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园艺学报 ›› 2018, Vol. 45 ›› Issue (7): 1261-1271.doi: 10.16420/j.issn.0513-353x.2017-0760

• 研究论文 • 上一篇    下一篇

越橘VcLon2蛋白酶在转化烟草衰老中的作用

陈文荣,陈侨月,尤式备,余 颖,李永强,杨 莉,郭卫东*   

  1. 陈文荣,陈侨月,尤式备,余 颖,李永强,杨 莉,郭卫东*
  • 出版日期:2018-07-25 发布日期:2018-07-25

The Functions of Blueberry VcLon2 Protease in Senescence of Transgenetic Tobacco

CHEN Wenrong,CHEN Qiaoyue,YOU Shibei,YU Ying,LI Yongqiang,YANG Li,and GUO Weidong*   

  1. CHEN Wenrong,CHEN Qiaoyue,YOU Shibei,YU Ying,LI Yongqiang,YANG Li,and GUO Weidong*
  • Online:2018-07-25 Published:2018-07-25

摘要: 植物衰老往往伴随着过氧化物酶体功能紊乱,Lon2蛋白酶对过氧化物酶体的功能和结构的维持具有重要作用,但目前未见其与衰老相关性的报道。从越橘(Vaccinium ssp.)中克隆了Lon2全长cDNA序列,命名为VcLon2(KX611379),并通过烟草中NbLon2基因沉默(Virus Induced Gene Silence,VIGS)技术处理和转VcLon2基因对该基因功能进行研究。结果表明,NbLon2沉默本氏烟植株矮小,叶片小且黄化严重,而VcLon2超表达本氏烟植株鲜绿且粗壮;NbLon2沉默植株叶绿素含量仅为对照的一半,H2O2含量及膜脂过氧化程度也显著高于对照,VcLon2超表达植株则相反。同时,抗氧化酶CAT基因的转录水平在本氏烟各株系中无显著差异,但NbLon2沉默植株内CAT酶活性仅为对照的62.62%,表明Lon2蛋白酶对过氧化物酶体内蛋白质结构和功能的维持发挥着重要作用。此外,NbLon2沉默植株蛋白羰基化程度显著高于对照,植物衰老标记基因SAG12表达量为对照的15 642.2倍。以上研究均表明,Lon2蛋白酶缺失显著加剧光合色素降解、破坏细胞膜系统、羰基化蛋白大量积累等;VcLon2蛋白酶对植物健康维系及衰老延缓等具有重要作用。

关键词: 越橘, 本氏烟, Lon2蛋白酶, 植物衰老

Abstract: Plant senescence always parallels peroxisome dysfunction,and Lon2 protease greatly involves in maintaining the function and structure of peroxisomes. However,little literature is available regarding Lon2 in plant senescence. In the present study,full length of Lon2 cDNA was cloned from blueberry(Vaccinium ssp.)and named VcLon2(KX611379),then the biological functions in plant senescence of VcLon2 were studied through Virus Induced Gene Silence(VIGS)and transgenetic techniques. Phenotypically,NbLon2-silenced Nicotiana benthamiana showed pygmyism with small and serious yellowing leaves,while VcLon2-overexpressed N. benthamiana plants grew well with bright green leaves. Moreover,chlorophyll content in NbLon2-silenced plants was about 50% of the control plants,and the H2O2 content as well as the membrane lipid peroxidation was more serious compared with the control. On the contrary,conversed change trend was observed in VcLon2- overexpressed plants. Furthermore,the transcription levels of catalase(CAT),a peroxisomal enzyme involved in the detoxification of ROS,showed no significant difference among different seedlings. However,CAT activity in NbLon2-silenced seedlings was reduced to 62.62% of the negative control,indicating that Lon2 plays an important role in structural and functional maintenance of peroxisomal protein. In addition,protein carbonylation degree in NbLon2-silenced plants were significantly higher than that in the control,and the expression level of SAG12 was 15 642.2 times the level in the control. It is suggested that Lon2 protease missing significantly accelerated degradation of photosynthetic pigment,destruction of membrane system and accumulation of carbonylated protein and Lon2 protease played an important role in delaying senescence in plants.

Key words: Vaccinium ssp., Nicotiana benthamiana, Lon2 protease, plant senescence

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