Development of genetically modified rust resistant wheat: A breakthrough by dinted introgression of novel DREB2C and HSFA2 genes under stress induced expression
Hina Firdous,
Arfan Ali,
Saira Saleem,
Abdul Razzaq,
Ghulam Mustafa,
Sezai Ercisli,
Khalid M. Elhindi,
Aqsa Ijaz,
Zunaira Anwar,
Muhammad Kashif,
Muhammad Hamza,
Muhammad Mubashar Zafar,
Wang Baotong,
Xuefei Jiang
Affiliations
Hina Firdous
State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China
Arfan Ali
Sanya Institute of Breeding and Multiplication, School of Tropical Agriculture and Forestry, Hainan University, Sanya, China; FB Genetics, Four Brothers Group, Lahore, Pakistan
Saira Saleem
Scientific Officer (PB&G), Oilseeds Research Station, Khanpur, Ayub Agricultural Research Institute, AARI, Faisalabad 38850, Pakistan
Abdul Razzaq
Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore 53700, Pakistan
Ghulam Mustafa
Center of Agricultural Biochemistry and Biotechnology, University of Agriculture, Faisalabad 38000, Pakistan
Sezai Ercisli
Department of Horticulture, Faculty of Agriculture, Ataturk University, Erzurum, Turkey
Khalid M. Elhindi
Plant Production Department, College of Food & Agriculture Sciences, King Saud University, P.O. Box 2460, Riyadh 11451, Saudi Arabia
Aqsa Ijaz
Nuclear Institute for Agriculture and Biology College (NIAB-C), Pakistan Institute of Engineering and Applied Sciences (PIEAS), Nilore, Islamabad 45650, Pakistan
Zunaira Anwar
Nuclear Institute for Agriculture and Biology College (NIAB-C), Pakistan Institute of Engineering and Applied Sciences (PIEAS), Nilore, Islamabad 45650, Pakistan
Muhammad Kashif
Department of Plant Breeding and Genetics, University of Agriculture, Faisalabad 38000, Pakistan
Muhammad Hamza
FB Genetics, Four Brothers Group, Lahore, Pakistan
Muhammad Mubashar Zafar
Sanya Institute of Breeding and Multiplication, School of Tropical Agriculture and Forestry, Hainan University, Sanya, China; Corresponding authors.
Wang Baotong
State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China; Corresponding authors.
Xuefei Jiang
Sanya Institute of Breeding and Multiplication, School of Tropical Agriculture and Forestry, Hainan University, Sanya, China; Corresponding authors.
Wheat is a major staple food worldwide yet numerous yield limiting agents affect its productivity. Stripe rust is a major culprit in this context and efforts have been made to culminate this pathogen using conventional as well as advanced innovative techniques. Transgenic technology is of significant importance in this context and numerous success stories are evident to prove its worth. In the current study, two novel genes HSFA2 and DREB2C were expressed in an elite wheat genotype Akbar, Fakhre-e-Bhakhar under constitutive CAMV35S promoter and stress inducible rd29 Promoters. The shoot cut method was used for the Agrobacterium-mediated transformation and putative transformants were selected on kanamycin 50 mg/L. The resultant transformants were tested through PCR for transgene integration whereas expression analysis was carried out through realtime qPCR. Expression of both of the aforementioned genes was found to be higher under rd29 promoter as compared with transgene(s) expression under CAMV35S promoter. In the bioassay, transgenic wheat plants demonstrated significant tolerance to stress, exhibiting only minor spotting under constitutive expression conditions. Upon exposure to stress, these plants showed exceptional resistance to stripe rust, producing large, bold grains compared to individual trait expressions and negative controls. Subsequently, the DREB2C gene was knocked out to determine if stripe rust control was specifically attributed to this gene. Following the knockout, the onset of stripe rust was comparable to that of the negative control. This led to the conclusion that pyramiding the DREB2C gene with HSFA2 through dual expression represents a novel and highly effective strategy for controlling the widespread stripe rust in wheat. This approach also offers resistance to high temperatures (above 32 °C) from the pollination stage through to maturity.
