摘要
【目的】蛋白酶抑制剂(protease inhibitor,PI)是一类广泛存在于植物中的蛋白质,具有抵御植食性昆虫取食危害的功能。然而,水稻(Oryza sativa)PI基因在二化螟(Chilo suppressalis)取食过程中的表达模式尚不清楚。本研究旨在分析水稻蛋白酶抑制剂基因OsLTPL164和OsLTPL151在二化螟取食及机械损伤下的表达模式,为明确OsLTPL164和OsLTPL151在水稻防御二化螟中的作用及今后利用这两个基因构建转基因水稻株系打下基础。【方法】以东北地区常规种植的3个水稻品系辽盐2号(1654)、辽星17号(1665)和长白17号(1688)为研究对象,在二化螟危害关键时期(分蘖期)通过机械损伤和接虫处理,在处理后不同时间(0、3、6、12、24、48、72 h)分别对根、茎、叶3个组织进行取样,利用实时荧光定量PCR(RT-q PCR)检测3个水稻品系中OsLTPL164和OsLTPL151的表达模式。【结果】OsLTPL164在3个品系中的组织表达模式一致,在叶片和茎中的表达量均高于根部;OsLTPL151在3个品系中的组织表达模式不一致,在1654品系中茎和叶片中的表达量显著高于根部,但在1688和1665品系中根部的表达量显著高于茎和叶片;此外,这两个基因在3个品系间的相同组织中都有不同的表达模式,OsLTPL164在根、茎、叶中均呈现在1665品系中表达量最高、在1688品系中表达量次之、在1654品系中表达量最低的表达模式,但OsLTPL151在不同组织中的表达模式与OsLTPL164不同:在根中,OsLTPL151在1665品系中的表达量明显高于1688和1654品系;在茎中,OsLTPL151在1654和1665品系中的表达量明显高于1688品系;在叶中,OsLTPL151在1654品系中的表达量明显高于1665和1688品系。二化螟取食诱导OsLTPL164和OsLTPL151在3个品系的叶片中表达上调幅度高于茎和根,且在1665品系中上调的幅度较1688和1654品系中大。在二化螟取食和机械损伤处理不同时间后,两个基因均呈现表达水平先上升后下降再上升的趋势;OsLTPL164和OsLTPL151在机械损伤6 h后才被显著诱导表达,而二化螟取食3 h后便可诱导OsLTPL164和OsLTPL151上调表达。【结论】OsLTPL164和OsLTPL151在3个水稻品系中二化螟取食以及机械损伤均能诱导其表达,推测这两个基因可能与水稻抗二化螟有关,但这两个基因在不同水稻品系中其他具体的功能可能有所不同。
【Objective】Protease inhibitors(PI) is a kind of protein widely existed in plants. It has the function of resisting herbivorous insects feeding. However, the expression patterns of rice(Oryza sativa) PI genes in the feeding process of the Chilo suppressalis are not clear. The objective of this study is to analyze the expression patterns of rice protease inhibitor genes OsLTPL164 and OsLTPL151 under C. suppressalis infestation and mechanical damage, which will lay a theoretical foundation for confirming the function of OsLTPL164 and OsLTPL151 in rice against C. suppressalis infestation and constructing transgenic rice strains using these two genes in the future. 【Method】 Three rice lines, Liaoyan 2(1654), Liaoxing 17(1665) and Changbai 17(1688), which were conventionally planted in Northeast China, were selected as the research materials. Artificial infestation and mechanical injury treatments were performed at the tillering stage of rice, which is the key period of C. suppressalis infestation. The root, stem and leaf of three rice lines were sampled after different times(0, 3, 6, 12, 24, 48, 72 h) of treatments, then the expression level of OsLTPL164 and OsLTPL151 of three rice lines was determined by RT-q PCR. 【Result】The tissue expression pattern of the OsLTPL164 was consistent in three rice lines. The expression level of OsLTPL164 in leaf and stem was higher than that in root. The tissue expression pattern of the OsLTPL151 was not consistent in three rice lines. The expression level of OsLTPL151 in the stem and leaf was significantly higher than in root in the 1654 line, but in the 1688 and 1665 lines, the expression level of OsLTPL151 in root was significantly higher than that in stem and leaf. In addition, the two genes in the same tissue among three rice lines had different expression patterns. The expression level of the OsLTPL164 was the highest in 1665 line, followed by 1688 line, and the lowest in 1654 line. The OsLTPL151 exhibited different expression patterns in different tissues among three rice lines: in the root, the expression level of OsLTPL151 in the 1665 line was significantly higher than that in the 1688 and 1654 lines; in the stem, the expression level of OsLTPL151 in the 1654 and 1665 lines was significantly higher than that in the 1688 line; in the leaf, the expression of OsLTPL151 in the 1654 line was significantly higher than that in the 1665 and 1688 lines. Up-regulation of OsLTPL164 and OsLTPL151 in leaf was more obvious than that of stem and root in the three lines by C. suppressalis infestation, and with a higher inducing effect in 1665 line than that in 1688 and 1654 lines. Both OsLTPL164 and OsLTPL151 showed a rise-decrease-rise expression tendency at different time points after artificial infestation and mechanical injury treatments. The expression level of OsLTPL164 and OsLTPL151 was significantly induced after 6 h of mechanical damage treatment, while the expression level of OsLTPL164 and OsLTPL151 could be significantly induced after 3 h of C. suppressalis infestation. 【Conclusion】The expression of OsLTPL164 and OsLTPL151 in three conventionally grown rice lines can be induced by C. suppressalis infestation and mechanical damage treatment, suggesting that both the two genes are involved in resistance to C. suppressalis. However, other specific functions of the two genes in different rice lines might be different.
作者
何雨娟
鞠迪
王悦
杨雪清
王小奇
HE YuJuan;JU Di;WANG Yue;YANG XueQing;WANG XiaoQi(College of Plant Protection, Shenyang Agricultural University/Key Laboratory of Economic and Applied Entomology of Liaoning Province, Shenyang 110866)
出处
《中国农业科学》
CAS
CSCD
北大核心
2018年第12期2311-2321,共11页
Scientia Agricultura Sinica
基金
国家自然科学基金(31501666)
"青年托举人才工程"项目(YESS20160085)