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Overexpression of the Heat Shock-Specific Transcription Factor HSFA1D Enhances Thermotolerance in Tobacco Plants

Overexpression of the Heat Shock-Specific Transcription Factor HSFA1D Enhances Thermotolerance in Tobacco Plants

Zamarud Shah, Safdar Hussain Shah, Asad Jan and Gul Shad Ali

Department of Biotechnology, University of Science and Technology Bannu, KP, Pakistan; 2Institute of Biotechnology and Genetic Engineering, The University of Agriculture Peshawar, KP, Pakistan; 3Mid-Florida Research and Education Centre and Department of Plant Pathology, Institute of Food and Agricultural Sciences, University of Florida, 2725 Binion Rd, Apopka, FL-32703, USA.

zamarud_gd@yahoo.com

Figure 1

Restriction digestion of cloned plant expression vector pGWB442-HsfA1d along with their empty control pGWB442, using enzyme NcoI. L, 1kb plus DNA ladder; 1, pGWB442; 2, pGWB442-HsfA1d; 3, pGWB442-HsfA1d 

Figure 2

Diagrammatic representation of pGWB442-HsfA1d vector.
CaMV35S= promotor; nptII= Plant selectable marker; neomycin phosphotransferase= Bacterial selectable marker: YFP= yellow florescent proteins, HsfA1d= Gene of interest 

Figure 3

Different phases of invitro tobacco plant regeneration and establishment in soil. A, Swelling of leaf discs; B, Callus formation; C, Shoot development; D, Root formation; E, Establishment in soil; F, Flower production 

Figure 4

Expression of yellow florescent proteins in transgenic plants. 1 to 5, Five independent transgenic plant expressing YFP, cloned in frame with HsfA1d; WT, Wild type 

Figure 5

PCR based amplification of genomic DNA extracted from transgenic plants along with control using HsfA1d specific primers. L, 1kb plus DNA ladder; P, positive control; Lane 1 to 5, PCR amplification of genomic DNA extracted from 5 independent transgenic lines with HsfA1d primers; WT, Wild type 

Figure 6

Effect of heat treatment on electrolyte leakage of HsfA1d overexpression lines, WT= Wild type 

Figure 7

Effect of heat treatment on Chlorophyll content of HsfA1d overexpression lines (OX1, OX2 and OX3), WT= Wild type 

Figure 8

Effect of heat treatment on Relative water content of HsfA1d overexpression lines (OX1, OX2 and OX3), WT= Wild type 

Figure 9

Effect of heat treatment on Proline content of overexpression lines (OX1, OX2 and OX3), WT= Wild type 

Sarhad Journal of Agriculture

September

Vol. 37, Iss. 3, Pages 714-1097

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