Characterization of a Rice Chlorophyll-Deficient Mutant Using the T-DNA Gene-Trap System

Jung, Ki Hong; Hur, Junghe; Ryu, Choong Hwan; Choi, Yoon E.; Chung, Yong-Yoon; Miyao, Akio; Hirochika, Hirohiko; An, Gynheung

doi:10.1093/pcp/pcg064

Cited by 286 publications

(179 citation statements)

References 50 publications

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“…Such mutations were previously reported often induced by tissue culture application (Palmer et al 2000). Such chlorophyll-deficient phenotypes were also reported from previous work using a T-DNA insertional sequences in rice, in about similar percentage (Jeon and An 2001;Jung et al 2003). One mutant line was identified having a T-DNA insertion in the chlorina (OsCHLH) gene that is highly homologous to XANTHA-F gene in barley and CHLH gene in Arabidopsis.…”

Section: Molecular Analysis By Southern Blotsupporting

confidence: 75%

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Untagged Mutation in Rice Gal4/Vp16 Transcriptional Activator Facilitated-Enhancer Trap Lines

Koerniati

2013

Indones. J. Agric. Sci.

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An enhancer trap system is an insertional mutagenesis based upon gene expression, instead of gene knock-out, so its insertion in genome is expected not linked to any dramatic changes in plant phenotypes. Gene knock-out, leading to lossof- function (LoF) mutation, is a dominant approach for rice functional genomic studies. The objective of this study was to find out whether Transcriptional Activator-Facilitated Enhancer Trap (TAFET) T-DNA insertion inducing mutant phenotypes in rice TAFET population. Materials used in this experiment were T1 generation of 270 rice TAFET lines. Eight plants of each were grown in the greenhouse and observed for any mutant phenotypes. Phenotypic, histochemical, Southern<br />blot analyses were carried out to define a mutant of pSKC66.1- 8e. Result showed that about 10% of the 270 lines produced chlorophyll-deficient leaves, ranged from yellowish green (viridis), white stripe green zebra-like stripe) to completely white (albino). Albino plants died after two weeks, whilst white stripe or viridis mutants became normal in the next generation<br />(T2). Another mutant was pSKC66.1-8e line which had floral dramatic phenotype change with various spikelet shapes and number of organs, and had a single twisted culm. The flower of mutant also had gus gene expression. Plants with wild type did not express gus gene and had six or more straight culms. Molecular, histochemical and phenotypic analyses of this particular line for three generations indicated that mutant phenotype was not due to the T-DNA insertion. Since there was approved that Tos17 is activated during tissue culture and induced mutant phenotype, this line might relate to Tos17 insertion, but it needs further investigation to gain such conclusion.

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Section: Molecular Analysis By Southern Blotsupporting

confidence: 75%

“…One mutant line was identified having a T-DNA insertion in the chlorina (OsCHLH) gene that is highly homologous to XANTHA-F gene in barley and CHLH gene in Arabidopsis. Two other mutants with chlorophylldeficient phenotypes had a Tos17 insertion in the OsCHLH gene (Jung et al 2003).…”

Section: Molecular Analysis By Southern Blotmentioning

confidence: 99%

Untagged Mutation in Rice Gal4/Vp16 Transcriptional Activator Facilitated-Enhancer Trap Lines

Koerniati

2013

Indones. J. Agric. Sci.

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“…1A). We previously identified this mutant as a knockout of OsCHLH, the gene encoding Mg 2ϩ -chelatase that is involved in chlorophyll biosynthesis (1). The OsCHLH mutant contains only 0.27% or less of the normal chlorophyll content (on a per leaf area basis) (1) and did not exhibit chlorophyll fluorescence in its intact leaves (Fig.…”

Section: The Oschlh Mutation Affects Chlorophyll Pigmentation Of the mentioning

confidence: 99%

“…The T-DNA-tagged lines could be therefore verified by ␤-glucuroidase assay (data not shown). This mutant carries a stable DNA mutation that affects chlorophyll synthesis in the thylakoid light-harvesting complexes (1,44).…”

Section: The Oschlh Mutation Affects Chlorophyll Pigmentation Of the mentioning

confidence: 99%

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Mitochondria Provide the Main Source of Cytosolic ATP for Activation of Outward-rectifying K+ Channels in Mesophyll Protoplast of Chlorophyll-deficient Mutant Rice (OsCHLH) Seedlings

Goh

Jung

Roberts

et al. 2004

Journal of Biological Chemistry

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The role of mitochondria in providing intracellular ATP that controls the activity of plasma membrane outward-rectifying K ؉ channels was evaluated. The Os-CHLH rice mutant, which lacks chlorophyll in the thy- The OsCHLH mutant is unable to fix CO 2 and exhibits reduced growth. Wild type and mutant plants exhibit similar rates of respiratory O 2 uptake in the dark, whereas the rate of photosynthetic O 2 evolution by the mutant was negligible during illumination. During dark respiration the wild type and mutant exhibited similar levels of cytoplasmic ATP. In the mutant oligomycin treatment (an inhibitor of mitochondrial F 1 F 0 -ATPase) drastically reduced ATP production. The fact that this was reversed by the addition of glucose suggested that the mutant produced ATP exclusively from mitochondria but not from chloroplasts. In whole cell patch clamp experiments, the activity of outward-rectifying K ؉ channels of rice mesophyll cells showed ATPdependent currents, which were 1.5-fold greater in wild type than in mutant cells. Channels in both wild type and mutant cells were deactivated by the removal of cytosolic ATP, whereas in the presence of ATP the channels remained active. We conclude that mesophyll cells in the OsCHLH rice mutant derive ATP from mitochondrial respiration, and that this is critical for the normal function of plasma membrane outward-rectifying K ؉ channels.Mitochondrial metabolism not only impacts on other cellular biosynthetic and catabolic processes, but also plays an important role in providing the overall cellular energy supply. Dark respiration and photosynthesis are metabolic pathways that produce redox equivalents and ATP to meet the energy requirements to support cell growth and maintenance (2). The central role of mitochondria in plants is reflected by the array of mitochondrial transporters and shuttles that transfer metabolites and reducing equivalents back and forth between mitochondria and the cytosol. Oxidative phosphorylation provides the cytosol with ATP, which is required for sucrose synthesis and other cytosolic processes. Studies with specific respiratory inhibitors have shown that oxidative phosphorylation occurs both in darkness and in light (2-4). However, very little is known about the regulation of mitochondrial respiration in photosynthetic mesophyll cells, and the role of mitochondrial ATP production as a source of cytosolic ATP in this process is not fully understood.Many biochemical reactions in plant and mammalian cells depend on a tightly controlled ratio of ATP to ADP. In mammalian mitochondria, this ratio is preserved by regulatory mechanisms that couple the rate of cellular ATP consumption to the rate of ATP production by oxidative phosphorylation (5-8). Hence, efficient communication between cellular energy stores and membrane metabolic sensors is necessary for regulation of membrane excitability and associated functions. In most mammalian cells, there is a class of K ϩ channels whose activity is closely coupled to metabolism by ATP (9, 10). Recent studies have...

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Map‐based cloning and characterization of YGL22, a new yellow‐green leaf gene in rice (Oryza sativa)

Yan

Dong

et al. 2020

Crop Science

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Leaf‐color mutants have been extensively studied in rice (Oryza sativa L.) and many corresponding genes have been identified and cloned. However, the mechanism of complex leaf‐color mutations requires further study. In this study, we obtained an introgression line (TIL22) from a set of introgression lines raised using African cultivated rice (O. glaberrima Steud.) as the donor parent and O. sativa subsp. indica Teqing as the recipient. Compared with Teqing, TIL22 showed a yellow‐green leaf phenotype at the seedling stage, but the leaves gradually changed to green after the five‐leaf stage. The photosynthetic pigment contents of yellow‐green leaves at the seedling stage were significantly reduced and chloroplast development was retarded compared with those of Teqing. Genetic analysis indicated that the introgression line phenotype was controlled by a single nuclear gene, temporarily designated YGL22. Map‐based cloning and sequence analysis suggested that the candidate gene was likely to be LOC_Os01g15390, which encodes a chloroplast protein. Real‐time quantitative polymerase chain reaction (qPCR) analysis revealed that the YGL22 transcript level was significantly lower in TIL22 than that of Teqing at the seedling stage. We generated the ygl22 knockout mutant using the CRISPR/Cas9 system. The ygl22 mutants showed a similar phenotype to that of TIL22 at the seedling stage, suggesting that YGL22 may be involved in the early development of chloroplasts in rice. In addition, plant height, number of panicles per plant, and grain yield per plant of ygl22 were not significantly affected. The results indicate that YGL22 can be used as a trait marker gene in hybrid rice production.

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Characterization of a Rice Chlorophyll-Deficient Mutant Using the T-DNA Gene-Trap System

Cited by 286 publications

References 50 publications

Untagged Mutation in Rice Gal4/Vp16 Transcriptional Activator Facilitated-Enhancer Trap Lines

Untagged Mutation in Rice Gal4/Vp16 Transcriptional Activator Facilitated-Enhancer Trap Lines

Mitochondria Provide the Main Source of Cytosolic ATP for Activation of Outward-rectifying K+ Channels in Mesophyll Protoplast of Chlorophyll-deficient Mutant Rice (OsCHLH) Seedlings

Map‐based cloning and characterization of YGL22, a new yellow‐green leaf gene in rice (Oryza sativa)

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