Genetic transformation and expression of transgenic lines of Populus x euramericana with insect-resistance and salt-tolerance genes

Abstract Background As important forest tree species, biological stress and soil salinization are important factors that restrict the growth of Populus × euramericana. WRKYs are important transcription factors in plants that can regulate plant responses to biotic and abiotic stresses. In this study, PeWRKY31 was isolated from Populus × euramericana, and its bioinformation, salt resistance and insect resistance were analyzed. This study aims to provide guidance for producing salt-resistant and insect-resistant poplars. Results PeWRKY31 has a predicted open reading frame (ORF) of 1842 bp that encodes 613 amino acids. The predicted protein is the unstable, acidic, and hydrophilic protein with a molecular weight of 66.34 kDa, and it has numerous potential phosphorylation sites, chiefly on serines and threonines. PeWRKY31 is a zinc-finger C2H2 type-II WRKY TF that is closely related to WRKY TFs of Populus tomentosa, and localizes to the nucleus. A PeWRKY31 overexpression vector was constructed and transformed into Nicotiana tabacum L. Overexpression of PeWRKY31 improved the salt tolerance and insect resistance of the transgenic tobacco. Transcriptome sequencing and KEGG enrichment analysis showed the elevated expression of genes related to glutathione metabolism, plant hormone signal transduction, and MAPK signaling pathways, the functions of which were important in plant salt tolerance and insect resistance in the overexpressing tobacco line. Conclusions PeWRKY31 was isolated from Populus × euramericana. Overexpression of PeWRKY31 improved the resistance of transgenic plant to salt stress and pest stress. The study provides references for the generation of stress-resistant lines with potentially great economic benefit.

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Genetic Transformation of Crops for Insect Resistance: Potential and Limitations

Critical Reviews in Plant Sciences ◽

10.1080/07352680490273400 ◽

2004 ◽

Vol 23 (1) ◽

pp. 47-72 ◽

Cited By ~ 95

Author(s):

Hari C. Sharma ◽

Kiran K. Sharma ◽

Jonathan H. Crouch

Keyword(s):

Genetic Transformation ◽

Insect Resistance

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Comparative Functional Analysis of Class II Potassium Transporters, SvHKT2;1, SvHKT2;2, and HvHKT2;1, on Ionic Transport and Salt Tolerance in Transgenic Arabidopsis

Plants ◽

10.3390/plants9060786 ◽

2020 ◽

Vol 9 (6) ◽

pp. 786

Author(s):

Yuichi Tada ◽

Aki Ohnuma

Keyword(s):

Salt Tolerance ◽

Transport Properties ◽

Plant Cells ◽

Ionic Transport ◽

Class Ii ◽

Phloem Sap ◽

Over Expression ◽

Potassium Transporters ◽

Transgenic Lines ◽

Reduced Salt

Class II high-affinity potassium transporters (HKT2s) mediate Na+–K+ cotransport and Na+/K+ homeostasis under K+-starved or saline conditions. Their functions have been studied in yeast and X. laevis oocytes; however, little is known about their respective properties in plant cells. In this study, we characterized the Na+ and K+ transport properties of SvHKT2;1, SvHKT2;2 and HvHKT2;1 in Arabidopsis under different ionic conditions. The differences were detected in shoot K+ accumulation and root K+ uptake under salt stress conditions, K+ accumulation in roots and phloem sap under K+-starved conditions, and shoot and root Na+ accumulation under K+-starved conditions among the HKT2s transgenic lines and WT plants. These results indicate the diverse ionic transport properties of these HKT2s in plant cells, which could not be detected using yeast or X. laevis oocytes. Furthermore, Arabidopsis expressing HKT2s showed reduced salt tolerance, while over-expression of HvHKT2;1 in barley, which has the ability to sequestrate Na+, showed enhanced salt tolerance by accumulating Na+ in the shoots. These results suggest that the coordinated enhancement of Na+ accumulation and sequestration mechanisms in shoots could be a promising strategy to confer salt tolerance to glycophytes.

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Improvement of a Genetic Transformation System and Preliminary Study on the Function of LpABCB21 and LpPILS7 Based on Somatic Embryogenesis in Lilium pumilum DC. Fisch

International Journal of Molecular Sciences ◽

10.3390/ijms21186784 ◽

2020 ◽

Vol 21 (18) ◽

pp. 6784

Author(s):

Shengli Song ◽

Rui Yan ◽

Chunxia Wang ◽

Jinxia Wang ◽

Hongmei Sun

Keyword(s):

Somatic Embryogenesis ◽

Genetic Transformation ◽

Somatic Embryo ◽

Asymmetric Distribution ◽

Embryo Induction ◽

Transgenic Lines ◽

Preliminary Study ◽

Genetic Transformation System ◽

Mutant Lines ◽

Somatic Embryo Induction

Auxin transport mediates the asymmetric distribution of auxin that determines the fate of cell development. Agrobacterium-mediated genetic transformation is an important technical means to study gene function. Our previous study showed that the expression levels of LpABCB21 and LpPILS7 are significantly up-regulated in the somatic embryogenesis (SE) of Lilium pumilum DC. Fisch. (L. pumilum), but the functions of both genes remain unclear. Here, the genetic transformation technology previously developed by our team based on the L.pumilum system was improved, and the genetic transformation efficiency increased by 5.7–13.0%. Use of overexpression and CRISPR/Cas9 technology produced three overexpression and seven mutant lines of LpABCB21, and seven overexpression and six mutant lines of LpPILS7. Analysis of the differences in somatic embryo induction of transgenic lines confirmed that LpABCB21 regulates the early formation of the somatic embryo; however, excessive expression level of LpABCB21 inhibits somatic embryo induction efficiency. LpPILS7 mainly regulates somatic embryo induction efficiency. This study provides a more efficient method of genetic transformation of L. pumilum. LpABCB21 and LpPILS7 are confirmed to have important regulatory roles in L. pumilum SE thus laying the foundation for subsequent studies of the molecular mechanism of Lilium SE.

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GENETIC TRANSFORMATION OF APPLE WITH BIBAC VECTORS TO INCREASE SCAB RESISTANCE IN TRANSGENIC LINES

Acta Horticulturae ◽

10.17660/actahortic.2007.738.43 ◽

2007 ◽

pp. 361-367

Author(s):

J. Zhu ◽

A. Zhou ◽

S. Granozio ◽

E. Andruccioli ◽

E. Belfanti ◽

...

Keyword(s):

Genetic Transformation ◽

Scab Resistance ◽

Transgenic Lines

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Enhanced Salt Tolerance Conferred by the Complete 2.3 kb cDNA of the Rice Vacuolar Na+/H+ Antiporter Gene Compared to 1.9 kb Coding Region with 5′ UTR in Transgenic Lines of Rice

Frontiers in Plant Science ◽

10.3389/fpls.2016.00014 ◽

2016 ◽

Vol 7 ◽

Cited By ~ 21

Author(s):

U. S. M. Amin ◽

Sudip Biswas ◽

Sabrina M. Elias ◽

Samsad Razzaque ◽

Taslima Haque ◽

...

Keyword(s):

Salt Tolerance ◽

Coding Region ◽

Transgenic Lines

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Expression of Multiple Exogenous Insect Resistance and Salt Tolerance Genes in Populus nigra L.

Frontiers in Plant Science ◽

10.3389/fpls.2020.01123 ◽

2020 ◽

Vol 11 ◽

Author(s):

Xinglu Zhou ◽

Yan Dong ◽

Qi Zhang ◽

Dandan Xiao ◽

Minsheng Yang ◽

...

Keyword(s):

Salt Tolerance ◽

Insect Resistance ◽

Populus Nigra

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Aplikasi Teknologi DNA untuk Akselerasi Program Pemuliaan Ketahanan Tanaman Kakao terhadap Hama dan Penyakit Utama

Jurnal Penelitian dan Pengembangan Pertanian ◽

10.21082/jp3.v35n4.2016.p155-166 ◽

2017 ◽

Vol 35 (4) ◽

pp. 155

Author(s):

I Made Tasma

Keyword(s):

Quantitative Trait Loci ◽

Genetic Transformation ◽

Insect Resistance ◽

Quantitative Trait ◽

Insect Pests ◽

Insect Pest ◽

Breeding Program ◽

Pests And Diseases ◽

Dna Technology ◽

Modern Genomic

ABSTRACT One of the main constraints on cacao cultivation is disease and insect pest attacks causing significant yield loss. The main insect pests and diseases on cacao plantation are cacao pod borer, cacao fruit rot, vascular streak dieback and cacao mirids (Helopeltis spp.). Conventional breeding method to obtain new cacao clones resistant to insect pests and diseases is a slow process. It may take 1520 years to obtain a new superior clone. Applying DNA technology should expedite cacao breeding program. The article described the application of DNA technology currently available to expedite cacao breeding program for disease and insect resistance. Many genes and quantitative trait loci (QTLs) of important traits have been discovered related to cacao plant productivity and yield quality, disease and insect pest resistance traits. Modern genomic technologies as well as DNA marker have also been applied in cacao breeding program. Genetic transformation technology has been explored its application for cacao improvement. With the development of modern genomic technology, important gene/QTL discoveries would be faster to accelerate insect pest and disease resistant cultivar development. All these new DNA technologies have been assessed their potential applications for coping important pest and disease and for yield improvement. DNA technologies, mainly MAS and genomic-data based breeding technologies are ready to be applied to support breeding programs for main pest and disease resistance to enhance Indonesian cacao productivity and quality. Keywords: Cacao, disease and insect resistance, genomics, DNA markers, genetic transformation, marker-assisted breeding AbstrakSalah satu kendala utama dalam budi daya kakao ialah serangan hama dan penyakit. Hama dan penyakit utama kakao adalah penggerek buah kakao (PBK), busuk buah kakao (BBK), vascular streak dieback (VSD), dan cacao mirids (Helopeltis spp.). Kegiatan pemuliaan tanaman kakao secara konvensional berjalan lambat dan perlu waktu panjang. Untuk menghasilkan satu varietas unggul diperlukan waktu 15-20 tahun. Aplikasi teknologi DNA (genomika melalui pemuliaan berbantuan marka dan rekayasa genetik) dapat mempercepat program pemuliaan tanaman kakao. Tulisan ini mengulas teknologi DNA yang tersedia saat ini dan potensi aplikasinya untuk mempercepat pemuliaan kakao tahan hama dan penyakit. Penemuan marka DNA dan gen/quantitative trait loci (QTL) kakao berkembang cukup pesat. Banyak gen dan QTL karakter penting telah diidentifikasi yang terkait ketahanan hama dan penyakit serta produktivitas tanaman. Teknologi genomika dan pemanfaatan teknik marker-assisted selection (MAS) juga telah diaplikasikan untuk pemuliaan kakao termasuk untuk karakter ketahanan terhadap hama dan penyakit. Teknologi rekayasa genetik telah diteliti untuk menganalisis potensi pemanfaatannya dalam perbaikan bahan tanam kakao. Dengan berkembangnya teknologi genomika modern, penemuan gen/QTL unggul dapat dipercepat, lebih efisien dan komprehensif untuk mempercepat perakitan varietas unggul kakao tahan hama dan penyakit. Teknologi DNA khususnya MAS dan pemuliaan berbasis data genom siap diaplikasikan untuk mendukung program perbaikan ketahanan tanaman kakao terhadap hama dan penyakit utama dalam rangka peningkatan produktivitas dan mutu kakao nasional.

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