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Soybean phyA Downstream Gene Prediction and Expression Analysis and Its Knockout Vector Construction(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2021年03期
Page:
309-318
Research Field:
Publishing date:

Info

Title:
Soybean phyA Downstream Gene Prediction and Expression Analysis and Its Knockout Vector Construction
Author(s):
ZHANG Ting; MA Li-xin; LIU Jun; LIN Xiao-ya; LIU Bao-hui
（Innovation Research Center of Molecular Genetics and Evolution, Guangzhou University/Guangzhou Key Laboratory, Guangzhou 510006, China）
Keywords:
Soybean;  PIF3;  LAF1;  FHY1;  FHL;  Tissue expression analysis;  CRISPR/Cas 9;  Gene knockout vector
PACS:
-
DOI:
10.11861/j.issn.1000-9841.2021.03.0309
Abstract:
To further study the downstream genes of E3 and E4 in the photoperiod regulatory pathway of soybean, and lay a foundation for obtaining its mutant soybean，we focusd on PIF3, LAF1, and FHY1/FHL, which are important signaling factors downstream of Arabidopsis far-red light receptor phyA. We used genome sequences of Arabidopsis PIF3, LAF1 and FHY1/FHL as references for searching soybean homologs in Phytozome 12 database, made sequence alignment and phylogenetic tree to analyze the homologs in soybean, and performed real-time quantitative PCR to detect the expression patterns of these soybean homologs in various tissues. We applied CRISPR/Cas 9 system to knockout these homologous genes, and verified the effectiveness of each target through the soybean hair root system. The results showed that: AtPIF3, AtLAF1, and AtFHL has six, four, and two homologs in soybean, respectively. And AtFHY1 doesn′t have homolog in soybean. Four LAF1 homologs were mainly expressed in shoot apical meristem, six PIF3 homologs and two FHL homologs were mainly expressed in pods. According to the CAISPR/Cas9 knockout results, a total of twelve effective targets were identified, which can successfully knock out six PIF3, four LAF1, and two FHL homologs in soybean. This research provides a theoretical basis for obtaining stable soybean knockout mutant, which pave a way for studying the functions of soybean phyA downstream genes.Keywords:

References:

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Last Update: 2021-07-20