水稻显性窄叶突变体Dnal1的鉴定与基因定位

doi:10.3864/j.issn.0578-1752.2014.09.017

摘要/Abstract

摘要： 【目的】对一个新的水稻显性窄叶突变体进行鉴定，为水稻叶片形态建成的分子机理和株型育种研究奠定基础。【方法】甲基磺酸乙酯（EMS）诱变水稻籼型恢复系缙恢10号，获得一个窄叶突变体Dnal1，连续7代种植，表型均稳定遗传。开花期，调查Dnal1和野生型剑叶、倒2叶和倒3叶的叶宽及卷曲度，对剑叶最宽处进行石蜡切片分析；灌浆期，测量剑叶的光合色素含量，并利用Li-6400便携式光合测定仪测量光合速率、气孔导度和蒸腾速率。配制西农1A/Dnal1和Dnal1/中花11杂交组合，利用F1和F2群体进行遗传分析，并利用Dnal1/中花11的F2隐性群体进行基因定位。田间小区种植，设置2个种植密度，株行距分别为16.7 cm×26.72 cm和13.36 cm×20.04 cm，3次重复，成熟期考查株高、有效穗数、穗实粒数、结实率、千粒重、籽粒长和籽粒宽等主要农艺性状，并估算理论产量。【结果】Dnal1的叶片宽度全生育期内均极显著变窄，剑叶、倒2叶和倒3叶的叶宽分别为0.96、0.89和0.88 cm，与野生型的19.6、19.41和18.42 cm相比，降低了一半左右。Dnal1大维管束数量与缙恢10号相比无显著变化，小维管束数量则下降了44.13%，达极显著差异水平。缙恢10号相邻大维管束之间一般有6个小维管束，Dnal1则只有3个。Dnal1的叶片微卷，剑叶、倒2叶和倒3叶的卷曲度分别为11.2、12.1和11.8，野生型的叶片卷曲度为零。此外，Dnal1的叶片颜色略深于野生型，叶绿素a的含量略有升高，但未达到显著差异水平。光合生理测定表明，与野生型相比，Dnal1的水分利用率略有升高，光合速率和蒸腾速率略有下降，气孔导度则显著降低。除叶片性状变化外，Dnal1还表现籽粒变窄和茎秆变细，株高和籽粒长无明显变化。Dnal1的籽粒宽度为2.33 mm，与缙恢10号的2.78 mm相比，极显著下降，导致Dnal1的千粒重仅为野生型的73.18%，极显著下降。低密度种植条件下，Dnal1的理论产量显著低于野生型；高密度种植条件下，Dnal1的理论产量则显著高于野生型，较高的结实率和单株有效穗是Dnal1产量提高的主要原因。西农1A/Dnal1与Dnal1/中花11杂交组合的F1群体，剑叶的叶片宽度分别为0.99 cm和0.94 cm，与Dnal1的0.96 cm相比，无显著差异；F2群体中出现窄叶植株和宽叶植株两种类型，窄叶和宽叶单株出现的频率呈双峰分布，且窄叶和宽叶的分离比符合3﹕1，表明Dnal1的窄叶性状受1个显性主效基因调控。利用分子标记最终将DNAL1定位在第2染色体上，位于SSR标记RM13646和RM1307之间，物理距离为391 kb。【结论】Dnal1是一个EMS诱变获得的显性窄叶水稻突变体，基因定位在第2染色体391 kb的物理范围内，在高密度种植条件下具有增产潜力。

关键词: 水稻（Oryza sativa） , 显性窄叶 , 基因定位 , 光合系统 , 株型育种

Abstract: 【Objective】Leaf blade is the main location of photosynthesis in high plants. A moderate narrowing could enhance crop quality and yield by fascinating the utilization of solar energy and has been paid wide attentions in crop breeding. A novel dominant narrower leaf mutant has been identified and the results provided a foundation in the research of leaf morphological formation and plant type breeding in Oryza sativa.【Method】A narrow leaf mutant (Dnal1) was discovered from the progeny of indica restorer line Jinhui10 with seeds treated by ethyl methanesulfonate and the trait of narrow leaf has inherited steadily after seven generations’ self-fertility. The flag, second and third leaf blades were utilized for detecting leaf widths and rolled indexes at the blooming stage, meanwhile, the middle position of flag leaf was used for paraffin section. At the filling stage, the characteristics of chloroplast pigment, photosynthetic rate (Pn), stomatal conductance (Gs) and transpiration rate (Tr) were measured. The Dnal1 was crossed with indica line Xinong1A and japonica line Zhonghua11, respectively, and the F1 and F2 generations were used for genetic analysis. Additionally, gene mapping was performed based on the F2 recessive individuals of Dnal1/Zhonghua11. The Dnal1 and the wild type were cultivated in paddy field with the spacing in the rows and between the rows 13.36 cm×20.04 cm and 16.7 cm×26.72 cm, respectively. At the maturity stage, agronomic traits were analyzed and they were plant height, efficient panicle per plant, filled grain number per panicle, seed setting ratio, 1000-seed weight, seed length and seed width. At last, theory yield per 667m2 was estimated.【Result】The Dnal1 displayed narrower leaf blades in the life and the width of flag leaf, second leaf and third leaf were 0.96, 0.89 and 0.88 cm, respectively; correspondingly, those of the wild type were 19.6, 19.41 and 18.42 cm, respectively. The number of large vascular bundles in the mutant had no significant difference with those of the wild type and the number of small vascular bundles decreased by 44.13%, leading to the significant difference. In detail, the number of small vascular bundles between the large vascular bundles was six in the wild type, and three in the Dnal1. The leaf blades of Dnal1 displayed slightly curling and the rolled indexes of flag, second and third leaf blades were 11.2, 12.1 and 11.8, respectively, while those of the wild type were zero. In addition, the colour of leaf blades of Dnal1 was darker than those of wild type with a slight higher content of chloroplast a. The increases were detected for the values of the water use efficiency and the decrease was identified for the parameters of the photosynthetic rate, transpiration rate and stomata conductance in the Dnal1, while only the alteration of stomata conductance led to the level of significant difference between the Dnal1 and the wild type. The culm and seed of Dnal1 were narrower than those of the wild type, while no changing in plant height and seed length was detected. The seed width of Dnal1 was 2.33 mm, which was significantly lower than those of the wild type (2.78 mm), therefore, the 1000-seed weight decreased significantly in the Dnal1. The yield of per unit area in the Dnal1 reduced under the lower plant density while increased under the higher plant density than those of the wild type, which was caused by higher efficient panicle per plant and seed setting ratio. For the combinations of Xinong1A/ Dnal1 and Dnal1/Zhonghua11, the widths of F1 flag leaves were 0.99 cm and 0.94 cm, respectively, both have no significant differences with the wild type’s 0.96 cm; in the F2 generations, the plants could be classified as two groups according to the leaf width, the narrow-leaf individuals to those with broad leaves has no significant difference with the ratio of 3:1, suggesting that the narrow leaf trait of Dnal1 was controlled by a dominant nuclear gene. Finally, the DNAL1 was mapped on chromosome 2 between SSR marker RM13646 and RM1307 with the physical distance 391 kb in this paper. 【Conclusion】 Dnal1 was a dominant narrow leaf mutant coming from EMS-inducement and showed higher yield potential when cultivated under the higher planting density. The DNAL1 was mapped on chromosome 2 with 391 kb physical distance.

Key words: rice (Oryza sativa) , dominant narrow leaf , gene mapping , photosystems , plant type breeding

桑贤春, 林婷婷, 何沛龙, 王晓雯, 廖红香, 张孝波, 马玲, 何光华. 水稻显性窄叶突变体Dnal1的鉴定与基因定位[J]. 中国农业科学, 2014, 47(9): 1819-1827.

SANG Xian-Chun, LIN Ting-Ting, HE Pei-Long, WANG Xiao-Wen, LIAO Hong-Xiang, ZHANG Xiao-Bo, MA Ling, HE Guang-Hua. Identification and Gene Mapping of a Dominant Narrow Leaf Mutant Dnal1 in Rice (Oryza sativa)[J]. Scientia Agricultura Sinica, 2014, 47(9): 1819-1827.

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