烟草叶形突变体的形态特征与关键基因表达差异研究

doi:10.13304/j.nykjdb.2019.0721

中国农业科技导报 ›› 2020, Vol. 22 ›› Issue (3): 38-45.DOI: 10.13304/j.nykjdb.2019.0721

烟草叶形突变体的形态特征与关键基因表达差异研究

周蕾1,2,张彦1,2,张娟1,2,吴晓颖1,2,马兴华1*

1.中国农业科学院烟草研究所，农业农村部烟草生物学与加工重点实验室, 山东青岛 266101; 2.中国农业科学院研究生院, 北京 100081

收稿日期:2019-09-02 出版日期:2020-03-15 发布日期:2019-10-10
通讯作者: *通信作者马兴华 E-mail：maxinghua@caas.cn
作者简介:周蕾 E-mail：1332696786@qq.com；
基金资助:
中国农业科学院基本科研业务费所级统筹项目（1610232017008）;中国农业科学院科技创新工程（ASTIP-TRIC03）;中国烟草总公司山东省公司重点科技项目（201803）。

Differences of Morphological Characterization and Key Genes Expression in Tobacco Leaf Shape Mutants

ZHOU Lei1,2, ZHANG Yan1,2, ZHANG Juan1,2, WU Xiaoying1,2, MA Xinghua1*

1.Key Laboratory of Tobacco Biology and Processing, Ministry of Agriculture and Rural Affairs; Institute of Tobacco Research, Chinese Academy of Agricultural Sciences, Shandong Qingdao 266101, China; 2.Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China

Received:2019-09-02 Online:2020-03-15 Published:2019-10-10

摘要/Abstract

摘要： 为分析烟草叶片大小差异的原因，以‘红花大金元’野生型和两个叶片突变体（突变体1、突变体2）为材料，对叶片形态、解剖结构以及叶发育关键基因的表达差异进行研究。结果表明：突变体1的叶长、叶宽、叶面积分别为野生型的61.88%、25.87%、10.12%，比叶重是野生型的1.91倍；突变体2的叶长、叶宽、叶面积分别为野生型的78.82%、106.63%、83.92%，比叶重是野生型的1.35倍。突变体1同一层海绵组织细胞总数为野生型的10.93%，海绵组织细胞体积较野生型减小，叶片厚度显著增加，增幅为23.15%；突变体2同一层海绵组织细胞总数为野生型的48.12%，海绵组织细胞体积与野生型相比增加，叶片厚度显著增加，增幅为18.03%。基因NtARF2-1、NtDA1、NtTOR1、NtARF10-1、NtEBP1、NtGRF8、NtGRF16在突变体1中的表达量与野生型相比差异显著，其中NtEBP1上调表达最显著，上调2.22倍；NtGRF8、NtGRF16下调最显著，表达量分别为野生型的16.06%、13.07%。与野生型相比，NtTOR1、NtTOR2、NtARF10-1、NtEBP1的表达量在突变体2中显著上调，其中NtTOR1表达量上调最显著，上调3.17倍；NtARF2-1、NtARF2-2、NtDA1、NtGRF8、NtGRF16的表达量在突变体2中显著下调， NtGRF8、NtGRF16下调最显著，表达量分别为野生型的13.43%、21.68%。以上结果表明，突变体1与突变体2叶面积的减小与细胞总数的减少密切相关，突变体1、突变体2叶片细胞大小与数目的改变可能与基因NtTOR1、NtEBP1、NtGRF8、NtGRF16表达有关。

关键词: 烟草, 叶片突变体, 叶片解剖, 关键基因表达

Abstract: In order to analyze the differences in tobacco leaf size, the morphological and anatomical changes, and some key leaf developmental genes were examined in wild type tobacco (Honghua Dajinyuan) and two leaf mutants (mutant 1, mutant 2). Results showed that the leaf length, leaf width and leaf area of mutant 1 was 61.88%, 25.87%, 10.12% of wild type respectively, while its LMA was 1.91 times more than that of wild type. Similarly, the leaf length, leaf width and leaf area of mutant 2 was 78.82%, 106.63%, and 83.92% of the wild type respectively, and its LMA was 1.35 times greater than that of the wild type tobacco. The total number of sponge tissue cells in the same layer of mutant 1 was 10.93% of the wild type with a reduction in the volume of sponge tissue cells as compared with the wild type. The leaf thickness of mutant 1 was significantly increased by 23.15% of that of the wild type. Similarly, we also witnessed 18.03% increment in the leaf thickness of the mutant 2 when compared with the wild type tobacco plant. Reduction in the number of spongy tissues cells of mutant 2 was also observed as the total number of the spongy tissue cells were only 48.12% of that of the wild type, and the volume of spongy tissue cells was larger than wild type. The expression of NtARF2-1, NtDA1, NtTOR1, NtARF10-1, NtEBP1, NtGRF8 and NtGRF16 genes in mutant 1 was significantly different compared with the wild type. NtEBP1 was 2.22 times up-regulated significantly, while NtGRF8 and NtGRF16 were down-regulated and its expression levels were 16.06% and 13.07% of the wild type, respectively. Similarly, NtTOR1, NtTOR2, NtARF10-1, and NtEBP1 were significantly up-regulated in mutant 2 in comparison with the wild type in which the up-regulation of NtTOR1 was 3.17 times more among the other genes expression. The expression of NtARF2-1, NtARF2-2, NtDA1, NtGRF8 and NtGRF16 was significantly down-regulated in mutant 2. NtGRF8 and NtGRF16 were the most significantly down-regulated genes and its expression levels were 13.43% and 21.68%, respectively, of that of the wild type. The above results indicated that the decrease in the leaf area of mutant 1 and mutant 2 was closely related to the decrease in the total cell number. The changes in the size and number of leaf cells of mutant 1 and mutant 2 maybe related to the expression of NtTOR1, NtEBP1, NtGRF8 and NtGRF16 genes.

Key words: tobacco, leaf mutant, leaf anatomy, key gene expression

周蕾1,2,张彦1,2,张娟1,2,吴晓颖1,2,马兴华1*. 烟草叶形突变体的形态特征与关键基因表达差异研究[J]. 中国农业科技导报, 2020, 22(3): 38-45.

ZHOU Lei1,2, ZHANG Yan1,2, ZHANG Juan1,2, WU Xiaoying1,2, MA Xinghua1*. Differences of Morphological Characterization and Key Genes Expression in Tobacco Leaf Shape Mutants[J]. Journal of Agricultural Science and Technology, 2020, 22(3): 38-45.

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烟草叶形突变体的形态特征与关键基因表达差异研究

Differences of Morphological Characterization and Key Genes Expression in Tobacco Leaf Shape Mutants

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