Optimization of Agrobacterium-mediated transformation in spring bread wheat using mature and immature embryos

2019 ◽  
Vol 46 (2) ◽  
pp. 1845-1853 ◽  
Author(s):  
Rakesh Kumar ◽  
Harohalli Masthigowda Mamrutha ◽  
Amandeep Kaur ◽  
Karnam Venkatesh ◽  
Davinder Sharma ◽  
...  
Plants ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 895
Author(s):  
Samira El Hanafi ◽  
Souad Cherkaoui ◽  
Zakaria Kehel ◽  
Ayed Al-Abdallat ◽  
Wuletaw Tadesse

Hybrid wheat breeding is one of the most promising technologies for further sustainable yield increases. However, the cleistogamous nature of wheat displays a major bottleneck for a successful hybrid breeding program. Thus, an optimized breeding strategy by developing appropriate parental lines with favorable floral trait combinations is the best way to enhance the outcrossing ability. This study, therefore, aimed to dissect the genetic basis of various floral traits using genome-wide association study (GWAS) and to assess the potential of genome-wide prediction (GP) for anther extrusion (AE), visual anther extrusion (VAE), pollen mass (PM), pollen shedding (PSH), pollen viability (PV), anther length (AL), openness of the flower (OPF), duration of floret opening (DFO) and stigma length. To this end, we employed 196 ICARDA spring bread wheat lines evaluated for three years and genotyped with 10,477 polymorphic SNP. In total, 70 significant markers were identified associated to the various assessed traits at FDR ≤ 0.05 contributing a minor to large proportion of the phenotypic variance (8–26.9%), affecting the traits either positively or negatively. GWAS revealed multi-marker-based associations among AE, VAE, PM, OPF and DFO, most likely linked markers, suggesting a potential genomic region controlling the genetic association of these complex traits. Of these markers, Kukri_rep_c103359_233 and wsnp_Ex_rep_c107911_91350930 deserve particular attention. The consistently significant markers with large effect could be useful for marker-assisted selection. Genomic selection revealed medium to high prediction accuracy ranging between 52% and 92% for the assessed traits with the least and maximum value observed for stigma length and visual anther extrusion, respectively. This indicates the feasibility to implement genomic selection to predict the performance of hybrid floral traits with high reliability.


aBIOTECH ◽  
2021 ◽  
Author(s):  
Xiaojuan Ran ◽  
Tengfei Tang ◽  
Meiyue Wang ◽  
Luhuan Ye ◽  
Yili Zhuang ◽  
...  

2018 ◽  
Vol 23 ◽  
pp. 120-125
Author(s):  
S. M. Sichkar ◽  
L. H. Velykozhon ◽  
O. V. Dubrovna ◽  
B. V. Morgun

Aim. Determination of the allelic composition of Glu-1 loci in samples of rare wheat species and their hybrids with spring bread wheat. Methods. PCR analysis. Results. In existing collection samples of the rare wheat species the alleles of the a/c loci Glu-A1 were found, while allele b was found only in T.dicoccum, var. volgense (Emmer Kokchetavskaya). An additional amplicon with a length of 450 bp was found in the Glu-A1 locus in the sample (T. dicoccum × Dasypyrum villosum), and in (Ae. ventricosa × T. dicoccum) another one a length of 700 bp in genome B, was found require further research. Among hexaploid wheat, it was revealed genotypes differing in the presence of alleles a and d of the locus Glu-D1. The hybridity of the received forms has been confirmed, as evidenced by the identification of both parent components in the hybrids or alleles of the Glu-D1 locus of bread wheat in hybrids with emmer. Conclusions. Analysis of the allele composition of Glu1 locuses in samples of rare wheat species and their hybrids with bread wheat allowed to select genotypes and hybrid combinations that may be promising for further breeding work. Keywords: Triticum spelta L., T. dicoccum, hybrids, PCR analysis, Glu-1 locus.


2018 ◽  
Vol 22 ◽  
pp. 222-227
Author(s):  
O. M. Honcharuk ◽  
O. V. Dubrovna

Aim. Receiving of genetically modified plants of bread wheat with heterologous ornithine‑δ‑aminotransferase gene. Methods. Agrobacterium-mediated transformation of callus cultures in vitro, PCR-analysis. Results. By Agrobacterium-mediated transformation of the morphogenic calluses of bread wheat (Triticum aestivum L.) using the AGLO strain containing the binary vector pBi-OAT with the target ornithine-δ-aminotransferase (oat) and selective neomycinphosphotransferase II (nptII), transgenic plants-regenerators have been obtained. Conclusions. As a result of the genetic transformation of Zimoyarka variety, 12 wheat regenerants were obtained in the genome which revealed a complete integration of the genetic construct containing the oat and nptII transgenes. Keywords: Triticum aestivum L., Agrobacterium-mediated transformation, ornithine‑δ‑aminotransferase gene, PCR-analysis.


2021 ◽  
Vol 28 ◽  
pp. 66-71
Author(s):  
O. V. Dubrovna ◽  
L. V. Slivka

Aim. Optimization of conditions for genetic transformation of new promising genotypes of winter bread wheat (T. aestivum L.) by in planta method. Methods. Agrobacterium-mediated transformation by in planta method using the strain AGL0 and vector construct pBi2E. Results. The influence of air temperature, optical density of cells of agrobacterial suspension, inoculation day and composition of inoculation medium on the frequency of obtaining transgenic plants of new winter wheat genotypes was studied. The dependence of the frequency obtaining of transgenic plants from environmental conditions, in particular temperature, has been established. It was found that the temperature regime of 20-22°C provided the largest number (4.8%) of wheat transformants, and when the temperature is reduced to 16-18°C there is a decrease in the efficiency of T-DNA transfer into the plant genome and the lowest frequency of transformation (0.7%). Conclusions. The largest number of transformants was obtained using a inoculation medium without sucrose, the optical density of cells of the agrobacterial suspension of 0.6 op.od. and inoculation on the third day after castration of ears. Keywords: T. aestivum, Agrobacterium-mediated transformation in planta, optimization of conditions.


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