Generation of diagnostic anti-teliospore antibodies for development of specific immunodiagnostic format for detection of Karnal bunt (Tilletia indica) of wheat

Figures & data

Figure 1. SDS-PAGE of solubilised teliosporic protein(s) of T. indica extracted by four different solubilising agents.

Table 1. Determination of extractability of teliosporic proteins using different solubilising agents.

S. No	Solubilising agent	Protein concentration (µg/µl)	Protein concentration in teliospores extracts (µg/mg teliospores)
1	I	1.66	149.4
2	II	0.83	74.7
3	III	0.67	60.3
4	IV	1.10	99.0

Figure 2. Antibody dilution curve for determination of titres of ATA generated against (a) intact teliospores, (b) pellet of crushed teliospores, (c) SY and (d) supernatant of crushed teliospores.

Table 2. Characteristics of ATA.

	Types of ATA
	ATA against
Characteristics of ATA	Intact teliospores	Sonicated teliospores	Teliospore pellet	Teliospore supernatant
Maximal reactivity (OD at 405)	3.21	2.85	3.50	3.08
Range of antibody reactivity	125:10,000	125:2500	125:10,000	125:5000
Maximum titre	1:10,000	1:2500	1:10,000	1:5000
Optimal teliospore number	1000	1000	1000	1000

Figure 3. Determination of immunoreactivity of solubilised teliospore proteins extracted by four different solubilising agents (SA1, SA2, SA3 and SA4) using four different types of ATA (anti-intact, anti-sonicated, anti-pellet and anti-supernatant) of T. indica by indirect ELISA.

Figure 4. SDS-PAGE protein profiling of teliospores/spores of fungal pathogens extracted by solubilising agent I; Lane M: Marker; Lane KB: T. indica; Lane CS: U. hordei; Lane HB: T. foetida; Lane FS: U. virens; Lane RB: T. barclayana; Lane WR: P. recondita.

Figure 5. Identification of shared immunoreactive epitopes in spores/teliospores walls of common fungal pathogens by western blotting using anti-intact teliospores antibody.

Figure 6. Preparative gel for extraction of unique solubilised protein from teliosporic protein(s).

Figure 7. Development of monospecific diagnostic antibody against solubilised teliosporic diagnostic antigen of 28 kDa (a) Western blot of solubilised teliosporic proteins with monospecific antibody and (b) Commasaie brilliant blue stained solubilised teliosporic protein of teliospore of T. indica.