Casein acid hydrolysate

The grapevine embryogenic cell line 41B was obtained from the most commonly used rootstock in the vineyards of Champagne (a hybrid between Vitis vinifera L. cv. Chasselas × Vitis berlandieri P.). The embryogenic cell suspension was subcultured every two weeks by transferring 0.3 mL of packed cell volume into 25 mL of a half-strength MS medium (Duchefa M0232) containing glycerol (4.6 G·L⁻¹) and maltose (18 g·L⁻¹) as carbon sources, as well as naphthoxyacetic acid (1 mg·L⁻¹) and casein acid hydrolysate (Sigma A2427). They were cultured under constant agitation (110 rpm), in darkness and at 21 °C. In order to silence the grape ASR gene (VvMSA), 41B embryogenic cells were transformed with the 35S::VvMSA-RNAi construct via Agrobacterium tumefaciens strain EHA 105, co-culture of the grape 41B cells with the bacteria, for 60 h on the above-mentioned solid medium, and selection of transformed cells on paromomycin (2 µg·mL⁻¹) [73 (link)]. The somatic embryogenesis of 41B cells was induced by their subculture into the same fresh medium depleted of auxin and at 26 °C.

Lactobacillus helveticus CICC22171 were isolated from traditional Chinese cheeses in the microbiology laboratory of Beijing Forestry University. The strain was deposited at the China Industrial Culture Collection (CICC). Glucose, 20 basic amino acids, vitamins (nicotinic acid, calcium pantothenate, pyridoxal, riboflavin, biotin, folic acid, and thiamine), sodium acetate, MgSO₄, K₂HPO₄, MnSO₄, Tween 80, and casein acid hydrolysate were purchased from Sigma-Aldrich Corp. (St. Louis, MO, United States).

Antibiotics, 5-aminolevulinic acid hydrochloride (5-ALA), ß-mercaptoethanol, ß-NADH, bovine xanthine oxidase, casein acid hydrolysate, cytochrome c from equine heart, deamino-NADH, desferoxamine mesylate (DFO), diethylenetriamine pentaacetic acid (DTPA), 2,2’-dipyridyl (DIP), ethyl acetate, ferric chloride, ferrous ammonium sulfate, 30% hydrogen peroxide, 8-hydroxyquinoline-5-sulphonic acid, E. coli manganese-containing superoxide dismutase, manganese (II) chloride tetrahydrate, 2-[N-morpholino]ethanesulfonic acid (MES), o-dianisidine dihydrochloride, o-nitrophenyl-ß-D-galactopyranoside (ONPG), potassium ferricyanide, potassium cyanide, tricine, protoporphyrin IX, and xanthine were purchased from Sigma. Ethylenediamine tetraacetic acid (EDTA), guanidine hydrochloride, hydrochloric acid, and 3-(N-morpholino) propane-sulfonic acid (MOPS) were purchased from Fisher Scientific; Coomassie protein assay reagent and albumin standard, from Thermo Scientific; sodium dithionite, from Fluka; and glacial acetic acid, from J.T.Baker.