Wang Xiao-Ran, Wang Yu-Hong, Li Wei-Xi, Li Zhen, Liu Fang-Ming, Zhang Zhong-Yi, Chen Xin-Jian
College of Life Sciences,Henan Agricultural University Zhengzhou 450002,China.
College of Forestry,Henan Agricultural University Zhengzhou 450002,China.
Zhongguo Zhong Yao Za Zhi. 2019 Feb;44(4):703-711. doi: 10.19540/j.cnki.cjcmm.20181204.003.
The consecutive monoculture obstacle is a major problem in the field of Rehmannia glutinosa( R. glutinosa),has severely declined the yield and quality of R. glutinosa. Here,using hi TAIL-PCR and RACE techniques,we have cloned the full-length transcript( 1 573 bp) of Unigene 29334＿All screened by DGE as a consecutive monoculture obstacle response gene of R. glutinosa. Based on ORF Finder prediction,all ORFs detected in the full-length transcript were less than 300 nt,which suggested that the above transcript was confirmed to be a long non-coding RNA( LncRNA). With alignment in R. glutinosa transcriptome,this LncRNA was partially homologous to alanine glyoxylate transaminase 2 gene( Rg AGT2),which was named LncRNA-RgATG2. To further explore the function of LncRNA-RgAGT2,we have examined expression patterns of LncRNA-RgAGT2 and Rg AGT2 at five critical development stages( seedling,elongation,pre-expanding,mid-expanding,late-expanding) in the first and second year replanting of R. glutinosa,respectively. The results indicated that LncRNA-RgAGT2,as a potential regulator,is possible to play a vital role in Rg AGT2 expression regulation. Meanwhile,LncRNA-RgAGT2 has presented significant variation in all development stages of R. glutinosa,which could be used as a " diagnostic label" to assess consecutive monoculture obstacle. This study,for the first time,showed that LncRNA was responsible for the response and regulation of consecutive monoculture obstacle,which would be a powerful supplement to reveal the molecular mechanisms of consecutive monoculture obstacle of R. glutinosa.
连作障碍是地黄种植领域的一个主要问题，已严重降低了地黄的产量和品质。在此，我们利用热不对称交错PCR（hiTAIL-PCR）和快速扩增cDNA末端（RACE）技术，克隆了Unigene 29334＿All的全长转录本（1573 bp），该转录本经数字基因表达谱（DGE）筛选，作为地黄连作障碍响应基因。基于开放阅读框查找器（ORF Finder）预测，在全长转录本中检测到的所有开放阅读框均小于300 nt，这表明上述转录本被确认为长链非编码RNA（LncRNA）。通过与地黄转录组比对，该LncRNA与丙氨酸乙醛酸转氨酶2基因（RgAGT2）部分同源，命名为LncRNA-RgAGT2。为进一步探究LncRNA-RgAGT2的功能，我们分别检测了LncRNA-RgAGT2和RgAGT2在地黄第一年和第二年重植的五个关键发育阶段（幼苗期、伸长期、膨大前期、膨大中期、膨大后期）的表达模式。结果表明，LncRNA-RgAGT2作为一种潜在的调节因子，可能在RgAGT2表达调控中发挥重要作用。同时，LncRNA-RgAGT2在地黄的所有发育阶段均呈现出显著差异，可作为评估连作障碍的“诊断标签”。本研究首次表明LncRNA参与了连作障碍的响应和调控，这将为揭示地黄连作障碍的分子机制提供有力补充。
