First release: June 0, 20221

Correspondent: Wang Guodong, Shaanxi Normal University.

It was found that CLE 1-CLE7 could be strongly induced to express in the bud regeneration stage, while the expression of WUS was inhibited by CLV 1 and BAM 1, thus regulating the bud regeneration.

On the phylogenetic tree, cle 1-CLE7 is on a branch, which is consistent with previous reports. The florescence of CLE is earlier than that of overexpression of CLE 1-7.

Furthermore, the overexpression phenotype of CLE 1-CLE7 is consistent with that of clv3, and similar to that of wus mutation. The overexpression of CLE 1, 4, 6 and 7 can completely complement CLV3, indicating that CLE 1-7 may participate in CLV3-CLV/kloc-7 in a functionally redundant way.

Knock out the CLE 1-CLE7 gene, the SAM of cle 1-cle7 has no phenotype, and clv3 has a severe phenotype. Further, eight processes are obtained, and it is found that there is no difference between clv3cle 1-7 and clv3.

Cle 1-cle7 single process has little effect on bud regeneration, indicating its functional redundancy; In 7-process cle 1-7sep and 8-process clv3cle 1-7oct, the explants showed considerable regeneration potential, which was much higher than that of WT and clv3 explants.

CLE 1-7 encodes four different CLE mature peptides? 10nM is close to the physiological concentration, and the displayed therapeutic concentration is 5uM or 10uM.

Inhibitory effect of exogenous application: CLE5p and CLE6p are weak, CLE2p is strong at nM level, and the remaining CLE 1/3/4/7p is useful at nM level but not obvious, but obvious at uM level.

Self-promoter can drive the precursor expression of CLE4 and CLE7, and can also inhibit bud regeneration.

The results showed that exogenous application and endogenous expression of CLE 1-7 were helpful for bud regeneration, but did not affect callus formation.

That is, the overexpression of CLE inhibits bud regeneration and promotes bud regeneration after knockout.

According to the expression profile of pCLE:GUS, CLE 1-7 was strongly induced and expressed in CIM except 5.

After transferring from CIM to SIM, pCLE4:GUS and pCLE7:GUS are gradually restricted to prominent positions.

Description: Spatio-temporal expression corresponds to the bud regeneration process.

CLV 1 does not exist in CIM, but is only expressed in the protrusion formed by SAM in SIM (previously reported). PBAM 1:GFP showed that BAM 1 was strongly and widely expressed in CIM and SIM.

Arabidopsis thaliana was treated with CLE 1-7, and the length of taproot was observed.

When CLE 1-7 was applied, short roots appeared in BAM1-4clv101mutant, but became insensitive in CLV1-kloc-0/mutant. Is it because BAM 1 is the main receptor of taproot inhibition, that is, the lack of BAM makes the root feel less inhibited by CLE 1-7?

In addition, it also shows that the inhibition of CLE 1-7 on root length depends on BAM 1.

Look again, whether the suspicious receptor is helpful for bud regeneration and whether it has phenotype:

Bam 1-3, monopulse, CLV1-kloc-0/monopulse and BAM 1-4 CLV 1 bipulse all showed strong bud regeneration ability. In addition, CLE 1-7 was applied to mutants, in which CLE could not inhibit bud regeneration, indicating that CLV 1 and BAM 1 participated in bud regeneration mediated by CLE 1-7. BAM2 and BAM3 don't work.

The expression of pWUS:GUS was observed by CLEp, and the transcription level of WUS in cle 1-7 was increased by QRT.

Through these two experiments, it is speculated that CLE 1-7 may regulate bud regeneration by limiting the expression of downstream WUS.