Diversity and extracellular enzyme activities of fungal endophytes isolated from medicinal plants of Western Ghats, Karnataka

Figures & data

Table 1 List of selected medicinal plants and their uses

Sl. No	Medicinal Plant	Family	Herbarium Accession no.	Medicinal uses of the plants
1	Tinospora cordifolia (Willd.) Hook. f. & Thomson	Menispermaceae	RRCBI-8976	Antipyretic and anti-asthmatic.
2	Piper nigrum L.	Piperaceae	RRCBI-MUS135	Treatment of vertigo, asthma and sinusitis.
3	Piper longum L.	Piperaceae	RRCBI-AP -2591	Arthritis and respiratory tract infections.
4	Zingiber officinale Roscoe	Zingiberaceae	RRCBI-AP.4046	Throat infections, common colds & fever.
5	Hedychium coronarium J. Koenig	Zingiberaceae	RRCBI-7331	Nausea, halitosis, vomiting, and stimulant
6	Hedychium flavescens Carey ex Roscoe	Zingiberaceae	RRCBI-MUS.145	Anti-rheumatic, stimulant and antipyretic

Table 2 Colonization and isolation rates of endophytic fungi in six medicinal plants.

Sl. no	Medicinal plant	Tissue segments	Colonization rate CR (%)	Isolation rate (IR)
1	Tinospora cordifolia (Willd.) Hook. f. & Thomson	Leaf	32	0.4
		Petiole	12	0.15
		Stem	36	0.45
		Roots	20	0.25
2	Piper nigrum L.	Leaf	15.38	0.1
		Petiole	15.38	0.1
		Stem	30.76	0.2
		Roots	38.46	0.25
3	Piper longum L.	Leaf	40.74	0.55
		Petiole	22.22	0.3
		Stem	29.62	0.4
		Roots	7.40	0.1
4	Zingiber officinale Roscoe	Leaf	35.71	0.25
		Petiole	14.28	0.1
		Stem	28.57	0.2
		Roots	21.42	0.15
5	Hedychium coronarium J. Koenig	Leaf	40	0.3
		Petiole	26.66	0.2
		Stem	20	0.15
		Roots	13.33	0.1
6	Hedychium flavescens Carey ex Roscoe	Leaf	44.44	0.4
		Petiole	27.77	0.25
		Stem	16.66	0.15
		Roots	11.11	0.1

Table 3 Absolute (f) and relative frequency (fr) of endophytic fungal isolates from selected medicinal plants.

Sl. no	Endophytic fungi	Tinospora cordifolia				Piper nigrum				Piper longum				Zingiber officinale				Hedychium flavescens				Hedychium coronarium				Absolute frequency (f)	Relative frequency fr (%)
		L	P	S	R	L	P	S	R	L	P	S	R	L	P	S	R	L	P	S	R	L	P	S	R
1	Acremonium sp.															1										1	0.892
2	Alternaria sp.													1			1		1	1	1				1	6	5.357
3	Aspergillus sp.	3	1	3				2	2	3	1	3	1			1							2			22	19.642
4	Bipolaris sp.									1								1						1		3	2.678
5	Chaetomium sp.																	1								1	0.892
6	Cladosporium sp.		1	2										2						1						6	5.357
7	Colletotrichum sp.				1			1		1						1		1	1							6	5.357
8	Curvularia sp.			1						1		1			1											4	3.571
9	Cylindrocephalum sp.																							1		1	0.892
10	Dreshclera sp.									1																1	0.892
11	Fusarium sp.	1		1	1							1		1				1		1			1			8	7.142
12	Lasiodiplodia sp.							1																		1	0.892
13	Mucor sp.	1			1					1		1					1		1							6	5.357
14	Mycelia sterilia	1	1	1	1		1				1	1	1				1		1			1				11	9.821
15	Myrothecium sp.																					1				1	0.892
16	Nigrospora sp.																						1	1		2	1.785
17	Paecilomyces sp.					1																				1	0.892
18	Penicillium sp.	1							1	2	1	1										1			1	8	7.142
19	Pestalotiopsis sp.										1					1		1								3	2.678
20	Phoma sp.					1	1																			2	1.785
21	Phomopsis sp.				1						1											1				3	2.678
22	Pithomyces sp.									1								1								2	1.785
23	Rhizopus sp.	1												1	1			1	1		1					6	5.357
24	Sordaria sp.								1		1															2	1.785
25	Torula sp.																	1								1	0.892
26	Trichoderma sp.			1					1													2				4	3.571
	TOTAL	8	3	9	5	2	2	4	5	11	6	8	2	5	2	4	3	8	5	3	2	6	4	3	2	112	99.984
	Species richness	6	3	6	5	2	2	3	4	8	6	6	2	4	2	4	3	8	5	3	2	5	3	3	2

L, Leaf; P, Petiole; S, Stem; R, Root.

Table 4 Diversity indices of endophytic fungi isolated from T. cordifolia, P. nigrum and P. longum.

Sl. No	Indices	Tinospora cordifolia				Piper nigrum				Piper longum
		L	P	S	R	L	P	S	R	L	P	S	R
1	Simpson's dominance	0.218	0.333	0.209	0.200	0.500	0.500	0.375	0.280	0.157	0.166	0.218	0.500
2	Simpson's diversity	0.781	0.666	0.790	0.800	0.500	0.500	0.625	0.720	0.843	0.833	0.781	0.500
3	Species Richness	6	3	6	5	2	2	3	4	8	6	6	2
4	Shannon–Wiener	1.667	1.097	1.676	1.609	0.693	0.693	1.039	1.331	1.971	1.791	1.667	0.693
5	Evenness	0.930	0.998	0.935	0.999	0.999	0.999	0.946	0.960	0.948	0.999	0.930	0.999

L, Leaf; P, Petiole; S, stem; R, root.

Table 5 Diversity indices for endophytic fungi isolated from Z. officinale, H. flavescens and H. coronarium.

Sl. No	Indices	Zingiber officinale				Hedychium flavescens				Hedychium coronarium
		L	P	Rh	Ad. R	L	P	S	R	L	P	S	R
1	Simpson's dominance	0.280	0.500	0.250	0.333	0.125	0.200	0.333	0.500	0.222	0.370	0.333	0.500
2	Simpson's diversity	0.720	0.500	0.750	0.666	0.875	0.800	0.666	0.500	0.777	0.625	0.666	0.500
3	Species Richness	4	2	4	3	8	5	3	2	5	3	3	2
4	Shannon–Wiener	1.331	0.693	1.386	1.098	2.079	1.604	1.096	0.693	1.560	1.039	1.098	0.693
5	Evenness	0.960	0.999	0.999	0.999	0.999	0.999	0.999	0.999	0.969	0.946	0.999	0.999

L, Leaf; P, Petiole; S, stem; R, root; Rh, Rhizome; Ad. R, Adventitious roots.

Table 6 Endophytic fungal isolates screened for extracellular enzymes on solid media.

Sl. No	Medicinal plant	Code	Endophytic fungal isolate	Amylase	Cellulase	Pectinase	Laccase	Asparaginase
1	Tinospora cordifolia (Willd.) Hook. f. & Thomson	Tc2	Fusarium sp.	29.00^d ± 0.51	0^a	0^a	0^a	25.00^b ± 0.59
2		Tc6	Aspergillus sp.	0^a	0^a	0^a	0^a	47.66^cd ± 1.30
3		Tc7	Penicillium sp.	36.33^f ± 0.59	0^a	0^a	0^a	43.66^cd ± 0.78
4		Tc8	Mycelia sterilia	34.33^e ± 0.29	55.33^f ± 1.07	72.33^f ± 0.78	0^a	34.66^bc ± 0.29
5		Tc9	Cladosporium sp.	0^a	15.33^c ± 0.78	69.33^e ± 0.59	0^a	54.00^d ± 0.51
6		Tc10	Aspergillus sp.	18.33^b ± 0.78	21.66^d ± 0.29	41.66^c ± 0.78	0^a	0^a
7		Tc12	Curvularia sp.	0^a	0^a	0^a	0^a	59.00^d ± 0.51
8		Tc15	Trichoderma sp.	0^a	0^a	0^a	0^a	75.00^e ± 0.29
9		Tc16	Cladosporium sp.	38.33^g ± 0.78	13.66^b ± 0.78	41.00^c ± 0.51	0^a	0^a
10		Tc17	Fusarium sp.	0^a	0^a	0^a	0^a	23.66^ab ± 0.78
11		Tc18	Cladosporium sp.	0^a	21.00^d ± 0.51	0^a	0^a
12		Tc19	Aspergillus sp.	0^a	24.00^e ± 0.23	53.00^d ± 0.51	0^a	24.00^b ± 0.78
13		Tc20	Aspergillus sp.	24.66^c ± 0.29	24.00^e ± 0.51	39.33^b ± 0.59	0^a	36.00^bc ± 0.51
14		Tc25	Fusarium sp.	29.00^d ± 0.51	0^a	0^a	0^a	79.66^e ± 0.29
15	Piper nigrum L.	Pn1	Phoma sp.	0^a	0^a	9.00^b ± 0.51	0^a	0^a
16		Pn2	Paecilomyces sp.	0^a	21.6^6b ± 0.29	0^a	0^a	0^a
17		Pn6	Aspergillus sp.	19.33^b ± 0.29	0^a	0^a	0^a	69.66^e ± 0.29
18		Pn7	Lasiodiplodia sp.	69.00^d ± 0.51	0^a	0^a	0^a	65.66^d ± 0.29
19		Pn8	Aspergillus sp.	0^a	0^a	0^a	0^a	34.00^b ± 0.51
20		Pn9	Aspergillus sp.	49.00^c ± 0.51	0^a	0^a	0^a	0^a
21		Pn11	Sordaria sp.	0^a	43.33^d ± 0.29	0^a	0^a	0^a
22		Pn12	Aspergillus sp.	0^a	26.00^c ± 0.51	23.66^c ± 0.29	0^a	69.00^e ± 0.51
23		Pn13	Trichoderma sp.	0^a	0^a	0^a	0^a	64.00^c ± 0.51
24	Piper longum L.	Pl1	Mucor sp.	44.00^i ± 0.51	46.00^f ± 0.51	0^a	0^a	39.00^c ± 0.51
25		Pl2	Penicillium sp.	0^a	49.00^g ± 0.51	0^a	0^a	0^a
26		Pl3	Aspergillus sp.	0^a	32.33^c ± 0.29	0^a	0^a	0^a
27		Pl4	Colletotrichum sp.	0^a	37.00^e ± 0.51	10.66^ab ± 0.29	0^a	0^a
28		Pl5	Penicillium sp.	0^a	34.00^d ± 0.51	0^a	0^a	47.66^gh ± 0.29
29		Pl7	Dreshclera sp.	31.66^f ± 0.29	0^a	0^a	0^a	0^a
30		Pl8	Aspergillus sp.	0^a	49.00^g ± 0.57	0^a	0^a	47.66^gh ± 0.33
31		Pl9	Pithomyces sp.	0^a	0^a	0^a	0^a	39.00c ± 0.57
32		Pl10	Bipolaris sp.	0^a	54.66^i ± 0.33	0^a	0^a	40.33^de ± 0.33
33		Pl11	Curvularia sp.	0^a	0^a	0^a	0^a	39.66^cd ± 0.29
34		Pl12	Fusarium sp.	0^a	52.33^h ± 0.29	0^a	0^a	39.33^cd ± 0.59
35		Pl13	Aspergillus sp.	52.66^j ± 0.29	71.00^k ± 0.51	0^a	0^a	41.00^e ± 0.51
36		Pl14	Curvularia sp.	24.00^bc ± 0.51	0^a	0^a	0^a	0^a
37		Pl15	Aspergillus sp.	0^a	54.66^i ± 0.29	0^a	0^a	45.00^f ± 0.48
38		Pl16	Aspergillus sp.	0^a	0^a	18.33^b ± 0.29	0^a	24.33^b ± 0.63
39		Pl17	Mycelia sterilia	29.33^e ± 0.59	0^a	0^a	0^a	0^a
40		Pl18	Mucor sp.	25.66^d ± 0.29	0^a	0^a	0^a	0^a
41		Pl19	Penicillium sp.	36.00^g ± 0.51	0^a	0^a	0^a	0^a
42		Pl20	Aspergillus sp.	0^a	54.00^i ± 0.51	0^a	0^a	0^a
43		Pl21	Pestalotiopsis sp.	0^a	56.33^j ± 0.29	0^a	0^a	48.00^h ± 1.03
44		Pl22	Mycelia sterilia	39.00^h ± 0.51	0^a	0^a	0^a	0^a
45		Pl24	Phomopsis sp.	29.66^e ± 0.29	21.66^d ± 0.29	61.66^c ± 0.29	0^a	0^a
46		Pl25	Penicillium sp.	0^a	0^a	0^a	0^a	39.66^cd ± 0.29
47		Pl26	Aspergillus sp.	23.33^b ± 0.59	0^a	0^a	0^a	46.66^g ± 0.29
48		Pl27	Mycelia sterilia	24.66^c ± 0.29	0^a	81.00^d ± 0	0^a	0^a
49	Zingiber officinale Roscoe	Zo1	Aspergillus sp.	0^a	0^a	0^a	0^a	32.00^b ± 0.51
50		Zo2	Rhizopus sp.	41.00^d ± 0.51	40.33^c ± 0.29	39.66^d ± 0.29	0^a	0^a
51		Zo3	Cladosporium sp.	53.33^f ± 0.78	0^a	63.33^e ± 0.78	0^a	0^a
52		Zo4	Cladosporium sp.	29.00^c ± 0.51	0^a	0^a	0^a	0^a
53		Zo5	Alternaria sp.	0^a	0^a	0^a	0^a	63.33^d ± 0.29
54		Zo6	Curvularia sp.	14.66^b ± 0.29	0^a	0^a	0^a	0^a
55		Zo7	Aspergillus sp.	0^a	35.33^b ± 0.29	0^a	0^a	72.33^e ± 0.29
56		Zo8	Mycelia sterilia	0^a	0^a	26.66^c ± 0.29	0^a	0^a
57		Zo11	Mycelia sterilia	0^a	0^a	0^a	0^a	41.66^c ± 0.29
58		Zo13	Mucor sp.	42.66^e ± 0.29	0^a	10.33^b ± 0.59	0^a	0^a
59		Zo14	Rhizopus sp.	0^a	0^a	0^a	0^a	72.33^e ± 1.30
60	Hedychium coronarium J. Koenig	Hc2	Trichoderma sp.	0^a	26.66^c ± 0.29	0^a	0^a	0^a
61		Hc3	Mycelia sterilia	0^a	65.00^d ± 0.51	0^a	0^a	23.66^c ± 0.78
62		Hc4	Penicillium sp.	29.00^c ± 0.51	0^a	0^a	0^a	0^a
63		Hc5	Mycelia sterilia	0^a	0^a	0^a	0^a	31.00^e ± 0.51
64		Hc7	Alternaria sp.	11.66^b ± 0.29	0^a	0^a	0^a	0^a
65		Hc8	Penicillium sp.	0^a	0^a	14.00^b ± 0.51	0^a	51.00^g ± 0.51
66		Hc9	Fusarium sp.	0^a	20.66^b ± 0.29	0^a	0^a	0^a
67		Hc11	Aspergillus sp.	0^a	0^a	19.00^c ± 0.51	0^a	15.66^b ± 0.29
68		Hc12	Bipolaris sp.	12.00^b ± 0.51	0^a	0^a	0^a	0^a
69		Hc13	Nigrospora sp.	0^a	0^a	0^a	0^a	24.66^d ± 0.29
70		Hc15	Aspergillus sp.	44.66^d ± 0.29	0^a	0^a	0^a	32.66^f ± 0.29
71	Hedychium flavescens Carey ex Roscoe	Hf3	Colletotrichum sp.	0^a	29.33^c ± 0.29	27.33^c ± 0.29	0^a	0^a
72		Hf4	Fusarium sp.	39.66^c ± 0.29	0^a	0^a	0^a	41.33^d ± 0.29
73		Hf7	Bipolaris sp.	0^a	0^a	0^a	0^a	65.00^e ± 1.03
74		Hf8	Pithomyces sp.	52.66^d ± 0.29	0^a	0^a	0^a	0^a
75		Hf9	Mucor sp.	0^a	0^a	62.66^d ± 0.29	0^a	0^a
76		Hf10	Alternaria sp.	0^a	17.66^b ± 0.29	0^a	0^a	0^a
77		Hf11	Mycelia sterilia	0^a	0^a	0^a	0^a	11.66^b ± 0.29
78		Hf12	Rhizopus sp.	19.66^b ± 0.29	0^a	0^a	0^a	0^a
79		Hf12	Fusarium sp.	0^a	0^a	15.66^b ± 0.29	0^a	23.66^c ± 0.29
80		Hf14	Cladosporium sp.	0^a	42.66^d ± 0.29	0^a	0^a	0^a

Values followed by lower case alphabets in the column are statistically equivalent (P < 0.05) according to the Duncan multiple range test.