The Agrobacterium-mediated floral dip protocol is the most extensively used transformation method for a model plant Arabidopsis thaliana. Several useful methods for Agrobacterium tumefaciens–mediated transformations of Arabidopsis are existing, but they are time consuming and with low transformation efficiency. Here, we developed a transgenic Arabidopsis lines TET12p::TET12-RFP in a short period of time and enhanced transformation efficiency by using a modified transformation method by applying drought stress after floral dip. In this protocol, Agrobacterium cells carrying TET12p::TET12-RFP recombinant vectors were resuspended in a solution of 5% sucrose, 0.05% (v/v) silwet L-77 to transform female gametes of developing Arabidopsis inflorescences. Treated Arabidopsis were then applied with different levels of drought stresses to stimulate plants for the utilization of maximum plant energy in seed maturation process. The applied stresses achieved the fast maturation of already treated inflorescences while stopped the growing of newly arising untreated inflorescence, thus decreased the chances of wrong collection of untransformed seeds. Consequently, the collected seeds were mostly transgenic with a transformation frequency of at least 10%, thus the screening for positive transformants selection was more advantageous on a selective medium as compared to a classical floral dip method. Within 2-3 months, two hundred of individual transgenic plants were produced from just 10 infiltrated plants. This study concludes that application of drought stresses in a specific stage of plant is a beneficial strategy for achieving the transgenic Arabidopsis in a short period of time with high transformation efficiency.

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The worldwide fossil fuel reserves are rapidly and continually being depleted as a result of the rapid increase in global population and rising energy sector needs. Fossil fuels should not be used carelessly since they produce greenhouse gases, air pollution, and global warming, which leads to ecological imbalance and health risks. The study aims to discuss the alternative renewable energy source that is necessary to meet the needs of the global energy industry in the future. Both microalgae and macroalgae have great potential for several industrial applications. Algae-based biofuels can surmount the inadequacies presented by conventional fuels, thereby reducing the ‘food versus fuel’ debate. Cultivation of algae can be performed in all three systems; closed, open, and hybrid frameworks from which algal biomass is harvested, treated and converted into the desired biofuels. Among these, closed photobioreactors are considered the most efficient system for the cultivation of algae. Different types of closed systems can be employed for the cultivation of algae such as stirred tank photobioreactor, flat panel photobioreactor, vertical column photobioreactor, bubble column photobioreactor, and horizontal tubular photobioreactor. The type of cultivation system along with various factors, such as light, temperature, nutrients, carbon dioxide, and pH affect the yield of algal biomass and hence the biofuel production. Algae-based biofuels present numerous benefits in terms of economic growth. Developing a biofuel industry based on algal cultivation can provide us with a lot of socio-economic advantages contributing to a publicly maintainable result. This article outlines the third-generation biofuels, how they are cultivated in different systems, different influencing factors, and the technologies for the conversion of biomass. The benefits provided by these new generation biofuels are also discussed. The development of algae-based biofuel would not only change environmental pollution control but also benefit producers' economic and social advancement. Graphical abstract

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