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

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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Agrobacterium rhizogenes-Mediated Transformation of Hypericum tomentosum L. and Hypericum tetrapterum Fries.

Zeitschrift für Naturforschung C ◽

10.1515/znc-2009-11-1218 ◽

2009 ◽

Vol 64 (11-12) ◽

pp. 864-868 ◽

Cited By ~ 14

Author(s):

Hedviga Komarovská ◽

Annalisa Giovannini ◽

Ján Košuth ◽

Eva Čellárová

Keyword(s):

Genetic Transformation ◽

Agrobacterium Rhizogenes ◽

Hairy Root ◽

Hairy Root Cultures ◽

Root Cultures ◽

Wild Type ◽

Root Segments ◽

Transgenic Lines ◽

Type Strains ◽

Hypericum Species

This is the fi rst evidence on successful Agrobacterium rhizogenes-mediated genetic transformation of two species from the genus Hypericum, H. tomentosum L. and H. tetrapterum Fries. Hairy root cultures were induced from root segments of both Hypericum species by two agropine wild-type strains of A. rhizogenes, ATCC 15834 and A4. The transgenic character of the hairy root cultures was proved by PCR amplifi cation of the rolABCD genes. In some H. tetrapterum transgenic lines aux genes were detected as well.

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Genetic transformation of Citrus sinensis cv. Hamlin with hrpN gene from Erwinia amylovora and evaluation of the transgenic lines for resistance to citrus canker

Scientia Horticulturae ◽

10.1016/j.scienta.2009.04.001 ◽

2009 ◽

Vol 122 (1) ◽

pp. 109-115 ◽

Cited By ~ 37

Author(s):

Janaynna Magalhães Barbosa-Mendes ◽

Francisco de Assis Alves Mourão Filho ◽

Armando Bergamin Filho ◽

Ricardo Harakava ◽

Steven V. Beer ◽

...

Keyword(s):

Genetic Transformation ◽

Citrus Sinensis ◽

Erwinia Amylovora ◽

Citrus Canker ◽

Transgenic Lines

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Genetic Transformation of Passionflower and Evaluation of R1 and R2 Generations for Resistance to Cowpea aphid borne mosaic virus

Plant Disease ◽

10.1094/pdis-12-10-0873 ◽

2011 ◽

Vol 95 (8) ◽

pp. 1021-1025 ◽

Cited By ~ 16

Author(s):

Alessandra C. B. A. Monteiro-Hara ◽

Adriana S. Jadão ◽

Beatriz M. J. Mendes ◽

Jorge A. M. Rezende ◽

Flavio Trevisan ◽

...

Keyword(s):

Genetic Transformation ◽

Mosaic Virus ◽

Transgenic Line ◽

Plant Genome ◽

Capsid Protein Gene ◽

Cowpea Aphid ◽

Myzus Nicotianae ◽

Transgenic Lines ◽

Resistance To Infection ◽

Inoculum Pressure

We report on the production and evaluation of passionflower transgenic lines for resistance to Cowpea aphid borne mosaic virus (CABMV). Genetic transformation was done using Agrobacterium tumefaciens and transgene integration was confirmed by Southern blot analyses, resulting in nine transgenic lines for ‘IAC 275’ and three for ‘IAC 277’. Transgenic lines were clonally propagated and evaluated for resistance to CABMV. After the third inoculation, under higher inoculum pressure, only propagated plants of the transgenic line T16 remained asymptomatic, indicating a high resistance to infection with CABMV. This transgenic line was self-pollinated and the R1 generation was evaluated together with the R1 generation of another resistant transgenic line (T2) identified previously. Plants were inoculated with CABMV by means of viruliferous Myzus nicotianae. All 524 T2R1 plants became infected, whereas 13 of 279 T16R1 remained asymptomatic after four successive inoculations. A T16R2 generation was obtained and plants were inoculated with CABMV mechanically or by aphids. After successive inoculations, 118 of 258 plants were symptomless, suggesting that the resistance to CABMV was maintained in the plant genome as the homozygous condition was achieved. Five selected resistant T16R2 plants which contained the capsid protein gene are being crossed for further analyses.

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Efficient Genetic Transformation and Regeneration of a Farmer-Preferred Cassava Cultivar From Ghana

Frontiers in Plant Science ◽

10.3389/fpls.2021.668042 ◽

2021 ◽

Vol 12 ◽

Author(s):

Wilfred Elegba ◽

Emily McCallum ◽

Wilhelm Gruissem ◽

Hervé Vanderschuren

Keyword(s):

Genetic Transformation ◽

Virus Resistance ◽

Agronomic Traits ◽

Mosaic Disease ◽

Cassava Mosaic Disease ◽

Local Cultivar ◽

Friable Embryogenic Callus ◽

Genetic Barriers ◽

Successful Induction ◽

Transgenic Lines

Cassava is an important staple crop that provides food and income for about 700 million Africans. Cassava productivity in Africa is limited by viral diseases, mainly cassava mosaic disease (CMD) and cassava brown streak disease (CBSD). Genetic barriers such as high heterozygosity, allopolyploidy, poor seed set, and irregular flowering constrain the development of virus-resistant cassava varieties via conventional breeding. Genetic transformation represents a valuable tool to circumvent several challenges associated with the development of virus resistance and other valuable agronomic traits in cassava. The implementation of genetic transformation in many local African cultivars is limited either by the difficulty to produce friable embryogenic callus (FEC), low transformation, and/or regeneration efficiencies. Here, we report the successful induction of organized embryogenic structures (OES) in 11 farmer-preferred cultivars locally grown in Ghana. The production of high quality FEC from one local cultivar, ADI 001, facilitated its genetic transformation with high shoot regeneration and selection efficiency, comparable to the model cassava cultivar 60444. We show that using flow cytometry for analysis of nuclear ploidy in FEC tissues prior to genetic transformation ensures the selection of genetically uniform FEC tissue for transformation. The high percentage of single insertion events in transgenic lines indicates the suitability of the ADI 001 cultivar for the introduction of virus resistance and other useful agronomic traits into the farmer-preferred cassava germplasm in Ghana and Africa.

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Expression of Endochitinase from Trichoderma harzianum in Transgenic Apple Increases Resistance to Apple Scab and Reduces Vigor

Phytopathology ◽

10.1094/phyto.2000.90.1.72 ◽

2000 ◽

Vol 90 (1) ◽

pp. 72-77 ◽

Cited By ~ 132

Author(s):

Jyothi Prakash Bolar ◽

John L. Norelli ◽

Kwai-Weng Wong ◽

Christopher K. Hayes ◽

Gary E. Harman ◽

...

Keyword(s):

Trichoderma Harzianum ◽

Venturia Inaequalis ◽

Apple Scab ◽

Significant Negative Correlation ◽

Scab Resistance ◽

Enzyme Linked Immunosorbent Assay ◽

Negative Effects ◽

Genes Encoding ◽

Transgenic Lines ◽

Polymerase Chain

The goal of this research was to improve scab resistance of apple by transformation with genes encoding chitinolytic enzymes from the bio-control organism Trichoderma harzianum. The endochitinase gene, as cDNA and genomic clones, was transferred into apple cv. Marshall McIntosh by Agrobacterium-transformation. A total of 15 lines were identified as transgenic by NPTII enzyme-linked immunosorbent assay and polymerase chain reaction and confirmed by Southern analysis. Substantial differences in endochitinase activity were detected among different lines by enzymatic assay and western analysis. Eight lines propagated as grafted and own-rooted plants were inoculated with Venturia inaequalis. Six of these transgenic lines expressing endochitinase were more resistant than nontransformed cv. Marshall McIntosh. Disease severity compared with cv. Marshall McIntosh was reduced by 0 to 99.7% (number of lesions), 0 to 90% (percentage of leaf area infected), and 1 to 56% (conidia recovered) in the transgenic lines tested. Endochitinase also had negative effects on the growth of both inoculated and uninoculated plants. There was a significant negative correlation between the level of endochitinase production and both the amount of disease and plant growth.

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Genetic transformation of Citrus sinensis with Citrus tristeza virus (CTV) derived sequences and reaction of transgenic lines to CTV infection

Biologia Plantarum ◽

10.1007/s10535-012-0035-0 ◽

2012 ◽

Vol 56 (1) ◽

pp. 162-166 ◽

Cited By ~ 11

Author(s):

F. R. Muniz ◽

A. J. Souza ◽

L. C. L. Stipp ◽

E. Schinor ◽

W. Freitas ◽

...

Keyword(s):

Genetic Transformation ◽

Citrus Sinensis ◽

Citrus Tristeza Virus ◽

Transgenic Lines ◽

Tristeza Virus ◽

Citrus Tristeza

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Establishment of Hevea brasiliensis lines overexpressing genes involved in ethylene signalling pathway

E-Journal Menara Perkebunan ◽

10.22302/ppbbi.jur.mp.v84i1.190 ◽

2016 ◽

Vol 84 (1) ◽

Author(s):

Retno LESTARI ◽

Maryannick RIO ◽

Florence MARTIN ◽

Julie LECLERCQ ◽

Florence DESSAILLY ◽

...

Keyword(s):

Agrobacterium Tumefaciens ◽

Genetic Transformation ◽

Hevea Brasiliensis ◽

Blot Hybridization ◽

Southern Blot Hybridization ◽

Signalling Pathway ◽

Response Factors ◽

Callus Line ◽

Transgenic Lines ◽

Ethylene Signalling

The gaseous plant hormone ethylene has a wide variety of applications in agriculture and horticulture. Ethylene Response Factors (ERF) are the last transcription factors of the ethylene signalling pathway and control a large number of ethylene-responsive genes. Two Hevea brasiliensis ERF, HbERF-IXc4 and HbERF-IXc5, are orthologs to ERF1 a key regulator at the crosstalk of ethylene and jasmonate signalling pathways. These genes were suggested to play an important role in regulating latex cell metabolism in response to tapping and ethephon stimulation. In this study, transgenic lines overexpressing HbERF-IXc4 and HbERF-IXc5 under control of 35S CaMV and HEV2.1 promoter have been conducted. Transgenic Hevea lines were obtained by Agrobacterium tumefaciens-mediated genetic transformation. The somatic embryogenesis process was affected by these modifications. Agrobacterium tumefaciens genetic transformation procedure has been developed from friable callus line for clone PB260. Hevea callus was sub-cultured as small aggregates on paromomycin selection medium. Transgenic callus lines were established from sub-aggregates showing full GFP activity. Ten transgenic lines were confirmed as transgenic by Southern blot hybridization. This result showed successfully establishment of H. brasiliensis transgenic lines. Further plant regeneration and characterization were necessary to understand the function HbERF-IXc4 and HbERF-IXc5 in latex.

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