Abstract
Green-revertible albino mutants are important sources for studying chloroplast structure, chloroplast development, chlorophyll biosynthesis, and plant photosynthesis. In the present study, we characterized a green-revertible albino mutant gra(k), which was obtained from the tissue-cultured rice Kitaake. The mutant gra(k) exhibited albino on its first three leaves. The leaf color started to turn green at the four-leaf stage. The chlorophyll contents were deeply reduced at the seedling stage, and the chloroplast development was delayed in gra(k). The green-revertible albino (gra) phenotype of the mutant gra(k) was temperature dependent. The main agronomic traits, including plant height, tilling number per plant, seed setting rate, and thousand-grain weight, slightly decreased in gra(k) comparing to those in the wild-type Kitaake. Genetic analysis showed that the gra phenotype was controlled by a single recessive nucleic gene. By using 5,168 recessive F 2 individuals derived from the cross of gra(k) × Jodan, the locus of the gene Gra(k) was delimited in a DNA region of 200 kb between the makers B-31 and P11 on chromosome 5. Sequencing analysis indicated that the three functionally annotated genes, LOC_Os05g23700, LOC_Os05g23720, and LOC_Os05g23740, were all deleted in the 200 kb region in the mutant gra(k). Transgenic test revealed that the gra(k) plants over-expressing LOC_Os05g23740CDS were restored to normal green as the wild-type Kitaake. Our results proved that the deletion of the DnaK protein gene LOC_Os05g23740 (encoding the chaperon protein OsHsp70CP1) led to the gra phenotype in the mutant gra(k).
摘要
白化转绿突变体是研究叶绿体结构、发育、叶绿素合成和植物光合作用的重要材料。白化转绿突变体gra(k)在三叶期前均表现白化,并受高温诱导,第四叶时开始转绿。突变体gra(k)的株高、分蘖数、结实率和千粒重等农艺性状与野生型差异不大。我们利用图位克隆策略成功克隆了Gra(k)基因。转基因实验发现过量表达LOC_Os05g23740基因能够使突变体gra(k)白化表型恢复成野生型Kitaake苗期的叶片颜色。我们的研究证实突变体gra(k)的白化转绿表型是由编码DnaK蛋白的基因LOC_Os05g23740的缺失引起的。
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (31171622, 31371705, 3140135, 31501627), the “Hundred Talents Plan” Foundation of Sichuan, and the Specialized Research Funds for Doctoral Program of Higher Education (20125103120011, 20135103120004), and the Key Project of Sichuan Education Department (15ZA0020). We thank Mr. Guo Yang from Sichuan University for the help on transmission electron microscopy.
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Weitao Li, Junjie Yin and Bangquan Ye contributed equally to this work.
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Li, W., Yin, J., Ye, B. et al. Deletion of a DnaK protein gene causes seedling green-revertible albino by retarding chloroplast development in rice. Sci. Bull. 60, 2054–2058 (2015). https://doi.org/10.1007/s11434-015-0897-6
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DOI: https://doi.org/10.1007/s11434-015-0897-6