The effects of enhanced methionine synthesis on amino acid and anthocyanin content of potato tubers.
BACKGROUND
Potato is a staple food in the diet of the world's population and also being used as animal feed. Compared to other crops, however, potato tubers are relatively poor in the essential amino acid, methionine. Our aim was to increase the methionine content of tubers by co-expressing a gene involved in methionine synthesis with a gene encoding a methionine-rich storage protein in potato plants.


RESULTS
In higher plants, cystathionine gamma-synthase (CgS) is the first enzyme specific to methionine biosynthesis. We attempted to increase the methionine content of tubers by expressing the deleted form of the Arabidopsis CgS (CgSDelta90), which is not regulated by methionine, in potato plants. To increase the incorporation of free methionine into a storage protein the CgSDelta90 was co-transformed with the methionine-rich 15-kD beta-zein. Results demonstrated a 2- to 6-fold increase in the free methionine content and in the methionine content of the zein-containing protein fraction of the transgenic tubers. In addition, in line with higher methionine content, the amounts of soluble isoleucine and serine were also increased. However, all of the lines with high level of CgSDelta90 expression were phenotypically abnormal showing severe growth retardation, changes in leaf architecture and 40- to 60% reduction in tuber yield. Furthermore, the colour of the transgenic tubers was altered due to the reduced amounts of anthocyanin pigments. The mRNA levels of phenylalanine ammonia-lyase (PAL), the enzyme catalysing the first step of anthocyanin synthesis, were decreased.


CONCLUSIONS
Ectopic expression of CgSDelta90 increases the methionine content of tubers, however, results in phenotypic aberrations in potato. Co-expression of the 15-kD beta-zein with CgSDelta90 results in elevation of protein-bound methionine content of tubers, but can not overcome the phenotypical changes caused by CgSDelta90 and can not significantly improve the nutritional value of tubers. The level of PAL mRNA and consequently the amount of anthocyanin pigments are reduced in the CgSDelta90 transgenic tubers suggesting that methionine synthesis and production of anthocyanins is linked.
Dancs
Gábor
G
Agricultural Biotechnology Center, P,O, Box 411, H-2101 Gödöllõ, Hungary. dancs@abc.hu
Kondrák
Mihály
M
Bánfalvi
Zsófia
Z
D000596:Amino Acids / Q000378:metabolism*; D000872:Anthocyanins / Q000378:metabolism*; D019757:Carbon-Oxygen Lyases / Q000235:genetics / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D018506:Gene Expression Regulation, Plant; D007532:Isoleucine / Q000378:metabolism; D008715:Methionine / Q000378:metabolism*; D008954:Models, Biological; D010650:Phenylalanine Ammonia-Lyase / Q000235:genetics / Q000378:metabolism; D035281:Plant Tubers / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D015398:Signal Transduction; D011198:Solanum tuberosum / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D015028:Zein / Q000235:genetics / Q000378:metabolism
D000596:Amino Acids; D000872:Anthocyanins; D007532:Isoleucine; D012694:Serine; D015028:Zein; D008715:Methionine; C022696:O-succinylhomoserine (thiol)-lyase; D019757:Carbon-Oxygen Lyases; D010650:Phenylalanine Ammonia-Lyase