G60 plates

Mycelia of the above 7 mutants and the wild-type isolate were harvested after 4 d incubation in PDB liquid medium with or without propamocarb, and the resulting mycelial mat were washed twice with double-distilled water. 0.5 g of fresh tissue was extracted for lipid by the method of Cartwright (1993) (link). Total lipid extracts were separated into lipid classes by thin-layer chromatography (TLC) on silica gel G60 plates (Merck, Darmstadt, Germany) (Kates, 1986 ). Polar lipids were separated by two-dimension TLC using chloroform/methanol/acetic acid/water [170:30:20:7 (v/v)] and chloroform/methanol/ammonium hydroxide [60:25:4 (v/v)], allowing phosphatidic acid (PtdOH) to be resolved from other phosphoglycerides. Neutral lipids were separated using petroleum ether/diethyl ether/acetic acid [80:20:1 (v/v)]. The lipid bands separated by TLC were routinely visualized by spraying the plates with 8-anilino-1-napthalenesulphonic acid (ANSA) in anhydrous methanol [0.2% (w/v)], and viewing under UV light. Routine identification was by comparison with standards, but further identification was made by using color stains and destructive reagents as described in Kates (1986) . All of chemical agents used in lipid analysis were purchased form Sigma-Aldrich, Shanghai.

Individual PLs were separated by two-dimensional TLC on silica gel G 60 plates (Merck, Darmstadt, Germany). In order to separate PI from PS, the TLC plates were pre-treated with a solution of 1.2% (w/v) boric acid in ethanol:water (1:1, v/v) [40] . Chromatography used solvent systems adapted from Katyal et al. [41] (link):•

Solvent system 1: chloroform:methanol:ammonium hydroxide (65:35:10, by volume),

•

Solvent system 2: n-butanol:acetic acid:water (90:20:20, by volume).

Plates were then sprayed with 0.2% (w/v) ANSA (8-anilino-1-naphthalene sulphonic acid) in dry methanol and lipids were visualised under UV light (2UV Transilluminator UVP) [40] . The PLs were identified routinely by comparison with phospholipid standards and confirmed using specific colour reagents [42] .

After transesterification, the FAMEs (fatty acid methyl esters (FAME)) and residual triacylglycerides (TAGs) were measured using 0.25 mm thick silica gel G-60 TLC plates (Merck, Darmstadt, Germany). The detailed method was as follows: the elution solvent was n-hexane : diethyl ether (90 : 10, v/v) and after full development to the detection of the FAME spots, the spots were visualized using an iodine vapor and spraying the plates with 10% phosphomolybdic acid (98% purity, Daejung Co., Ltd, Shi Heung, Korea) in ethanol, and then drying them in the oven at 105°C [26 ]. The mono-, di-, and triglyceride mixture (TAG STD, Supelco, Bellefonte, PA, USA) and commercial biodiesel (FAME, S Company Houston, TX, USA) were used as standards [26 ]. Additionally, the pigments that remained in the FAME fraction were also separated by TLC, using the same 0.25 mm thick silica gel G-60 plates (Merck, Darmstadt, Germany) that were developed using petroleum ether (99.5%, Daejung Co., Ltd, Shi Heung, Korea) : cyclo hexane (%, Wako, Osaka, Japan) : ethyl acetate (99%, Daejung Co., Ltd, Shi Heung, Korea) : acetone (99.5%, Daejung Co., Ltd, Shi Heung, Korea) : methanol (99.5%, Daejung Co., Ltd, Shi Heung, Korea) (60 : 16 : 10 : 10 : 4, v/v) [27 ].