Sudhir Shukla

High frequency somatic embryogenesis, regeneration and correlation of alkaloid biosynthesis with gene expression in Papaver somniferum

Opium poppy (Papaver somniferum) remains the most important source for several pharmaceutical benzylisoquinoline alkaloids including the narcotic analgesic morphine, the anti-tussive drug codeine. Selection, breeding as well as other molecular practices have produced large number of germplasm with modulated biosynthesis and altered accumulation of specific alkaloids. However, there is need to modulate biosynthetic pathways of these alkaloids through transgenic approach, which requires development of an efficient regeneration system for genetic transformation. We studied response of different explants for somatic embryogenesis and developed an efficient protocol for P. somniferum somatic embryogenesis, regeneration as well as successful transplantation in fields. In our system, embryogenic callus from root explants was induced by 2,4-D and kinetin followed by maturation with gibberellin and abscisic acid and regeneration by hormone free half-strength MS media. Though root regeneration from somatic embryos has been shown as limiting factor in established protocols, we observed efficient root and shoot regeneration through somatic embryos in our study. We analyzed alkaloid content in in vitro callus and field grown plants and concluded developmental control of alkaloid biosynthesis. Absence of transcripts of some of the genes and specific alkaloids also suggests regulation at transcriptional level of alkaloid biosynthesis pathway.

Biplot Approach for Identification of Heterotic Crosses in Linseed (Linum usitatissimum L.)

In India, among nonedible oilseed crops, linseed is a commercial crop having tremendous economic and industrial importance. The seed production is low due to limited resources, so the development of high-yielding lines with desirable characters is urgently needed. In the present study seven parents half diallel data was subjected to biplot analysis to identify the heterotic crosses, genetically similar parents, and to study their interrelationship. Parent Sln-Ys with A-79 and A-03 with A-79 for capsules per plant and seed yield, parent Mukta with Sln-Ys and Mukta with BAU-45 for seeds per capsule, and parents Mukta, A-103, A-79 and A-94 for test weight had lowest correlation. Parent B [Mukta] and F [A-79] were good general combiner for all the traits. The crosses F [A-79] × A [Sln-Ys] and D [A-03] × F [A-79] for capsules per plant, test weight and seed yield per plant, cross D [A-03] × A [Sln-Ys] for capsules per plant and test weight and cross D [A-03] × F [A-79] for test weight and seed yield per plant were heterotic. None of the crosses were heterotic for seeds per capsule.

Opium Poppy: Genetic Upgradation Through Intervention of Plant Breeding Techniques

Opium poppy (Papaver somniferum L.) has its importance as a plant based natural pain re‐ liever from the time dating back to early civilization till today. Its pain relieving properties had been described in various books of unani, allopathy and ayurvedic medication system. Today our pharmaceutical industries solely depend on opium poppy for their crude resour‐ ces for manufacturing of pain killing drugs. The medical practitioners around the world routinely prescribe important life saving drugs, are the secondary metabolites produced as a result of complex plant metabolism. The important life saving drugs are mostly derived from five major alkaloids viz., morphine, codeine, thebaine, noscapine and papaverine which are present in opium latex in ample amount [1]. According to a report from an inter‐ national organization i.e. WHO (World Health Organization), about 85% of the population in developing countries depend on herbal plants for curatives, medicinal and other medico related applications. India being one of the twelve mega biodiversity centers of the world is fully fledged with diverse array of herbal and medicinal plants which makes it “Botanical Garden of World”. About 10,000 different medicinal plant species are found in India among which opium poppy occupies the highest place in terms of food (seeds) and pharmaceuticals (alkaloids). These valuable alkaloids are mainly extracted in India from green unripe capsu‐ les by making incision upto 1-2 mm in the epidermal wall of the capsule (Figure 1), but glob‐ ally it is extracted from the dried capsule which is called CPS (Concentrated poppy straw) system. In CPS system, the dried capsules along with eight inches of peduncle are harvested and seeds are threshed. The remaining husk is used to extract various alkaloids. The whole plant parts of opium poppy are valuable in terms of food, medicine, vegetable and as brew‐

Genetic Relationship Between Foliage Yield and its Biochemical Components in Vegetable Amaranth

ABSTRACT Vegetable amaranth (Amaranthus tricolor L.) is an alternative source of protein and nutrients in developing countries. The investigation was undertaken to examine genetic variability and genetic and phenotypic associations for foliage yield and quality traits to improve vegetable amaranth. The objective was to obtain high foliage-yielding varieties rich in nutritional components. There was considerable natural and genetic variability in the quality traits. The genotypic (GCV) and phenotypic (PCV) coefficients of variation were high for Cu and Mn and moderate for chlorophyll (Chl) a and b, Fe, Ni, and Zn. High heritability coupled with high genetic gain occurred for all traits except for moisture, fiber, total phenol content, and carbohydrate content. The phenotypic correlation between foliage yield and carotenoid content was significant and positive. Chl a, protein, carbohydrate, total phenol, and Fe had positive direct effects on foliage yield. The elements Cu, Fe, Mn, Ni, and Zn and Chl a and b were major contributing traits toward foliage yield. Strains AV-55, AV-61, and AV-62 were best for foliage yield or quality traits in vegetable amaranth. Selection strategies based on these traits could enhance foliage yield in vegetable amaranth.

Conservation of Plant Genetic Resources and Their Utilisation in Global Perspective

Plant genetic resources such as recent cultivars and breeds, traditional cultivars and breeds, special genetic livestocks (breeding lines, mutants, etc.), wild relatives of domesticated species are commonly used to study the genetical, evolutionary, physiological, cytological, breeding, pathological, environmental or ecological aspects of plants. Likewise, crop genetic resources are used by plant breeders to develop new and improved varieties having significant economic benefits. In the past due to overexploitation of naturally existing variability in a plant species exhausted substantially leading to a narrow genetic base of the plant species. Land races with narrow genetic base having a high risk of elimination by diseases or any other natural calamity. So, the present day need is to check the depletion of existing variability of a species of a gene pool. According to FAO, genetic erosion or the loss of genetic diversity is an issue of serious concern in relation to sustainable global food security. If we do not counteract with the rising genetic impoverishment it may have severe consequences in near future. Almost all the ecosystems and genetic resources are facing threats due to increasing pollution, invasive alien species, overexploitation by humans and climate change. Therefore, there is an urgent need for conservation, sustainable utilisation and management of plant genetic resources so as to meet the growing requirement of food, fodder, fibre, health and other needs of present as well as future generation. The conservation of wild relatives of crop plants and the offsite conservation of crop varieties and culture of microorganisms is a very necessary aspect. Some of the short-term and long-term programs for plant genetic conservation along with other measures such as sustainable consumption, taking pro-active steps to control climate changes, are needed to be emphasised. By adopting a number of the methodologies discussed in the review, plant genetic resources are being tried to conserve for future genetic advancement and an insurance against unpredicted threats to future agricultural production. The detailed and exhaustive scenario of the genetic resources, its conservation (ex situ and in situ) and application is also discussed in depth in this review.

Ageratum enation virus Infection Induces Programmed Cell Death and Alters Metabolite Biosynthesis in Papaver somniferum

A previously unknown disease which causes severe vein thickening and inward leaf curl was observed in a number of opium poppy (Papaver somniferum L.) plants. The sequence analysis of full-length viral genome and associated betasatellite reveals the occurrence of Ageratum enation virus (AEV) and Ageratum leaf curl betasatellite (ALCB), respectively. Co-infiltration of cloned agroinfectious DNAs of AEV and ALCB induces the leaf curl and vein thickening symptoms as were observed naturally. Infectivity assay confirmed this complex as the cause of disease and also satisfied the Koch’s postulates. Comprehensive microscopic analysis of infiltrated plants reveals severe structural anomalies in leaf and stem tissues represented by unorganized cell architecture and vascular bundles. Moreover, the characteristic blebs and membranous vesicles formed due to the virus-induced disintegration of the plasma membrane and intracellular organelles were also present. An accelerated nuclear DNA fragmentation was observed by Comet assay and confirmed by TUNEL and Hoechst dye staining assays suggesting virus-induced programmed cell death. Virus-infection altered the biosynthesis of several important metabolites. The biosynthesis potential of morphine, thebaine, codeine, and papaverine alkaloids reduced significantly in infected plants except for noscapine whose biosynthesis was comparatively enhanced. The expression analysis of corresponding alkaloid pathway genes by real time-PCR corroborated well with the results of HPLC analysis for alkaloid perturbations. The changes in the metabolite and alkaloid contents affect the commercial value of the poppy plants.