Cloning of TaPRP from Wheat and Its Cold Tolerance Analysis in Transgenic Tobacco

TaPRP, a proline rich protein (PRP) cDNA, was cloned by RT-PCR from winter wheat. Nucleotide sequence analysis showed TaPRP is composed of 1137 bp (378 amino acid residues with a Mr of 42.19 kD). TaPRP shows 92.6%, 89.3%, 73.0%, and 73.3% sequence homologies with PRP genes from wheat, sorghum, rice, and maize, respectively. The deduced protein includes 170 prolines, presenting a normal PRP primary structure. Expression vector pBI-TaPRP was constructed, in which TaPRP was driven by CaMV35S promoter and stopped by NospolyA. Tobaccos were transformed by Agrobacterium containing the constructed vectors. Three transgenic lines were confirmed by PCR detection and Southern blot. Under the same low temperature stress conditions, transgenic plants had lower conductivity rate compared with the non-transgenic plants, suggesting that cold tolerance in transgenic tobacco plants was improved. However, the different transgenic plants showed significant differences in cold resistant, and there also existed significant interactions between plant and treatment temperature. TaPRP might have an important role in wheat in cold adaptation process.

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Regeneration of vgb-transgenic poplar (Populus alba×P. glandulosa) and the primary observation of growth

Chinese Journal of Agricultural Biotechnology ◽

10.1079/cjb200584 ◽

2006 ◽

Vol 3 (1) ◽

pp. 59-64 ◽

Cited By ~ 2

Author(s):

Zhang Bing-Yu ◽

Su Xiao-Hua ◽

Li Yi-Liang ◽

Huang Qin-Jun ◽

Zhang Xiang-Hua ◽

...

Keyword(s):

Transgenic Plants ◽

Blot Hybridization ◽

Pcr Amplification ◽

First Year ◽

Populus Alba ◽

Rt Pcr ◽

Dot Blot ◽

Transgenic Lines ◽

Polymerase Chain ◽

Morphological Differences

AbstractIncreasing the growth rate is especially important for low-quality wood applications, so this has become an important goal in poplar breeding. The present study describes the transfer of Vitreoscilla haemoglobin (VHb) gene (vgb) driven by constitutive promoters, by Agrobacterium tumefaciens into poplar (Populus alba×P. glandulosa). From about 450 leaf discs used for transformation, 60 Kan-resistant plants were obtained, and 52 proved to be true transgenic plants. The transgenic nature of these plants was confirmed by polymerase chain reaction (PCR) amplification and Southern dot blot hybridization. The expression of vgb gene in transgenic plants was confirmed by reverse transcriptase-PCR (RT-PCR). The performance of the transgenic lines was evaluated during the first year of growth in a greenhouse. These plants showed no significant stable morphological differences from the untransformed plants. Among them, three vgb-transgenic lines exhibited noticeably higher growth rates in terms of height and diameter.

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Geminivirus Resistance in Transgenic Tobacco Expressing Mutated BC1 Protein

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.1997.10.5.617 ◽

1997 ◽

Vol 10 (5) ◽

pp. 617-623 ◽

Cited By ~ 24

Author(s):

Y.-P. Duan ◽

C. A. Powell ◽

S. E. Webb ◽

D. E. Purcifull ◽

E. Hiebert

Keyword(s):

Transgenic Plants ◽

Transgenic Tobacco ◽

Movement Protein ◽

Enzyme Linked Immunosorbent Assay ◽

Continuous Exposure ◽

Symptom Development ◽

Dot Blot ◽

Disease Symptoms ◽

Transgenic Lines ◽

Antisense Constructs

Tobacco explants were transformed by Agrobacterium-mediated transformation with sense and antisense constructs of the movement protein genes (BC1 and BV1) of tomato mottle geminivirus (TMoV). Transgenic plants were tested for virus resistance either by exposure to viruliferous whiteflies carrying TMoV or cabbage leaf curl geminivirus (CabLCV) for a 72-h inoculation period or by continuous exposure to TMoV during the life of the plants. The transgenic lines were scored for disease symptoms, and virus replication and distribution were determined by enzyme-linked immunosorbent assay and dot blot hybridizations. Transgenic plants which expressed a mutated form (identified in a previous study) of the BC1 gene showed TMoV and CabLCV resistance. Three resistant phenotypes were observed: a delay in symptom development, a recovery from early symptoms, and an absence of virus symptoms at all stages. Geminivirus was detected in inoculated leaves but was not readily detected in leaves beyond the inoculation sites in the highly resistant plants. The geminivirus resistance conferred by expression of the spontaneously mutated BC1 gene (detected after transformation) in transgenic tobacco may involve transdominant negative interference.

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The Occurrence of CMV-Specific Short RNAs in Transgenic Tobacco Expressing Virus-Derived Double-Stranded RNA is Indicative of Resistance to the Virus

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.2002.15.8.826 ◽

2002 ◽

Vol 15 (8) ◽

pp. 826-833 ◽

Cited By ~ 133

Author(s):

Kriton Kalantidis ◽

Stavros Psaradakis ◽

Martin Tabler ◽

Mina Tsagris

Keyword(s):

Transgenic Plants ◽

Gene Silencing ◽

Transgenic Tobacco ◽

Virus Resistance ◽

Northern Analysis ◽

Specific Gene ◽

Posttranscriptional Gene Silencing ◽

Short Rnas ◽

Transgenic Lines ◽

Viral Sequences

Expression or introduction of double-stranded (ds)RNA in eukaryotic cells can trigger sequence-specific gene silencing of transgenes, endogenes, and viruses. Transgenic plants producing dsRNAs with homology to viral sequences are likely to exhibit pathogen-derived resistance to the virus. Cucumber mosaic virus (CMV), a very widespread virus with over 1,000 host species, has the natural ability to suppress silencing in order to establish infection. Here, we report the generation of transgenic tobacco lines, where a DNA transgene containing an inverted repeat of CMV cDNA had been introduced. Expression of this DNA construct delivered an RNA transcript that is able to form an intramolecular double strand. Transgenic plants were challenged with CMV. Three categories of plants could be discriminated: susceptible plants, which typically reacted with milder symptoms than the wild-type control; a “recovery” phenotype, in which newly emerging leaves were free of symptoms; and plants that showed complete resistance. Northern analysis showed that the expression of CMV dsRNA caused, in some transgenic lines, the generation of short RNAs characteristic of posttranscriptional gene silencing. Those lines were CMV resistant. The correlation between the detection of short RNAs and virus resistance provides a molecular marker that makes it possible to predict success in attempts to engineer virus resistance by dsRNA.

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Constitutive Overexpressing At.TC Improves Drought-mediated Oxidative Tolerance in Transgenic Brassica Napus L.

10.21203/rs.3.rs-751269/v1 ◽

2021 ◽

Author(s):

Sajjad Sobhanverdi ◽

Atefeh Majidi ◽

Alireza Abbasi ◽

Zahra Asghari Mollabashi ◽

Manijeh Sabokdast ◽

...

Keyword(s):

Drought Stress ◽

Transgenic Plants ◽

Water Shortage ◽

Ros Scavenging ◽

Rt Pcr ◽

Brassica Napus L ◽

Transgenic Lines ◽

Oxidative Tolerance ◽

Scavenging Enzymes ◽

Relative Gene

Abstract Background: Environmental stresses are the most important factors limiting crops production in worldwide. Tocopherol, belonging to family of vitamin E compounds, is an amphipathic antioxidants involved in oxidative responses. In the current study, we generated transgenic canola plants overexpressing Arabidopsis VTE1 gene (At.TC) through Agrobacterium tumefaciens system. Methods and results: The putative transgenic plants were successfully regenerated and acclimated in greenhouse conditions. The transcriptional activity of the At.TC gene was evaluated by RT-PCR. In addition, the relative gene expression analysis by qRT-PCR confirmed an increased expression pattern of the transformed gene in canola transgenic lines, with the highest level in R. Line1. Given the results, the transgenic plants, particularly H. Line1 and R. Line2 showed a lower lipid peroxidation compared to WTs under FC 30%. Moreover, two ROS scavenging enzymes including CAT and PPO were up-regulated in transgenic lines; however, no significant pattern was observed for Ascorbate Peroxidase. Also, the amount of leaf tocopherol was significantly more in all T1 lines under drought stress (FC 30%). Conclusion: Taken together, here we successfully developed transgenic lines overexpressing At.TC gene constituently throughout the plant. The results confirmed that the generated transgenic plants are resistant to drought stress, thereby paving the way toward introducing canola plants to deal with the climate change and water shortage.

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Co-expression of AGPS and AGPL genes encoded for AGPase from cassava to enhance starch accumulation in transgenic tobacco

Vietnam Journal of Biotechnology ◽

10.15625/1811-4989/14/2/9354 ◽

2016 ◽

Vol 14 (2) ◽

pp. 287-293

Author(s):

Nguyễn Văn Đoài ◽

Nguyễn Minh Hồng ◽

Lê Thu Ngọc ◽

Nguyễn Thị Thơm ◽

Nguyễn Đình Trọng ◽

...

Keyword(s):

Transgenic Tobacco ◽

Higher Plants ◽

Starch Content ◽

Genetically Modified Crops ◽

Starch Accumulation ◽

35S Promoter ◽

Effective Strategies ◽

Plant Expression Vector ◽

Transgenic Lines ◽

Cassava Varieties

The AGPase (ADP-Glucose pyrophosphorylase) is one of the ubiquitous enzymes catalyzing the first step in starch biosynthesis. It plays an important role in regulation and adjusts the speed of the entire cycle of glycogen biosynthesis in bacteria and starch in plants. In higher plants, it is a heterotetramer and tetrameric enzyme consisting two large subunits (AGPL) and two small subunits (AGPS) and encoded by two genes. In this paper, both AGPS and AGPL genes were sucessfully isolated from cassava varieties KM140 and deposited in Genbank with accession numbers KU243124 (AGPS) and KU243122 (AGPL), these two genes were fused with P2a and inserted into plant expression vector pBI121 under the control of 35S promoter. The efficient of this construct was tested in transgenic N. tabacum. The presence and expression of AGPS and AGPL in transgenic plants were confirmed by PCR and Western hybridization. The starch content was quantified by the Anthrone method. Transgenic plant analysis indicated that that two targeted genes were expressed simultaneously in several transgenic tobacco lines under the control of CaMV 35S promoter. The starch contents in 4 analyzed tobacco transgenic lines displays the increase 13-116% compared to WT plants. These results indicated that the co-expression of AGPS and AGPL is one of effective strategies for enhanced starch production in plant. These results can provide a foundation for developing other genetically modified crops to increase starch accumulation capacity.

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Cold Tolerance of Transgenic Tobacco Carrying Gene Encoding Insect Antifreeze Protein

ACTA AGRONOMICA SINICA ◽

10.3724/sp.j.1006.2008.00397 ◽

2008 ◽

Vol 34 (3) ◽

pp. 397-402 ◽

Cited By ~ 2

Author(s):

Yan WANG

Keyword(s):

Cold Tolerance ◽

Transgenic Tobacco ◽

Antifreeze Protein ◽

Gene Encoding

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MaMAPK3-MaICE1-MaPOD P7 pathway, a positive regulator of cold tolerance in banana

BMC Plant Biology ◽

10.1186/s12870-021-02868-z ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Jie Gao ◽

Tongxin Dou ◽

Weidi He ◽

Ou Sheng ◽

Fangcheng Bi ◽

...

Keyword(s):

Transgenic Plants ◽

Cold Tolerance ◽

Molecular Breeding ◽

Cold Resistance ◽

Oxidoreductase Activity ◽

Tropical Fruit ◽

Over Expression ◽

Cavendish Banana ◽

Cold Tolerant ◽

Necrotic Symptoms

Abstract Background Banana is a tropical fruit with a high economic impact worldwide. Cold stress greatly affects the development and production of banana. Results In the present study, we investigated the functions of MaMAPK3 and MaICE1 involved in cold tolerance of banana. The effect of RNAi of MaMAPK3 on Dajiao (Musa spp. ‘Dajiao’; ABB Group) cold tolerance was evaluated. The leaves of the MaMAPK3 RNAi transgenic plants showed wilting and severe necrotic symptoms, while the wide-type (WT) plants remained normal after cold exposure. RNAi of MaMAPK3 significantly changed the expressions of the cold-responsive genes, and the oxidoreductase activity was significantly changed in WT plants, while no changes in transgenic plants were observed. MaICE1 interacted with MaMAPK3, and the expression level of MaICE1 was significantly decreased in MaMAPK3 RNAi transgenic plants. Over-expression of MaICE1 in Cavendish banana (Musa spp. AAA group) indicated that the cold resistance of transgenic plants was superior to that of the WT plants. The POD P7 gene was significantly up-regulated in MaICE1-overexpressing transgenic plants compared with WT plants, and the POD P7 was proved to interact with MaICE1. Conclusions Taken together, our work provided new and solid evidence that MaMAPK3-MaICE1-MaPOD P7 pathway positively improved the cold tolerance in monocotyledon banana, shedding light on molecular breeding for the cold-tolerant banana or other agricultural species.

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Overexpression of the pitaya phosphoethanolamine N-methyltransferase gene (HpPEAMT) enhanced simulated drought stress in tobacco

Plant Cell Tissue and Organ Culture (PCTOC) ◽

10.1007/s11240-021-02040-3 ◽

2021 ◽

Author(s):

Ai-Hua Wang ◽

Lan Yang ◽

Xin-Zhuan Yao ◽

Xiao-Peng Wen

Keyword(s):

Drought Stress ◽

Stress Response ◽

Transgenic Plants ◽

Drought Tolerance ◽

Transgenic Tobacco ◽

Sequence Similarity ◽

Amino Acid Sequence Similarity ◽

Transgenic Tobacco Plants ◽

Wild Type ◽

Tobacco Plants

AbstractPhosphoethanolamine N-methyltransferase (PEAMTase) catalyzes the methylation of phosphoethanolamine to produce phosphocholine and plays an important role in the abiotic stress response. Although the PEAMT genes has been isolated from many species other than pitaya, its role in the drought stress response has not yet been fully elucidated. In the present study, we isolated a 1485 bp cDNA fragment of HpPEAMT from pitaya (Hylocereus polyrhizus). Phylogenetic analysis showed that, during its evolution, HpPEAMT has shown a high degree of amino acid sequence similarity with the orthologous genes in Chenopodiaceae species. To further investigate the function of HpPEAMT, we generated transgenic tobacco plants overexpressing HpPEAMT, and the transgenic plants accumulated significantly more glycine betaine (GB) than did the wild type (WT). Drought tolerance trials indicated that, compared with those of the wild-type (WT) plants, the roots of the transgenic plants showed higher drought tolerance ability and exhibited improved drought tolerance. Further analysis revealed that overexpression of HpPEAM in Nicotiana tabacum resulted in upregulation of transcript levels of GB biosynthesis-related genes (NiBADH, NiCMO and NiSDC) in the leaves. Furthermore, compared with the wild-type plants, the transgenic tobacco plants displayed a significantly lower malondialdehyde (MDA) accumulation and higher activities of the superoxide dismutase (SOD) and peroxidase (POD) antioxidant enzymes under drought stress. Taken together, our results suggested that HpPEAMT enhanced the drought tolerance of transgenic tobacco.

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