Isolation, characterization, and plant growth-promoting activities of endophytic fungi from a wild orchid Vanda cristata

Figures & data

Table 1. Endophytic fungi isolated from different parts of Vanda cristata along with their homology percentage and class.

ID	Name of fungi	Identity (%)	Query Coverage (%)	Class	Accession number
PVS2	Agaricus bisporus	99	98	Agaricomycetes	CP015465
PVS3	Unidentified I	-		-	-
PVL2	Unidentified II	-		-	-
PVL3	Alternaria tenuissima	100	96	Dothideomycetes	HG798721
PVL4	Fusarium tricinctum	99	100	Sordariomycetes	KJ188669
PVL5	Alternaria sp.	-		Dothideomycetes	-
PVL6	Pseudochaetosphaeronema sp.	95	97	Dothideomycetes	KU365986
CVS4	Fusarium oxysporium	-		Sordariomycetes	-
PVR1	Fusarium solani	-		Sordariomycetes	-
PVR2	Fusarium sp.	98	99	Sordariomycetes	KU747889
PVR3	Mycoleptodiscus sp.	99	94	Sordariomycetes	JQ388252
CVL1	Paraconiothyrium hawaiiense	99	100	Dothideomycetes	KF881749

Figure 1. CVS4 colony: white, yellow in color (a); mycelium appeared long, branched, and septate; conidia curved shaped long, septate with pointed end (b). PVR1 colony: cottony white in early stage whereas yellowish with maturation (c); mycelium was long, septate, and branched; conidiophores were found attached with mycelium; conidia was long, septate, curved, and pointed at both end (d).

Figure 2. Amount of IAA synthesized by different strains of endophytic fungi in growth medium with or without tryptophan condition. PVL2 synthesized maximum amount of IAA and PVR2 was the least IAA synthesizer. Experiments were performed in biologically independent replicate (n = 3) with ±SE. Tested values are significantly different at p ≤ 0.05 (one way ANOVA).

Figure 3. Soluble phosphate estimation for different endophytic fungi isolated from V. cristata. PVL2 was highest phosphate solubilizer and PVS2 was least. Experiments were performed in a biologically independent replicate (n = 3) with ±SE. Tested values are significantly different at p ≤ 0.05 (one way ANOVA).

Table 2. Different endophytic fungi showing intensity of ammonia synthesis by color intensity according to their strength.

S.N.	Endophytic fungi	Intensity of ammonia	S.N.	Endophytic fungi	Intensity of ammonia
1	Control	-	8	PVL6	++
2	PVS2	+++	9	PVR1	+
3	PVS3	+	10	PVR2	++
4	PVL2	+	11	PVR3	+++
5	PVL3	++	12	CVL1	+
6	PVL4	+	13	CVS4	++
7	PVL5	+

(+++) high; (++) medium; (+) low; (-) no observation.

Figure 4. Qualitative test of ammonia according to color strength (left to right).

Table 3. The plant growth assay for 90 days with the fungal elicitor supplemented versus IAA supplemented MS medium and MS basal media (control). Data: Mean ± SE (n = 10), significant at p ≤ 0.05.

Isolates	Root number		Root length		Shoot number		Shoot length
	Without Tryptophan	With Tryptophan	Without Tryptophan	With Tryptophan	Without Tryptophan	With Tryptophan	Without Tryptophan	With Tryptophan
Control	2.75 ± 0.62^e	2.00 ± 0.40^de	2.75 ± 0.12^g	2.75 ± 0.59^k	2.5 ± 0.28^a	2.75 ± 0.62^b	2.50 ± 0.35g	2.75 ± 0.52^i
MS+IAA (1 µg/ml)	1.00 ± 0.57^f	2.50 ± 0.64^ab	1.00 ± 0.28^b	2.00 ± 0.08^ij	3 ± 0.86^b	2 ± 0.40^a	2.63 ± 0.94^h	2.20 ± 0.26^f
PVS2	2.50 ± 0.5^h	3.75 ± 0.47^d	2.50 ± 0.125^f	3.75 ± 0.75^l	3.25 ± 0.86^c	4.25 ± 0.95^g	1.00 ± 0.00^b	4.13 ± 0.85^k
PVS3	4.50 ± 0.64^f	2.50 ± 0.86^f	4.50 ± 0.375^i	2.00 ± 0.73^ij	4.5 ± 0.86^g	3.5 ± 0.86^e	3.13 ± 0.31^i	2.00 ± 0.54^d
PVL2	3.00 ± 0.81^f	2.50 ± 0.5^e	3.00 ± 0.14^h	1.38 ± 0.125^e	3.5 ± 0.64^d	4.25 ± 0.47^g	3.68 ± 0.85^l	2.50 ± 0.46^h
PVL3	2.75 ± 0.47^h	3.75 ± 0.75^de	2.75 ± 0.94^g	1.88 ± 0.125^g	3.25 ± 0.47^c	7.25 ± 0.91^k	4.25 ± 0.75^m	3.88 ± 0.52^j
PVL4	2.50 ± 0.28^g	3.00 ± 0.70^d	2.50 ± 0.40^f	1.88 ± 0.42^gh	4 ± 0.70^f	4.75 ± 0.81^h	3.38 ± 0.94^j	1.88 ± 0.13^c
PVL5	1.75 ± 0.47^f	2.50 ± 0.65^cd	1.75 ± 0.23^e	1.43 ± 0.41^ef	5 ± 0.95^i	5 ± 0.85^i	1.50 ± 0.5^d	1.75 ± 0.48^b
PVL6	1.50 ± 0.64^g	3.00 ± 0.40^c	1.50 ± 0.86^d	1.95 ± 0.33^hi	4 ± 0.94^f	5.75 ± 0.25^j	3.50 ± 0.83^k	2.38 ± 0.24^g
PVR1	1.25 ± 0.25^a	0.50 ± 0.5^b	1.25 ± 0.125^c	0.50 ± 0.5^b	2.5 ± 0.8^a	3 ± 0.57^c	1.13 ± 0.63^c	1.40 ± 0.55^a
PVR2	1.00 ± 0.40^c	1.00 ± 0.40^ab	1.00 ± 0.375^b	0.38 ± 0.125^a	4.5 ± 0.5^g	3.25 ± 0.47^d	1.63 ± 0.24^e	1.38 ± 0.13^a
PVR3	1.75 ± 0.75^b	0.75 ± 0.25^cd	1.75 ± 0.23^e	0.88 ± 0.42^c	3.75 ± 0.47^e	3.75 ± 0.81^f	0.88 ± 0.24^a	2.13 ± 0.13^e
CVL1	0.25 ± 0.25^i	4.25 ± 0.85^a	0.25 ± 0.25^a	1.50 ± 0.45^ef	5 ± 0.64^i	5.75 ± 0.91^j	2.38 ± 0.47^f	2.50 ± 0.54^i
CVS4	6.00 ± 0.70^d	1.25 ± 0.25^g	7.25 ± 0.59^j	1.00 ± 0.35^d	4.75 ± 0.85^h	8.75 ± 0.47^l	4.25 ± 0.25^h	1.88 ± 0.13^c

Figure 5. The in-vitro plant growth assay of Cymbidium aliofolium from the protocorms stage for 90 days on Murashige and Skoog (MS) medium control (a), MS + IAA (1 µg per ml) (b), MS media supplemented with CVS4 fungal elicitors (c).