Improved plastid transformation efficiency in Scoparia dulcis L.

Sadanandam Abbagani,Srinivas Kota,Qiang Hao,Muralikrishna Narra,Vaishnavi Anumula,A.V Rao,Zanmin Hu 한국식물생명공학회 2019 JOURNAL OF PLANT BIOTECHNOLOGY Vol.46 No.4

The high expression level of industrial and metabolically important proteins in plants can be achieved by plastid transformation. The CaIA vector, a Capsicum- specific vector harboring aadA (spectinomycin resistance), is a selectable marker controlled by the PsbA promoter, and the terminator is flanked by the trnA and trnI regions of the inverted repeat (IR) region of the plastid. The CaIA vector can introduce foreign genes into the IR region of the plastid genome. The biolistic method was used for chloroplast transformation in Scoparia dulcis with leaf explants followed by antibiotic selection on regeneration medium. Transplastomes were successfully screened, and the trans-formation efficiency of 3 transgenic lines from 25 bombarded leaf explants was determined. Transplastomic lines were evaluated by PCR and Southern blotting for the confirmation of aadA insertion and its integration into the chloroplast genome. Seeds collected from transplastomes were analyzed on spectinomycin medium with wild types to determine genetic stability. The increased chloroplast transformation efficiency (3 transplastomic lines from 25 bombarded explants) would be useful for expressing therapeutically and industrially important genes in Scoparia dulcis L.

Highly efficient production of transgenic Scoparia dulcis L. mediated by Agrobacterium tumefaciens: plant regeneration via shoot organogenesis

Aileni, Mahender,Abbagani, Sadanandam,Zhang, Peng The Korean Society of Plant Biotechnology 2011 Plant biotechnology reports Vol.5 No.2

Efficient Agrobacterium-mediated genetic transformation of Scoparia dulcis L. was developed using Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pCAMBIA1301 with ${\beta}$-glucuronidase (GUS) (uidA) and hygromycin phosphotransferase (hpt) genes. Two-day precultured leaf segments of in vitro shoot culture were found to be suitable for cocultivation with the Agrobacterium strain, and acetosyringone was able to promote the transformation process. After selection on shoot organogenesis medium with appropriate concentrations of hygromycin and carbenicillin, adventitious shoots were developed on elongation medium by twice subculturing under the same selection scheme. The elongated hygromycin-resistant shoots were subsequently rooted on the MS medium supplemented with $1mg\;l^{-1}$ indole-3-butyric acid and $15mg\;l^{-1}$ hygromycin. Successful transformation was confirmed by PCR analysis using uidA- and hpt-specific primers and monitored by histochemical assay for ${\beta}$-GUS activity during shoot organogenesis. Integration of hpt gene into the genome of transgenic plants was also verified by Southern blot analysis. High transformation efficiency at a rate of 54.6% with an average of $3.9{\pm}0.39$ transgenic plantlets per explant was achieved in the present transformation system. It took only 2-3 months from seed germination to positive transformants transplanted to soil. Therefore, an efficient and fast genetic transformation system was developed for S. dulcis using an Agrobacterium-mediated approach and plant regeneration via shoot organogenesis, which provides a useful platform for future genetic engineering studies in this medicinally important plant.

Highly Convenient and Large Scale Synthesis of 5-chloroindole and its 3-substituted Analogues

Keetha, Laxminarayana,Palle, Sadanandam,Ramanatham, Vinodkumar,Khagga, Mukkanti,Chinnapillai, Rajendiran Korean Chemical Society 2011 대한화학회지 Vol.55 No.2

A large scale and commercially feasible synthesis of 5-chloroindole and its 3-substituted analogues has been described via a halogen - halogen exchange reaction from 5-bromoindole and its derivatives using cuprous chloride and dipolar aprotic solvent N-methyl-2-pyrrolidone in one pot with good yields. CuCl과 N-methyl-2-pyrrolidone을 이용하여 5-bromoindole을 halogen - halogen 교환반응을 통하여 5-chloroindole계 화합물을 합성하는 one-pot대량합성방법을 개발하였다.

Intraspecific genetic variation in Corynandra chelidonii (Angiosperms: Cleomaceae) as revealed by SCoT, ISSR and RAPD analyses

Sirangi, Subash,Jogam, Phanikanth,Nemali, Gandhi,Ajmeera, Ragan,Abbagani, Sadanandam,Raju, Vatsavaya S. The Korean Society of Plant Biotechnology 2020 JOURNAL OF PLANT BIOTECHNOLOGY Vol.47 No.4

The genetic diversity of two subpopulations of Corynandra chelidonii, one of terrestrial and the other of aquatic environments, was measured with molecular markers, such as start codon targeted (SCoT), inter simple sequence repeats (ISSR), and random amplification of polymorphic DNA (RAPD). The traditional morphological traits such as habitat, habit, leaf morphology, the colour of the sepals and petals, number of stamens, and seed morphology formed the base for their realization as two varieties, C. chelidonii var. pallae and C. chelidonii var. chelidonii. The polymorphism between the two variants was 100% with the primers SCoT-2 and OPA-1 and 4, while maximum polymorphism was detected with ISSR-2, SCoT-3, and OPA-3. The study used, for the first time, more than one molecular marker to assess the genetic variation underscoring the morphological variation in Corynandra chelidonii (L.f.) Cochrane & Iltis. The study justifies the recognition of the two subpopulations of Corynandra chelidonii from aquatic and terrestrial environments as two distinct varieties, C. chelidonii var. pallae (Reddy & Raju) V.S.Raju and C. chelidonii var. chelidonii, respectively, based on the traditional taxonomic evidence.

Nanostructured KTaTeO6 and Ag‑doped KTaTeO6 Defect Pyrochlores: Promising Photocatalysts for Dye Degradation and Water Splitting

Perala Venkataswamy,CH. Sudhakar Reddy,Ravi Gundeboina,Gullapelli Sadanandam,Naveen Kumar Veldurthi,M. Vithal 대한금속·재료학회 2018 ELECTRONIC MATERIALS LETTERS Vol.14 No.4

In this study, the nanostructured parent KTaTeO6(KTTO) and Ag-doped KTaTeO6(ATTO) catalysts with defect pyrochlorestructure were prepared by solid-state and ion-exchange methods, respectively. The synthesized materials were characterizedby various techniques to determine their chemical composition, morphology and microstructural features. The XRD studiesshow that both KTTO and ATTO have cubic structure (space group Fd3m) with high crystallinity. The doping of Ag alteredthe BET surface area of parent KTTO. The nano nature of the samples was studied by TEM images. A considerable redshiftin the absorption edge is observed for ATTO compared to KTTO. Incorporation of Ag+in the KTTO lattice is clearlyidentified from EDX, elemental mapping and XPS results. Degradation of methyl violet and solar water splitting reactionswere used to access the photocatalytic activity of KTTO and ATTO. The results obtained suggest that compared to KTTO,the ATTO showed higher photocatalytic activity in both cases. The favourable properties such as high surface area, moresurface hydroxyl groups, stronger light absorption in visible region and narrower band gap energy were supposed to be thereasons for the high activity observed in ATTO.

Adventitious Shoot Regeneration from Immature Zygotic Embryos of Indian Kino Tree (Pterocarpus marsupium Roxb.) and Genetic Integrity Analysis of in Vitro Derived Plants Using ISSR Markers

Radhika Tippani,Anil Kumar Vemunoori,Rajesh Yarra,Rama Swamy Nanna,Sadanandam Abbagani,Christopher Thammidala 한국원예학회 2013 Horticulture, Environment, and Biotechnology Vol.54 No.6

A simple and highly efficient protocol has been developed for in vitro regeneration of Pterocarpus marsupium using immature zygotic embryo (IZE) as explant. The highest frequency of shoot regeneration (93.8%) was obtained on Murashige and Skoog (MS) medium supplemented with 3.0 mg·L-1 6-benzylaminopurine (BA) and 0.5 mg·L-1 indole-3-acetic acid (IAA), wherein maximum of 17.3 ± 0.9 shoots per explant was induced. When these cultures were sub-cultured on MS medium supplemented with 1.0 mg·L-1 BA, more shoots (27.2 ± 1.1) with an average shoot length of 4.5 cm were observed. The highest rooting (70.8%) and maximum number of roots (3.2 ± 0.3) per shoot were obtained when shoots were dipped in 3.0 mg·L-1 indol-3-butyric acid (IBA) solution for 24 hours and further cultured on half-strength MS medium. Plantlets obtained in vitro were transferred to the field after being hardened with a 74%survival rate. Analysis of regenerated plantlets using inter-simple sequence repeat (ISSR) markers confirmed that there was no genetic variability. All ISSR banding profiles from regenerated plantlets were monomorphic and similar to those of the mother plant. This protocol might be helpful for the mass multiplication and in vitro conservation of P. marsupium.

Improved plastid transformation efficiency in Scoparia dulcis L.

Kota, Srinivas,Hao, Qiang,Narra, Muralikrishna,Anumula, Vaishnavi,Rao, A.V,Hu, Zanmin,Abbagani, Sadanandam The Korean Society of Plant Biotechnology 2019 식물생명공학회지 Vol.46 No.4

The high expression level of industrial and metabolically important proteins in plants can be achieved by plastid transformation. The CaIA vector, a Capsicum-specific vector harboring aadA (spectinomycin resistance), is a selectable marker controlled by the PsbA promoter, and the terminator is flanked by the trnA and trnI regions of the inverted repeat (IR) region of the plastid. The CaIA vector can introduce foreign genes into the IR region of the plastid genome. The biolistic method was used for chloroplast transformation in Scoparia dulcis with leaf explants followed by antibiotic selection on regeneration medium. Transplastomes were successfully screened, and the transformation efficiency of 3 transgenic lines from 25 bombarded leaf explants was determined. Transplastomic lines were evaluated by PCR and Southern blotting for the confirmation of aadA insertion and its integration into the chloroplast genome. Seeds collected from transplastomes were analyzed on spectinomycin medium with wild types to determine genetic stability. The increased chloroplast transformation efficiency (3 transplastomic lines from 25 bombarded explants) would be useful for expressing therapeutically and industrially important genes in Scoparia dulcis L.