Table 1.

Genetic material to study MP/ARF5 PB1 domain function.

Line name	Characterization	References
Chemical and insertional mutants
mp abn chemical mutagenesis	A nonsense codon at position 837 corresponds to the mpabn mutation.	Garrett et al. (2012)
mp-14 (SALK_001058) insertional mutagenesis	SALK_001058 has a T-DNA insertion in the 11th exon of MP at the beginning of the sequence coding for the C-terminal domain. Mutant plants have completely penetrant defects only in some developmental processes that depend on MP.	Odat et al. (2014)
Transgenic line
MPΔ MPΔ-2 MPΔ-3	Driven by the endogenous MP promoter (3.3 kb upstream of the start codon). MPΔ encodes amino acids 1–813 of MP, followed by a cloning artifact of 20 extra amino acids (DLEELARISPIVQTFGNKVS). MPΔ-2 and MPΔ-3 encode amino acids 1–794 and 1–813 of MP, respectively, and lack extra residues.	Krogan et al. (2012)
pMP::MPΔ pAS1::MPΔ	pMP::MPΔ contains the endogenous MP promoter (3302 kb upstream of the start codon) and the endogenous MP transcriptional sequence truncated 730 bp downstream of the stop codon. MPΔ encodes amino acids 1–813 of MP without extra residues. In pAS2::MPΔ, MPΔ is controlled by the AS2 promoter containing 3303 bp upstream of the AS2 start codon and 18 bp of the N-terminal AS2 coding region.	Qi et al. (2014)
pUBQ10::MP794-YPet	A 794 amino acid truncated version of MP cDNA was amplified with primer set 648f/675r as a translational fusion to C-terminal YPet to generate UBQ10>> MP794-YPet.	Bhatia et al. (2016)
pMP::MP∆-GR pMP::MP∆-EAR-GR	To construct pMP::MP∆-GR, a 6695 bp MP genomic DNA fragment (containing 3231 bp upstream and 3461 bp downstream of the start codon, respectively) was amplified and fused in-frame to GR. To construct pMP::MP∆-EAR-GR, a 120 bp fragment coding for amino acids 183–222 of AtERF4 was amplified and fused to the C-terminus of pMP::MP∆.	Guan et al. (2017)
pPXY::GR-ARF5ΔIII/IV p35S::ARF5ΔIII/IV	To generate pPXY::GR-ARF5ΔIII/IV (pKB25), the GR-ARF5ΔIII/IV fragment, including a stop codon, was amplified from pKB17 using the MP_for18/MP_rev16 primer pair and inserted in pTOM50 using NcoI/Cfr9I restriction sites. To generate p35S::ARF5ΔIII/IV (pKB40), the ARF5ΔIII/IV CDS was amplified from pKB25 using MP_for17/MP_rev15 and introduced into pGreen0229-35S using XbaI/EcoRI restriction sites.	Brackmann et al. (2018)
pHMG::MP∆	The pHMG promoter corresponds to a 1347 bp fragment upstream of the At1g76110 locus. To generate MPΔ, a fragment of the MP cDNA coding for a truncated protein before domain III (amino acids 1–794) was amplified.	Ma et al. (2019)
XVE::ARF5Δ DR5rev::GFP	The first 2382 nucleotides of MP/ARF5 (encoding the first 794 amino acids and lacking the C-terminal PB1 domain) were cloned into the β-estradiol-inducible, gateway-compatible vector pMDC7. This vector was used to transform plants carrying the DR5rev::GFP reporter.	Gonzalez et al. (2021)
pUBQ10::MPΔ-TagRFP	pMOA34-pUBQ10-loxP-GUS-35S-polyA-loxP-MPΔ-TagRFP construct contains a 2389 bp UBQ10 promoter fragment up to the start codon and a 3461 bp genomic fragment for the MP coding region.	Guan et al. (2022)
Other approaches
GD-ARF5M Protoplast transfection assays	The first and last amino acids from MP/ARF5 in ARF5M, were 349 and 766.	Tiwari et al. (2003)
MPΔC Y3H/BiFC	To generate estradiol-inducible MP, a truncated version of MP was missing the C-terminal PB1 domain (amino acids 795–902) and was amplified from cDNA, cloned into pENTR/D-TOPO (Thermo Fisher). The clone was shuttled into the estradiol-inducible expression vector pMDC7.	Wu et al. (2015)
MP ΔIII,IV GR::MP ΔIII,IV Protoplast transfections Dual-luciferase reporter assay system	Truncated MP protein lacking domains III and IV.	Lau et al. (2011) Herud et al. (2016)
pUAS::MPΔ-GUS p35S::lox-MPΔ-YFP pUBQ10::XVE-amiMP-3AT	pSm43GW-pUAS::MPΔ-GUS-OcsT [OS 30.1] combines pENTR41R-6xUAS2, p221z-cMPdelta, p2R3e-GUS-OcsT into pSm43GW. pBm43GW-p35S::lox-MPΔ-YFP [OS 74.1] combines p1R4z-p35S:lox, p221z-cMPdelta, p2R3a-venYFP-3AT into pBm43GW. pBm43GW-pUBQ10::XVE-amiMP-3AT [RM25.1] combines p1R4a-pUBQ10::XVE, pENTR-AtMIR167a-MP; p2R3a-3AT into pBm43GW	Smetana et al. (2019)

A list of mutants after chemical mutagenesis, insertional mutagenesis, and transgenic lines used to study the truncated MP/ARF5 protein, which acts independently on Aux/IAA repressors.