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硫苷介导的白菜抵御甜菜夜蛾幼虫取食胁迫的分子机制

郝娇娇 马永华 陆彦池 许丽爱 朱祝军 郁有健

郝娇娇, 马永华, 陆彦池, 许丽爱, 朱祝军, 郁有健. 硫苷介导的白菜抵御甜菜夜蛾幼虫取食胁迫的分子机制[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20220172
引用本文: 郝娇娇, 马永华, 陆彦池, 许丽爱, 朱祝军, 郁有健. 硫苷介导的白菜抵御甜菜夜蛾幼虫取食胁迫的分子机制[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20220172
HAO Jiaojiao, MA Yonghua, LU Yanchi, XU Li’ai, ZHU Zhujun, YU Youjian. Molecular mechanism of glucosinolate-mediated Brassica campestris ssp. chinensis against feeding stress of Spodoptera exigua larvae[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20220172
Citation: HAO Jiaojiao, MA Yonghua, LU Yanchi, XU Li’ai, ZHU Zhujun, YU Youjian. Molecular mechanism of glucosinolate-mediated Brassica campestris ssp. chinensis against feeding stress of Spodoptera exigua larvae[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20220172

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硫苷介导的白菜抵御甜菜夜蛾幼虫取食胁迫的分子机制

doi: 10.11833/j.issn.2095-0756.20220172
基金项目: 国家自然科学基金面上资助项目(32072557,31572115),浙江省自然科学基金重点资助项目(LZ14C150001)
详细信息
    作者简介: 郝娇娇(ORCID: 0000-0001-5049-799x),从事园艺作物栽培生理与品质调控研究。E-mail: 2407605896@qq.com
    通信作者: 朱祝军(ORCID: 0000-0001-8551-7751 ),教授,从事园艺作物栽培生理与品质调控研究。E-mail: zhuzj@zafu.edu.cn
  • 中图分类号: S603

Molecular mechanism of glucosinolate-mediated Brassica campestris ssp. chinensis against feeding stress of Spodoptera exigua larvae

  • 摘要:   目的  探究甜菜夜蛾Spodoptera exigua幼虫取食对白菜Brassica campestris ssp. chinensis硫代葡萄糖苷质量摩尔浓度及组分的影响,初步明确硫苷介导的白菜抵御甜菜夜蛾幼虫取食胁迫的分子机制。  方法  使用高效液相色谱法检测甜菜夜蛾幼虫取食对白菜硫苷质量摩尔浓度和组分的影响,实时荧光定量聚合酶链式反应(RT-qPCR)分析硫苷代谢关键基因表达模式的变化,谷胱甘肽硫转移酶试剂盒检测取食白菜后甜菜夜蛾幼虫体内谷胱甘肽硫转移酶活性的变化。  结果  白菜中8种硫苷组分在甜菜夜蛾幼虫取食后显著增加(P<0.05),其中吲哚族的4-羟基吲哚-3-甲基硫苷(4OH)、1-甲氧基-3-吲哚基甲基硫苷(NEO)、3-吲哚基甲基(GBC)和脂肪族的4-戊烯基硫苷(GBN)对甜菜夜蛾幼虫取食反应最为强烈;RT-qPCR结果显示:甜菜夜蛾幼虫取食后,白菜脂肪族硫苷合成相关基因BcBCAT4、BcMAM1的显著上调可能与脂肪族硫苷的增加有关(P<0.05),而BcSOT16的上调与4OH、NEO、GBC呈正相关;取食后,甜菜夜蛾幼虫体内谷胱甘肽硫转移酶活性显著增强(P<0.05),并呈现与总硫苷和吲哚族硫苷质量摩尔浓度变化规律的一致性。  结论  甜菜夜蛾幼虫的取食能诱导白菜硫苷的合成,而硫苷质量摩尔浓度的升高会诱导幼虫体内谷胱甘肽硫转移酶活性的增强。甜菜夜蛾幼虫取食胁迫下,白菜BcSOT16、BcBCAT4、BcMAM1等硫苷合成关键基因被激活,4OH、NEO、GBC和GBN显著增加,从而更好地抵御甜菜夜蛾幼虫取食胁迫。图2表3参44
  • 图  1  甜菜夜蛾幼虫取食对白菜硫苷代谢关键基因表达模式的影响

    不同字母表示不同的时间间差异显著(P<0.05)

    Figure  1  Effects of S. exigua larvae feeding on expression patterns of key genes related to glucosinolates metabolism in B. campestris ssp. chinensis

    图  2  取食白菜后甜菜夜蛾幼虫GSTs活性的变化     

    不同小写字母表示不同时间间差异显著(P<0.05)

    Figure  2  Changes of GSTs activity in S. exigua larve after feeding on B. campestris ssp. chinensis

    表  1  实时荧光定量PCR(RT-qPCR)表达分析引物序列

    Table  1.   Primer sequences for RT-qPCR analysis

    基因名称上游引物(5 →3 )下游引物(5 →3 )
    BcMAM1 CCAGAGTACATACCGCACAA AGAGGACACCGGAAAACCAA
    BcCYP83A1 AGTTCTCCTCTTCTTCCTCT CCATTGTTTGACTTCCTATC
    BcCYP83B1 ACACTTCCTCTTTCGTCTCT CATCGTTTGTTGCCCCTTCA
    BcMYB28 GAGAATTTGCATTCCCTTGC TGGTGTCCCATCTTTGTTGG
    BcSOT16 GGGTTATGGGTTTAGTGCTG CTCCAACCTTCCCTTTCCTA
    BcUBC10 GGGTCCTACAGACAGTCCTTAC ATGGAACACCTTCGTCCTAAA
    BcAPK2 CAACACCGTCTGGGATCTGC AACCGACATCGACACCTGGA
    BcSUR1 GGCGTTATCTACATGCTGTTCG CAGGTGCGGAAGCAAGGGTA
    BcAOP2 TGGATTTGCACCAAAGGAAA AGATAGATCACTGGAAGTTG
    BcBCAT4 AAGCAACTCGACTCAAACACT CGATAAAACCCGAATCCTAAT
    下载: 导出CSV

    表  2  白菜硫苷组分

    Table  2.   Components of glucosinolate in B. campestris ssp. chinensis

    化合物名称缩写所属类别分子量保留时间/min
    2-羟基-3-丁烯基硫苷progoitrin PRO 脂肪族 388 7.790
    3-丁烯基硫苷gluconapin GNA 脂肪族 372 14.480
    4-羟基吲哚-3-甲基硫苷4-OH-glucobrassicin 4OH 吲哚族 463 17.067
    4-戊烯基硫苷glucobrassicanapin GBN 脂肪族 386 22.087
    3-吲哚基甲基硫苷glucobrassicin GBC 吲哚族 447 26.753
    2-苯乙基硫苷gluconasturtiin NAS 芳香族 422 31.497
    4-甲氧基-3-吲哚基甲基硫苷4-methoxy-glucobrassicin 4ME 吲哚族 477 32.433
    1-甲氧基-3-吲哚基甲基硫苷1-methoxy-glucobrassicin NEO 吲哚族 477 40.793
    下载: 导出CSV

    表  3  甜菜夜蛾幼虫取食对白菜硫苷质量摩尔浓度的影响

    Table  3.   Effects of feeding on the content of glucosinolate in B. campestris ssp. chinensis by S. exigua larvae

    类型化合物缩写硫苷质量摩尔浓度/(µmol·g−1)
    0 (CR)1244872 h
    脂肪族GNA0.154±0.010 b0.174±0.005 b0.220±0.003 c0.132±0.003 a0.132±0.002 a
    PRO0.152±0.003 a0.150±0.002 a0.176±0.001 b0.182±0.002 c0.179±0.001 bc
    GBN0.434±0.032 a0.466±0.009 a0.550±0.018 b0.819±0.004 c0.781±0.001 c
    总脂肪族 0.724±0.009 a0.792±0.001 b0.959±0.008 c1.132±0.002 e1.116±0.004 d
    吲哚族4OH0.157±0.012 a0.177±0.003 b0.222±0.004 c0.257±0.006 d0.234±0.004 c
    GBC0.044±0.013 a0.062±0.000 b0.105±0.003 c0.130±0.001 d0.209±0.005 e
    4ME0.089±0.026 a0.69±0.000 a0.140±0.003 b0.154±0.002 b0.135±0.000 b
    NEO0.054±0.011 a0.067±0.001 b0.092±0.001 c0.162±0.003 d0.263±0.001 e
    总吲哚族0.351±0.001 a0.375±0.001 b0.565±0.005 c0.703±0.010 d0.840±0.008 e
    芳香族NAS0.096±0.004 ab0.109±0.002 c0.142±0.001 d0.086±0.001 a0.101±0.002 bc
    总硫苷1.154±0.003 a1.277±0.002 b1.664±0.014 c1.921±0.011 d2.032±0.014 e
      说明:同行不同小写字母表示同一指标不同时间差异显著(P<0.05)
    下载: 导出CSV
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    https://zlxb.zafu.edu.cn/article/doi/10.11833/j.issn.2095-0756.20220172

    https://zlxb.zafu.edu.cn/article/zjnldxxb/2022//1

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  • 文章访问数:  27
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出版历程
  • 收稿日期:  2022-02-23
  • 录用日期:  2022-09-14
  • 修回日期:  2022-09-05

硫苷介导的白菜抵御甜菜夜蛾幼虫取食胁迫的分子机制

doi: 10.11833/j.issn.2095-0756.20220172
    基金项目:  国家自然科学基金面上资助项目(32072557,31572115),浙江省自然科学基金重点资助项目(LZ14C150001)
    作者简介:

    郝娇娇(ORCID: 0000-0001-5049-799x),从事园艺作物栽培生理与品质调控研究。E-mail: 2407605896@qq.com

    通信作者: 朱祝军(ORCID: 0000-0001-8551-7751 ),教授,从事园艺作物栽培生理与品质调控研究。E-mail: zhuzj@zafu.edu.cn
  • 中图分类号: S603

摘要:   目的  探究甜菜夜蛾Spodoptera exigua幼虫取食对白菜Brassica campestris ssp. chinensis硫代葡萄糖苷质量摩尔浓度及组分的影响,初步明确硫苷介导的白菜抵御甜菜夜蛾幼虫取食胁迫的分子机制。  方法  使用高效液相色谱法检测甜菜夜蛾幼虫取食对白菜硫苷质量摩尔浓度和组分的影响,实时荧光定量聚合酶链式反应(RT-qPCR)分析硫苷代谢关键基因表达模式的变化,谷胱甘肽硫转移酶试剂盒检测取食白菜后甜菜夜蛾幼虫体内谷胱甘肽硫转移酶活性的变化。  结果  白菜中8种硫苷组分在甜菜夜蛾幼虫取食后显著增加(P<0.05),其中吲哚族的4-羟基吲哚-3-甲基硫苷(4OH)、1-甲氧基-3-吲哚基甲基硫苷(NEO)、3-吲哚基甲基(GBC)和脂肪族的4-戊烯基硫苷(GBN)对甜菜夜蛾幼虫取食反应最为强烈;RT-qPCR结果显示:甜菜夜蛾幼虫取食后,白菜脂肪族硫苷合成相关基因BcBCAT4、BcMAM1的显著上调可能与脂肪族硫苷的增加有关(P<0.05),而BcSOT16的上调与4OH、NEO、GBC呈正相关;取食后,甜菜夜蛾幼虫体内谷胱甘肽硫转移酶活性显著增强(P<0.05),并呈现与总硫苷和吲哚族硫苷质量摩尔浓度变化规律的一致性。  结论  甜菜夜蛾幼虫的取食能诱导白菜硫苷的合成,而硫苷质量摩尔浓度的升高会诱导幼虫体内谷胱甘肽硫转移酶活性的增强。甜菜夜蛾幼虫取食胁迫下,白菜BcSOT16、BcBCAT4、BcMAM1等硫苷合成关键基因被激活,4OH、NEO、GBC和GBN显著增加,从而更好地抵御甜菜夜蛾幼虫取食胁迫。图2表3参44

English Abstract

郝娇娇, 马永华, 陆彦池, 许丽爱, 朱祝军, 郁有健. 硫苷介导的白菜抵御甜菜夜蛾幼虫取食胁迫的分子机制[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20220172
引用本文: 郝娇娇, 马永华, 陆彦池, 许丽爱, 朱祝军, 郁有健. 硫苷介导的白菜抵御甜菜夜蛾幼虫取食胁迫的分子机制[J]. 浙江农林大学学报. doi: 10.11833/j.issn.2095-0756.20220172
HAO Jiaojiao, MA Yonghua, LU Yanchi, XU Li’ai, ZHU Zhujun, YU Youjian. Molecular mechanism of glucosinolate-mediated Brassica campestris ssp. chinensis against feeding stress of Spodoptera exigua larvae[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20220172
Citation: HAO Jiaojiao, MA Yonghua, LU Yanchi, XU Li’ai, ZHU Zhujun, YU Youjian. Molecular mechanism of glucosinolate-mediated Brassica campestris ssp. chinensis against feeding stress of Spodoptera exigua larvae[J]. Journal of Zhejiang A&F University. doi: 10.11833/j.issn.2095-0756.20220172

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