A quick, easy and cost-effective in planta method to develop direct transformants in wheat

Abstract

Agrobacterium mediated in planta method was used to transform Indian elite wheat genotype HD2894 with herbicidetolerant CP4-EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) gene. The apical meristems of germinated seeds were targeted for introgression of transgene. The obtained T 1 plants were screened by spraying 1% glyphosate and only positive transformants survived. The presence of transgene was also confirmed by PCR and Southern hybridization. Using this method, 3.07% transformation rate was observed. To identify transgenic lines carrying stably integrated CP4-EPSPS gene, the transgenic populations were screened in T 3 generation using 1% glyphosate and lines with 100% survival were considered as homozygous. No significant morpho-physiological variations were observed within the transgenic lines as compared to non-transgenic plants. The present study resulted in herbicide-tolerant transgenic wheat and provides a valuable tool for development of wheat genetic transformation.

Auxin independent1 (Axi1) as an endogenous gene for copy number determination in transgenic tobacco

“…Antibiotic or herbicide selection is a conventional method that is used to differentiate between hemizygous and homozygous lines if the transgene is a dominant trait and follows the Mendelian first law of segregation (Passricha et al 2016, Tarafdar et al 2019. To validate the data obtained for the copy number by real-time PCR, a segregation test was performed on the five independent pMCG1005 events.…”

A simple and efficient Agrobacterium-mediated in planta transformation protocol for horse gram (Macrotyloma uniflorum Lam. Verdc.)

“…Correspondingly, nominal cell density with increased co-cultivation duration has the lowest stress response with maximum survivability of explants [21,34,35]. On the contrary, herbicide-resistant wheat and cotton plants raised by in planta method resulted in a maximum survival rate with 3.07% and 0.12% transformation efficiency [30,36].…”

“…There are several recent reports on the seeds as explants for the in planta transformation [27][28][29][30]. Selection of appropriate strain and cell density are the prerequisites for transformation in dicot species.…”