Identification of <italic style="font-style: italic">Rehmannia glutinosa</italic> miR166 Family in Response to Endophytic Fungal Infection and Expression Analysis Under Stresses

ZHU Yunhao; XU Jiao; ZHANG Mengjia; DONG Chengming; SHI Bo

doi:10.13422/j.cnki.syfjx.20221618

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Identification of Rehmannia glutinosa miR166 Family in Response to Endophytic Fungal Infection and Expression Analysis Under Stresses

更新时间：2022-12-21

- Identification of Rehmannia glutinosa miR166 Family in Response to Endophytic Fungal Infection and Expression Analysis Under Stresses
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 2, Pages: 133-140(2023)
- 作者机构：
  
  河南中医药大学药学院，呼吸疾病诊疗与新药研发河南省协同创新中心，郑州 450046
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20221618
  CLC： R284.2;R285;R289;R22;R2-031;R33
- Received：17 August 2022，
  
  Published Online：10 November 2022，
  
  Published：20 January 2023
- 稿件说明：
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朱畇昊,许姣,张梦佳等.响应内生真菌侵染的地黄miR166家族鉴定及胁迫下的表达分析[J].中国实验方剂学杂志,2023,29(02):133-140.

ZHU Yunhao,XU Jiao,ZHANG Mengjia,et al.Identification of Rehmannia glutinosa miR166 Family in Response to Endophytic Fungal Infection and Expression Analysis Under Stresses[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(02):133-140.
朱畇昊,许姣,张梦佳等.响应内生真菌侵染的地黄miR166家族鉴定及胁迫下的表达分析[J].中国实验方剂学杂志,2023,29(02):133-140. DOI： 10.13422/j.cnki.syfjx.20221618.

ZHU Yunhao,XU Jiao,ZHANG Mengjia,et al.Identification of Rehmannia glutinosa miR166 Family in Response to Endophytic Fungal Infection and Expression Analysis Under Stresses[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(02):133-140. DOI： 10.13422/j.cnki.syfjx.20221618.

摘要

目的

为了鉴定地黄miR166基因家族的成员，明确其家族成员在逆境下的响应模式。

方法

采用高通量测序技术获得小RNA数据库，筛选地黄miR166家族成员，利用RNAfold分析其前体结构，DNAMAN和MEGA分别进行保守性和进化性分析；采用TargetFinder软件预测地黄miR166家族成员的靶基因；实时荧光定量聚合酶链式反应（Real-time PCR）分析其家族成员在响应非生物胁迫中的表达模式。

结果

鉴定到5条miR166，前体都具有完整的茎环结构；序列比对结果显示前体和成熟体保守性均较高；预测到48个miR166的靶基因，主要注释到HD-ZIP Ⅲ家族转录因子；表达特性分析表明，地黄经过内生真菌侵染后，miR160均呈上调表达，在非生物胁迫下其家族成员的表达情况不同，其中rgl-miR166b-5p在旱涝处理组及高低温处理中表达量分别都较空白组显著下调，与在内生真菌侵染下表达模式相反。

结论

该研究结果初步明确了地黄miR166家族在响应生物胁迫和非生物胁迫下的表达模式，为地黄今后育种改良及相关研究提供理论依据。

Abstract

Objective

To identify the members of the

Rehmannia glutinosa

miR166 gene family and clarify the response mode under adversity.

Method

High-throughput sequencing technology was employed to obtain a small RNA database and the miR166 family members of

R. glutinosa

were screened out. The precursor structures were analyzed by RNAfold. DNAMAN and MEGA were used for conservative and evolutionary analyses， respectively. TargetFinder software was used to predict the target genes of

R. glutinosa

miR166 family members. The expression of miR166 family members in response to abiotic stress was analyzed by real-time polymerase chain reaction（Real-time PCR）.

Result

Five miR166s were identified with precursors possessing complete stem-loop structures. As revealed by sequence alignment results， the precursors and matures were both highly conserved. Forty-eight target genes of miR166s were predicted， which were mainly annotated to the HD-ZIP Ⅲ family transcription factors. The expression characteristics showed that the expression of miR160s was up-regulated after

R. glutinosa

was infected by endophytic fungi， which was different from the expression of the family members under abiotic stress. The expression level of rgl-miR166b-5p in the drought-flood treatment group and the high-low temperature treatment group was significantly down-regulated compared with that in the control group， and the expression pattern was opposite under the endophytic fungal infection.

Conclusion

The results of this study preliminarily clarified the expression patterns of

R. glutinosa

in response to biotic and abiotic stresses and provided a theoretical basis for future breeding and improvement of

R. glutinosa

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references

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Identification of Rehmannia glutinosa miR166 Family in Response to Endophytic Fungal Infection and Expression Analysis Under Stresses

DOI：10.13422/j.cnki.syfjx.20221618

摘要

Abstract

关键词

Keywords

references