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  • YUCCA-Mediated Biosynthesis of the Auxin IAA Is Required during the Somatic Embryogenic Induction Process in Coffea canephora.

YUCCA-Mediated Biosynthesis of the Auxin IAA Is Required during the Somatic Embryogenic Induction Process in Coffea canephora.

International journal of molecular sciences (2020-07-09)
Miguel A Uc-Chuc, Cleyre Pérez-Hernández, Rosa M Galaz-Ávalos, Ligia Brito-Argaez, Víctor Aguilar-Hernández, Víctor M Loyola-Vargas
ABSTRACT

Despite the existence of considerable research on somatic embryogenesis (SE), the molecular mechanism that regulates the biosynthesis of auxins during the SE induction process remains unknown. Indole-3-acetic acid (IAA) is an auxin that is synthesized in plants through five pathways. The biosynthetic pathway most frequently used in this synthesis is the conversion of tryptophan to indol-3-pyruvic acid (IPA) by tryptophan aminotransferase of Arabidopsis (TAA) followed by the conversion of IPA to IAA by enzymes encoded by YUCCA (YUC) genes of the flavin monooxygenase family; however, it is unclear whether YUC-mediated IAA biosynthesis is involved in SE induction. In this study, we report that the increase of IAA observed during SE pre-treatment (plants in MS medium supplemented with 1-naphthaleneacetic acid (NAA) 0.54 µM and kinetin (Kin) 2.32 µM for 14 days) was due to its de novo biosynthesis. By qRT-PCR, we demonstrated that YUC gene expression was consistent with the free IAA signal found in the explants during the induction of SE. In addition, the use of yucasin to inhibit the activity of YUC enzymes reduced the signal of free IAA in the leaf explants and dramatically decreased the induction of SE. The exogenous addition of IAA restored the SE process in explants treated with yucasin. Our findings suggest that the biosynthesis and localization of IAA play an essential role during the induction process of SE in Coffea canephora.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Calcofluor White Stain, suitable for microbiology
Sigma-Aldrich
Anti-Auxin antibody, Mouse monoclonal, clone 1E11-C11, purified from hybridoma cell culture
Sigma-Aldrich
N-(3-Indolylacetyl)-L-alanine, 98%
Sigma-Aldrich
myo-Inositol, ≥99%
Sigma-Aldrich
Nicotinic acid, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Kinetin, suitable for plant cell culture, crystalline
Supelco
3-Indoleacetic acid, PESTANAL®, analytical standard
Sigma-Aldrich
Acetic acid, glacial, ACS reagent, ≥99.7%
Sigma-Aldrich
Potassium phosphate monobasic, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
Sigma-Aldrich
Indole-3-acetic acid sodium salt, BioReagent, suitable for plant cell culture, ≥98%
Sigma-Aldrich
L-Tryptophan, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
Sodium phosphate dibasic, for molecular biology, ≥98.5% (titration)
(2-Butylbenzofuran-3-yl)(4-hydroxy-3,5-diiodophenyl)methanone, British Pharmacopoeia (BP) Reference Standard