Toxic tetrapyrrole accumulation in protoporphyrinogen IX oxidase-overexpressing transgenic rice plants.
We generated transgenic rice plants (Oryza sativa cv. Dongjin) over-expressing human protoporphyrinogen IX oxidase (PPO) with the aim to increase mitochondrial PPO activity and confer herbicide resistance (Lee et al., Pestic Biochem Physiol 80:65-74, 2004). The transgenic plants showed during further leaf development the formation of severe necrotic spots and growth retardation. Several experiments were performed to examine the reasons for the formation of necrotic leaf lesions. Human PPO is normally located in mitochondria. An in vitro organellar import experiment revealed translocation of human PPO into pea chloroplasts, but not into mitochondria. Using a specific antibody raised against human PPO confirmed its plastidic localisation. The heme and chlorophyll contents were lower in necrotic leaves than wild-type leaves. Interestingly, mature and necrotic leaves of 12-week-old transgenic plants contained up to 14- and 24-fold more protoporphyrin IX, respectively, than mature wild-type leaves. Enhanced levels of Mg-Protoporphyrin IX, Mg-Protoporphyrin IX monomethyl ester and protochlorophyllide were concurrently observed in transgenic plants relative to wild type. Accumulated porphyrins and Mg-porphyrins likely act as photosensitizers and cause high formation of the reactive oxygen species. These high levels of tetrapyrrole intermediates correlated with increased rates of 5-aminolevulinic acid synthesis in transgenic plants. Tetrapyrrole-induced photooxidation was confirmed by increased lipid peroxidation and subsequent cell death. The transgenic phenotype is the consequence of a highly modified tetrapyrrole metabolism due to additional expression of human PPO. A possible regulatory role of PPO in graminaceous seedlings is discussed.
Jung
Sunyo
S
School of Life Sciences and Biotechnology, Kyungpook National University, Daegu 702-701, Republic of Korea.
Lee
Hye-Jung
HJ
Lee
Yonghyuk
Y
Kang
Kiyoon
K
Kim
Young Soon
YS
Grimm
Bernhard
B
Back
Kyoungwhan
K
D000595:Amino Acid Sequence; D012275:Oryza / Q000201:enzymology / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics*; D050768:Protoporphyrinogen Oxidase / Q000235:genetics*; D013347:Subcellular Fractions / Q000201:enzymology; D045725:Tetrapyrroles / Q000378:metabolism*
D045725:Tetrapyrroles; D050768:Protoporphyrinogen Oxidase