Fingerprinting and molecular comparison among two parental strains of Trichoderma spp. and their corresponding fusants produced by protoplast fusion

Figures & data

Table

Content	Quantity (µL)	Concentration
Taq buffer (10 x) with 15 mmol/L MgCl2	2.50	10 mmol/L Tris 1.5 mmol/L MgCl2
dNTPs mix (2.5 mmol/L each)	2.00	0.2 mmol/L
Taq DNA polymerase (3 U/µL)	0.5	2.25 U
Primer (10 pmoles/µL)	2.00	20 pmoles
Template DNA (25 ng/µL)	2.00	50 ng
Sterile Millipore water	16
Total reaction mixture	25

Table 1. Polymorphic bands of each RAPD primers and percentage of polymorphism in parent's strains and their corresponding fusants.

Primers	Total bands	No. of monomorphic bands	No. of polymorphic bands	% of monomorphic bands	% of polymorphic bands
OPA-1	14	11	3	78.5	21.5
OPA-2	10	9	1	90.0	10.0
OPA-7	10	10	0	100	0.00
OPA-12	10	10	0	100	0.00
OPB-07	13	12	1	92.3	7.70
OPB-08	18	15	3	83.3	16.7
OPB-18	8	7	1	87.5	12.5
OPB-19	9	8	1	88.9	11.1
OPC-04	12	7	5	58.3	61.7
OPC-05	12	11	1	91.7	8.30
OPC-09	10	9	1	90.0	10.0
OPD-02	11	11	0	100	0.00
OPD-03	8	8	0	100	0.00
OPD-07	8	7	1	87.5	12.5
OPF-06	8	7	1	87.5	12.5
OPF-15	9	7	2	77.8	22.2
OPH-15	7	6	1	85.7	14.3
OPO-06	15	10	5	66.7	33.3
OPO-15	10	10	0	100	0.00
OPZ-15	8	8	0	100	0.00
Total	210	183	27	87.2	12.8

Figure 1. RAPD-PCR profile of two parent's strains (P1 and P2) (T. harzianum NBAII Th 1 and T. viride NBAII Tv 23) and their 21 corresponding fusants (lane 1 to lane 21). M: is 100 bp DNA ladder.

Figure 2. RAPD-PCR profile of two parent's strains (P1 and P2) (T. harzianum NBAII Th 1 and T. viride NBAII Tv 23) and their 21 corresponding fusants (lane 1 to lane 21). M: is 100 bp DNA ladder.

Figure 3. ISSR-PCR profile of two parent's strains (P1 and P2) (T. harzianum NBAII Th 1 and T. viride NBAII Tv 23) and their 21 corresponding fusants (lane 1 to lane 21). M: is 100 bp DNA ladder.

Figure 4. ISSR-PCR profile of two parent's strains (P1 and P2) (T. harzianum NBAII Th 1 and T. viride NBAII Tv 23) and their 21 corresponding fusants (lane 1 to lane 21). M: is 100 bp DNA ladder.

Table 2. Polymorphic bands of each ISSR primers and percentage of polymorphism in parent's strains and their corresponding fusants.

Primers	Total bands	No. of monomorphic bands	No. of polymorphic bands	% of monomorphic bands	% of polymorphic bands
UBC-809	10	7	3	70.0	30.0
UBC-810	9	6	3	66.7	33.3
UBC-812	6	6	0	100	0.00
UBC-823	8	8	0	100	0.00
UBC-824	9	9	0	100	0.00
UBC-825	12	9	3	75.0	25.0
UBC-830	12	12	0	100	0.00
UBC-834	11	10	1	91.0	9.00
UBC-835	7	3	4	42.8	57.2
UBC-836	8	6	2	75.0	25.0
UBC-840	8	7	1	87.5	12.5
UBC-844	13	12	1	92.3	7.70
UBC-845	9	9	0	100	0.00
UBC-855	8	8	0	100	0.00
UBC-857	11	10	1	91.0	9.00
UBC-864	6	5	1	83.3	16.7
UBC-876	7	6	1	85.7	14.3
UBC-878	9	6	3	66.7	33.3
UBC-881	6	6	0	100	0.00
UBC-884	5	5	0	100	0.00
Total	174	150	24	86.2	13.8

Table 3. Polymorphic bands of each ITS primers and percentage of polymorphism in parent's strains and their corresponding fusants.

Primers	Total bands	No. of monomorphic bands	No. of polymorphic bands	% of monomorphic bands	% of polymorphic bands
BOXA1	10	9	1	90.0	10.0
ERIC	10	10	0	100	0.00
REP	10	9	1	90.0	10.0
ITS1	5	5	0	100	0.00
Total	35	33	2	94.2	5.80

Figure 5. Dendrogram analysis among the two parent's strains (P1 and P2) (T. harzianum NBAII Th 1 and T. viride NBAII Tv 23) and their 21 corresponding fusants (fusant-1 to fusant-21) based on RAPD primers that have been used for fingerprinting.

Figure 6. Dendrogram analysis among the two parent's strains (P1 and P2) (T. harzianum NBAII Th 1 and T. viride NBAII Tv 23) and their 21 corresponding fusants (fusant-1 to fusant-21) based on ISSR and ITS primers that have been used for fingerprinting.

Figure 7. ITS-PCR profile of two parent's strains (P1 and P2) (T. harzianum NBAII Th 1 and T. viride NBAII Tv 23) and their 21 corresponding fusants (lane 1 to lane 21). M: is 100 bp DNA ladder.