Brachypodium distachyon Transformation

Brachypodium distachyon is an annual herbaceous plant, native to central Asia, Africa and southern Europe. Brachypodium distachyon is a dwarf plant widely grown in temperate regions. The application of plant tissue culture in the research of Brachypodium distachyon allows it to be produced without seasonal control and achieve year-round production, which has important practical significance for the actual production and application of plants. Based on plant tissue culture technology, a genetic transformation system can be effectively established. Genetic transformation research on Brachypodium distachyon can promote the research and application.

Lifeasible is a comprehensive plant genetic transformation service unit and has established happy, cooperative relationships with hundreds of universities and scientific research institutes. It has a powerful plant transformation platform and is capable of high-throughput transformation of various plants. Based on the cutting-edge field of modern agriculture, we continuously introduce new technologies and products of excellent quality that are in line with industry development trends by integrating research results from different disciplines. After years of research, we have established an efficient and reproducible Brachypodium distachyon regeneration system. We have also developed an efficient genetic transformation method for Brachypodium distachyon.

What do we offer?

Acquisition of young embryos

We used young embryos from seeds of Brachypodium distachyon as explants, which were cleaned ,sterilized and stripped of young embryos with the help of a microscope in an ultra-clean bench.

Induction of healing tissue

After sterilizing the explants, the plates were inoculated with a healing tissue induction medium and incubated in a dark incubator. When the healing tissues matured, they were cut off and transferred to the successor medium.

Healing tissue differentiation and plant regeneration

Transfer the different sources of guiding tissues into the differentiation medium, wait until the adventitious buds appear on the differentiation medium, and at the same time, transfer the adventitious buds into the successor medium to grow. The rooted seedlings were inoculated into the histoculture flasks for further cultivation, and when the seedlings grew up, they were potted for cultivation.

Genetic transformation of healing tissue

In the genetic transformation of Brachypodium distachyon, we utilized the Agrobacterium transformation method, inoculated Agrobacterium sap in YEB medium, selected the healing tissues with vigorous growth and bright yellow color, and placed them in Agrobacterium sap for a few minutes, and then sucked off the sap with absorbent paper, and sucked off the surface of the healing tissues with sterilized filter paper.

Screening of resistant healing wounds and plants

After a few weeks of transformation, the healing tissues were transferred to a new resistance-inducing medium. After two successive cultures, the healing tissues were transferred to a differentiation medium for culture. When the resistant regenerated green seedlings came out, they were transferred to a rooting medium for screening. The resistant seedlings would be rooted one after another in the following weeks.

Our services of Brachypodium distachyon transformation

• Vector construction.
• Induction and succession of Brachypodium distachyon recipient healing tissues.
• Genetic transformation and screening of Brachypodium distachyon mediated by Agrobacterium tumefaciens.
• Differentiation of positive healing tissues into seedlings.
• Rooting and strengthening of regenerated green seedlings.
• PCR detection of exogenous genes in T0 generation transgenic seedlings.

Our experimental period

We can carry out experiments throughout the year. Generally, it takes us 3-5 months from constructing the vector to getting the test results of T0 generation transgenic seedlings.

Experimental results we provide

• Some pictures of genetic transformation and screening.
• Pictures of some samples of differentiation and regeneration of healing tissue.
• PCR test results of exogenous gene of T0 generation transgenic seedlings.
• T0 generation Brachypodium distachyon transgenic seedlings, more than 10 T0 generation positive seedlings for each transformation experiment.

Our service workflow

Lifeasible is committed to providing high-quality plant genetic transformation services; we use the Agrobacterium-mediated method to realize the transformation of Brachypodium distachyon. Customers can provide plasmids, Agrobacterium, etc., so that we can complete the whole experimental process, from constructing vectors to obtaining transgenic seedlings for you. If you are interested in us, please feel free to contact us.