Segregation analysis and floral phenotype of T2 transgenic yellow cosmos (Cosmos sulphureus Cav.) carrying neomycin phosphotransferase II gene in second generation
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Abstract. Muchyiddin MGAA, Sawitri WD, Irsyadi MB, Swandari T, Purwantoro A. 2024. Segregation analysis and floral phenotype of T2 transgenic yellow cosmos (Cosmos sulphureus Cav.) carrying neomycin phosphotransferase II gene in second generation. Biodiversitas 25: 1711-1717. The seed of transgenic yellow cosmos (Cosmos sulphureus Cav.) lines harbouring neomycin phosphotranferase II (nptII) gene have been obtained from our previous studies. The nptII selectable marker gene is commonly used for developing a method to estimate the number of targeted gene integrated into the genome. In order to produce successful clones, it is important to ensure that transgenes are inherited sexually as the dominant trait in T2 progeny. This study aimed to obtain the segregation patterns of nptII gene in two populations derived from open- and self-pollinated and evaluated floral phenotypic of T2 transgenic yellow cosmos. The molecular analysis of transgenic yellow cosmos was conducted by PCR and the obtained data was analyzed using a Chi-Square test. The results showed that transgenes were inherited in 1:1 for transgenic and wild-type in both self- and open-pollinated populations. Therefore, it indicated that T2 transgenic plants were still unstable and evolved continuously to segregate independently in the over generation. Surprisingly, the morphological changes were detected in transgenic plants. The mixture of ligulate and tubular types in ray floret was found in some of the flower types in T2 yellow cosmos. In this case, nptII gene may diverge in apparently random genome insertion. This insertion would induce morphological changes in the ray floret type of flower since recombination estimates varied among transformation events.
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References
Aftab N, Saleem K, Khan AHA, Butt TA, Mirza CR, Hussain J, Farooq G, Tahir A, Yousaf S, Zafar MI, Nawaz I, Iqbal M. 2021. Cosmos sulphureus Cav. is more tolerant to lead than copper and chromium in hydroponics system. Intl J Environ Sci Technol 18 (8): 2325-2334. DOI: 10.1007/s13762-020-02981-w.
Ahuja MR, Fladung M. 2014. Integration and inheritance of transgenes in crop plants and trees. Tree Genet Genomes.10. DOI:/10.1007/s11295-014-0724-2.
Breygina M, Klimenko E, Schekaleva O. 2021. Pollen germination and pollen tube growth in gymnosperms. Plants 10 (7): 1301.
Doyle J.J. and Doyle J.L. 1990. Isolation of plant DNA from fresh tissue. Focus. 12. 13-15.
Fibriani S, Agustien I, Sawitri WD, Sugiharto B. 2019. Genetic transformation and expression of sucrose phosphate synthase mutant in tomato plant. Jurnal Bioteknologi Biosains Indonesia 6 (1): 134- 143. DOI: 10.29122/jbbi.v6i1.3341.
Fu J and Ristic Z. 2010. Analysis of transgenic wheat (Triticum aestivum L.) harboring a maize (Zea mays L.) gene for plastid EF-Tu: segregation pattern, expression and effects of the transgene. Plant Mol Biol 73, 339–347. DOI:10.1007/s11103-010-9622-7
Fu X, Kohli A, Twyman RM, Christou P. 2000. Alternative silencing effects involve distinct types of non-spreading cytosine methylation at a three-gene, single-copy transgenic locus in rice. Molecular and General Genetics MGG 263(1): 106-118.
Gahakwa D, Maqbool SB, Fu X, Sudhakar D, Christou P, Kohli A. 2000. Transgenic rice as a system to study the stability of transgene expression: multiple heterologous transgenes show similar behaviour in diverse genetic backgrounds. Theoretical and Applied Genetics 101 (3): 388-399.
Hussain T, Bakhsh A, Munir B, Hassan S, Rao AQ, Shahid AA, et al. 2014. Mendelian segregation pattern and expression studies of insecticidal gene (cry1Ac) in insect resistant cotton progeny. Emirates J Food Agric. 26: 706–715. DOI: 10.9755/ejfa.v26i8.17794.
Irsyadi MB, Sawitri WD, Purwantoro A. 2022. Molecular and phenotype characteristics of T1 transgenic yellow cosmos (Cosmos sulphureus) carrying neomycinphosphotransferase II gene. Biodiversitas 23 (12): 6097-6105. DOI: 10.13057/biodiv/d231202.
Kishimoto S, Ohmiya A. 2006. Regulation of carotenoid biosynthesis in petals and leavesof chrysanthemum (Chrysanthemum morifolium). Physiol Plant 128: 436-447. DOI: 10.1111/j.1399-3054.2006.00761.x.
Kowalczyk, T., Merecz-Sadowska, A, Picot, L., Br?ci´c Kara?conji, I., Wieczfinska, J., Sliwi ´nski, T., Sitarek, P. 2022. Genetic ´Manipulation and Bioreactor Culture of Plants as a Tool for Industry and Its Applications. Molecules 2022, 27, 795. DOI: 10.3390/molecules 27030795.
Lebedev, V. 2022. Stability of Transgene Inheritance in Progeny of Field-Grown Pear Trees over a 7-Year Period. Plants 2022, 11, 151. DOI: 10.3390/plants11020151.
Liao P, Chen X, Wang M, Bach TJ, Chye ML. 2018. Improved fruit ?-tocopherol, carotenoid, squalene and phytosterol contents through manipulation of Brassica juncea 3-HYDROXY-3-METHYLGLUTARYL-COA SYNTHASE1 in transgenic tomato. Plant Biotechnol J. 16(3): 784-796. DOI: 10.1111/pbi.12828.
Limanton-Grevet A. and Jullien M. 2001. Agrobacterium-mediated transformation of Asparagus officinalis L.: molecular and genetic analysis of transgenic plants. Molecular Breeding 7 (2): 141-150.
Ma M, Chen X, Yin Y, Fan R, Li B, Zhan Y and Zeng F. 2020. DNA Methylation Silences Exogenous Gene Expression in Transgenic Birch Progeny. Front. Plant Sci. 11:523748. DOI: 10.3389/fpls.2020.523748.
Mahajan, S., Sharma, N., Kaur, R., Shilpa, C., and Kumar, K. 2017. In vitro propagation and analysis of genetic stability of in vitro propagated plants of jaspi- a clonal rootstock of prunus. Adv. Res. 12, 1–12. DOI: 10.9734/AIR/2017/37784.
Marenkova TV, Sidorchuk YV, Kusnetsov VV, Deineko EV. 2020. Effect of changes in genome ploidy on the mosaic character of nptii gene expression in epialleles of the transgenic tobacco line Nu21. Russ J Genet 56 (2): 204-212. DOI: 10.1134/S1022795420020088.
Purwantoro A, Irsyadi MB, Sawitri WD, Fatumi NC, Fajrina SN. 2023. Efficient floral dip transformation method using Agrobacterium tumefaciens on Cosmos sulphureus Cav. Saudi Journal of Biological Sciences. 30 (7): 103702. DOI:10.1016/j.sjbs.2023.103702
Raldugina GN, Maree M, Mattana M, Shumkova G, Mapelli S, Kholodova VP, Karpichev IV, Kuznetsov VV. 2018. Expression of rice OsMyb4 transcription factor improves tolerance to copper or zinc in canola plants. Biol. Plant. 62: 511-520. DOI 10.1007/s10535- 018-0800-9.
Rashid A. 2017. Comparison of a kanamycin versus hygromycin resistance gene in transgenic plant selection of Arabidopsis thalianaL. Adv Cell Sci Tissue Cul 1 (1): 1-2. DOI: 10.35841/Cell- Science.1000101.
Ren Y, Zhang J, Liang H, Wang J, Yang M. 2017. Inheritance and expression stability of exogenous genes in insect-resistant transgenic poplar. Plant Cell Tissue Organ Cult. 130: 567–576. DOI:10. 1007/s11240-017-1247-y.
Respatie DW, Yudono P, Purwantoro A, Trisyono YA. 2019. The potential of Cosmos sulphureus flower extract as a bioherbicide for Cyperus rotundus. Biodiversitas 20 (12): 3568-3574. DOI: 10.13057/biodiv/d201215.
Saleem M, Ali HA, Furqan MA, Uzma S, Ammara S, Irshad I. 2017. Chemical characterisation and hepatoprotective potential of Cosmos sulphureus Cav. and Cosmos bipinnatus Cav. Nat Prod Res 33 (6): 897-900. DOI: 10.1080/14786419.2017.1413557.
Sun, Y., Chen, Z., Chen, He, H., Peng X. 2023. Analysis of the Genetic Stability of Insect and Herbicide Resistance Genes in Transgenic Rice Lines: A Laboratory and Field Experiment. DOI: 10.1186/s12284-023-00624-5.
Tizaoui K, Kchouk ME. 2012. Genetic approaches for studying transgene inheritance and genetic recombination in three successive generation of transformed tobacco. Genetics and Molecular Biology 35: 640-649. DOI: 10.1590/SN1415-47572012000400015.
Tong J., Han Z., Han A. 2021. Genetic analysis and molecular mapping of Rp, a mutant gene encoding red pericarp in rice (Oryza sativa L.). Czech J. Genet. Plant Breed., 57: 51?57. DOI: 10.17221/70/2020-CJGPB.
Wang, T. D., Huang, T. D., Huang, H. S., and Hua, Y. W. 2017. Origin of secondary somatic embryos and genetic stability of the regenerated plants in Hevea brasiliensis. J. Rubber Res. 20, 101–116. DOI: 10.1007/BF03449145.
Yu Q, Tian X, Lin C, Specht CD, Liao J. 2020. Expression and function studies of CYC/TB1-Like genes in the asymmetric flower (cannaceae, zingiberales). Frontiers in Plants Science 11: 580576.
Zhou G, Guo J, Yang J. 2018. Effect of fertilizers on Cd accumulation and subcellular distribution of two cosmos species (Cosmos sulphureus and Cosmos bipinnata). Intl J Phytoremediat 20: 930-938. DOI: 10.1080/15226514.2018.1448362.
Zhu, H., Li, C. & Gao, C. 2020. Applications of CRISPR–Cas in agriculture and plant biotechnology. Nat Rev Mol Cell Biol (21): 661–677. DOI: 10.1038/s41580-020-00288-9.
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