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杨树PsnGA20ox1过表达对烟草叶片发育的影响

刘露露 鲁婷婷 王爽 李美靓 赵双菁 刘莹莹 魏志刚

刘露露, 鲁婷婷, 王爽, 李美靓, 赵双菁, 刘莹莹, 魏志刚. 杨树PsnGA20ox1过表达对烟草叶片发育的影响[J]. 北京林业大学学报, 2018, 40(2): 22-30. doi: 10.13332/j.1000-1522.20170245
引用本文: 刘露露, 鲁婷婷, 王爽, 李美靓, 赵双菁, 刘莹莹, 魏志刚. 杨树PsnGA20ox1过表达对烟草叶片发育的影响[J]. 北京林业大学学报, 2018, 40(2): 22-30. doi: 10.13332/j.1000-1522.20170245
Liu Lu-lu, Lu Ting-ting, Wang Shuang, Li Mei-liang, Zhao Shuang-jing, Liu Ying-ying, Wei Zhi-gang. Effects of poplar PsnGA20ox1 overexpression on leaf development of tobacco[J]. Journal of Beijing Forestry University, 2018, 40(2): 22-30. doi: 10.13332/j.1000-1522.20170245
Citation: Liu Lu-lu, Lu Ting-ting, Wang Shuang, Li Mei-liang, Zhao Shuang-jing, Liu Ying-ying, Wei Zhi-gang. Effects of poplar PsnGA20ox1 overexpression on leaf development of tobacco[J]. Journal of Beijing Forestry University, 2018, 40(2): 22-30. doi: 10.13332/j.1000-1522.20170245

杨树PsnGA20ox1过表达对烟草叶片发育的影响

doi: 10.13332/j.1000-1522.20170245
基金项目: 

黑龙江省留学归国基金项目 LC201512

国家自然科学基金项目 31770640

详细信息
    作者简介:

    刘露露。主要研究方向:植物次生细胞壁。Email: 1398314698@qq.com 地址:150040黑龙江省哈尔滨市香坊区和兴路26号东北林业大学林学院

    责任作者:

    魏志刚,教授。主要研究方向:植物次生细胞壁。Email:zhigangwei@nefu.edu.c 地址:同上

  • 中图分类号: S718.46;S792.11

Effects of poplar PsnGA20ox1 overexpression on leaf development of tobacco

  • 摘要: 目的赤霉素(Gibberellins, GA)参与植物多个生长发育过程,然而赤霉素在植物叶片发育中的生物学功能尚无研究报道。GA20ox是GAs生物合成途径中关键限速酶(GA20-oxidases)的编码基因,本项研究目的是揭示PsnGA20ox1在植物叶片发育中的生物学功能。方法本项研究分析了PsnGA20ox1在烟草中过表达后对烟草叶片大小、超微结构、生物量、叶绿素及生物活性GAs含量,以及细胞分裂与气孔发育相关的基因表达的影响。结果PsnGA20ox1过表达烟草叶片变大、叶绿素与GA4含量上升,叶片表皮细胞变小。而且,转基因烟草叶片栅栏组织与海绵组织厚度、表皮细胞与气孔数目、光合作用和叶片生物量与野生型烟草相比均显著增加。此外,转基因烟草叶片气孔与表皮细胞发育相关基因表达也发生了显著变化,而且变化趋势与叶片表型的改变相吻合。结论PsnGA20ox1过表达影响植物叶片的结构与功能,从而提高了叶片的生物量。

     

  • 图  1  PsnGA20ox1在小黑杨不同组织中表达模式

    A.顶端分生组织; B.初生区叶片; C.过渡区叶片; D.次生区叶片; E.韧皮部; F.过渡区木质部; G.次生木质部;H.次生区节间。

    Figure  1.  PsnGA20ox1 expression patterns in different tissues of Populous simonii×Populus nigra

    A, top meristem; B, newborn leaves; C, transition zone leaves; D, secondary zone leaves; E, phloem; F, transition zone xylem; G, secondary xylem; H, secondary zone internals.

    图  2  转基因烟草PsnGA20ox1表达量、GAs含量与表型

    WT为野生型烟草;L为转基因株系;L2、L5、L7、L11、L16为PsnGA20ox1过表达烟草的5个株系。下同。A.3个月生烟草表型; B.1个月生烟草功能叶片表型; C.PsnGA20ox1相对表达量; D.1个月生烟草功能叶片中GAs含量; E.3个月生烟草叶片长和宽; F.3个月生烟草叶片鲜质量与干质量。

    Figure  2.  PsnGA20ox1 expression level, GAs content and phonotype of tobacco

    WT is wild type tobacco; L is transgenic line; L2, L5, L7, L11 and L16 are PsnGA20ox1 overexpressing tobacco. The same below. A, 3-month-old tobacco phenotype; B, 1-month tobacco leaf functional phenotype; C, PsnGA20ox1 relative expression level; D, 1-month tobacco leaf functional GAs content; E, the length and width of 3-month-old tobacco leaves; F, the fresh mass and dry mass of 3-month-old tobacco leaves.

    图  3  过表达PsnGA20ox1对烟草叶片超微结构影响

    A、C、E.野生型烟草; B、D、F.PsnGA20ox1过表达烟草;A、B.放大100倍; C、D.放大300倍; E、F.放大1000倍。

    Figure  3.  Effects of PsnGA20ox1 overexpression on leaf ultrastructure of tobacco

    A, C, E, wild type tobacco; B, D, F, PsnGA20ox1 overexpression tobacco; A, B, magnification 100 times; C, D, magnification 300 times; E, F, magnification 1000 times.

    图  4  PsnGA20ox1过表达对烟草叶绿素含量与光合作用影响

    A.PsnGA20ox1过表达与野生型烟草叶绿素含量; B.PsnGA20ox1过表达与野生型烟草光合速率。

    Figure  4.  Effects of PsnGA20ox1 overexpression on chlorophyll and photosynthetic rate of tobacco

    A, PsnGA20ox1 overexpression and wild-type tobacco chlorophyll content; B, PsnGA20ox1 overexpression and wild-type tobacco photosynthetic rate.

    图  5  过表达PsnGA20ox1对烟草表皮与气孔发育相关基因表达影响

    Figure  5.  Effects of PsnGA20ox1 overexpression on the expression level of genes involved in epidermal cell and stomatal development of tobacco

    表  1  烟草气孔发育相关基因qRT-PCR引物

    Table  1.   qRT-PCR primers of genes involved in stomatal and epidermal cell development of tobacc

    基因名
    Gene name
    登录号
    Accession No.
    引物名称
    Primer name
    引物序列(5′-3′)
    Primer sequence(5′-3′)
    Actin2 AB158612.1 Actin-S
    Actin-A
    GATCTTGCTGGTCGTGATCT
    ACTTCCGGACATCTGAACCT
    ER JQ267739.1 ER-S
    ER-A
    GAAGGTTTTGGTAGTTACACGTAGC
    AGGGTGAGTCGTTGTCTAGAAGG
    ERL1 XM_009764054.1 ERL1-S
    ERL1-A
    GACTCCATTGCTTTTTGTTCTTAA
    CTCAGAAGTAAAATCCCAAGAAGC
    ERL2 XM_009785106.1 ERL2-S
    ERL2-A
    CAGGCTTTCAGGTACTCTACCATC
    GCCCTCTGAGTCTTGAAGAATTTG
    MPK3 XM_009804071.1 MPK3-S
    MPK3-A
    TGCTCGGTGTTGAATACGG
    GGTCTAAATGGCGGAGGAG
    MKK4 XM_009760486.1
    MPK4-S
    MPK4-A
    GCCTTCAGATGTGCCGTG
    CCCTTCGAGAGAGCCTTTATC
    EPF1 XM_009760610.1 EPF1-S
    EPF1-A
    GTGATGATATTGGCGGCG
    ACTTGTAGTTTCTGGCATCGG
    EPF2 XM_009791751.1 EPF2-S
    EPF2-A
    TTACCAGATTGTTCCCATGC
    TCAATTGGAAGGGACATGG
    STOMAGEN XM_009767439.1 STOMAGEN-S
    STOMAGEN-A
    ATGAGGGAAAAAGATGGGTG
    CCTCCACTGGGACTTGCTC
    SPCH XP_009801585.1 SPCH-F
    SPCH-R
    GTTCTACAATCTCTGGAGGCG
    TGGACTTTCAGGTGTGGGAC
    SCRM XP_009769541.1 SCRM-F
    SCRM-R
    GCAGCAGATGTAGACTGGTTCA
    AGTTACTATTGTTTTCAGTTTCGG
    SCRM2 XP_009769541.1 SCRM2-F
    SCRM2-R
    GCAAAACCAAAACCATACTGAC
    GAAGCTGAAGATGAAGATGAGG
    MUTE XP_009765536.1 MUTE-F
    MUTE-R
    TGCACCACAAACTGGCTTC
    GTTCCCCGTGGCCCTC
    FAMA XP_009795997.1 FAMA-F
    FAMA-R
    ACAACAACAACAACAACACGAG
    TTCCTCTGATACCTTGGCTTG
    CYCD2;1 XP_009789780.1 CYCD2;1-F
    CYCD2;1-R
    TGGAGTTTTTGCCTAAAGATG
    TCAAAATCCAATCAAGAGCC
    CYCD3;1 XP_009788121.1 CYCD3;1-F
    CYCD3;1-R
    TATTTTGGGAAGATGAAGAGC
    CACAGAATCAACACGGGC
    下载: 导出CSV
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