Evaluation of glufosinate gene transfer to safflower (Carthamus tinctorius L.) through floral dipping in Agrobacterium suspension

Document Type : Research Paper


1 Graduated MSc student, Department of Agricultural Biotechnology, Faculty of Agriculture, Shahid Bahona University of Kerman, Kerman, Iran

2 Molecular physiology/Faculty of Biology/University of Freibburg

3 BiologyIII/Faculty of Biology/university of Freiburg

4 Professor, Genetics and Plant Breeding, Research & Technology Institute of Plant Production (RTIPP(, Shahid Bahonar University of Kerman, Kerman, Iran.


Among Iran's climate-well-adapted oily plants, safflower (Carthamus tinctorius L.) is considered as a top-ranked plant since it is well-adapted to areas with water shortage. Safflower is known as a model crop with a variety of fatty acids with a relatively high tolerance to salinity and drought, and also due to its high nutritional value that is related to the composition of 90 percent unsaturated fatty acids in its oil content. It has always been considered as a desirable and valuable oilseed plant and can play an important role in expanding the area under planting of oilseeds in the country. Although, Iran with its diverse climate, is one of the richest regions in the world in terms of safflower genetic resources, safflower planting is not very common in our country. One of the main reasons is probably the lack of promotion and low grain yield of its cultivars. In this study, For the first time the floral dipping was performed in Agrobacterium suspension in order that the glufosinate gene would be transferred to the safflower plant. Actually, the access to appropriate engineered cultivars will play a critical role in the development of safflower planting.
Materials and methods
Nine safflower lines were selected from a world-wide collection genotypes based on the different agronomic and phenotypic traits. They were submerged in Agrobacterium suspensions with optical density (OD600 nm) of 1 and in the presence of two different amounts of silwet-77.
The genomic analysis of T1 generations using glufosinate resistance gene primers indicated the success of glufosinate transfer in safflower plant through flower immersion.
Among 9 studied genotypes and 17 transformed plants using the floral dipping method, 3 transgenic plants were obtained with sequencing confirmation: one seedling belongs to Mahali Yazd genotype, one seedling belongs to the Mahali Kerman and one seedling belongs to the Paeize 12 genotype. All transgenic plants were inoculated with 40μl concentration of silwet-77.


معتمدی جواد، زبرجدی علیرضا، کهریزی دانیال؛ سلمانیان علی­هاتف (1391) انتقال ژن جهش یافته aroA باکتریایی به کمک Agrobacterium tumefaciens به گیاه گلرنگ. ژنتیک نوین 7، 63 - 57.
سلطانی لیدا (1397) خاموشی ژن FAD2-1 با استفاده از سیستم CRISPR-CAS 9 به منظور افزایش اولئیک اسید در گیاه گلرنگ (Carthamus tinctorius). رساله کارشناسی ارشد بیوتکنولوژی کشاورزی، دانشگاه شهید باهنر کرمان.
Bechtold N, Ellis J, Pelletier G (1993) In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. proceedings of the French Academy of Sciences: Life Science 316, 1194–1199.
Belide S, Hac L, Singh SP et al. (2011) Agrobacterium-mediated transformation of safflower and the efficient recovery of transgenic plants via grafting. Plant Methods 7,12.
Birch, RG (1997) Plant transformation: problems and strategies for practical Application. Annual Review of plant Physiology and Plant molecular biology 48, 297-326.
Clough SJ (1998) Floral dip, a simple method for agrobacterium-mediated transformation of Arabidopsis thaliana. Plant journal 16, 735-743.
Glevin, SB (2003) Agrobacterium-mediated plant transformation: the biology behind the “gene-jockeying” tool. Microbiology and Molecular Biology Reviews 67, 16-37.
Ji Q, Xu X, Wang K (2013) Genetic transformation of major cereal crops. The International Journal of Developmental Biology 57, 495-508.
Mrizova K, Holaskova E, Tufan M et al. (2014) Transgenic barley: A prospective tool for biotechnology and agriculture. Biotechnology Advances 32, 137-157.
Motamedi J, Zabarjadi AR, Kahrizi D, Salmanian AH (2012) Transfer of bacterial aroA mutant gene by Agrobacterium tumefaciens to Safflower. Modern Genet 7, 57-63 (In Persian).
Orlikowska TK, Cranston HJ, Dyer WE (1995) Factors influencing Agrobacterium tumefaciens mediated transformation and regeneration of the safflower cultivar centennial. Plant Cell Tissue and Organ Culture 40, 85–91.
Rao SK, Rohini VK (1999) Gene transfer into Indian cultivars of safflower (Carthamus tinctorius L.)  using   Agrobacterium tumefaciens.Plant Biotechnology16:201-206.
Rohini VK, Sankara Rao K (2000) Embryo transformation, a practical approach for realizing transgenic plants of safflower (Carthamus tinctorius L.). Annals of Botany 86, 1043-1049.
Saghai-Maroof MA, Soliman,KM, Jorgensen RA, Allard RW (1984) Ribosomal spacer length polymorphism in barley: mendelian inheritance, chromosomal location and population dynamics. Proceedings of the National Academy of Sciences, USA 83, 1757-1761.
Sanford JC (2000) The development of the biolistic process. In vitro Cellular & Developmental Biology-Plant 36, 303-308.
Sankara Rao K, Rohini VK (1999) Gene transfer into Indian cultivars of safflower (Carthamus tinctorius L.) using Agrobacterium tumefaciens. Plant Biotechnology 16, 201-206.
Singh V, Nimbkar N (2006) Safflower (Carthamus tinctorius L.) Resources, Chromosome Engineering and Crop Improvement. Oil Seed Crops, Chapter 6, 167-194.
Soltani L (2018) Extinction of FAD2-1 gene using CRISPR-CAS 9 system to increase oleic acid in safflower (Carthamus tinctorius). MSc Thesis, Shahid Bahonar University of Kerman (In Persian).
Southgate EM, Davey MR, Power JB, Marchant R (1995) Factors affecting the enetic engineering of plants by Microprojectile bombardment Biotechnology Advances 13, 631-651.
Sujatha M (2002) Current status and future prospects of in vitro techniques and biotechnology in safflower breeding. Sesame and safflower Newsletter 17, 92-97.
Taylor NJ, Fauquet, CM (2002) Microparticle bombardment as a tool in plant science and agricultural biotechnology. DNA and Cell Biology 21, 963-977.
Ying M, Dyer WE, Bergman JW (1992) Agrobacterium tumefaciens-mediated transformation of safflower (Carthamus tinctorius L.) cv. ‘Centennial’. Plant cell Reports 11, 581–585.
Zhu J, Oger PM, Schrammeijer B et al. (2000) The bases of crown gall tumorigenesis. Journal of Bacteriology 182, 3885 3895.