水稻淡绿叶基因PGL11的鉴定与精细定位
收稿日期: 2017-04-11
修回日期: 2017-05-12
网络出版日期: 2017-09-10
基金资助
中央级公益性科研院所专项;中国农科院科技创新工程资助项目;国家自然科学基金资助项目(31661143006,91435105)
Identification and Fine Mapping of Pale Green Leaf PGL11 in Rice
Received date: 2017-04-11
Revised date: 2017-05-12
Online published: 2017-09-10
目的 叶片是水稻进行光合作用的主要场所,叶片颜色的变化与水稻的生长发育直接相关。发掘水稻叶色突变体,是水稻功能基因组学研究的重要遗传基础。方法 利用EMS诱变日本晴获得一个能稳定遗传的淡绿叶突变体,暂命名为pgl11(pale green leaf 11)。在不同生育期测定野生型与突变体的叶绿素含量。在苗期,取野生型与突变体叶片进行叶绿体结构的透射电镜观察。在分蘖期,测定野生型与突变体的光合参数并观察气孔结构。在成熟期,测定野生型和pgl11的主要农艺性状。以pgl11为母本,南京6号为父本构建相应的F2群体,采用图位克隆的方法,对该基因进行定位。结果 从苗期开始,突变体pgl11的每一片新叶均表现为淡绿色,叶绿素含量显著降低,叶绿体发育异常。随着叶片的生长,叶色由淡绿逐渐转绿,至抽穗期时叶绿素含量亦无明显差异。pgl11还表现光合速率、气孔导度明显下降,胞间CO2浓度上升。扫描电镜观察发现,突变体pgl11的气孔发育异常。与野生型相比,突变体的农艺性状如株高、剑叶宽、二次枝梗数、每穗粒数、粒长、粒宽、千粒重以及结实率等均显著降低。对叶绿素合成、光合作用以及质体发育相关基因的表达量测定表明,突变体pgl11中参与叶绿体转录和翻译相关基因的表达量显著升高,而叶绿素合成和光合作用相关基因的表达量显著下降。遗传分析表明,该突变表型受一对隐性核基因控制。通过图位克隆的方法将该基因定位于第1染色体上的C6和C8标记之间,物理距离约为110 kb。结论 该定位区间内未见有叶色相关基因报道,推测PGL11基因可能是一个新的水稻叶色基因。
涂政军, 邹国兴, 黄李超, 陈龙, 代丽萍, 高易宏, 冷语佳, 朱丽, 张光恒, 胡江, 任德勇, 高振宇, 董国军, 陈光, 郭龙彪, 钱前, 曾大力 . 水稻淡绿叶基因PGL11的鉴定与精细定位[J]. 中国水稻科学, 2017 , 31(5) : 489 -499 . DOI: 10.16819/j.1001-7216.2017.7043 489
【Objective】 Leaf is the main site of photosynthesis in rice. The change of leaf color is directly related to the growth and development of rice. The research of leaf color mutants is an important genetic basis for the study of rice functional genomics. 【Method】We identified a pale green leaf mutant termed pgl11(pale green leaf 11) from japonica cultivar Nipponbare by ethyl methylsulfonate (EMS) treatment. The chlorophyll content of wild type(WT) and mutant was measured at different growth stages. At the seedling stage, the chloroplast structure of the leaves of the WT and the mutant were observed with a transmission electron microscopy. At the tillering stage, the photosynthetic parameters of WT and pgl11 were measured and the stomatal structure was observed. At the mature stage, the main agronomic traits of WT and pgl11 were determined. F2 population derived from pgl11/Nanjing 6 was used to map this gene by position cloning approach.【Result】At seedling stage, every new leaf of the mutant pgl11 was pale green. As the leaves matured, the leaf color gradually turned green. Compared with wild type, the chlorophyll content in pgl11 decreased at seedling stage. However, there was no significant difference at heading stage. In addition, the photosynthetic rate and stomatal conductance of pgl11 were significantly decreased, but the intercellular CO2 concentration was apparently increased. The stomata was abnormal in pgl11. The agronomic traits including plant height, flag leaf width, secondary rachis branch number, grain number per panicle, grain length, grain width, 1000-grain weight and seed setting rate decreased significantly in pgl11 compared with wild type. The expression of genes associated with chloroplast transcription and translation were upregulated in pgl11, while the expression of chlorophyll synthesis and photosynthesis related genes were downregulated. Genetic analysis showed that the mutant phenotype was controlled by a single recessive nuclear gene, and PGL11 was mapped to a 110kb region between the markers C6 and C8 on the short arm of chromosome 1. 【Conclusion】It was suggested that PGL11 gene would be a putative novel pale green leaf gene.
Key words： rice; pale green leaf; chloroplast; genetic analysis; gene mapping.
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