Genetic variability within n-rDNA region of ectomycorrhizal isolates originating from temperate ecosystems

Identification of Ectomycorrhiza Fungi (ECM) based on morphological characters has been laborious and time consuming, especially samples collected from the environment. Additionally, due to its microscopic nature and limited morphological characters, intraspecies variation is difficult to detect. In view of this, this study aimed at confirming the earlier identification of these fungi, which was based on morphological characters, and also to find suitable molecular restriction fragment length polymorphism (RFLP) markers for the identification of ECM fungi up to the level of species or isolates as part of an expandable database of RFLP patterns of the internal transcribed spacers (ITS) region of ECM fungi. Mycelia of 14 species of ectomycorrhizal fungi representing five genera were isolated in pure culture and characterized by morphological and molecular methods. Molecular identification was performed by analysis of the internal transcribed spacers of the nuclear encoded ribosomal RNA (n-RNA) gene region using restriction fragment length polymorphism (RFLP). The region was first amplified by polymerase chain reaction with specific primers and then cleaved with different restriction enzymes. The degree of polymorphism, although extensive, proved inadequate for proper identification of most of the isolates. Depending on the restriction enzymes used, the genera or species could be grouped on the basis of common fragment patterns, thereby confirming the potential of the small subunit (SSU)-ITS region in PCR-RFLP in molecular characterization and identification of ectomycorrhizal fungi. Based on the results of this study, congruent of morphology and molecular RFLP analysis is recommended for characterization of species/strain of ectomycorrhizal fungi.Keywords: Classification, diversity, ectomycorrhizal fungi, ecosystem, ribosomal DNAAfrican Journal of Biotechnology Vol. 12(22), pp. 3390-339

INFLUENCE OF MEDIA GELLING AGENTS ON ROOT BIOMASS AND IN VITRO VA-MYCORRfflZAL SYMBIOSIS OF CARROT WITH GIGASPORA MARGARITA

An in vitro study with Ri-TDNA transformed roots of carrot  (Daucus carota) was carried out  to evaluate  the role of macro-elements contributed as impurities in  the gelling agent (phytagel) over and above  those present  in the minimal (M) medium. Production of root biomass was taken as a measure to quantify the influence of macro-elements added to the minimal medium. The levels of phosphorus when adjusted to 1.19 mg/1 and 1.09 mg/l, lead to dry root biomass production at par with the control. Attempts made to lower the amount of impurities in phytagel by de-ionization using different alkalies, proved NaOH to give the best results in terms of relatively high amount of root biomass. In an in vitro dual culture system with carrot as host and Gigaspora margarita as the vesicular-arbuscular mycorrhizal fungus, phytagel impurities helped to produce maximum number of infection units and auxiliary cells when phytagel was added to the minimal medium.Key words:    Agrobacterium    rhiiogenesfDaucus    caro/a/Gelling    agents/diaspora    margarita/Macro- elements/Vesicular-arbuscular mycorrhiza/Transformed roots

Effects of mycorrhiza and plant growth promoting rhizobacteria inoculants on rice crops in Northern India

Mutualistic root microorganisms such as arbuscular mycorrhizal fungi (AMF) and plant growth promoting rhizobacteria (PGPR) can ameliorate plant nutrition through an extended extra-radical hyphal network and by nutrient mobilisation. Running under the Indo-Swiss Collaboration in Biotechnology (ISCB), our project focuses on the integration of AMF and PGPR as biofertilisers in wheat-rice and wheat-black gram systems

A novel in vitro whole plant system for analysis of polyphenolics and their antioxidant potential in cultivars of Ocimum basilicum

Plants are an important source for medicinal compounds. Chemical screening and selection is critical for identification of compounds of interest. Ocimum basilicum (Basil) is a rich source of polyphenolics and exhibits high diversity, therefore bioprospecting of a suitable cultivar is a necessity. This study reports on the development of a true to type novel "in vitro system" and its comparison with a conventional system for screening and selection of cultivars for high total phenolics, individual polyphenolics, and antioxidant content. We have shown for the first time using online acidic potassium permanganate chemiluminescence that extracts from Ocimum basilicum showed antioxidant potential. The current study identified the cultivar specific composition of polyphenolics and their antioxidant properties. Further, a distinct relationship between plant morphotype and polyphenolic content was also found. Of the 15 cultivars examined, "Holy Green", "Red Rubin", and "Basil Genovese" were identified as high polyphenolic producing cultivars while "Subja" was determined to be a low producer. The "in vitro system" enabled differentiation of the cultivars in their morphology, polyphenolic content, and antioxidant activity and is a cheap and efficient method for bioprospecting studies

Influence of Media Gelling Agents on Root Biomass and in Vitro Va-mycorrfflzal Symbiosis of Carrot with Gigaspora Margarita

An in vitro study with Ri-TDNA transformed roots of carrot (Daucus carota) was carried out to evaluate the role of macro-elements contributed as impurities in the gelling agent (phytagel) over and above those present in the minimal (M) medium. Production of root biomass was taken as a measure to quantify the influence of macro-elements added to the minimal medium. The levels of phosphorus when adjusted to 1.19 mg/1 and 1.09 mg/l, lead to dry root biomass production at par with the control. Attempts made to lower the amount of impurities in phytagel by de-ionization using different alkalies, proved NaOH to give the best results in terms of relatively high amount of root biomass. In an in vitro dual culture system with carrot as host and Gigaspora margarita as the vesicular-arbuscular mycorrhizal fungus, phytagel impurities helped to produce maximum number of infection units and auxiliary cells when phytagel was added to the minimal medium

A review on the assessment of stress conditions for simultaneous production of microalgal lipids and carotenoids

Microalgal species are potential resource of both biofuels and high-value metabolites, and their production is growth dependent. Growth parameters can be screened for the selection of novel microalgal species that produce molecules of interest. In this context our review confirms that, autotrophic and heterotrophic organisms have demonstrated a dual potential, namely the ability to produce lipids as well as value-added products (particularly carotenoids) under influence of various physico-chemical stresses on microalgae. Some species of microalgae can synthesize, besides some pigments, very-long-chain polyunsaturated fatty acids (VL-PUFA,>20C) such as docosahexaenoic acid and eicosapentaenoic acid, those have significant applications in food and health. Producing value-added by-products in addition to biofuels, fatty acid methyl esters (FAME), and lipids has the potential to improve microalgae-based biorefineries by employing either the autotrophic or the heterotrophic mode, which could be an offshoot of biotechnology. The review considers the potential of microalgae to produce a range of products and indicates future directions for developing suitable criteria for choosing novel isolates through bioprospecting large gene pool of microalga obtained from various habitats and climatic conditions

Expression of apoplast-targeted plant defensin \u3ci\u3eMtDef4.2\u3c/i\u3e confers resistance to leaf rust pathogen \u3ci\u3ePuccinia triticina\u3c/i\u3e but does not affect mycorrhizal symbiosis in transgenic wheat

Rust fungi of the order Pucciniales are destructive pathogens of wheat worldwide. Leaf rust caused by the obligate, biotrophic basidiomycete fungus Puccinia triticina (Pt) is an economically important disease capable of causing up to 50 % yield losses. Historically, resistant wheat cultivars have been used to control leaf rust, but genetic resistance is ephemeral and breaks down with the emergence of new virulent Pt races. There is a need to develop alternative measures for control of leaf rust in wheat. Development of transgenic wheat expressing an antifungal defensin offers a promising approach to complement the endogenous resistance genes within the wheat germplasm for durable resistance to Pt. To that end, two different wheat genotypes, Bobwhite and Xin Chun 9 were transformed with a chimeric gene encoding an apoplast-targeted antifungal plant defensin MtDEF4.2 from Medicago truncatula. Transgenic lines from four independent events were further characterized. Homozygous transgenic wheat lines expressing MtDEF4.2 displayed resistance to Pt race MCPSS relative to the non-transgenic controls in growth chamber bioassays. Histopathological analysis suggested the presence of both pre- and posthaustorial resistance to leaf rust in these transgenic lines. MtDEF4.2 did not, however, affect the root colonization of a beneficial arbuscular mycorrhizal fungus Rhizophagus irregularis. This study demonstrates that the expression of apoplast-targeted plant defensin MtDEF4.2 can provide substantial resistance to an economically important leaf rust disease in transgenic wheat without negatively impacting its symbioti

Artificial intelligence and real-world data for drug and food safety - A regulatory science perspective

In 2013, the Global Coalition for Regulatory Science Research (GCRSR) was established with members from over ten countries (www.gcrsr.net). One of the main objectives of GCRSR is to facilitate communication among global regulators on the rise of new technologies with regulatory applications through the annual conference Global Summit on Regulatory Science (GSRS). The 11th annual GSRS conference (GSRS21) focused on "Regulatory Sciences for Food/Drug Safety with Real-World Data (RWD) and Artificial Intelligence (AI)." The conference discussed current advancements in both AI and RWD approaches with a specific emphasis on how they impact regulatory sciences and how regulatory agencies across the globe are pursuing the adaptation and oversight of these technologies. There were presentations from Brazil, Canada, India, Italy, Japan, Germany, Switzerland, Singapore, the United Kingdom, and the United States. These presentations highlighted how various agencies are moving forward with these technologies by either improving the agencies' operation and/or preparing regulatory mechanisms to approve the products containing these innovations. To increase the content and discussion, the GSRS21 hosted two debate sessions on the question of "Is Regulatory Science Ready for AI?" and a workshop to showcase the analytical data tools that global regulatory agencies have been using and/or plan to apply to regulatory science. Several key topics were highlighted and discussed during the conference, such as the capabilities of AI and RWD to assist regulatory science policies for drug and food safety, the readiness of AI and data science to provide solutions for regulatory science. Discussions highlighted the need for a constant effort to evaluate emerging technologies for fit-for-purpose regulatory applications. The annual GSRS conferences offer a unique platform to facilitate discussion and collaboration across regulatory agencies, modernizing regulatory approaches, and harmonizing efforts