scholarly journals Pyramiding of Cry Toxins and Methanol Producing Genes to Increase Insect Resistance in Cotton

2020 ◽  
Author(s):  
Yuan Youlu ◽  
Abdul Razzaq ◽  
Arfan Ali ◽  
Muhammad Zafar ◽  
Deng Xiaoying ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Expression Vector ◽  
Insect Pests ◽  
Transgenic Cotton ◽  
35S Promoter ◽  
Cry Genes ◽  
Methanol Production ◽  
Transformed Plants ◽  
Plant Expression Vector ◽  
Cotton Plants

Abstract The idea of enhanced methanol production from cell wall by pectin methyl esterase enzymes (PME) combined with expression of cry genes from Bacillus thuringiensis as a strategy to improve pests control in cotton is presented. We constructed a cassette containing two cry genes (cry1Fa and Cry32Aa) and two pme genes, one from Arabidopsis thaliana (AtPME), and other from Aspergillus niger (AnPME) in pCAMBIA1301 plant expression vector using CAMV-35S promoter. This construction was transformed in Eagle-2 cotton variety using shoot apex-cut Agrobacterium-mediated transformation. The expression of cry genes and pme genes was confirmed by qPCR. Methanol production was measured in control and in the cry and pme transformed plants showing methanol production only in transformed plants, then the non-transgenic cotton plants. Finally, insect bioassays performed with transgenic plants expressing cry and pme genes, showed 100 % mortality for Helicoverpa armigera (cotton bollworm) larvae, 70% mortality for pectinophore gossypiella (pink bollworm) larvae and 95% mortality of Earias fabia, (spotted bollworm) larvae, that was higher than the transgenic plants expressing only cry genes that showed 84%, 49% and 79% mortality, respectively. These results demonstrate that Bt. cry-genes coupled with pme genes is an effective strategy to improve the control of different insect pests.

Expression of a Chicken Ovalbumin Gene in Three Lucerne Cultivars

1991 ◽  
Vol 18 (5) ◽  
pp. 495 ◽  
Author(s):  
HE Schroeder ◽  
MRI Khan ◽  
WR Knibb ◽  
D Spencer ◽  
TJV Higgins
Keyword(s):  
Transgenic Plants ◽  
Alternative Interpretation ◽  
Cauliflower Mosaic Virus ◽  
Vector System ◽  
35S Promoter ◽  
Restriction Enzyme Digestion ◽  
Flanking Sequence ◽  
Gene Encoding ◽  
Transformed Plants ◽  
Chicken Ovalbumin

Routine procedures have been developed for the transformation of lucerne (Medicago sativa cv. Rangelander) with foreign genes using the Agrobacterium tumefaciens binary vector system and for the regeneration of transgenic plants from tissue culture, via somatic embryogenesis. Lucerne transformation was carried out with a gene encoding neomycin phosphotransferase (npt), which conferred resistance to the antibiotic kanamycin, together with a cDNA clone encoding chicken ovalbumin which was modified for expression in plant cells. The ovalbumin cDNA protein coding sequence was combined with the cauliflower mosaic virus 35S promoter and the nopaline synthase 3' flanking sequence to make a chimeric ovalbumin gene. A DNA construct containing both these genes was transferred to lucerne, and ovalbumin was detected in leaves of regenerated plants using protein immunoblots. Pulse-chase labelling experiments and analysis of leaves from the top to bottom of the transformed plants indicated that ovalbumin, once formed, was stable in the leaves of transgenic lucerne. A wide variation in ovalbumin level was frequently observed in plants regenerated from multiple embryos on a single transformed callus. This variation correlated with changes in the restriction enzyme digestion pattern of the ovalbumin DNA from the transgenic plants. These results indicate that each transformed callus may have arisen from more than one transformation event. An alternative interpretation is that the callus may have arisen from a single transformed cell but during cell proliferation the DNA in some cells may have undergone rearrangement prior to embryogenesis. Transformation and regeneration procedures were also developed for two Australian commercial cultivars of lucerne. Although the frequency of recovery of transformed plants was lower than with cv. Rangelander, these protocols open the way for a relatively rapid

scholarly journals Gene Expression Evaluation of Multigenic Cotton (Gossypium hirsutum) against Cotton Leaf Curl Virus

2021 ◽  
Vol 26 (01) ◽  
pp. 169-176
Author(s):  
Muhammad Rizwan Afzal
Keyword(s):  
Transgenic Plants ◽  
Transgenic Cotton ◽  
Resistant Variety ◽  
Cotton Leaf ◽  
Cotton Leaf Curl Virus ◽  
Cotton Plants ◽  
Total Dna ◽  
Expression Evaluation ◽  
And Control ◽  
Leaf Curl

One of the most crucial threats limiting the sustainable production of cotton is cotton leaf curl disease (CLCuD). There is dire need to produce a resistant variety that can combat CLCuD. For this purpose, virus resistant transgenic cotton plants (MNH-786) with C4 gene construct at T3 generation were selected and sown. Young fresh leaves of multigenic variety of MNH-786 were collected to confirm the transformed construct. Infected whiteflies were used for spreading on transgenic cotton MNH-786 variety with C4 construct to check percentage of infection. Whiteflies were collected from infected cotton plants showing CLCuD and reared in lab to increase the population of whiteflies. After 15 days of feeding, infected leaves of transgenic plants were collected and total DNA of infected leaves of transgenic cotton plant with virus load was extracted. At maturity, data of morphological characteristic was taken from the transgenic cotton plants of MNH-786 and control plants. Resistant transgenic cotton plants showed < 0.5% disease index and recorded more plant height in field condition. Total number of bolls per plant was 20% more in tolerant plants and 40% more in resistant plants as compared to susceptible plants. Molecular analysis of transgenic plants showed clear evidence that expression of construct 4 virus resistant gene against begomoviruses in resistant and tolerant group of transgenic plants was more as compared to susceptible group and control. © 2021 Friends Science Publishers

Construction of plant expression vector and analysis of herbicide resistance and salt tolerance of transgenic tobacco

2004 ◽  
Vol 1 (2) ◽  
pp. 85-91 ◽  
Author(s):  
Wang Xian-Yan ◽  
Shan Xiao-Yi ◽  
Yang Ai-Fang ◽  
Zhang Ju-Ren
Keyword(s):  
Salt Tolerance ◽  
Transgenic Plants ◽  
Transgenic Tobacco ◽  
Herbicide Resistance ◽  
Expression Vector ◽  
Gene Encoding ◽  
Plant Expression ◽  
Plant Expression Vector ◽  
Herbicide Resistance Gene ◽  
Nacl Concentration

AbstractThe plant expression vector pCAMBIA1300-AtNHX1-als was constructed by inserting the herbicide resistance gene als of Arabidopsis thaliana into the plasmid pCAMBIA1300-AtNHX1, which contains the AtNHX1 gene encoding the Na+/H+ antiport from the vacuolar membrane of A. thaliana. Transgenic tobacco plants were obtained via Agrobacterium-mediated transformation. PCR and Southern blot assay indicated that genes als and AtNHX1 had been integrated into the genome of the transgenic plants. The herbicide resistance and salt tolerance of transgenic plants increased by about 1000-fold and by 1.5% NaCl concentration, respectively, compared with controls. Herbicide resistance of the T1 progeny was evaluated by spraying transgenic plants with different concentrations of Luhuanglong at the four-leaf stage. Controls gradually died under 100 mg/l Luhuanglong whereas 73% of the T1 plants still survived at 500 mg/l Luhuanglong. Thus the plant expression vector pCAMBIA1300-AtNHX1-als could be used to improve the herbicide resistance and salt tolerance of crops.

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

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.

Co-overexpression of AVP1 and AtNHX1 in Cotton Further Improves Drought and Salt Tolerance in Transgenic Cotton Plants

2014 ◽  
Vol 33 (2) ◽  
pp. 167-177 ◽  
Author(s):  
Guoxin Shen ◽  
Jia Wei ◽  
Xiaoyun Qiu ◽  
Rongbin Hu ◽  
Sundaram Kuppu ◽  
...  
Keyword(s):  
Salt Tolerance ◽  
Transgenic Cotton ◽  
Drought And Salt Tolerance ◽  
Cotton Plants

scholarly journals The Construction of Plant Expression Vector harbouring Carica Papaya L. WRKY Gene in Escherichia coli

2020 ◽  
Vol 28 (S2) ◽  
Author(s):  
Fauziah Abu Bakar ◽  
Pavitra Paramalingam ◽  
Kamariah Hasan
Keyword(s):  
Abiotic Stress ◽  
Stress Responses ◽  
Expression Vector ◽  
Carica Papaya ◽  
Sequencing Analysis ◽  
Wrky Gene ◽  
Limited Information ◽  
Abiotic Stress Response ◽  
Plant Expression ◽  
Plant Expression Vector

Carica papaya is a well-liked and economically important fruit with outstanding nutritional and medicinal values. Its susceptibility to abiotic stress which affects the growth and harvest, causes significant yield loss to farmers. In recent years, significant progress has been made to understand the genes that play critical roles in abiotic stress response, especially some transcription factor (TF) encoding genes. Among all TFs, WRKY TF gene family is one of the best-studied TFs involved in various stress responses. To date, only limited information on functionally characterised WRKY TFs is available for C. papaya. The aim of this study was to produce a recombinant construct harbouring WRKY gene in pGEM®-T Easy cloning vector. The presence of a DNA band of the expected size of 465 bp on agarose gel electrophoresis indicated that WRKY gene was successfully amplified from all treated samples. DNA sequencing analysis revealed that the amplified sequence isolated from the treated samples were closely related to Carica papaya species with 97% similarity. Following transformation, 4 out of 5 colonies that were randomly selected showed the WRKY gene had been successfully inserted into pGEM®-T Easy vector and transformed into E. coli. In future, the WRKY gene from pGEMT-WRKY recombinant construct will be cloned into the plant expression vector pCAMBIA 1304 prior to transformation in the plant. The success of demonstrating the WRKY gene towards the response in abiotic stress will enable us to produce stress tolerant transgenic crops under unfavourable conditions via genetic engineering for sustained growth.

scholarly journals Transfer of human proinsulin gene into Cucumber (Cucumis sativus L.) via agrobacterium method

Genetika ◽  
2017 ◽  
Vol 49 (2) ◽  
pp. 717-728
Author(s):  
Kameh Abookazemi ◽  
Javaran Jalali ◽  
Mehdi Mohebodini ◽  
Akbar Vaseghi
Keyword(s):  
Transgenic Plants ◽  
Molecular Farming ◽  
Adult Population ◽  
Advanced Technology ◽  
High Rate ◽  
35S Promoter ◽  
Dot Blot ◽  
Hormone Production ◽  
Human Proinsulin ◽  
Number Of Patients

Nowadays, approximately 5.8% in adult population around the world are suffering by diabetes. It can be caused by an increase in risk factors such as being overweight. Also it has been estimated that the number of patients will be doubled in near future and the demands for insulin hormone will be growing up by 3 to 4 % annually. Therefore, it?s necessary to develop new methods for hormone production with high rate of capacity in future. By advanced technology of transgenic DNA, the transgenic plants are introduced as an attractive system for expression and production of many kinds of pharmaceutical proteins. In this study, we investigated transfer of Human Proinsulin Gene into the Cucumber (Cucumissativus L.). Transgenic cucumber could be a great prospect for future source of eatable insulin pharmaceutical drugs to be taken by patients.Agrobacterium tumefaciensstrain LBA4404 carrying proinsulin genes with CaMV 35S promoter was used for the transformation purpose. The transgenic plants were analyzed by PCR, RT-PCR, SDS-PAGE, Dot blot and Electrochemiluminescence techniques. Production of proinsulin in cucumber could be a great prospect in molecular farming of human proinsulin.

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