Journal of Integrative Plant Biology, 60 (5) : 382-396, May 2018     Research Article

OPEN ACCESS ARTICLE

Method for hull©\less barley transformation and manipulation of grain mixed©\linkage beta©\glucan[OA]

Wai Li Lim, Helen M. Collins, Rohan R. Singh, Natalie A. J. Kibble, Kuok Yap, Jillian Taylor, Geoffrey B. Fincher and Rachel A. Burton
ARC Centre of Excellence in Plant Cell Walls, School of Agriculture, Food and Wine, University of Adelaide, Glen Osmond, South Australia 5064, Australia

These authors contributed equally to the work
Current address: Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, 4072, Australia

Correspondence:
Email: Rachel A Burton (rachel.burton@adelaide.edu.au)

Online on 16 Dec 2017 at www.jipb.net and www.wileyonlinelibrary.com/journal/jipb

Abstract

Hull©\less barley is increasingly offering scope for breeding grains with improved characteristics for human nutrition; however, recalcitrance of hull©\less cultivars to transformation has limited the use of these varieties. To overcome this limitation, we sought to develop an effective transformation system for hull©\less barley using the cultivar Torrens. Torrens yielded a transformation efficiency of 1.8%, using a modified Agrobacterium transformation method. This method was used to over©\express genes encoding synthases for the important dietary fiber component, (1,3;1,4)©\β©\glucan (mixed©\linkage glucan), primarily present in starchy endosperm cell walls. Over©\expression of the HvCslF6 gene, driven by an endosperm©\specific promoter, produced lines where mixed©\linkage glucan content increased on average by 45%, peaking at 70% in some lines, with smaller increases in transgenic HvCslH1 grain. Transgenic HvCslF6 lines displayed alterations where grain had a darker color, were more easily crushed than wild type and were smaller. This was associated with an enlarged cavity in the central endosperm and changes in cell morphology, including aleurone and sub©\aleurone cells. This work provides proof©\of©\concept evidence that mixed©\linkage glucan content in hull©\less barley grain can be increased by over©\expression of the HvCslF6 gene, but also indicates that hull©\less cultivars may be more sensitive to attempts to modify cell wall composition.


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Received 15 Nov 2017   Accepted 13 Dec 2017
© 2017 Institute of Botany, Chinese Academy of Sciences


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