Transient expression of marker genes in immature zygotic embryos of spring wheat (Triticum aestivum) through microprojectile bombardment

Genome ◽  
1991 ◽  
Vol 34 (3) ◽  
pp. 453-460 ◽  
Author(s):  
R. N. Chibbar ◽  
K. K. Kartha ◽  
N. Leung ◽  
J. Qureshi ◽  
K. Caswell
Keyword(s):  
Triticum Aestivum ◽  
Transient Expression ◽  
Microprojectile Bombardment ◽  
Triticum Aestivum L ◽  
Cauliflower Mosaic Virus ◽  
Expression Vectors ◽  
Marker Genes ◽  
Zygotic Embryos ◽  
Coding Region ◽  
Immature Zygotic Embryos

Transient expression of marker genes (cat and uidA) delivered by the Biolistics microprojectile bombardment technique has been detected in immature zygotic embryos of wheat (Triticum aestivum L.). The DNA expression vectors that gave maximal expression of both cat (pCaMVI1CN) and uidA (pCaMVI1GusN) genes had an alcohol dehydrogenase (Adh1) intron 1 cloned in between the cauliflower mosaic virus (CaMV35S) promoter and the coding region of the gene. Detection of chloramphenicol acetyltransferase (CAT) activity in response to cat gene was complicated by the presence of an inhibitor of CAT activity as well as an endogenous CAT-Iike activity. The results of enzymatic assays were confirmed by an ELISA technique using CAT-specific antibodies, whereas the β-glucuronidase (GUS) activity following the introduction of the uidA gene was confirmed by both histochemical and fluorometric techniques.Key words: cereal transformation, gene expression, ELISA, wheat.

scholarly journals Triticum Aestivum L. cv. Guizi 1 ANS-6D Positively Regulates Leaf Senescence Through the Abscisic Acid Mediated Chlorophyll Degradation in Tobacco

2021 ◽  
Author(s):  
Luhua Li ◽  
Chang An ◽  
Zhongni Wang ◽  
Fumin Xiong ◽  
Yingxi Wang ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Abscisic Acid ◽  
Leaf Senescence ◽  
Triticum Aestivum L ◽  
Degradation Pathway ◽  
Chlorophyll Degradation ◽  
Anthocyanin Synthesis ◽  
Marker Genes ◽  
Wild Type ◽  
Transgenic Lines

Abstract Anthocyanidin synthase (ANS) is involved in the synthesis of anthocyanins, which are important phytonutrients because of their beneficial effects on human health. Here, we identified ANS-6D of purple-colored Triticum aestivum L. cv. Guizi 1 (Gz) that is involved in leaf senescence through the abscisic acid (ABA) mediated chlorophyll degradation pathway in tobacco. After characterizing the leaf-senescence phenotype in GzANS-6D overexpression (OxGzANS-6D) lines, we found that the increased anthocyanin accumulation and decreased chlorophyll content in OxGzANS-6D lines were closely correlated with the expression levels of anthocyanin synthesis-related structural genes and senescence marker genes, as well as the accumulation of reactive oxygen species. The endogenous ABA content increased and ethylene content decreased in OxGzANS-6D transgenic lines compared with wild type. Additionally, the levels of the abscisic acid-responsive transcription factors ABF1 and ABF2, as well as those of chlorophyll degradation-related genes (PAO, NYC, SGR and CHL), were significantly higher in OxGzANS-6D transgenic lines than in wild type. Furthermore, we found that GzABF1 and NtABF1 binds to the promoter of GzANS-6D, and NtABF2 binds to the promoter of NtSGR. Thus, GzANS-6D participated in leaf senescence through ABA-mediated chlorophyll degradation, and ABF1/2 play important role in GzANS-6D functions.

scholarly journals Transgenic sorghum plants via microprojectile bombardment

1993 ◽  
Vol 90 (23) ◽  
pp. 11212-11216 ◽  
Author(s):  
A M Casas ◽  
A K Kononowicz ◽  
U B Zehr ◽  
D T Tomes ◽  
J D Axtell ◽  
...  
Keyword(s):  
Microprojectile Bombardment ◽  
Dna Delivery ◽  
Regulatory Elements ◽  
Zygotic Embryos ◽  
Bar Gene ◽  
Immature Zygotic Embryos ◽  
Regenerated Plantlets ◽  
Embryogenic Tissues ◽  
Callus Lines ◽  
Independent Transformation

Transgenic sorghum plants have been obtained after microprojectile bombardment of immature zygotic embryos of a drought-resistant sorghum cultivar, P898012. DNA delivery parameters were optimized based on transient expression of R and C1 maize anthocyanin regulatory elements in scutellar cells. The protocol for obtaining transgenic plants consists of the delivery of the bar gene to immature zygotic embryos and the imposition of bialaphos selection pressure at various stages during culture, from induction of somatic embryogenesis to rooting of regenerated plantlets. One in about every 350 embryos produced embryogenic tissues that survived bialaphos treatment; six transformed callus lines were obtained from three of the eight sorghum cultivars used in this research. Transgenic (T0) plants were obtained from cultivar P898012 (two independent transformation events). The presence of the bar and uidA genes in the T0 plants was confirmed by Southern blot analysis of genomic DNA. Phosphinothricin acetyltransferase activity was detected in extracts of the T0 plants. These plants were resistant to local application of the herbicide Ignite/Basta, and the resistance was inherited in T1 plants as a single dominant locus.

scholarly journals GENETIC TRANSFORMATION OF EGYPTIAN WHEAT (Triticum aestivum L.) WITH CHITINASE GENE VIA MICROPROJECTILE BOMBARDMENT

2013 ◽  
Vol 42 (2) ◽  
pp. 233-246 ◽  
Author(s):  
A. FAHMY ◽  
K EL MANGOURY ◽  
WALAA ABOU EL-WAFA ◽  
M. BARAKAT ◽  
SOHEIR EL-KHODARY ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Genetic Transformation ◽  
Microprojectile Bombardment ◽  
Triticum Aestivum L ◽  
Chitinase Gene
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