[Characteristics and molecular mapping of a novel chlorophyll-deficient yellow-leaf mutant in rice].

A yellow-leaf mutant (yl11) with chlorophyll-deficient in rice (Oryza sativa L.) was selected from the progeny of a japonica rice variety "Jiahua 1" treated with 60Co γ-radiation. In comparison with the wild-type parent, "Jiahua 1", the mutant had yellow-leaves at whole growth stages and displayed significantly decreased in chlorophyll content and net photosynthetic rate. Underdeveloped chloroplast and alterations of the major agronomic traits, such as plant-heights, were also observed in the mutant. Genetic analysis confirmed that the yellow-leaf mutant trait was controlled by a single recessive nuclear gene (yl11). Using SSR and In/Del molecular markers and 920 F2 and F3 plants from the cross of yl11 with the indica variety Peiai 64S, the yl11 was mapped between the molecular markers MM2199 and InDel21039 with a physical distance of 110 kb on the long arm of chromosome 11, in which no known functional genes for chlorophyll synthesis or chloroplast development in rice has been found. These findings will provide a foundation for the cloning and functional analysis of this gene in the future.

The rice TCD11 encoding plastid ribosomal protein S6 is essential for chloroplast development at low temperature.

Plastid ribosome proteins (PRPs) are important components for chloroplast biogenesis and early chloroplast development. Although it has been known that chloroplast ribosomes are similar to bacterial ones, the precise molecular function of ribosomal proteins remains to be elucidated in rice. Here, we identified a novel rice mutant, designated tcd11 (thermo-sensitive chlorophyll-deficient mutant 11), characterized by the albino phenotype until it died at 20°C, while displaying normal phenotype at 32°C. The alteration of leaf color in tcd11 mutants was aligned with chlorophyll (Chl) content and chloroplast development. The map-based cloning and molecular complementation showed that TCD11 encodes the ribosomal small subunit protein S6 in chloroplasts (RPS6). TCD11 was abundantly expressed in leaves, suggesting its different expressions in tissues. In addition, the disruption of TCD11 greatly reduced the transcript levels of certain chloroplasts-associated genes and prevented the assembly of ribosome in chloroplasts at low temperature (20°C), whereas they recovered to nearly normal levels at high temperature (32°C). Thus, our data indicate that TCD11 plays an important role in chloroplast development at low temperature. Upon our knowledge, the observations from this study provide a first glimpse into the importance of RPS6 function in rice chloroplast development.

Importance of the rice TCD9 encoding α subunit of chaperonin protein 60 (Cpn60α) for the chloroplast development during the early leaf stage.

The chloroplast development governs plant growth and metabolism. This study characterized a novel rice thermo-sensitive chloroplast development 9 (tcd9) mutant, which exhibited the albino phenotype before the 3-leaf stage grown below 24 °C whereas displayed normal green at over 28 °C or even at 20 °C after 5-leaf stage. The obvious decrease in Chl levels, abnormal chloroplasts with few thylakoid lamella and abnormal thylakoids were observed for the albino mutant seedlings at low temperature, but the mutant was apparently normal green at high temperature, suggesting the thermo-sensitivity of albino phenotype. Genetic analysis showed that the albino phenotype was controlled by a single recessive nuclear gene (tcd9). The map-based cloning and molecular complementation tests revealed that the mutation of TCD9 encoding α subunit of Cpn60 protein (Cpn60α), localized in chloroplasts, was responsible for albino phenotype. In addition, TCD9 exhibited the high expression in all tested tissues, especially in young leaves. The transcriptional analysis indicated that all expression levels of the studied genes related to chloroplast development in tcd9 mutant were seriously affected in the albino seedlings at 20 °C, whereas some of them recovered into normal levels in green-seedlings at 32 °C. Our observations suggest that the nuclear-encoded Cpn60α protein TCD9 plays a crucial role in chloroplast development at early leaf stage of rice.