Induksi kalus daun binahong merah (Basella rubra L.) dengan pe,berian 2,4-D dan kinetin

Tanaman binahong merah (Basella rubra L.) merupakan salah satu tanaman yang mengandung senyawa metabolit sekunder berkhasiat obat. Kultur kalus adalah salah satu solusi dalam menghasilkan senyawa metabolit sekunder dengan jumlah yang besar. Penelitian ini bertujuan untuk mengetahui pengaruh pemberian 2,4-D dan kinetin dalam menginduksi kalus daun binahong merah. Penelitian dilakukan di Laboratorium Bioteknologi Tanaman Fakultas Pertanian Universitas Riau pada bulan November 2019 sampai Maret 2020. Percobaan menggunakan rancangan acak kelompok yang terdiri dari dua faktor yaitu 2,4-D dengan empat taraf konsentrasi yaitu 0 ppm, 0,5 ppm, 1 ppm dan 2 ppm dan kinetin dengan empat taraf konsentrasi yaitu 0 ppm, 0,5 ppm, 1 ppm dan 2 ppm dengan tiga ulangan. Hasil penelitian menunjukkan bahwa kombinasi perlakuan tanpa 2,4-D dan 0,5 ppm kinetin memberikan respon terbaik pada saat muncul kalus 11,67 HST dan perlakuan 1 ppm 2,4-D dan 2 ppm kinetin memberikan respon terbaik pada persentase keberhasilan induksi kalus 62,50 %. Red binahong (Basella rubra L.) is a plant that contains medicinal secondary metabolites. Callus culture is one solution in producing secondary metabolites in large quantities. This research aimed to determine the effect of 2,4-D and kinetin in inducing callus on red binahong leaves. The research was conducted at the Laboratory of Plant Biotechnology, Faculty of Agriculture, the University of Riau from November 2019 to March 2020. The experiment used a randomized block design with two factors, namely four levels of 2,4-D 0, 0.5, 1, and 2 ppm and four levels of kinetin namely 0, 0.5, 1, and 2 ppm with three replications. The results showed that a combination of 0 ppm 2,4-D and 0.5 ppm kinetin and 1 ppm 2,4-D and 2 ppm had the fastest callus formation 11.67 DAP and a combination of 1 ppm 2,4-D and 2 ppm kinetin produced weigher callus 6.4 mg and had a percentage of callus formation 62.50%.

Highly Variable Dietary RNAi Sensitivity Among Coleoptera

Many herbivorous beetles (Order Coleoptera) contribute to serious losses in crop yields and forest trees, and plant biotechnology solutions are being developed with the hope of limiting these losses. Due to the unprecedented target-specificity of double-stranded RNA (dsRNA), and its utility in inducing RNA interference (RNAi) when consumed by target pest species, dsRNA-based plant biotechnology approaches represent the cutting edge of current pesticide research and development. We review dietary RNAi studies in coleopterans and discuss prospects and future directions regarding RNAi-based management of coleopteran plant pests. Herein, we also provide a balanced overview of existing studies in order to provide an accurate re-assessment of dietary RNAi sensitivity in coleopterans, despite the limitations to the existing body of scientific literature. We further discuss impediments to our understanding of RNAi sensitivity in this important insect order and identify critical future directions for research in this area, with an emphasis on using plant biotechnology approaches.

A plant-biotechnology approach for producing highly potent anti-HIV antibodies for antiretroviral therapy consideration

Despite a reduction in global HIV prevalence the development of a pipeline of new therapeutics or pre-exposure prophylaxis to control the HIV/AIDS epidemic are of high priority. Antibody-based therapies offer several advantages and have been shown to prevent HIV-infection. Plant-based production is efficient for several biologics, including antibodies. We provide a short review on the work by Singh et al., 2020 who demonstrated the transient production of potent CAP256-VRC26 broadly neutralizing antibodies. These antibodies have engineered posttranslational modifications, namely N-glycosylation in the fragment crystallizable region and O-sulfation of tyrosine residues in the complementary-determining region H3 loop. The glycoengineered Nicotiana benthamiana mutant (ΔXTFT) was used, with glycosylating structures lacking β1,2-xylose and/or α1,3-fucose residues, which is critical for enhanced effector activity. The CAP256-VRC26 antibody lineage targets the first and second variable region of the HIV-1 gp120 envelope glycoprotein. The high potency of this lineage is mediated by a protruding O-sulfated tyrosine in the CDR H3 loop. Nicotiana benthamiana lacks human tyrosyl protein sulfotransferase 1, the enzyme responsible for tyrosine O-sulfation. The transient coexpression of the CAP256-VRC26 antibodies with tyrosyl protein sulfotransferase 1 in planta had restored the efficacy of these antibodies through the incorporation of the O-sulfation modification. This approach demonstrates the strategic incorporation of posttranslational modifications in production systems, which may have not been previously considered. These plant-produced CAP256-VRC26 antibodies have therapeutic as well as topical and systemic pre-exposure prophylaxis potential in enabling the empowerment of young girls and women given that gender inequalities remain a major driver of the epidemic.

Peculiarities of the Transformation of Asteraceae Family Species: The Cases of Sunflower and Lettuce

The Asteraceae family is the largest and most diversified family of the Angiosperms, characterized by the presence of numerous clustered inflorescences, which have the appearance of a single compound flower. It is estimated that this family represents around 10% of all flowered species, with a great biodiversity, covering all environments on the planet, except Antarctica. Also, it includes economically important crops, such as lettuce, sunflower, and chrysanthemum; wild flowers; herbs, and several species that produce molecules with pharmacological properties. Nevertheless, the biotechnological improvement of this family is limited to a few species and their genetic transformation was achieved later than in other plant families. Lettuce (Lactuca sativa L.) is a model species in molecular biology and plant biotechnology that has easily adapted to tissue culture, with efficient shoot regeneration from different tissues, organs, cells, and protoplasts. Due to this plasticity, it was possible to obtain transgenic plants tolerant to biotic or abiotic stresses as well as for the production of commercially interesting molecules (molecular farming). These advances, together with the complete sequencing of lettuce genome allowed the rapid adoption of gene editing using the CRISPR system. On the other hand, sunflower (Helianthus annuus L.) is a species that for years was considered recalcitrant to in vitro culture. Although this difficulty was overcome and some publications were made on sunflower genetic transformation, until now there is no transgenic variety commercialized or authorized for cultivation. In this article, we review similarities (such as avoiding the utilization of the CaMV35S promoter in transformation vectors) and differences (such as transformation efficiency) in the state of the art of genetic transformation techniques performed in these two species.

Improved the Activity of Phosphite Dehydrogenase and its Application in Plant Biotechnology

Phosphorus (P) is a nonrenewable resource, which is one of the major challenges for sustainable agriculture. Although phosphite (Phi) can be absorbed by the plant cells through the Pi transporters, it cannot be metabolized by plant and unable to use as P fertilizers for crops. However, transgenic plants that overexpressed phosphite dehydrogenase (PtxD) from bacteria can utilize phosphite as the sole P source. In this study, we aimed to improve the catalytic efficiency of PtxD from Ralstonia sp.4506 (PtxDR4506), by directed evolution. Five mutations were generated by saturation mutagenesis at the 139th site of PtxD R4506 and showed higher catalytic efficiency than native PtxDR4506. The PtxDQ showed the highest catalytic efficiency (5.83-fold as compared to PtxDR4506) contributed by the 41.1% decrease in the Km and 2.5-fold increase in the kcat values. Overexpression of PtxDQ in Arabidopsis and rice showed increased efficiency of phosphite utilization and excellent development when phosphite was used as the primary source of P. High-efficiency PtxD transgenic plant is an essential prerequisite for future agricultural production using phosphite as P fertilizers.

Genomic Designing of Climate Smart Turmeric

Turmeric is highly tolerant to several climatic changes and can grow under high temperatures and moderate drought conditions. This herb is very much dependant on optimum rainfall, optimum heat with less chilling or freezing conditions. These conditions if are more than normal would tend to reduce the yields of the crops and also effect the productivity. To reduce such drastic yield losses certain conventional plant breeding methods were employed but were very less effective compared to plant biotechnology. To reduce these loses by stresses, extensive and effective molecular biology methods were employed which identifies the genes that are stress responsive along with certain methods like gene transfer, genetic engineering was also known to be effective. All these methods are quite helpful in mitigating the yield losses and promoting healthy growth in the plants. The maintenance of rhizome size, curcumin content, essential oils etc. is very much necessary for the turmeric crop because of its role, especially in the medical field. Therefore, the yield losses are reduced to a maximum extent so that development of smart turmeric is easy and crop designing is possible only with the advanced techniques involved in agriculture biotechnology.

Nanotechnology applications in plant tissue culture and molecular genetics: A holistic approach

: Nanotechnology is one of the most important modern sciences that has integrated all sectors of science. Nanotechnology has been applied in the agricultural sector in the last ten years in pursuit of increasing agricultural production and ensuring food security. Plant biotechnology is an essential science that is concerned with plant production. The use of nanotechnology in plant biotechnology under controlled conditions has facilitated the understanding of important internal mechanisms of the plant biological system. The application of nanoparticles (NPs) in plant biotechnology has demonstrated an interesting impact on in vitro plant growth and development. This includes the positive effect of the NPs on micropropagation, callus induction, somatic embryogenesis, cell suspension culture, and plant disinfection. In addition, other biotechnology processes, including the genetic transformation of plants, plant conservation, and secondary metabolite production have improved by the use of NPs. Furthermore, nanotechnology is used to improve plant tolerance to different stress conditions that limit plant production. In this review article, we attempt to consolidate the achievements of nanotechnology and plant biotechnology and discuss advances in the applications of nanotechnology in plant biotechnology. It has been concluded that more research is needed to understand the mechanism of nanoparticle delivery and translocation in plants in order to avoid any future hazardous effects of nanomaterials. This will be key to the achievement of magnificent progress in plant nanobiotechnology.

High-efficiency retron-mediated single-stranded DNA production in plants

ABSTRACT Background: Retrons are a class of retroelements that produce multicopy single-stranded DNA (msDNA) and participate in anti-phage defenses in bacteria. Retrons have been harnessed for the over-production of single-stranded DNA (ssDNA), genome engineering, and directed evolution in bacteria, yeast, and mammalian cells. However, no studies have shown retron-mediated ssDNA production in plants, which could unlock potential applications in plant biotechnology. For example, ssDNA can be used as a template for homology-directed repair (HDR) in several organisms. However, current gene editing technologies rely on the physical delivery of synthetic ssDNA, which limits their applications. Main methods and major results: Here, we demonstrated retron-mediated over-production of ssDNA in Nicotiana benthamiana. Additionally, we tested different retron architectures for improved ssDNA production and identified a new retron architecture that resulted in greater ssDNA abundance. Furthermore, co-expression of the gene encoding the ssDNA-protecting protein VirE2 from Agrobacterium tumefaciens with the retron systems resulted in a 10.7-fold increase in ssDNA production in vivo. We also demonstrated CRISPR-retron-coupled ssDNA over-production and targeted HDR in N. benthamiana. Conclusion: We present an efficient approach for in vivo ssDNA production in plants, which can be harnessed for biotechnological applications.

Transcriptomic Changes in Internode Explants of Stinging Nettle during Callogenesis

Callogenesis, the process during which explants derived from differentiated plant tissues are subjected to a trans-differentiation step characterized by the proliferation of a mass of cells, is fundamental to indirect organogenesis and the establishment of cell suspension cultures. Therefore, understanding how callogenesis takes place is helpful to plant tissue culture, as well as to plant biotechnology and bioprocess engineering. The common herbaceous plant stinging nettle (Urtica dioica L.) is a species producing cellulosic fibres (the bast fibres) and a whole array of phytochemicals for pharmacological, nutraceutical and cosmeceutical use. Thus, it is of interest as a potential multi-purpose plant. In this study, callogenesis in internode explants of a nettle fibre clone (clone 13) was studied using RNA-Seq to understand which gene ontologies predominate at different time points. Callogenesis was induced with the plant growth regulators α-napthaleneacetic acid (NAA) and 6-benzyl aminopurine (BAP) after having determined their optimal concentrations. The process was studied over a period of 34 days, a time point at which a well-visible callus mass developed on the explants. The bioinformatic analysis of the transcriptomic dataset revealed specific gene ontologies characterizing each of the four time points investigated (0, 1, 10 and 34 days). The results show that, while the advanced stage of callogenesis is characterized by the iron deficiency response triggered by the high levels of reactive oxygen species accumulated by the proliferating cell mass, the intermediate and early phases are dominated by ontologies related to the immune response and cell wall loosening, respectively.