林业科学 ›› 2021, Vol. 57 ›› Issue (3): 108-116.doi: 10.11707/j.1001-7488.20210311
周心怡,闫丽琼,吕云彤,孙丽丽,朱靖闻,曹传旺*
收稿日期:
2019-12-24
出版日期:
2021-03-01
发布日期:
2021-04-07
通讯作者:
曹传旺
基金资助:
Xinyi Zhou,Liqiong Yan,Yuntong Lü,Lili Sun,Jingwen Zhu,Chuanwang Cao*
Received:
2019-12-24
Online:
2021-03-01
Published:
2021-04-07
Contact:
Chuanwang Cao
摘要:
目的: 探究舞毒蛾取食对小黑杨苯丙氨酸代谢途径中关键酶诱导防御的影响,为明确杨树抗虫性分子机制提供理论依据,并为利用基因工程手段提高寄主抗虫性提供基因材料。方法: 通过RT-PCR法获得小黑杨C4H和4CL基因全长序列,并进行基因特性分析;采用邻接法构建C4H和4CL系统进化树;利用分光光度计法比较舞毒蛾取食和机械损伤处理下小黑杨C4H和4CL活性,并运用实时荧光定量RT-PCR技术测定2种损伤处理对C4H和4CL基因表达量的影响。结果: 从小黑杨转组文库中克隆鉴定获得2个属于p450家族的C4H基因(命名为C4H-1和C4H-2)以及1个多结构域蛋白超家族(AFD_class_I)4CL基因,全长基因开放阅读框大小为1 518~1 740 bp,编码505~579个氨基酸,蛋白质分子质量为58.00~63.03 kDa,理论等电点pI为5.63~9.09。系统进化树分析表明,小黑杨C4H-1与银白杨亲缘关系近聚为一类;而C4H-2与毛白杨、欧洲山杨、毛果杨×美洲黑杨和毛果杨具有较高同源性聚为一类,小黑杨4CL与毛果杨×美洲黑杨和毛果杨亲缘关系较近。舞毒蛾取食诱导小黑杨C4H-1、C4H-2和4CL基因表达量增加,分别为对照组的6.86、1.15和1.50倍;而机械损伤诱导C4H-1基因表达上调,抑制C4H-2和4CL基因表达下调,分别为对照组的1.80、0.71和0.60倍。取食和机械损伤均能诱导小黑杨C4H和4CL活性增加,且取食诱导显著高于机械损伤。结论: 舞毒蛾取食显著诱导小黑杨C4H和4CL活性增加,可能分别来源于C4H-1、C4H-2和4CL基因表达量增加。舞毒蛾取食参与诱导小黑杨苯丙氨酸生物合成途径防御反应,可为进一步明确小黑杨诱导抗性机制以及探讨小黑杨木质素合成防御昆虫取食提供基础理论。
中图分类号:
周心怡,闫丽琼,吕云彤,孙丽丽,朱靖闻,曹传旺. 舞毒蛾取食诱导小黑杨苯丙烷代谢酶活性及其相关基因的表达[J]. 林业科学, 2021, 57(3): 108-116.
Xinyi Zhou,Liqiong Yan,Yuntong Lü,Lili Sun,Jingwen Zhu,Chuanwang Cao. Activities and Gene Expressions of Phenylpropane Metabolic Enzymes in Populus simonii×P. nigra by Herbivore Induction of Lymantria dispar (Lepidoptera: Lymantriidae)[J]. Scientia Silvae Sinicae, 2021, 57(3): 108-116.
表1
实时荧光定量RT-PCR引物序列"
基因 Gene | 正向引物序列 Forward primer sequence (5′-3′) | 反向引物序列 Reverse primer sequence (5′-3′) |
C4H-1 | CGACCCTGAACTAGCCAACC | GGTCATCGACCACAAGGTCC |
C4H-2 | TGGGGAATTGCTGAGCTTG | TGCTCTCAGCTGGAATGTCG |
4CL | GGCGATGTAATCATGACCCTGC | CTCTGGTGGGTCATCAATGGTG |
EF1β | AAGAGGACAAGAAGGCAGCA | CTAACCGCCTTCTCCAACAC |
UBQ | GTTGATTTTTGCTGGGAAGC | GATCTTGGCCTTCACGTTGT |
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