NDLI: Engineering cyanogen synthesis and turnover in cassava (Manihot esculenta)

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Engineering cyanogen synthesis and turnover in cassava (Manihot esculenta)

Content Provider	Springer Nature Link
Author	Siritunga, Dimuth Sayre, Richard
Copyright Year	2004
Abstract	Cassava is the major root crop for a quarter billion subsistence farmers in sub-Saharan Africa. It is valued for its ability to grow in adverse environments and the food security it provides. Cassava contains potentially toxic levels of cyanogenic glycosides (linamarin) which protect the plant from herbivory and theft. The cyanogens, including linamarin and its deglycosylated product, acetone cyanohydrin, can be efficiently removed from the root by various processing procedures. Short-cuts in processing, which may occur during famines, can result in only partial removal of cyanogens. Residual cyanogens in cassava foods may cause neurological disorders or paralysis, particularly in nutritionally compromised individuals. To address this problem and to further understand the function of cyanogenic glycosides in cassava, we have generated transgenic cassava in which cyanogenic glycoside synthesis has been selectively inhibited in leaves and roots by antisense expression of CYP79D1/D2 gene fragments. The CYP79D1/D2 genes encode two highly similar cytochrome P450s that catalyze the first-dedicated step in cyanogenic glycoside synthesis. Transgenic plants in which the expression of these genes was selectively inhibited in leaves had substantially reduced (60– 94% reduction) linamarin leaf levels. Surprisingly, these plants also had a greater than a 99% reduction in root linamarin content. In contrast, transgenic plants in which the CYP79D1/D2 transcripts were reduced to non-detectable levels in roots had normal root linamarin levels. These results demonstrate that linamarin synthesized in leaves is transported to the roots and accounts for nearly all of the root linamarin content. Importantly, transgenic plants having reduced leaf and root linamarin content were unable to grow in the absence of reduced nitrogen (NH$_{3}$) . Cassava roots have previously been demonstrated to have an active cyanide assimilation pathway leading to the synthesis of amino acids. We propose that cyanide derived from linamarin is a major source of reduced nitrogen for cassava root protein synthesis. Disruption of linamarin transport from leaves in CYP79D1/D2 anti-sense plants prevents the growth of cassava roots in the absence of an alternate source of reduced nitrogen. An alternative strategy for reducing cyanogen toxicity in cassava foods is to accelerate cyanogenesis and cyanide volatilization during food processing. To achieve this objective, we have expressed the leaf-specific enzyme hydroxynitrile lyase (HNL) in roots. HNL catalyzes the breakdown of acetone cyanohydrin to cyanide. Expression of HNL in roots accelerated cyanogenesis by more than three-fold substantially reducing the accumulation of acetone cyanohydrin during processing relative to wild-type roots.
Starting Page	661
Ending Page	669
Page Count	9
File Format	PDF
ISSN	01674412
Journal	Plant Molecular Biology
Volume Number	56
Issue Number	4
e-ISSN	15735028
Language	English
Publisher	Kluwer Academic Publishers
Publisher Date	2004-01-01
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	cyanogenesis cyanogenic glycoside cytochrome P450 hydroxynitrile lyase linamarin Biochemistry Plant Sciences Plant Pathology
Content Type	Text
Resource Type	Article
Subject	Genetics Plant Science Medicine Agronomy and Crop Science

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Generation of cyanogen-free transgenic cassava

Identification and characterization of CYP79D16 and CYP71AN24 catalyzing the first and second steps in l-phenylalanine-derived cyanogenic glycoside biosynthesis in the Japanese apricot, Prunus mume Sieb. et Zucc.

Isoforms of Linamarase in Cassava (Manihot esculenta)

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Engineering cyanogen synthesis and turnover in cassava (Manihot esculenta)