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园艺学报 ›› 2019, Vol. 46 ›› Issue (3): 486-498.doi: 10.16420/j.issn.0513-353x.2018-0497

• 研究论文 • 上一篇    下一篇

蕹菜耐受长时间高温后的miRNA分析

王杏茹1,李文静1,2,陈冰星1,2,刘 涛1,尚 维1,赖钟雄2,*,郭容芳1,2,*   

  1. (1福建农林大学戴尔豪西大学联合实验室,福州 350002;2 福建农林大学园艺植物生物工程研究所,福州 350002)
  • 出版日期:2019-03-25 发布日期:2019-03-25

Analysis of miRNA in Water Spinach(Ipomoea aquatica)Under Long-time High Temperature

WANG Xingru1,LI Wenjing1,2,CHEN Bingxing 1,2,LIU Tao1,SHANG Wei1,LAI Zhongxiong2,*,and GUO Rongfang 1,2,*   

  1. (1Joint FAFU-Dalhousie Lab,Fujian Agriculture and Forestry University,Fuzhou 350002,China;2Institute of Horticultural Biotechnology,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
  • Online:2019-03-25 Published:2019-03-25

摘要:

miRNA是广泛存在于真核生物体内的一类长度为20 ~ 24 nt的小分子非编码RNA,在植物生长发育和逆境适应过程中发挥重要作用。蕹菜(Ipomoea aquatica)可以耐受长时间高温,而其miRNA还未被鉴定,其在耐受长时间高温后的作用还未知。以蕹菜耐热性强的品种‘泰国三叉’和耐热性差的品种‘柳叶’为材料,经42 ℃高温处理15 d后分别构建两个高质量sRNA库,经检测两个库中碱基质量值大于或等于30的序列数量均超过17.3 Mb,且‘泰国三叉’中24 nt的序列数量远远高于‘柳叶’。与蕹菜转录组比对分析后,分别得到1 363 258和1 629 209条序列,两个库共有的序列仅有1 047 133条(11.36%),说明耐热性不同的蕹菜中sRNA有很大差别。共鉴定出蕹菜中存在71个miRNA,其中差异表达的有22个,通过TargetFinder预测其靶基因,共得到233个靶基因。进一步以耐热性强的品种‘本地三叉’和耐热性弱的品种‘竹叶’为材料研究其在长时间高温后miR160、miR166、miR172、miR393和miR3627前体的差异表达量,结果表明,高温后两品种中pre-miR160-2的表达都显著上调,但是不耐热的‘竹叶’中上调的倍数更多;miR166在耐热性强的‘本地三叉’中表达量下降,在耐热性较差的‘竹叶’中表达量显著上升;miR172可以被长时间高温诱导,但在耐热性不同的两个品种蕹菜中并没有差异;miR3627的表达量在耐热蕹菜‘本地三叉’中下降,在不耐热蕹菜‘竹叶’中没有明显变化。miRNA前体表达模式的差异说明其在蕹菜耐受长时间高温过程中的作用机制不同。

关键词: 蕹菜, 长时间高温, miR160, miR166, miR393

Abstract:

miRNAs are small non-coding RNAs of 20–24 nt widely present in eukaryotes and play important roles in plant growth and stress adaptation. Water spinach(Ipomoea aquatica)can tolerate prolonged high temperature. However,miRNAs in water spinach have not yet been identified and their role in long-time high temperature is still unknown. In this paper,taking highly heat-resistant‘Taiguo Sancha’and poorly heat-resistant‘Liuye’as material,two high-quality sRNA pools were constructed after treatment with 42 ℃ for 15 days. The number of high-quality sequences both exceeded 17.3 Mb,and the number of 24 nt clean reads in‘Taiguo Sancha’was much higher than that of‘Liuye’. After comparison with water spinach’s transcriptome,1 363 258 and 1 629 209 sequences were obtained respectively. The two libraries shared 1 047 133(11.36%)unique reads,indicating that sRNAs varied in cultivars with different heat resistance. A total of 71 miRNAs were identified in water spinach,of which 22 were differentially expressed. The target genes were predicted by TargetFinder and a sum of 233 target genes were obtained. In addition,water spinach‘Bendi Sancha’with strong-heat-resistance and weakly heat-resistant‘Zhuye’were used as materials to study the expression levels of pre-miR160,pre-miR166,pre-miR172,pre-miR393,and pre-miR3627 that were differentially expressed after long-time high temperature treatment. The results showed that the expression of pre-miR160-2 in‘Bendi Sancha’and‘Zhuye’increased significantly,but in different levels;the expression of miR166 was decreased in‘Bendi Sancha’but increased significantly in‘Zhuye’;miR172 could be induced by high temperature,however,there was no difference in the two cultivars;the expression of miR3627 was decreased in‘Bendi Sancha’after high temperature treatment while there was no obvious change in‘Zhuye’. The difference in the expression patterns of miRNA precursors suggests that the mechanism of miRNA is different during long-term high temperature in water spinach.

Key words: Ipomoea aquatica, long-time high temperature, miR160, miR166, miR393

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