Journal
JOURNAL OF EXPERIMENTAL BOTANY
Volume 67, Issue 14, Pages 4285-4296Publisher
OXFORD UNIV PRESS
DOI: 10.1093/jxb/erw209
Keywords
Field performance; grain quality; lysine; metabolic engineering; pyramid transgenic line; rice (Oryza sativa L.)
Categories
Funding
- Ministry of Science and Technology of China [2012CB944803]
- Ministry of Agriculture of China [2016ZX08001006, 2009ZX08009-001B]
- National Nature and Science Foundation of China [31471459]
- Jiangsu PAPD Project
- Qinglan Project
- State Key Laboratory of Agrobiotechnology (CUHK)
- Lo Kwee Seong Foundation
- Lee Hysan Foundation
- Graduate Students Research and Innovation Program from Jiangsu Government of China [CXLX_1421]
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Marker-free transgenic rice that over-accumulates the essential amino acid lysine has been generated by combining lines with endosperm-specific and constitutive engineering of lysine, and has been evaluated in the field.Rice (Oryza sativa L.), a major staple crop worldwide, has limited levels of the essential amino acid lysine. We previously produced engineered rice with increased lysine content by expressing bacterial aspartate kinase and dihydrodipicolinate synthase and inhibiting rice lysine ketoglutarate reductase/saccharopine dehydrogenase activity. However, the grain quality, field performance, and integration patterns of the transgenes in these lysine-enriched lines remain unclear. In the present study, we selected several elite transgenic lines with endosperm-specific or constitutive regulation of the above key enzymes but lacking the selectable marker gene. All target transgenes were integrated into the intragenic region in the rice genome. Two pyramid transgenic lines (High Free Lysine; HFL1 and HFL2) with free lysine levels in seeds up to 25-fold that of wild type were obtained via a combination of the above two transgenic events. We observed a dramatic increase in total free amino acids and a slight increase in total protein content in both pyramid lines. Moreover, the general physicochemical properties were improved in pyramid transgenic rice, but the starch composition was not affected. Field trials indicated that the growth of HFL transgenic rice was normal, except for a slight difference in plant height and grain colour. Taken together, these findings will be useful for the potential commercialization of high-lysine transgenic rice.
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