A Combinational Approach of Enhanced Methanol Production and Double Bt Genes for Broad Spectrum Insect Resistance in Transgenic Cotton

Zubair, Mahrukh; Latif, Ayesha; Rao, Abdul Qayyum; Azam, Saira; Shahid, Naila; Samiullah, Tahir Rehman; Yasmeen, Aneela; Shahid, Ahmad Ali; Nasir, Idrees Ahmad; Husnain, Tayyab

doi:10.1007/s12033-019-00192-4

Cited by 14 publications

(7 citation statements)

References 30 publications

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“…The genetic makeup of the cotton explant's wounding treatment, Agrobacterium strain and cell density, length of the transgene, and finally, the selection pressure of the antibiotic 37 all contribute to transformation efficiency. The results are consistent with other studies where the transformation efficiency was recorded as, 1.25 % 38 . Higher transformation efficiency reported in other studies as (9%) 39 and (20%) 40 were only due to difference in efficiency calculation formula from current study.…”

Section: Phenotypic Analysis Of Transgenic Cotton Lines Though the Esupporting

confidence: 93%

Constitutive expression of Asparaginase in Gossypium hirsutum triggers insecticidal activity against Bemisia tabaci

Gul¹,

Hussain²,

Iqbal³

et al. 2020

Sci Rep

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Whitefly infestation of cotton crop imparts enormous damage to cotton yield by severely affecting plant health, vigour and transmitting Cotton Leaf Curl Virus (CLCuV). Genetic modification of cotton helps to overcome both the direct whitefly infestation as well as CLCuV based cotton yield losses. We have constitutively overexpressed asparaginase (ZmASN) gene in Gossypium hirsutum to overcome the cotton yield losses imparted by whitefly infestation. We achieved 2.54% transformation efficiency in CIM-482 by Agrobacterium-mediated shoot apex transformation method. The relative qRT-PCR revealed 40-fold higher transcripts of asparaginase in transgenic cotton line vs. non-transgenic cotton lines. Metabolic analysis showed higher contents of aspartic acid and glutamic acid in seeds and phloem sap of the transgenic cotton lines. Phenotypically, the transgenic cotton lines showed vigorous growth and height, greater number of bolls, and yield. Among six representative transgenic cotton lines, line 14 had higher photosynthetic rate, stomatal conductance, smooth fiber surface, increased fiber convolutions (SEM analysis) and 95% whitefly mortality as compared to non-transgenic cotton line. The gene integration analysis by fluorescence in situ hybridization showed single copy gene integration at chromosome number 1. Collectively, asparaginase gene demonstrated potential to control whitefly infestation, post-infestation damages and improve cotton plant health and yield: a prerequisite for farmer's community. L-asparaginases play an important role in bacteria, fungi, plants, and the tissues of some animals excluding humans 1. In plants, asparaginases are key players in regulating asparagine (primary N source) by releasing ammonia and aspartate via the deamidation pathway, which is the preferred route for developing plant organs. The aspartic acid resulting from this pathway is incorporated into the aspartate family of amino acids 2,3. There are two reported categories of asparaginases: plant and microbial. Plant asparaginases are further subdivided into two subclasses based on their dependency on potassium (K +): K +-dependent and K +-independent. These are isolated from various plants including Arabidopsis thaliana, Lupinus albus, Lupinus polyphyllus, Phaseolus vulgaris 4 , Lupinus arboreus, Lupinus angustifolius 5 , Lotus japonicus 6 , Glycine max 7 , Withania somnifera 8,9 , and Pisum sativum 10. K +-dependent asparaginases exhibit asparaginase activity 11,12 whereas K +-independent asparaginases (isolated from Lupinus luteus (LlA)) possess both isoaspartyl peptidase and asparaginase activity and recognize β-aspartyl-His as a substrate 3,13. Under nitrogen-limiting conditions, the stored asparagine is the main source of nitrogen for the growing parts of plants, i.e., developing leaves and seeds 14. A total of 81 % of the global cotton fiber stems from genetically modified cotton; GM cotton helps combat various biotic and abiotic stresses 15 that negatively affect the quality and quantity of fiber 16. In 2017, Rahman et al. (2...

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Section: Phenotypic Analysis Of Transgenic Cotton Lines Though the Esupporting

confidence: 93%

Constitutive expression of Asparaginase in Gossypium hirsutum triggers insecticidal activity against Bemisia tabaci

Gul¹,

Hussain²,

Iqbal³

et al. 2020

Sci Rep

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“…Herein, the transcription factor AGL42 (FYF), a repressor protein cloned into the plant expression vector pCAMBIA1301 under the 35S promoter with Nco1 and BglII restriction enzymes, was used to delay leaf senescence in cotton because of its ability to delay floral senescence in Arabidopsis 14 , 16 and fruit, sepal, and leaf senescence in tomato 18 . The pCAMBIA 1301 high-copy noplant expression vector for cotton transformation was used as previously described 21 – 24 . The transformation efficiency in G. hirsutum was 1.47%, which was almost similar to 1.32% 24 , and this may be attributed to similar methodology and crop response 24 .…”

Section: Discussionmentioning

confidence: 99%

Overexpression of the AGL42 gene in cotton delayed leaf senescence through downregulation of NAC transcription factors

Latif

Azam

Shahid

et al. 2022

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Premature leaf senescence negatively influences the physiology and yield of cotton plants. The conserved IDLNL sequence in the C-terminal region of AGL42 MADS-box determines its repressor potential for the down regulation of senescence-related genes. To determine the delay in premature leaf senescence, Arabidopsis AGL42 gene was overexpressed in cotton plants. The absolute quantification of transgenic cotton plants revealed higher mRNA expression of AGL42 compared to that of the non-transgenic control. The spatial expression of GUS fused with AGL42 and the mRNA level was highest in the petals, abscission zone (flower and bud), 8 days post anthesis (DPA) fiber, fresh mature leaves, and senescenced leaves. The mRNA levels of different NAC senescence-promoting genes were significantly downregulated in AGL42 transgenic cotton lines than those in the non-transgenic control. The photosynthetic rate and chlorophyll content were higher in AGL42 transgenic cotton lines than those in the non-transgenic control. Fluorescence in situ hybridization of the AG3 transgenic cotton line revealed a fluorescent signal on chromosome 1 in the hemizygous form. Moreover, the average number of bolls in the transgenic cotton lines was significantly higher than that in the non-transgenic control because of the higher retention of floral buds and squares, which has the potential to improve cotton fiber yield.

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“…Alternatively, to circumvent resistance, several novel strategies like gene pyramiding and generation of chimeric toxins ( Zghal et al, 2017 ; Khabbazi et al, 2018 ; Sellami et al, 2018 ; Rathinam et al, 2019 ; Zubair et al, 2019 ; Salim et al, 2020 ); RNAi-mediated knockdown ( Tian et al, 2015 ; Han et al, 2017 ; Luo et al, 2017 ; Ni et al, 2017 ; Shen et al, 2017 ); CRISPR/Cas9 mediated genome editing ( Zhang et al, 2018 ; Bisht et al, 2019 ; Tyagi et al, 2020 ) and multiomic approaches ( Du et al, 2018 ; Peng et al, 2020 ) for the management of insect resistance have also become an integral part of pest management through biotechnology.…”

Section: Introductionmentioning

confidence: 99%

Exploitation of Novel Bt ICPs for the Management of Helicoverpa armigera (Hübner) in Cotton (Gossypium hirsutum L.): A Transgenic Approach

et al. 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.

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A Combinational Approach of Enhanced Methanol Production and Double Bt Genes for Broad Spectrum Insect Resistance in Transgenic Cotton

Cited by 14 publications

References 30 publications

Constitutive expression of Asparaginase in Gossypium hirsutum triggers insecticidal activity against Bemisia tabaci

Constitutive expression of Asparaginase in Gossypium hirsutum triggers insecticidal activity against Bemisia tabaci

Overexpression of the AGL42 gene in cotton delayed leaf senescence through downregulation of NAC transcription factors

Exploitation of Novel Bt ICPs for the Management of Helicoverpa armigera (Hübner) in Cotton (Gossypium hirsutum L.): A Transgenic Approach

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