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1.
Plant Biotechnol J ; 15(11): 1465-1477, 2017 Nov.
Article in English | MEDLINE | ID: mdl-28378532

ABSTRACT

Drought stress has often caused significant decreases in crop production which could be associated with global warming. Enhancing drought tolerance without a grain yield penalty has been a great challenge in crop improvement. Here, we report the Arabidopsis thaliana galactinol synthase 2 gene (AtGolS2) was able to confer drought tolerance and increase grain yield in two different rice (Oryza sativa) genotypes under dry field conditions. The developed transgenic lines expressing AtGolS2 under the control of the constitutive maize ubiquitin promoter (Ubi:AtGolS2) also had higher levels of galactinol than the non-transgenic control. The increased grain yield of the transgenic rice under drought conditions was related to a higher number of panicles, grain fertility and biomass. Extensive confined field trials using Ubi:AtGolS2 transgenic lines in Curinga, tropical japonica and NERICA4, interspecific hybrid across two different seasons and environments revealed the verified lines have the proven field drought tolerance of the Ubi:AtGolS2 transgenic rice. The amended drought tolerance was associated with higher relative water content of leaves, higher photosynthesis activity, lesser reduction in plant growth and faster recovering ability. Collectively, our results provide strong evidence that AtGolS2 is a useful biotechnological tool to reduce grain yield losses in rice beyond genetic differences under field drought stress.


Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Droughts , Edible Grain/growth & development , Galactosyltransferases/genetics , Oryza/genetics , Stress, Physiological , Arabidopsis Proteins/metabolism , Edible Grain/genetics , Gene Expression Regulation, Plant , Oryza/growth & development , Photosynthesis , Plant Leaves/metabolism , Plants, Genetically Modified , Seeds/genetics , Seeds/growth & development , Stress, Physiological/genetics , Transcription Factors/genetics , Transcription Factors/metabolism
2.
Plant Biotechnol J ; 15(6): 775-787, 2017 Jun.
Article in English | MEDLINE | ID: mdl-27889933

ABSTRACT

Nitrogen (N) fertilizers are a major input cost in rice production, and its excess application leads to major environmental pollution. Development of rice varieties with improved nitrogen use efficiency (NUE) is essential for sustainable agriculture. Here, we report the results of field evaluations of marker-free transgenic NERICA4 (New Rice for Africa 4) rice lines overexpressing barley alanine amino transferase (HvAlaAT) under the control of a rice stress-inducible promoter (pOsAnt1). Field evaluations over three growing seasons and two rice growing ecologies (lowland and upland) revealed that grain yield of pOsAnt1:HvAlaAT transgenic events was significantly higher than sibling nulls and wild-type controls under different N application rates. Our field results clearly demonstrated that this genetic modification can significantly increase the dry biomass and grain yield compared to controls under limited N supply. Increased yield in transgenic events was correlated with increased tiller and panicle number in the field, and evidence of early establishment of a vigorous root system in hydroponic growth. Our results suggest that expression of the HvAlaAT gene can improve NUE in rice without causing undesirable growth phenotypes. The NUE technology described in this article has the potential to significantly reduce the need for N fertilizer and simultaneously improve food security, augment farm economics and mitigate greenhouse gas emissions from the rice ecosystem.


Subject(s)
Nitrogen/metabolism , Oryza/metabolism , Alanine Transaminase/genetics , Alanine Transaminase/metabolism , Genotype , Oryza/enzymology , Oryza/genetics , Plants, Genetically Modified/enzymology , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Transformation, Genetic/genetics
3.
Biol. Res ; 44(4): 369-375, 2011. ilus, tab
Article in English | LILACS | ID: lil-626737

ABSTRACT

Adiponectin is a polypeptide specifically secreted from human adipocytes, and its deficiency is closely linked to increased obesity and type II diabetes. There is an urgent demand for large-scale production of human adiponectin for pharmaceutical applications. Here, we report that we have successfully obtained a high-level of expression of modified genes encoding human adiponectin in transgenic rice. The 735 bp cDNA of the native human sequence was adopted to rice codon usage, fused to the translation initiation sequence in the N terminus and to the KDEL signal sequence in the C terminus. An amplification promoting sequence acting as an enhancer of transcription was also introduced to enhance gene expression. The presence of the transgene and mRNA transcripts was confirmed by PCR, Southern blot and RT-PCR. Western blot analysis revealed that a protein of approximately 30 kDa was produced in rice leaves. ELISA analysis was used to determine the amount of recombinant adiponectin in transformants with the modified gene in up to 0.32% of total soluble leaf protein. Our results establish the feasibility of high-level expression of recombinant human adiponectin in transgenic rice.


Subject(s)
Adiponectin/genetics , Oryza/genetics , Plants, Genetically Modified/genetics , Adiponectin/metabolism , Blotting, Southern , Codon , DNA, Complementary , Gene Expression Regulation, Plant , Oryza/chemistry , Reverse Transcriptase Polymerase Chain Reaction
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