Findings Will Accelerate Efforts To Harness The Vast Bioenergetic Potential Of This Highly Yielding But Genetically Complex Perennial Grass – ScienceDaily

For the first time, researchers have successfully demonstrated the precision of gene modification in miscanthus, a promising perennial crop for sustainable bioenergy production.

A team at the Center for Advanced Bioenergy and Bioproduct Innovation (CABBI), a bioenergy research center (BRC) funded by the US Department of Energy, modified the genomes of three staghorn species using CRISPR/Cas9 technology – a more targeted and effective method. To develop new varieties of previous methods.

The findings will accelerate efforts to harness the enormous potential of this highly productive yet genetically complex grass as a source of biofuels, renewable bioproducts and carbon sequestration. The study published in biotechnology for biofuels and bioproducts, Led by three CABBI researchers at the HudsonAlpha Institute of Biotechnology in Alabama–faculty investigator Kankshita Swaminathan, research associate Anthony Trieu and former postdoctoral researcher Muhammad Pleviv–and Nancy Reichert, professor of biological sciences at Mississippi State University.

Swaminathan co-led an international team that sequenced the genome of angiosperms in 2020. This work provided a roadmap for researchers exploring new ways to maximize plant productivity and decipher the genetic basis of its desirable traits. Miscanthus is very adaptable and easy to grow. It can thrive on marginal lands, requires limited fertilization, is highly tolerant of drought and cold temperatures, and uses the more efficient C4 form of photosynthesis.

To date, efforts to genetically improve misbehavior have focused on transforming plants by inserting exogenous genes at random locations in their genomes, rather than targeting specific sites or modifying existing genes.

The CABBI team has developed gene-editing procedures using CRISPR/Cas9 technology that will allow researchers to selectively target existing genes within slut-sham plants to knock out or modify their function and insert new genes at specific locations. This targeting ability offers a new avenue for the genetic improvement of this important biomass crop.

The study showed gene editing in three species of Misanthus – the most productive Miscanthus x giganteuswhich is grown commercially for bioenergy, and its parents, M. sacchariflorus And M. sinensis. Because these plants are ancient polyploids—with ancient sorghum-like DNA repeats and multiple sets of chromosomes—designing a guide RNA that locates the genetic material for the editing needed to target all copies of the gene is necessary to account for replication and ensure complete “knockouts.” “.

The CABBI researchers built on similar gene editing in Zia Miss (maize), which identified the white lemon 1 (lw1) gene as a useful target for visual confirmation of genetic alterations. This gene is involved in the biosynthesis of chlorophyll and a carotenoid, which affects leaf color, and previous studies have shown that deregulation of lw1 via CRISPR/Cas9 resulted in dull green/yellow, striped or white leaf phenotypes.

Using sequence information from both miscanthus and sorghum, the researchers identified directive RNAs that could target homologues, or duplicated gene copies, of lw1 in the tissues of the fluff plant. Leaves on the modified whipping plants showed the same phenotypes as on maize, with pale green/yellow, streaked, or white leaves instead of the typical green.

The work advances CABBI’s mission to develop sustainable bioenergy production and engineer select feedstocks (peaches, sorghum, and sugarcane) to produce new bioproducts, such as oils and specialty chemicals. Prior to this study, bioengineering work was limited to sorghum and cane because microengineering methods were not developed in miscanthus.

“Identification of transferable germplasm, development of reliable transformation methods, and demonstration of gene modification in Miscanthus are all critical steps toward engineering pathways in bacillus,” said Swaminathan. “The ability to finely tune bugs to enhance productivity, enable continued growth on marginal lands, and produce specialty chemicals such as oils will help remove the ‘potential’ from its status as a viable bioenergy crop.

“This research helps us get a few steps closer to reducing our dependence on petroleum-based energy.”

To identify the bug strains that turned out well, the researchers screened germplasm from commercial vendors and study collaborators. Most of the lines were provided by co-author Eric Sachs, professor of crop sciences at the University of Illinois Urbana-Champaign, who collected germplasms from around the world. Sacks and Swaminathan are deputy subject leaders for feedstock production research at CABBI.

“This research project was a collaborative, multi-institutional effort with researchers working across disciplines towards an important goal. It promoted a ‘big picture’ approach to research within CABBI, as well as at other BRCs,” said Reichert.

Other CABBI co-authors on the study include Steve Moss, professor of crop sciences at Illinois; Tom Clemente, Eugene W. Price Distinguished Professor of Biotechnology at the University of Nebraska Plant Science Innovation Center; postdoctoral researcher Pradipa Hiranyah, technologist Shilpa Manjunatha, intern Rebekah Wood, and workforce development specialist Yokshetha Patola, all from Hudson Alpha; and research associate Rebecca Billingsley and graduate student Anjali Urban from Mississippi.

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