Allah Bakhsh

The myth of plant transformation.

Technology development is innovative to many aspects of basic and applied plant transgenic science. Plant genetic engineering has opened new avenues to modify crops, and provided new solutions to solve specific needs. Development of procedures in cell biology to regenerate plants from single cells or organized tissue, and the discovery of novel techniques to transfer genes to plant cells provided the prerequisite for the practical use of genetic engineering in crop modification and improvement. Plant transformation technology has become an adaptable platform for cultivar improvement as well as for studying gene function in plants. This success represents the climax of years of efforts in tissue culture improvement, in transformation techniques and in genetic engineering. Plant transformation vectors and methodologies have been improved to increase the efficiency of transformation and to achieve stable expression of transgenes in plants. This review provides a comprehensive discussion of important issues related to plant transformation as well as advances made in transformation techniques during three decades.

Gossypium arboreum GHSP26 enhances drought tolerance in Gossypium hirsutum

Heat‐shock proteins (HSP) are molecular chaperones for protein molecules. These proteins play an important role in protein–protein interactions such as, folding and assisting in the establishment of proper protein conformation and prevention of unwanted protein aggregation. A small HSP gene GHSP26 present in Gossypium arboreum responds to dehydration. In the present study, an attempt was made to overcome the problem of drought stress in cotton. A cDNA of GHSP26 was isolated from G. arboreum, cloned in plant expression vector, pCAMBIA‐1301 driven by the cauliflower mosaic virus 35S promoter and introduced into Gossypium hirsutum. The integration and expression studies of putative transgenic plants were performed through GUS assay; PCR from genomic DNA, and quantitative real‐time PCR analysis. Transgenic cotton plants showed an enhanced drought tolerance, suggesting that GHSP26 may play a role in plant responsiveness to drought.

A molecular approach to combat spatio-temporal variation in insecticidal gene (Cry1Ac) expression in cotton

Spatio-temporal expression of an insecticidal gene (Cry1Ac) in pre existing transgenic lines of transgenic cotton was studied. Seasonal decline in expression of Cry1Ac differed significantly among different cotton lines tested in the field conditions. The leaves of the Bt cotton plants were found to have the highest levels of toxin expression followed by squares, bolls, anthers and petals. Expression of the gene decreased consistently with the age of plants. Toxin expression in fruiting parts was not enough to confer full resistance against bollworms. The reduction in efficacy of transgenic cotton plants late in the season was attributed to reduction in promoter activity. For this purpose, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) small subunit (rbcS) promoter was isolated from Gossypium arboreum that was further cloned upstream of an insecticidal gene (Cry1Ac) in expression vector pCAMBIA 1301. A local cotton cultivar NIAB-846 was transformed with Cry1Ac driven by rbcS promoter. The same cotton cultivar was also transformed with Cry1Ac gene driven by 35SCaMV promoter to compare the expression pattern of insecticidal gene under two different promoters. The results showed that rbcS is an efficient promoter to drive the expression of Cry1Ac gene consistent throughout the life of cotton plant as compared to 35S promoter. The use of tissue specific promoter is also useful for addressing the biosafety issues as the promoter activity is limited to green parts of plants, hence no gene expression in roots, cotton seed and other cotton products and by products.

Aquatic Plants as Human Food

Water is one of the main components of the earth and almost 70% of the earth is surrounded by water found in oceans, seas, lakes, rivers and below the Earth’s surface. All ancient human civilizations were established and developed near the water bodies in order to fill their stomach. These water bodies provided food source like fish for consumption and plants as vegetables or for medicinal purposes. Since ancient times, the aquatic plants have had significant economic and social impact on humans on the basis of their traditional knowledge. However, localized consumption of these aquatic plants as food or as medicinal plants limits their expansion to the world market. A large number of aquatic or semiaquatic plants have potential for commercialization as food plants due to their use and nutritional value but lack of knowledge hinders exploitation of their potential.

Inheritance of an insecticidal gene ( cry1Ab ) in genetically modified cotton

The present study was conducted to evaluate inheritance pattern of insecticidal gene (Cry1Ab) in F1 and F2 generations derived from six combinations of crosses made between two transgenic lines (CEMB-3 and CEMB-11) and two non transgenic lines (MNH-93 and CIM-482). PCR, southern blot, western dot blot assay and lab biotoxicity assay were used to confirm the gene integration and expression of insecticidal gene in successive generations. The insecticidal gene was stably integrated in the genome of all F1 plants showing the dominant nature of introduced gene (cry1Ab). Furthermore, heterosis, heterobeltiosis, heritability and genetic advance studies of Bt gene were also conducted. Heterosis and heterobeltiosis were estimated for five characters i.e. cotton yield per plant, no. of bolls per plant, boll weight, ginning out turn % age and laboratory bioassay results. The heterosis and heterobeltiosis ranged from—15.19 to 107.07% and 18.58 to 98.79%, respectively for yield per plant; from −20.34 to 81.36% and −20.34 to 81.36%, respectively for number of bolls per plant; from -6.96 to 21.38% and −9.30 to 9.99%, respectively for boll weight; from 13.02 to 26.44% and −0.52 to 26.17%, respectively for ginning outturn; and from −8.11 to 36.23% and −5.56 to 23.68%, respectively for mortality % age of Heliothis larvae in laboratory bioassays. The Broad Sense Heritability and Genetic Advance for insect resistance in Bt versus non-Bt crosses were calculated. Both of these were high in four out of six hybrids. Our data recorded showed that these transgenic lines are an excellent source of germplasm to be used in conventional breeding programme.