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Rohini Sreevathsa
ICAR-National Institute for Plant Biotechnology, India

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Journal article
Published: 29 April 2021 in Frontiers in Microbiology
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Cotton is a commercial crop of global importance. The major threat challenging the productivity in cotton has been the lepidopteron insect pest Helicoverpa armigera or cotton bollworm which voraciously feeds on various plant parts. Biotechnological interventions to manage this herbivore have been a universally inevitable option. The advent of plant genetic engineering and exploitation of Bacillus thuringiensis (Bt) insecticidal crystal proteins (ICPs) marked the beginning of plant protection in cotton through transgenic technology. Despite phenomenal success and widespread acceptance, the fear of resistance development in insects has been a perennial concern. To address this issue, alternate strategies like introgression of a combination of cry protein genes and protein-engineered chimeric toxin genes came into practice. The utility of chimeric toxins produced by domain swapping, rearrangement of domains, and other strategies aid in toxins emerging with broad spectrum efficacy that facilitate the avoidance of resistance in insects toward cry toxins. The present study demonstrates the utility of two Bt ICPs, cry1AcF (produced by domain swapping) and cry2Aa (produced by codon modification) in transgenic cotton for the mitigation of H. armigera. Transgenics were developed in cotton cv. Pusa 8–6 by the exploitation of an apical meristem-targeted in planta transformation protocol. Stringent trait efficacy-based selective screening of T1 and T2 generation transgenic plants enabled the identification of plants resistant to H. armigera upon deliberate challenging. Evaluation of shortlisted events in T3 generation identified a total of nine superior transgenic events with both the genes (six with cry1AcF and three with cry2Aa). The transgenic plants depicted 80–100% larval mortality of H. armigera and 10–30% leaf damage. Molecular characterization of the shortlisted transgenics demonstrated stable integration, inheritance and expression of transgenes. The study is the first of its kind to utilise a non-tissue culture-based transformation strategy for the development of stable transgenics in cotton harbouring two novel genes, cry1AcF and cry2Aa for insect resistance. The identified transgenic events can be potential options toward the exploitation of unique cry genes for the management of the polyphagous insect pest H. armigera.

ACS Style

Kesiraju Karthik; Jyotsana Negi; Maniraj Rathinam; Navinder Saini; Rohini Sreevathsa. Exploitation of Novel Bt ICPs for the Management of Helicoverpa armigera (Hübner) in Cotton (Gossypium hirsutum L.): A Transgenic Approach. Frontiers in Microbiology 2021, 12, 1 .

AMA Style

Kesiraju Karthik, Jyotsana Negi, Maniraj Rathinam, Navinder Saini, Rohini Sreevathsa. Exploitation of Novel Bt ICPs for the Management of Helicoverpa armigera (Hübner) in Cotton (Gossypium hirsutum L.): A Transgenic Approach. Frontiers in Microbiology. 2021; 12 ():1.

Chicago/Turabian Style

Kesiraju Karthik; Jyotsana Negi; Maniraj Rathinam; Navinder Saini; Rohini Sreevathsa. 2021. "Exploitation of Novel Bt ICPs for the Management of Helicoverpa armigera (Hübner) in Cotton (Gossypium hirsutum L.): A Transgenic Approach." Frontiers in Microbiology 12, no. : 1.

Microbiology
Published: 29 April 2021
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Cotton is a commercial crop of global importance. The major threat challenging the productivity in cotton has been the lepidopteron insect pest Helicoverpa armigera or cotton bollworm which voraciously feeds on various plant parts. Biotechnological interventions to manage this herbivore have been a universally inevitable option. The advent of plant genetic engineering and exploitation of Bacillus thuringiensis (Bt) insecticidal crystal proteins (ICPs) marked the beginning of plant protection in cotton through transgenic technology. Despite phenomenal success and widespread acceptance, the fear of resistance development in insects has been a perennial concern. To address this issue, alternate strategies like introgression of a combination of cry protein genes and protein-engineered chimeric toxin genes came into practice. The utility of chimeric toxins produced by domain swapping, rearrangement of domains, and other strategies aid in toxins emerging with broad spectrum efficacy that facilitate the avoidance of resistance in insects toward cry toxins. The present study demonstrates the utility of two Bt ICPs, cry1AcF (produced by domain swapping) and cry2Aa (produced by codon modification) in transgenic cotton for the mitigation of H. armigera. Transgenics were developed in cotton cv. Pusa 8–6 by the exploitation of an apical meristem-targeted in planta transformation protocol. Stringent trait efficacy-based selective screening of T1 and T2 generation transgenic plants enabled the identification of plants resistant to H. armigera upon deliberate challenging. Evaluation of shortlisted events in T3 generation identified a total of nine superior transgenic events with both the genes (six with cry1AcF and three with cry2Aa). The transgenic plants depicted 80–100% larval mortality of H. armigera and 10–30% leaf damage. Molecular characterization of the shortlisted transgenics demonstrated stable integration, inheritance and expression of transgenes. The study is the first of its kind to utilise a non-tissue culture-based transformation strategy for the development of stable transgenics in cotton harbouring two novel genes, cry1AcF and cry2Aa for insect resistance. The identified transgenic events can be potential options toward the exploitation of unique cry genes for the management of the polyphagous insect pest H. armigera.

ACS Style

Kesiraju Karthik; Jyotsana Negi; Maniraj Rathinam; Navinder Saini; Rohini Sreevathsa. Exploitation of Novel Bt ICPs for the Management of Helicoverpa armigera (Hübner) in Cotton (Gossypium hirsutum L.): A Transgenic Approach. 2021, 12, 1 .

AMA Style

Kesiraju Karthik, Jyotsana Negi, Maniraj Rathinam, Navinder Saini, Rohini Sreevathsa. Exploitation of Novel Bt ICPs for the Management of Helicoverpa armigera (Hübner) in Cotton (Gossypium hirsutum L.): A Transgenic Approach. . 2021; 12 ():1.

Chicago/Turabian Style

Kesiraju Karthik; Jyotsana Negi; Maniraj Rathinam; Navinder Saini; Rohini Sreevathsa. 2021. "Exploitation of Novel Bt ICPs for the Management of Helicoverpa armigera (Hübner) in Cotton (Gossypium hirsutum L.): A Transgenic Approach." 12, no. : 1.

Research article
Published: 31 March 2021 in Pest Management Science
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BACKGROUND Occurrence of multiple biotic stresses on crop plants result in drastic yield losses which may have severe impact on the food security. It is a challenge to design strategies for simultaneous management of these multiple stresses. Hence, establishment of innovative approaches that aid in their management is critical. Here, we have introgressed a micro RNA‐induced gene silencing (MIGS) based combinatorial gene construct containing seven target gene sequences of cotton leaf curl disease (CLCuD), cotton leaf hopper (Amrasca biguttula biguttula), cotton whitefly (Bemisia tabaci) and root‐knot nematode (Meloidogyne incognita). RESULTS Stable transgenic lines of Nicotiana benthamiana were generated with the T‐DNA harbouring Arabidopsis miR173 target site fused to fragments of Sec23 and ecdysone receptor (EcR) genes of cotton leaf hopper and cotton whitefly. It also contained C2/replication associated protein (C2/Rep) and C4 (movement protein) along with βC1 gene of betasatellite to target CLCuD, and two FMRFamide‐like peptide (FLP) genes, Mi‐flp14 and Mi‐flp18 of M. incognita. These transgenic plants were assessed for the amenability of MIGS approach for pest control by efficacy evaluation against M. incognita. Results showed successful production of siRNA through the tasiRNA pathway in the transgenic plants corresponding to Mi‐flp18 gene. Furthermore, we observed reduced Mi‐flp14 and Mi‐flp18 transcripts (up to 2.37 ± 0.12‐fold) in females extracted from transgenic plants. The average number of galls, total endoparasites, egg masses and number of eggs per egg mass reduced were in the range of 27–62%, 39–70%, 38–65% and 34–49% respectively. More importantly, MIGS transgenic plants showed 80% reduction in the nematode multiplication factor (MF). CONCLUSION This study demonstrates successful validation of the MIGS approach in the model plant, N. benthamiana for efficacy against M. incognita, as a prelude to translation to cotton.

ACS Style

Alkesh Hada; Basavaprabhu L. Patil; Akansha Bajpai; Karthik Kesiraju; Savithramma Dinesh‐Kumar; Bheema Paraselli; Rohini Sreevathsa; Uma Rao. Micro RNA ‐induced gene silencing strategy for the delivery of siRNAs targeting Meloidogyne incognita in a model plant Nicotiana benthamiana. Pest Management Science 2021, 77, 3396 -3405.

AMA Style

Alkesh Hada, Basavaprabhu L. Patil, Akansha Bajpai, Karthik Kesiraju, Savithramma Dinesh‐Kumar, Bheema Paraselli, Rohini Sreevathsa, Uma Rao. Micro RNA ‐induced gene silencing strategy for the delivery of siRNAs targeting Meloidogyne incognita in a model plant Nicotiana benthamiana. Pest Management Science. 2021; 77 (7):3396-3405.

Chicago/Turabian Style

Alkesh Hada; Basavaprabhu L. Patil; Akansha Bajpai; Karthik Kesiraju; Savithramma Dinesh‐Kumar; Bheema Paraselli; Rohini Sreevathsa; Uma Rao. 2021. "Micro RNA ‐induced gene silencing strategy for the delivery of siRNAs targeting Meloidogyne incognita in a model plant Nicotiana benthamiana." Pest Management Science 77, no. 7: 3396-3405.

Original article
Published: 27 March 2021 in 3 Biotech
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The polyphagous spotted pod borer, Maruca vitrata is an important agricultural pest that causes extensive damage on various food crops. Though the pest is managed by synthetic chemicals, exploration of biotechnological approaches for its control is important. RNAi-based gene silencing is one such tool that has been extensively used for functional genomics and is highly variable in insects. In view of this, we have attempted to demonstrate RNAi in M. vitrata through exogenous double-stranded RNA (dsRNA) administration targeting seven genes associated with midgut, chemosensory, cell signalling and development. Two modes of exogenous dsRNA delivery by either haemolymph injection and/or ingestion into third and late third instar larval stages respectively exhibited efficient silencing of specific transcripts. Furthermore, dsRNA injection into the haemolymph showed significant reduction of target gene expression compared to negative controls establishing this mode of delivery to be more efficient. Interestingly, haemolymph injection required lesser dsRNA and led to higher reduction of transcript level vis-à-vis ingestion as demonstrated in dsRNA Serine Protease 33 (ds-SP33)-fed larvae. Over-expression of key RNAi component DICER and detection of siRNA authenticated the presence of RNAi in M. vitrata. Additionally, we have identified inhibitor molecules like morpholine, piperidine, carboxamide and piperidine–carboxamide through in silico analysis for blocking the function of SP33 to demonstrate the utility of functional genomics. Thus, the present study establishes the usefulness of injection and ingestion approaches for exogenous dsRNA delivery into M. vitrata larvae for effective RNAi.

ACS Style

Madhurima Chatterjee; Jyoti Yadav; Maniraj Rathinam; Abhishek Mandal; Gopal Chowdhary; Rohini Sreevathsa; Uma Rao. Exogenous administration of dsRNA for the demonstration of RNAi in Maruca vitrata (lepidoptera: crambidae). 3 Biotech 2021, 11, 1 -12.

AMA Style

Madhurima Chatterjee, Jyoti Yadav, Maniraj Rathinam, Abhishek Mandal, Gopal Chowdhary, Rohini Sreevathsa, Uma Rao. Exogenous administration of dsRNA for the demonstration of RNAi in Maruca vitrata (lepidoptera: crambidae). 3 Biotech. 2021; 11 (4):1-12.

Chicago/Turabian Style

Madhurima Chatterjee; Jyoti Yadav; Maniraj Rathinam; Abhishek Mandal; Gopal Chowdhary; Rohini Sreevathsa; Uma Rao. 2021. "Exogenous administration of dsRNA for the demonstration of RNAi in Maruca vitrata (lepidoptera: crambidae)." 3 Biotech 11, no. 4: 1-12.

Original research article
Published: 28 January 2021 in Frontiers in Plant Science
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Efficient regeneration of explants devoid of intrinsic somaclonal variations is a cardinal step in plant tissue culture, thus, a vital component of transgenic technology. However, recalcitrance of economically important crops to tissue culture-based organogenesis ensues a setback in the use of transgenesis in the genetic engineering of crop plants. The present study developed an optimized, genotype-independent, nonconventional tissue culture-independent in planta strategy for the genetic transformation of flax/linseed. This apical meristem-targeted in planta transformation protocol will accelerate value addition in the dual purpose industrially important but recalcitrant fiber crop flax/linseed. The study delineated optimization of Agrobacterium tumefaciens-mediated transformation and stable T-DNA (pCambia2301:GUS:nptII) integration in flax. It established successful use of a stringent soilrite-based screening in the presence of 30 mg/L kanamycin for the identification of putative transformants. The amenability, authenticity, and reproducibility of soilrite-based kanamycin screening were further verified at the molecular level by GUS histochemical analysis of T0 seedlings, GUS and nptII gene-specific PCR, genomic Southern hybridization for stable integration of T-DNA, and expression analysis of transgenes by sqRT-PCR. This method resulted in a screening efficiency of 6.05% in the presence of kanamycin, indicating amenability of in planta flax transformation. The strategy can be a promising tool for the successful development of transgenics in flax.

ACS Style

Karthik Kesiraju; Shaily Tyagi; Soumyadeep Mukherjee; Rhitu Rai; Nagendra K. Singh; Rohini Sreevathsa; Prasanta K. Dash. An Apical Meristem-Targeted in planta Transformation Method for the Development of Transgenics in Flax (Linum usitatissimum): Optimization and Validation. Frontiers in Plant Science 2021, 11, 1 .

AMA Style

Karthik Kesiraju, Shaily Tyagi, Soumyadeep Mukherjee, Rhitu Rai, Nagendra K. Singh, Rohini Sreevathsa, Prasanta K. Dash. An Apical Meristem-Targeted in planta Transformation Method for the Development of Transgenics in Flax (Linum usitatissimum): Optimization and Validation. Frontiers in Plant Science. 2021; 11 ():1.

Chicago/Turabian Style

Karthik Kesiraju; Shaily Tyagi; Soumyadeep Mukherjee; Rhitu Rai; Nagendra K. Singh; Rohini Sreevathsa; Prasanta K. Dash. 2021. "An Apical Meristem-Targeted in planta Transformation Method for the Development of Transgenics in Flax (Linum usitatissimum): Optimization and Validation." Frontiers in Plant Science 11, no. : 1.

Research article
Published: 09 January 2021 in Pest Management Science
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Background Pigeonpea is one of the economically important legume food crops and a major source of dietary proteins. Management of pod borer, Helicoverpa armigera has been prominent among crop improvement programs. Lack of resistance sources in the cultivated germplasm and crossing incompatibility with the pod borer‐resistant wild relatives have prompted biotechnological interventions. Identification and exploitation of genes from pigeonpea wild relatives in host plant resistance towards the pod borer assumes pertinence. Dynamic transcriptome analysis of the wild relative vis a vis cultivated pigeonpea identified a CHI4 chitinase as one of the putative insect resistance genes. Results The study presents variations in important amino acids in CHI4 chitinases from Cajanus cajan and Cajanus platycarpus. Comparative protein modeling and docking analysis of the two proteins demonstrated differences in substrate binding efficacy of the chitinase from C. platycarpus, which resulted in a minimum binding energy of ‐8.7 Kcal/mol. Further, we successfully evaluated the insecticidal activity of the chitinase from C. platycarpus against H. armigera challenge through heterologous expression in tobacco. Molecular characterization of transgenic plants confirmed that their efficacy against pod borer was due to the integration of CHI4 from C. platycarpus. Conclusion Docking analysis demonstrated effective substrate interaction as a possible reason for efficacy against pod borer in the chitinase from C. platycarpus. This was authenticated by successful overexpression and bioefficacy assessment against H. armigera in tobacco. CHI4 gene from C. platycarpus can be useful in the mitigation of H. armegira in pigeonpea as well as in other crops.

ACS Style

Maniraj Rathinam; Sathish Kumar Marimuthu; Shaily Tyagi; Karthik Kesiraju; Lakshmi Prabha Alagiamanavalan; Uma Rao; Rohini Sreevathsa. Characterization and in planta validation of a CHI4 chitinase from Cajanus platycarpus (Benth.) Maesen for its efficacy against pod borer, Helicoverpa armigera (Hübner). Pest Management Science 2021, 77, 2337 -2349.

AMA Style

Maniraj Rathinam, Sathish Kumar Marimuthu, Shaily Tyagi, Karthik Kesiraju, Lakshmi Prabha Alagiamanavalan, Uma Rao, Rohini Sreevathsa. Characterization and in planta validation of a CHI4 chitinase from Cajanus platycarpus (Benth.) Maesen for its efficacy against pod borer, Helicoverpa armigera (Hübner). Pest Management Science. 2021; 77 (5):2337-2349.

Chicago/Turabian Style

Maniraj Rathinam; Sathish Kumar Marimuthu; Shaily Tyagi; Karthik Kesiraju; Lakshmi Prabha Alagiamanavalan; Uma Rao; Rohini Sreevathsa. 2021. "Characterization and in planta validation of a CHI4 chitinase from Cajanus platycarpus (Benth.) Maesen for its efficacy against pod borer, Helicoverpa armigera (Hübner)." Pest Management Science 77, no. 5: 2337-2349.

Short communication
Published: 19 October 2020 in Physiology and Molecular Biology of Plants
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Cotton (Gossypium hirsutum L.), a mercantile crop plant, is grown worldwide for fiber and seed oil. As with other economically important crops, cotton is bogged down with many biotic and abiotic stress factors. Towards this, genetic engineering offers numerous protocols to engineer plants for better resilience. However, recalcitrance of cotton to plant tissue culture has been the major constraint for successful in vitro regeneration. Hence, alternate methods that evade tissue culture regeneration have been envisaged. Non tissue culture-based in planta transformation strategies are in vogue due to amenability and ease in the generation of transgenic plants. In the present study, we demonstrate the utility of an in planta transformation protocol and establishment of a stringent selection agent-based screening for the identification of transgenics. The genotype independent nature of the protocol was validated in cotton cv. Pusa 8–6 using GFP. Preliminary transformation efficiency of 28% was achieved with a screening efficiency of 20% in the presence of hygromycin. The proof of T-DNA integration by various molecular and expression analysis in T1 and T2 generations proved that this technique can be employed to generate transgenic cotton.

ACS Style

Karthik Kesiraju; Pragya Mishra; Akansha Bajpai; Manju Sharma; Uma Rao; Rohini Sreevathsa. Agrobacterium tumefaciens-mediated in planta transformation strategy for development of transgenics in cotton (Gossypium hirsutum L.) with GFP as a visual marker. Physiology and Molecular Biology of Plants 2020, 26, 1 -9.

AMA Style

Karthik Kesiraju, Pragya Mishra, Akansha Bajpai, Manju Sharma, Uma Rao, Rohini Sreevathsa. Agrobacterium tumefaciens-mediated in planta transformation strategy for development of transgenics in cotton (Gossypium hirsutum L.) with GFP as a visual marker. Physiology and Molecular Biology of Plants. 2020; 26 (11):1-9.

Chicago/Turabian Style

Karthik Kesiraju; Pragya Mishra; Akansha Bajpai; Manju Sharma; Uma Rao; Rohini Sreevathsa. 2020. "Agrobacterium tumefaciens-mediated in planta transformation strategy for development of transgenics in cotton (Gossypium hirsutum L.) with GFP as a visual marker." Physiology and Molecular Biology of Plants 26, no. 11: 1-9.

Review article
Published: 11 August 2020 in ACS Omega
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Plants are challenged incessantly by several biotic and abiotic stresses during their entire growth period. As with other biotic stress factors, insect pests have also posed serious concerns related to yield losses due to which agricultural productivity is at stake. In plants, trait modification for crop improvement was initiated with breeding approaches followed by genetic engineering. However, stringent regulatory policies for risk assessment and lack of social acceptance for genetically modified crops worldwide have incited researchers toward alternate strategies. Genome engineering or genome editing has emerged as a new breeding technique with the ability to edit the genomes of plants, animals, microbes, and human beings. Several gene editing strategies are being executed with continuous emergence of variants. The scientific community has unraveled the utility of various editing tools from endonucleases to CRISPR/Cas in several aspects related to plant growth, development, and mitigation of stresses. The categorical focus on the development of tools and techniques including designing of binary vectors to facilitate ease in genome engineering are being pursued. Through this Review, we embark upon the conglomeration of various genome editing strategies that can be and are being used to design insect pest resistance in plants. Case studies and novel crop-based approaches that reiterate the successful use of these tools in insects as well as in plants are highlighted. Further, the Review also provides implications for the requirement of a specific regulatory framework and risk assessment of the edited crops. Genome editing toward insect pest management is here to stay, provided uncompromising efforts are made toward the identification of amiable target genes.

ACS Style

Shaily Tyagi; Karthik Kesiraju; Manjesh Saakre; Maniraj Rathinam; Venkat Raman; Debasis Pattanayak; Rohini Sreevathsa. Genome Editing for Resistance to Insect Pests: An Emerging Tool for Crop Improvement. ACS Omega 2020, 5, 20674 -20683.

AMA Style

Shaily Tyagi, Karthik Kesiraju, Manjesh Saakre, Maniraj Rathinam, Venkat Raman, Debasis Pattanayak, Rohini Sreevathsa. Genome Editing for Resistance to Insect Pests: An Emerging Tool for Crop Improvement. ACS Omega. 2020; 5 (33):20674-20683.

Chicago/Turabian Style

Shaily Tyagi; Karthik Kesiraju; Manjesh Saakre; Maniraj Rathinam; Venkat Raman; Debasis Pattanayak; Rohini Sreevathsa. 2020. "Genome Editing for Resistance to Insect Pests: An Emerging Tool for Crop Improvement." ACS Omega 5, no. 33: 20674-20683.

Journal article
Published: 20 July 2020 in Applied Microbiology and Biotechnology
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The pigeonpea wild relative Cajanus platycarpus is resistant to Helicoverpa armigera, one of the major pests responsible for yield losses in Cajanus cajan. Deciphering the molecular mechanism underlying host plant resistance is pertinent to identify proteins that aid in the mitigation of the insect pest. The present study adopted comparative proteomics as a tool to interpret the resistance mechanism(s) in C. platycarpus vis-à-vis C. cajan during continued herbivory (up to 96 h). Over-representation analysis of the differentially expressed proteins implicated a multi-dimensional resistance response accomplished by both physical and chemical barriers in C. platycarpus. While the chemical basis for resistance was depicted by the upregulation of proteins playing a rate limiting role in the phenylpropanoid pathway, the physical basis was provided by the regulation of proteins involved in microtubule assembly and synthesis of lignins. Upregulation of proteins in the polyamine pathway indicated the role of metabolite conjugates to be negatively affecting herbivore growth. Reallocation of resources and diversion of metabolic flux to support the production of secondary metabolites could be the probable approach in the wild relative against herbivory. Our study provided deeper insights into the pod borer resistance mechanism in C. platycarpus for utility in crop improvement. • Pod borer resistance in Cajanus platycarpus is multi-dimensional. • Pod borer resistance has been arbitrated to cell wall rigidity and secondary metabolites. • Phenylpropanoid pathway derivatives apparently shaped the plant chemical defense against pod borer.

ACS Style

Maniraj Rathinam; Bernd Roschitzki; Jonas Grossmann; Pragya Mishra; Laura Kunz; Witold Wolski; Christian Panse; Shaily Tyagi; Uma Rao; Ralph Schlapbach; Rohini Sreevathsa. Unraveling the proteomic changes involved in the resistance response of Cajanus platycarpus to herbivory by Helicoverpa armigera. Applied Microbiology and Biotechnology 2020, 104, 7603 -7618.

AMA Style

Maniraj Rathinam, Bernd Roschitzki, Jonas Grossmann, Pragya Mishra, Laura Kunz, Witold Wolski, Christian Panse, Shaily Tyagi, Uma Rao, Ralph Schlapbach, Rohini Sreevathsa. Unraveling the proteomic changes involved in the resistance response of Cajanus platycarpus to herbivory by Helicoverpa armigera. Applied Microbiology and Biotechnology. 2020; 104 (17):7603-7618.

Chicago/Turabian Style

Maniraj Rathinam; Bernd Roschitzki; Jonas Grossmann; Pragya Mishra; Laura Kunz; Witold Wolski; Christian Panse; Shaily Tyagi; Uma Rao; Ralph Schlapbach; Rohini Sreevathsa. 2020. "Unraveling the proteomic changes involved in the resistance response of Cajanus platycarpus to herbivory by Helicoverpa armigera." Applied Microbiology and Biotechnology 104, no. 17: 7603-7618.

Original research article
Published: 07 July 2020 in Frontiers in Plant Science
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Weeds burden plant growth as they compete for space, sunlight, and soil nutrients leading to 25–80% yield losses. Glyphosate [N-(phosphonomethyl)glycine] is a widely used broad spectrum non-selective herbicide that controls weeds by inhibiting 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) enzyme and interfering with the shikimate biosynthesis pathway. Cotton (Gossypium hirsutum L.) is one of the most important commercial crops grown worldwide for its fiber. We have developed herbicide tolerant transgenic cotton (cv. P8-6) by introgression of a codon-optimized and modified EPSPS gene (CP4-EPSPS) possessing an N-terminal chloroplast targeting peptide from Petunia hybrida. Because of the recalcitrant nature of cotton, a genotype-independent non-tissue culture-based apical meristem-targeted in planta transformation approach was used to develop transformants. Although in planta transformation methodologies are advantageous in developing a large number of transgenic plants, effective screening strategies are essential for initial identification of transformants. In the present study, the use of a two-level rigorous screening strategy identified 2.27% of T1 generation plants as tolerant to 800 and 1,500 mg/L of commercially available glyphosate (Roundup). Precise molecular characterization revealed stable integration, expression, and inheritance of CP4-EPSPS in advanced generations of the promising transgenic events. Further, superiority of selected transgenic plants in tolerating increasing levels of glyphosate (500–4,000 mg/L) was ascertained through reduced accumulation of shikimate. This report is the first of its kind where cotton transformants tolerating high levels of glyphosate (up to 4,000 mg/L) and accumulating low levels of shikimate have been identified. This study not only reiterated the genotype-independent nature of the transformation strategy but also reiterated the translational utility of the CP4-EPSPS gene in management of weeds.

ACS Style

Kesiraju Karthik; Muralimohan Nandiganti; Arulprakash Thangaraj; Shweta Singh; Pragya Mishra; Maniraj Rathinam; Manju Sharma; Nagendra Kumar Singh; Prasanta K. Dash; Rohini Sreevathsa. Transgenic Cotton (Gossypium hirsutum L.) to Combat Weed Vagaries: Utility of an Apical Meristem-Targeted in planta Transformation Strategy to Introgress a Modified CP4-EPSPS Gene for Glyphosate Tolerance. Frontiers in Plant Science 2020, 11, 768 .

AMA Style

Kesiraju Karthik, Muralimohan Nandiganti, Arulprakash Thangaraj, Shweta Singh, Pragya Mishra, Maniraj Rathinam, Manju Sharma, Nagendra Kumar Singh, Prasanta K. Dash, Rohini Sreevathsa. Transgenic Cotton (Gossypium hirsutum L.) to Combat Weed Vagaries: Utility of an Apical Meristem-Targeted in planta Transformation Strategy to Introgress a Modified CP4-EPSPS Gene for Glyphosate Tolerance. Frontiers in Plant Science. 2020; 11 ():768.

Chicago/Turabian Style

Kesiraju Karthik; Muralimohan Nandiganti; Arulprakash Thangaraj; Shweta Singh; Pragya Mishra; Maniraj Rathinam; Manju Sharma; Nagendra Kumar Singh; Prasanta K. Dash; Rohini Sreevathsa. 2020. "Transgenic Cotton (Gossypium hirsutum L.) to Combat Weed Vagaries: Utility of an Apical Meristem-Targeted in planta Transformation Strategy to Introgress a Modified CP4-EPSPS Gene for Glyphosate Tolerance." Frontiers in Plant Science 11, no. : 768.

Original article
Published: 15 May 2020 in Journal of Plant Biochemistry and Biotechnology
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Concurrent expression of multiple insecticidal toxins as pyramided genes in the same host plant is one of the tangible strategies to delay the development of resistance in insects. In this study, an ideal combination of Insecticidal Cry Proteins (ICP) of Bacillus thuringiensis (Bt) viz., chimeric Cry1AcF (produced by domain swapping) and Cry2Aa (a synthetic protein with modified codon usage for plants) were selected. The main aim of this study was to determine whether the combination of two cry genes can confer better protection against Helicoverpa armigera over single a cry gene. Transgenic tobacco plants harbouring single and double ICP genes were generated and analysed for their insecticidal activity against cotton bollworm (H. armigera) through detached leaf insect bioassay. Efficacy studies showed that transgenic plants carrying the combinatorial construct had higher insecticidal activity compared to the individual genes and required lesser time for effective and maximum mortality of the pest. Molecular analysis and protein expression analysis of transgenic tobacco plants also correlated with the bioassay results. This study suggests that transgenic tobacco plants having both the toxins can be an effective option against H. armigera. However, the toxin combination should be carefully selected depending upon the most important pest species present in each geographical area.

ACS Style

N. Muralimohan; Ravi Prakash Saini; Karthik Kesiraju; Debasis Pattanayak; P. Ananda Kumar; K. Kasturi; Rohini Sreevathsa. Molecular stacking of two codon-modified genes encoding Bt insecticidal proteins, Cry1AcF and Cry2Aa for management of resistance development in Helicoverpa armigera. Journal of Plant Biochemistry and Biotechnology 2020, 29, 518 -527.

AMA Style

N. Muralimohan, Ravi Prakash Saini, Karthik Kesiraju, Debasis Pattanayak, P. Ananda Kumar, K. Kasturi, Rohini Sreevathsa. Molecular stacking of two codon-modified genes encoding Bt insecticidal proteins, Cry1AcF and Cry2Aa for management of resistance development in Helicoverpa armigera. Journal of Plant Biochemistry and Biotechnology. 2020; 29 (3):518-527.

Chicago/Turabian Style

N. Muralimohan; Ravi Prakash Saini; Karthik Kesiraju; Debasis Pattanayak; P. Ananda Kumar; K. Kasturi; Rohini Sreevathsa. 2020. "Molecular stacking of two codon-modified genes encoding Bt insecticidal proteins, Cry1AcF and Cry2Aa for management of resistance development in Helicoverpa armigera." Journal of Plant Biochemistry and Biotechnology 29, no. 3: 518-527.

Mini review
Published: 27 January 2020 in Applied Microbiology and Biotechnology
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Global climate change and combinatorial environmental stresses pose grave challenges to food security and agricultural sustainability. This calls for diverse and futuristic approaches for the development of crops with increased resilience to natural vagaries. Though innumerable strategies involving diverse genes/pathways are being deciphered in plants to aid stress mitigation, the hunt is still on. Furthermore, strategies that work to alleviate a combination of stresses are always pertinent. In this review, we discuss polygalacturonase inhibitor (PGIP) proteins as a plausible option to mitigate multiple biotic stresses. These are ubiquitous cell wall proteins that inhibit the pectin-depolymerizing activity of cell wall loosening enzymes, polygalacturonases (PGs). While plant PGs are those responsible for developmental activities like fruit ripening, pollen tube elongation, etc., PGs from various biotic stress factors like insects, fungal and bacterial pathogens aid in invasion by reducing the plant cell wall rigidity. To counteract, plants secrete PGIPs, which inhibit the pectin hydrolyzing activity of PGs from the attacking pests and pathogens. Multiple approaches in diverse crop species have demonstrated PGIP-based protection against pathogens and insect pests. Additionally, effectual interaction between PGs-PGIP is an important aspect for successful utilization of this approach. Molecular strategies leading to improved PG-PGIPs interaction is a highlight to demonstrate the use of PGIPs as an amenable stress mitigation approach. The review focuses on a comprehensive update on phylogeny of PGIPs, natural variation of resistance as well as their emerging translational utility towards mitigation of various biotic stresses.

ACS Style

Maniraj Rathinam; Uma Rao; Rohini Sreevathsa. Novel biotechnological strategies to combat biotic stresses: polygalacturonase inhibitor (PGIP) proteins as a promising comprehensive option. Applied Microbiology and Biotechnology 2020, 104, 2333 -2342.

AMA Style

Maniraj Rathinam, Uma Rao, Rohini Sreevathsa. Novel biotechnological strategies to combat biotic stresses: polygalacturonase inhibitor (PGIP) proteins as a promising comprehensive option. Applied Microbiology and Biotechnology. 2020; 104 (6):2333-2342.

Chicago/Turabian Style

Maniraj Rathinam; Uma Rao; Rohini Sreevathsa. 2020. "Novel biotechnological strategies to combat biotic stresses: polygalacturonase inhibitor (PGIP) proteins as a promising comprehensive option." Applied Microbiology and Biotechnology 104, no. 6: 2333-2342.

Research article
Published: 16 December 2019 in Pest Management Science
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BACKGROUND Pigeonpea is a source of quality proteins and the main constituent of a well‐balanced diet for majority of Indian population. One of the major constraints in the production of pigeonpea is a polyphagous insect pest, Helicoverpa armigera. Non‐availability of resistant sources in the germplasm and limitations in conventional breeding have been key factors for continued yield losses. Additionally, hazards of chemical fertilizers on the environment have prompted the scientific community to develop alternative strategies. Bacillus thuringiensis (Bt) insecticidal proteins (ICPs) have emerged as the most reliable source for the control of insect pests through transgenics. RESULTS Transgenic pigeonpea plants harboring validated Bt ICPs, Cry2Aa and Cry1AcF were developed by a non‐tissue culture based in planta transformation strategy and assessed for integration of Transfer‐DNA (T‐DNA) and efficacy against pod borer under in vitro conditions. For the first time this study demonstrates the successful evaluation of 19 transgenic pigeonpea events (11 with cry2Aa and 8 with cry1AcF) under soil and pot conditions in a nethouse containment. The stability in the performance was assessed stringently by deliberate H. armigera larval challenging. The trial identified ten promising events of both the genes that portrayed reduced damage to the herbivore. CONCLUSION We present the first ever successful evaluation of pigeonpea transgenics with the ability to mitigate pod borer under nethouse conditions. The transgenics depicted molecular evidence for the stability of T‐DNA integration, consistency in the expression of Cry proteins and resistance against H. armigera. These events can form a pool of useful transgenics to manage the devastating pod borer. © 2019 Society of Chemical Industry

ACS Style

Nikhil Ram Kumar; Maniraj Rathinam; Shweta Singh; Karthik Kesiraju; Vikraman Muniyandi; Nagendra Kumar Singh; Prasanta K. Dash; Rohini Sreevathsa. Assessment of Pigeonpea ( Cajanus cajan L.) transgenics expressing Bt ICPs, Cry2Aa and Cry1AcF under nethouse containment implicated an effective control against herbivory by Helicoverpa armigera (Hübner). Pest Management Science 2019, 76, 1902 -1911.

AMA Style

Nikhil Ram Kumar, Maniraj Rathinam, Shweta Singh, Karthik Kesiraju, Vikraman Muniyandi, Nagendra Kumar Singh, Prasanta K. Dash, Rohini Sreevathsa. Assessment of Pigeonpea ( Cajanus cajan L.) transgenics expressing Bt ICPs, Cry2Aa and Cry1AcF under nethouse containment implicated an effective control against herbivory by Helicoverpa armigera (Hübner). Pest Management Science. 2019; 76 (5):1902-1911.

Chicago/Turabian Style

Nikhil Ram Kumar; Maniraj Rathinam; Shweta Singh; Karthik Kesiraju; Vikraman Muniyandi; Nagendra Kumar Singh; Prasanta K. Dash; Rohini Sreevathsa. 2019. "Assessment of Pigeonpea ( Cajanus cajan L.) transgenics expressing Bt ICPs, Cry2Aa and Cry1AcF under nethouse containment implicated an effective control against herbivory by Helicoverpa armigera (Hübner)." Pest Management Science 76, no. 5: 1902-1911.

Journal article
Published: 01 November 2019 in Plant Physiology and Biochemistry
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Soybeans are known for its good source of protein (40%), oil (20%) and also serve as a source of nutraceutical compounds including tocopherols (toc). To know the molecular basis of differential α-toc accumulation in two contrasting soybean genotypes: DS74 (low α-toc - 1.36 μg/g and total-toc -29.72 μg/g) and Bragg (high α-toc - 10.48 μg/g and total-toc 178.91 μg/g), the analysis of γ-TMT3 promoter activity and its methylation patterns were carried out. The sequencing results revealed nucleotide variation between Bragg:γ-TMT3-P and DS74:γ-TMT3-P, however none of the variations were found in core-promoter region or in cis-elements. The histochemical GUS assay revealed higher promoter activity of Bragg:γ-TMT3-P than that of DS74:γ-TMT3-P and correlated with significantly higher and lower (P < 0.05) expression of γ-TMT3 gene respectively. To know the molecular basis of differential accumulation of α-toc in these contrasting soybean genotypes, the DNA methylation pattern of γ-TMT3 gene body and its promoter was studied in both varieties. The results showed higher percentage (62.5%) of methylation in DS74:γ-TMT3-P than in Bragg:γ-TMT3-P (50%). Out of all the methylation sites in the promoter region, one of methylation site was found at CAAT box (-190 bp) of DS74:γ-TMT3-P. Further gene body methylation patterns revealed lowest % (40%) of CG methylation in DS74:γ-TMT3 gene as compared to Bragg:γ-TMT3 (64.2%). Thus our study revealed that, expression of γ-TMT3 gene was influenced by its promoter activity and methylation patterns in cis-elements of γ-TMT3 promoter and gene body. This study will help us to understand the possible role of methylation and promoter activity in determining the α-toc content in soybean seeds.

ACS Style

Khushboo Kumari; Monika Prakash Rai; Navita Bansal; G. Rama Prashat; Sweta Kumari; Veda Krishnan; Rohini Srivathsa; Anil Dahuja; Archana Sachdev; Shelly Praveen; T. Vinutha. Analysis of γ-Tocopherol methyl transferase3 promoter activity and study of methylation patterns of the promoter and its gene body. Plant Physiology and Biochemistry 2019, 144, 375 -385.

AMA Style

Khushboo Kumari, Monika Prakash Rai, Navita Bansal, G. Rama Prashat, Sweta Kumari, Veda Krishnan, Rohini Srivathsa, Anil Dahuja, Archana Sachdev, Shelly Praveen, T. Vinutha. Analysis of γ-Tocopherol methyl transferase3 promoter activity and study of methylation patterns of the promoter and its gene body. Plant Physiology and Biochemistry. 2019; 144 ():375-385.

Chicago/Turabian Style

Khushboo Kumari; Monika Prakash Rai; Navita Bansal; G. Rama Prashat; Sweta Kumari; Veda Krishnan; Rohini Srivathsa; Anil Dahuja; Archana Sachdev; Shelly Praveen; T. Vinutha. 2019. "Analysis of γ-Tocopherol methyl transferase3 promoter activity and study of methylation patterns of the promoter and its gene body." Plant Physiology and Biochemistry 144, no. : 375-385.

Comparative study
Published: 04 July 2019 in Plant Molecular Biology
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Deeper insights into the resistance response of Cajanus platycarpus were obtained based on comparative transcriptomics under Helicoverpa armigera infestation. Devastation by pod borer, Helicoverpa armigera is one of the major factors for stagnated productivity in Pigeonpea. Despite possessing a multitude of desirable traits including pod borer resistance, wild relatives of Cajanus spp. have remained under-utilized due to linkage drag and cross-incompatibility. Discovery and deployment of genes from them can provide means to tackle key pests like H. armigera. Transcriptomic differences between Cajanus platycarpus and Cajanus cajan during different time points (0, 18, 38, 96 h) of pod borer infestation were elucidated in this study. For the first ever time, we demonstrated captivating variations in their response; C. platycarpus apparently being reasonably agile with effectual transcriptomic reprogramming to deter the insect. Deeper insights into the differential response were obtained by identification of significant GO-terms related to herbivory followed by combined KEGG and ontology analyses. C. platycarpus portrayed a multilevel response with cardinal involvement of SAR, redox homeostasis and reconfiguration of primary metabolites leading to a comprehensive defense response. The credibility of RNA-seq analyses was ascertained by transient expression of selected putative insect resistance genes from C. platycarpus viz., chitinase (CHI4), Alpha-amylase/subtilisin inhibitor (IAAS) and Flavonoid 3_5 hydroxylase (C75A1) in Nicotiana benthamiana followed by efficacy analysis against H. armigera. qPCR validated results of the study provided innovative insights and useful leads for development of durable pod borer resistance.

ACS Style

Maniraj Rathinam; Pragya Mishra; Ajay Kumar Mahato; Nagendra Kumar Singh; Uma Rao; Rohini Sreevathsa. Comparative transcriptome analyses provide novel insights into the differential response of Pigeonpea (Cajanus cajan L.) and its wild relative (Cajanus platycarpus (Benth.) Maesen) to herbivory by Helicoverpa armigera (Hübner). Plant Molecular Biology 2019, 101, 163 -182.

AMA Style

Maniraj Rathinam, Pragya Mishra, Ajay Kumar Mahato, Nagendra Kumar Singh, Uma Rao, Rohini Sreevathsa. Comparative transcriptome analyses provide novel insights into the differential response of Pigeonpea (Cajanus cajan L.) and its wild relative (Cajanus platycarpus (Benth.) Maesen) to herbivory by Helicoverpa armigera (Hübner). Plant Molecular Biology. 2019; 101 (1-2):163-182.

Chicago/Turabian Style

Maniraj Rathinam; Pragya Mishra; Ajay Kumar Mahato; Nagendra Kumar Singh; Uma Rao; Rohini Sreevathsa. 2019. "Comparative transcriptome analyses provide novel insights into the differential response of Pigeonpea (Cajanus cajan L.) and its wild relative (Cajanus platycarpus (Benth.) Maesen) to herbivory by Helicoverpa armigera (Hübner)." Plant Molecular Biology 101, no. 1-2: 163-182.

Research article
Published: 03 July 2019 in PLOS ONE
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Pigeonpea is a major source of dietary protein to the vegetarian population of the Indian sub-continent. Crop improvement to mitigate biotic and abiotic stresses for realization of its potential yield and bridging yield gap is the need of the hour. Availability of limited genomic resources in the cultivated germplasm, however, is a serious bottleneck towards successful molecular breeding for the development of superior genotypes in pigeonpea. In view of this, improvement of pigeonpea can be attempted through transgenesis or by exploiting genetic resources from its wild relatives. Pigeonpea wild relatives are known to be bestowed with agronomic traits of importance; discovery and deployment of genes from them can provide a lucrative option for crop improvement. Understanding molecular signatures of wild relatives would not only provide information about the mechanism behind desired traits but also enable us to extrapolate the information to cultivated pigeonpea. The present study deals with the characterization of leaf transcriptomes of Cajanus cajan and one of its wild relatives, Cajanus platycarpus. Illumina sequencing revealed 0.11 million transcripts in both the species with an annotation of 0.09 million (82%) transcripts using BLASTX. Comparative transcriptome analyses on the whole, divulged cues about the wild relative being vigilant and agile. Gene ontology and Mapman analysis depicted higher number of transcripts in the wild relative pertaining to signaling, transcription factors and stress responsive genes. Further, networking between the differentially expressed MapMan bins demonstrated conspicuous interactions between different bins through 535 nodes (512 Genes and 23 Pathways) and 1857 edges. The authenticity of RNA-seq analysis was confirmed by qRT-PCR. The information emanating from this study can provide valuable information and resource for future translational research including genome editing to alleviate varied stresses. Further, this learning can be a platform for in-depth investigations to decipher molecular mechanisms for mitigation of various stresses in the wild relative.

ACS Style

Maniraj Rathinam; Pragya Mishra; Madavan Vasudevan; Roli Budhwar; Ajay Mahato; A. Lakshmi Prabha; Nagendra Kumar Singh; Uma Rao; Rohini Sreevathsa. Comparative transcriptome analysis of pigeonpea, Cajanus cajan (L.) and one of its wild relatives Cajanus platycarpus (Benth.) Maesen. PLOS ONE 2019, 14, e0218731 .

AMA Style

Maniraj Rathinam, Pragya Mishra, Madavan Vasudevan, Roli Budhwar, Ajay Mahato, A. Lakshmi Prabha, Nagendra Kumar Singh, Uma Rao, Rohini Sreevathsa. Comparative transcriptome analysis of pigeonpea, Cajanus cajan (L.) and one of its wild relatives Cajanus platycarpus (Benth.) Maesen. PLOS ONE. 2019; 14 (7):e0218731.

Chicago/Turabian Style

Maniraj Rathinam; Pragya Mishra; Madavan Vasudevan; Roli Budhwar; Ajay Mahato; A. Lakshmi Prabha; Nagendra Kumar Singh; Uma Rao; Rohini Sreevathsa. 2019. "Comparative transcriptome analysis of pigeonpea, Cajanus cajan (L.) and one of its wild relatives Cajanus platycarpus (Benth.) Maesen." PLOS ONE 14, no. 7: e0218731.

Review article
Published: 21 May 2019 in Seminars in Cell & Developmental Biology
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Crop productivity in rice is harshly limited due to high concentration of salt in the soil. To understand the intricacies of the mechanism it is important to unravel the key pathways operating inside the plant cell. Emerging state-of-the art technologies have provided the tools to discover the key components inside the plant cell for salt tolerance. Among the molecular entities, transcription factors and/or other important components of sensing and signaling cascades have been the attractive targets and the role of NHX and SOS1 transporters amply described. Not only marker assisted programs but also transgenic approaches by using reverse genetic strategies (knockout or knockdown) or overexpression have been extensively used to engineer rice crop. CRISPR/Cas is an attractive paradigm and provides the feasibility for manipulating several genes simultaneously. Here, in this review we highlight some of the molecular entities that could be potentially targeted for generating rice amenable to sustain growth under high salinity conditions by employing CRISPR/Cas. We also try to address key questions for rice salt stress tolerance other than what is already known.

ACS Style

Sufia Farhat; Neha Jain; Nisha Singh; Rohini Sreevathsa; Prasanta K. Dash; Rhitu Rai; Sandeep Yadav; Pramod Kumar; Ananda K. Sarkar; Ajay Jain; Nagendra K Singh; Vandna Rai. CRISPR-Cas9 directed genome engineering for enhancing salt stress tolerance in rice. Seminars in Cell & Developmental Biology 2019, 96, 91 -99.

AMA Style

Sufia Farhat, Neha Jain, Nisha Singh, Rohini Sreevathsa, Prasanta K. Dash, Rhitu Rai, Sandeep Yadav, Pramod Kumar, Ananda K. Sarkar, Ajay Jain, Nagendra K Singh, Vandna Rai. CRISPR-Cas9 directed genome engineering for enhancing salt stress tolerance in rice. Seminars in Cell & Developmental Biology. 2019; 96 ():91-99.

Chicago/Turabian Style

Sufia Farhat; Neha Jain; Nisha Singh; Rohini Sreevathsa; Prasanta K. Dash; Rhitu Rai; Sandeep Yadav; Pramod Kumar; Ananda K. Sarkar; Ajay Jain; Nagendra K Singh; Vandna Rai. 2019. "CRISPR-Cas9 directed genome engineering for enhancing salt stress tolerance in rice." Seminars in Cell & Developmental Biology 96, no. : 91-99.

Journal article
Published: 02 March 2019 in Toxins
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Bacillus thuringiensis insecticidal proteins (Bt ICPs) are reliable and valuable options for pest management in crops. Protein engineering of Bt ICPs is a competitive alternative for resistance management in insects. The primary focus of the study was to reiterate the translational utility of a protein-engineered chimeric Cry toxin, Cry1AcF, for its broad spectrum insecticidal efficacy using molecular modeling and docking studies. In-depth bioinformatic analysis was undertaken for structure prediction of the Cry toxin as the ligand and aminopeptidase1 receptors (APN1) from Helicoverpa armigera (HaAPN1) and Spodoptera litura (SlAPN1) as receptors, followed by interaction studies using protein-protein docking tools. The study revealed feasible interactions between the toxin and the two receptors through H-bonding and hydrophobic interactions. Further, molecular dynamics simulations substantiated the stability of the interactions, proving the broad spectrum efficacy of Cry1AcF in controlling H. armigera and S. litura. These findings justify the utility of protein-engineered toxins in pest management.

ACS Style

Maniraj Rathinam; Karthik Kesiraju; Shweta Singh; Vinutha Thimmegowda; Vandna Rai; Debasis Pattanayak; Rohini Sreevathsa. Molecular Interaction-Based Exploration of the Broad Spectrum Efficacy of a Bacillus thuringiensis Insecticidal Chimeric Protein, Cry1AcF. Toxins 2019, 11, 143 .

AMA Style

Maniraj Rathinam, Karthik Kesiraju, Shweta Singh, Vinutha Thimmegowda, Vandna Rai, Debasis Pattanayak, Rohini Sreevathsa. Molecular Interaction-Based Exploration of the Broad Spectrum Efficacy of a Bacillus thuringiensis Insecticidal Chimeric Protein, Cry1AcF. Toxins. 2019; 11 (3):143.

Chicago/Turabian Style

Maniraj Rathinam; Karthik Kesiraju; Shweta Singh; Vinutha Thimmegowda; Vandna Rai; Debasis Pattanayak; Rohini Sreevathsa. 2019. "Molecular Interaction-Based Exploration of the Broad Spectrum Efficacy of a Bacillus thuringiensis Insecticidal Chimeric Protein, Cry1AcF." Toxins 11, no. 3: 143.

Journal article
Published: 10 August 2018 in International Journal of Current Microbiology and Applied Sciences
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G. Keshavareddy; A.R.V. Kumar; Vemanna S. Ramu; Rohini Sreevathsa; M. Udayakumar. Synthetic cry1AcF Gene Imparts Resistance to Spodoptera litura (F.) and Amsacta albistriga Walker in Peanut (Arachis hypogaea L.) cv. VRI-2. International Journal of Current Microbiology and Applied Sciences 2018, 7, 2567 -2583.

AMA Style

G. Keshavareddy, A.R.V. Kumar, Vemanna S. Ramu, Rohini Sreevathsa, M. Udayakumar. Synthetic cry1AcF Gene Imparts Resistance to Spodoptera litura (F.) and Amsacta albistriga Walker in Peanut (Arachis hypogaea L.) cv. VRI-2. International Journal of Current Microbiology and Applied Sciences. 2018; 7 (8):2567-2583.

Chicago/Turabian Style

G. Keshavareddy; A.R.V. Kumar; Vemanna S. Ramu; Rohini Sreevathsa; M. Udayakumar. 2018. "Synthetic cry1AcF Gene Imparts Resistance to Spodoptera litura (F.) and Amsacta albistriga Walker in Peanut (Arachis hypogaea L.) cv. VRI-2." International Journal of Current Microbiology and Applied Sciences 7, no. 8: 2567-2583.

Journal article
Published: 11 June 2018 in Scientific Reports
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Pigeon pea is an important legume infested by a plethora of insect pests amongst which gram pod borer Helicoverpa armigera is very prominent. Imparting resistance to this insect herbivore is of global importance in attaining food security. Expression of insecticidal crystal proteins (ICP) in diverse crops has led to increased resistance to several pests. We report in this paper, expression of Cry2Aa in transgenic pigeon pea and its effectiveness towards H. armigera by employing Agrobacterium-mediated in planta transformation approach. Approximately 0.8% of T1 generation plants were identified as putative transformants based on screening in the presence of 70 ppm kanamycin as the selection agent. Promising events were further recognized in advanced generations based on integration, expression and bioefficacy of the transgenes. Seven T3 lines (11.8% of the selected T1 events) were categorized as superior as these events demonstrated 80-100% mortality of the challenged larvae and improved ability to prevent damage caused by the larvae. The selected transgenic plants accumulated Cry2Aa in the range of 25-80 µg/g FW. The transgenic events developed in the study can be used in pigeon pea improvement programmes for pod borer resistance.

ACS Style

Shweta Singh; Nikhil Ram Kumar; R. Maniraj; R. Lakshmikanth; K. Y. S. Rao; N. Muralimohan; T. Arulprakash; K. Karthik; N. B. Shashibhushan; T. Vinutha; Debasis Pattanayak; Prasanta K. Dash; P. Ananda Kumar; Rohini Sreevathsa. Expression of Cry2Aa, a Bacillus thuringiensis insecticidal protein in transgenic pigeon pea confers resistance to gram pod borer, Helicoverpa armigera. Scientific Reports 2018, 8, 8820 .

AMA Style

Shweta Singh, Nikhil Ram Kumar, R. Maniraj, R. Lakshmikanth, K. Y. S. Rao, N. Muralimohan, T. Arulprakash, K. Karthik, N. B. Shashibhushan, T. Vinutha, Debasis Pattanayak, Prasanta K. Dash, P. Ananda Kumar, Rohini Sreevathsa. Expression of Cry2Aa, a Bacillus thuringiensis insecticidal protein in transgenic pigeon pea confers resistance to gram pod borer, Helicoverpa armigera. Scientific Reports. 2018; 8 (1):8820.

Chicago/Turabian Style

Shweta Singh; Nikhil Ram Kumar; R. Maniraj; R. Lakshmikanth; K. Y. S. Rao; N. Muralimohan; T. Arulprakash; K. Karthik; N. B. Shashibhushan; T. Vinutha; Debasis Pattanayak; Prasanta K. Dash; P. Ananda Kumar; Rohini Sreevathsa. 2018. "Expression of Cry2Aa, a Bacillus thuringiensis insecticidal protein in transgenic pigeon pea confers resistance to gram pod borer, Helicoverpa armigera." Scientific Reports 8, no. 1: 8820.