Efficient, reproducible Agrobacterium-mediated transformation of sorghum using heat treatment of immature embryos.

Plant Cell Reports
Songul GurelPeggy G Lemaux

Abstract

A number of parameters related to Agrobacterium-mediated infection were tested to optimize transformation frequencies of sorghum (Sorghum bicolor L.). A plasmid with a selectable marker, phosphomannose isomerase, and an sgfp reporter gene was used. First, storing immature spikes at 4 degrees C before use decreased frequency of GFP-expressing calli, for example, in sorghum variety P898012 from 22.5% at 0 day to 6.4% at 5 days. Next, heating immature embryos (IEs) at various temperatures for 3 min prior to Agrobacterium infection increased frequencies of GFP-expressing calli, of mannose-selected calli and of transformed calli. The optimal 43 degrees C heat treatment increased transformation frequencies from 2.6% with no heat to 7.6%. Using different heating times at 43 degrees C prior to infection showed 3 min was optimal. Centrifuging IEs with no heat or heating at various temperatures decreased frequencies of all tissue responses; however, both heat and centrifugation increased de-differentiation of tissue. If IEs were cooled at 25 degrees C versus on ice after heating and prior to infection, numbers with GFP-expressing cells increased from 34.2 to 49.1%. The most optimal treatment, 43 degrees C for 3 min, cooling at 25 degrees...Continue Reading

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Citations

Jan 12, 2012·Plant Cell Reports·Guoquan Liu, Ian D Godwin
Nov 23, 2011·Functional & Integrative Genomics·Surinder SinghJaswinder Singh
Oct 24, 2012·Applied Biochemistry and Biotechnology·Shweta Mehrotra, Vinod Goyal
Jun 11, 2010·Plant Cell Reports·Anshu Raghuwanshi, Robert G Birch
Dec 7, 2011·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Moses BirumaChristina Dixelius
Aug 22, 2014·Molecular Plant·So-Yon ParkZhanyuan J Zhang
Jan 9, 2013·Plant Biotechnology Journal·Maria N SomlevaKristi D Snell
Jan 6, 2011·Journal of Integrative Plant Biology·Chunshan GuoGuihua Lu
Nov 27, 2014·Frontiers in Plant Science·Yukoh HieiToshihiko Komari
Oct 3, 2018·International Journal of Molecular Sciences·Rana Imtiaz AhmedYingzhen Kong
Oct 13, 2018·Current Protocols in Plant Biology·Phat Tien DoZhanyuan J Zhang
Oct 1, 2009·Physiology and Molecular Biology of Plants : an International Journal of Functional Plant Biology·V Girijashankar, V Swathisree
Jan 1, 2020·Biotechnology for Biofuels·Christine A Ondzighi-AssoumeC Neal Stewart
Jun 6, 2020·Current Protocols in Plant Biology·Si Nian CharBing Yang
Jan 1, 2014·In Vitro Cellular & Developmental Biology. Plant : Journal of the Tissue Culture Association·Emily WuZuo-Yu Zhao
Jul 6, 2019·Plant Biotechnology·Tomoko KuriyamaMinami Matsui
Jun 13, 2017·Biotechnology for Biofuels·Supriya MathurManoj K Sharma
Nov 6, 2021·Planta·Zhanguo XinGeoffrey Morris

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