Efficient agrobacterium-mediated transformation of Shiraia bambusicola and activation of a specific transcription factor for hypocrellin production

Figures & data

Figure 1. Construction of the binary vectors pAg1-OE (a) and pAg1-TF1 (b).

Note: The plasmids were constructed for the ATMT of Shiraia bambusicola with the SbTF1 gene placed under the strong promoter PgpdA from Aspergillus nidulans and the terminator TtrpC.

Table 1. Primers used in this investigation.

Primer	Sequence(5'-3')
SbTF1-F	TTGAGCAGACATCACCCCCCATGGCCACTCAACTCCCTACC
SbTF1-R	AACGTTAAGTGGATCCGCCCCTATCCCG ATCCGTTCAAATGTCTAGC
PgpdA-F	AACGACGGCCAGTGAATTCGGAATTCCCTTGTATCTCTACA
PgpdA-R	TCGAGGCCTGATCATCGATGCACTGTGGGT GATGTCTGCTCAAGCG
TrpC-F	CGCTTGAGCAGACATCACCCCACGTGCCAC TTAACGTTACTGAAAT
TrpC-R	GCCTGATCATCGATGAAGAAGGATTACCTC TAAACAAGTGTACCTG
PgpdAtest-1	GAATTCCCTTGTATCTCTACA
TrpCtest-R	GGATTACCTCTAAACAAGTGTACCTG
Hyg500-F	ATCGTTATGTTTATCGGCACT
Hyg500-R	TGGCGACCTCGTATTGG
PKSIq-F2	ACCCTCACTACAACGCAACC
PKSIq-R2	AGCACCTTTCCGTATGTTGG
SbTF1q-F1	TTGCACGCCCCATTTTCATG
SbTF1q-R1	TTGGAAGTGGAGTCTGCGTC
SbSAS1q-F	GCGATTCAGCGAAGACTCCT
SbSAS2q-R	ATGGTCCTGAAACGCTCCTG

Table 2. Diameter of colonies incubated in increasing concentrations of hygromycin and nourseothricin.

Concentration (mg/L)	Diameter of colonies, x̄±SD (mm)
	Hygromycin	Nourseothricin
0	4.65 ± 0.05	4.65 ± 0.05
100	1.98 ± 0.11	1.51 ± 0.04
150	1.23 ± 0.08	0.65 ± 0.03
200	0.98 ± 0.07	0.40 ± 0.04
250	1.79 ± 0.04	–
300	0.44 ± 0.01	–

^a x̄±SD, mean diameter (mm) with standard deviation.

“–”= not active..

Figure 2. Amount of hypocrellins (mg/L) and elsinochrome (mg/L) produced by the wild-type strain, and the overexpression mutants.

Note: Each sample was detected after 96 h and 144 h of incubation.

Figure 3. Relative expression levels of SbTF1 and SbPKSI by real-time PCR.

Note: Each sample was detected after 96 h and 144 h of incubation.