end, 197 recombinant inbred lines from a cross were grown over two seasons to characterize variability for seven biomass and 23 terpenoid indole alkaloids content-traits and yield-traits. The recombinant inbred lines were genotyped for 178 DNA markers which formed a framework genetic map of eight linkage groups (LG), spanning 1786.5 cM, with 10.0 cM average intermarker distance. Estimates of correlations between traits allowed selection of seven relatively more important traits for terpenoid indole alkaloids yields. QTL analysis was performed on them using single marker (regression) analysis, simple interval mapping and composite interval mapping procedures. A total of 20 QTL were detected on five of eight LG, 10 for five traits on LG1, five for four traits on LG2, three for one trait on LG3 and one each for different traits on LG three and four. QTL for the same or different traits were found clustered on three LG. Co-location of two QTL for biomass traits was in accord of correlation between them. The QTL were validated for use in marker assisted selection by the recombinant inbred line which transgressively expressed 16 traits contributory to the yield vinblastine, vindoline and catharanthine from leaves and roots that possessed favourable alleles of 13 relevant QTL.
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.
Correspondence between flowers and leaves in terpenoid indole alkaloid metabolism of the phytoplasma-infected Catharanthus roseus plants
(Springer, 2014) Srivastava, Suchi; Pandey, Richa; Kumar, Sushil; Nautiyal, Chandra Shekhar
Several plants of Catharanthus roseus cv 'leafless inflorescence (lli)' showing phenotype of phytoplasma infection were observed for symptoms of early flowering, virescence, phyllody, and apical clustering of branches. Symptomatic plants were studied for the presence/absence and identity of phytoplasma in flowers. Transcription levels of several genes involved in plants' metabolism and development, accumulation of pharmaceutically important terpenoid indole alkaloids in flowers and leaves and variation in the root-associated microbial flora were examined. The expression profile of 12 genes studied was semi-quantitatively similar in control leaves and phytoplasma-infected leaves and flowers, in agreement with the symptoms of virescence and phyllody in phytoplasma-infected plants. The flowers of phytoplasma-infected plants possessed the TIA profile of leaves and accumulated catharanthine, vindoline, and vincristine and vinblastine in higher concentrations than leaves. The roots of the infected plants displayed lower microbial diversity than those of normal plants. In conclusion, phytoplasma affected the biology of C. roseus lli plants multifariously, it reduced the differences between the metabolite accumulates of the leaves and flowers and restrict the microbial diversity of rhizosphere.
Establishment of artemisinin combination therapy as first line treatment for combating malaria: Artemisia annua cultivation in India needed for providing sustainable supply chain of artemisinin
(Indian Academy of Sciences, 2005) Kumar, Sushil; Srivastava, Suchi
The Plasmodium falciparum malaria is responsible for about 500 million clinical attacks and one million deaths each year, mostly in Africa and southeast Asia. On account of the parasite having become resistant to the conventional antimalarials, artemisinin-based combination therapy (ACT) is the only reliable treatment option. The Roll Back Malaria (RBM) and Millennium Development Goals (MDG) campaigns of WHO and UNDP, respectively, have made little headway, mainly because of paucity of artemisinin in world market. Here we show that the cultivation and processing of Artemisia annua, presently the only resource of artemisinins, in India, in partnerships, can help build a sound supply chain of artemisinin.
Perspectives of the Artemisia annua dry leaf therapy (ALT) for malaria and of its re-purposement as an affordable cure for artemisinin-treatable illnesses
(Indian National Science Academy, 2018) Goel, Richa; Kumari, Raj; Singh, Vijender; Kumari, Renu; Srivastava, Suchi; Kumar, Sushil
Malarial diseases continue to risk the lives of more than 3 billion people in 97 countries in the world, causing sickness in several million people and death to half a million patients. The preponderate malaria causing apicomplexan protozoan parasite species Plasmodium falciparum and Plasmodium vivax have become genetically resistant to most of the approved antimalarial drugs, including the artemisinin-based combination therapies (ACTs). At this time, there is a vigorous need to make enough efforts to meet the challenge of combating multi-drug resistant malaria by (a) speeding up the trials in progress on relatively more effective, new and mechanistically different antimalarial pharmaceuticals, (b) production of effective vaccines against falciparum and vivax malaria, (c) devising of new ways to use the presently available anti-malarials such as by using three-drugs ACTs and by using the different two-drug and three-drug ACTs sequentially, and (d) induction of Artemisia annua dry leaf therapy (ALT) of recent origin, but of ancient precedent, as an effective treatment for acute and complicated malaria. Here, a perspective type review is presented of the: pre-ALT antimalarial drugs, methodology of their usage and consequences of resistance development; safety, efficacy, affordability, quality maintenance and resilience to resistance development aspects of ALT; and possibilities of ALT re-purposement for treating many infectious-metabolic disorder related- and cancerous-diseases. In conclusion, an urgent need is emphasized for pilot studies and clinical trials on ALT to attest its deployment as anti-malarial and cure for diseases beyond malaria.
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.
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, Swati
Pisum 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.
Regulation of unipinnate character in the distal tendrilled domain of compound leaf-blade by the gene MULTIFOLIATE PINNA (MFP) in pea Pisum sativum
(Elsevier B.V., 2004) Kumar, Sushil; Rai, Sanjay Kumar; Pandey-Rai, Shashi; Srivastava, Suchi; Singh, Digvijay
The wild type compound leaf-blade of Pisum sativum has one to three pairs of simple leaflet pinnae in its petiole proximal domain, one to four pairs of simple tendril pinnae in the distal domain and a simple tendril pinna in the apical domain. A novel ethyl methane sulfonate induced dominant mutant was isolated and characterized whose leaf-blades formed MULTIFOLIATE PINNA pairs in the distal domain. The distal multifoliate pinnae or compound pinna-blades had three tendrilled-leaflets as pinnules. The pinnules had a bifacial elliptic-lanceolate leaflet body and arc shaped apex that mimicked the ringlet shaped apex of tendrils. The TL/tl, mfp/mfp and tl/tl, mfp/mfp leaf-blades also produced multifoliate (compound) pinna-blades in distal positions; the pinnules of these genotypes had elliptic shape. The pinnae were branched tendrils in TL/TL, MFP/mfp plants. The leaf-blade rachis was more ramified in af mfp double mutants than in af mutant. In the af mfp double mutant, the multifoliate pinna-blades were present on tertiary and secondary branches of the rachis in the proximal domain and on secondary branches and the primary rachis in the distal domain. The leaf-blades of the af tl mfp triple mutant genotype were an order of magnitude more ramified than those of af tl and af mfp genotypes in proximal as well as distal domains. The leaf-blade phenotypes of various genotypes revealed in this study and those known from previous work have allowed the following conclusions about the nature of mfp mutation and mfp function(s). (a) The presence of mfp mutation or mfp function changes the identity of distal primordia, from tendrils in the wildtype (MFP/MFP) leaf-blades to multifoliate pinna-blades in mfp/mfp mutant. (b) A pathway for the lamination of pinnules of multifoliate blades formed in distal and terminal domains in the mfp mutant and all domains in af mfp double mutant is activated by the mfp mutation. (c) The leaflet-/pinnule-lamination pathway activated by the tl mutation interacts with the mfp-directed pathway. (d) The mfp mutation intensifies rachis ramification in proximal and distal domains activated by the af mutation. This process is distinct from analogous rachis ramification that occurs in the af tl double mutant.
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.