Fungal phytotoxins used as ecofriendly bioherbicides are becoming efficient alternatives to chemical herbicides for sustainable weed management. Previous study found that cultures of the pathogenic fungus Colletotrichum gloeosporioides BWH-1 showed phytotoxic activity. This study further isolated the major phytotoxin from cultures of the strain BWH-1 using bioactivity-guided isolation, by puncturing its host plant for an activity test and analyzing on the HPLC-DAD-3D mode for a purity check. Then, the active and pure phytotoxin was characterized as a dirhamnolipid (Rha-Rha-C10-C10) using the NMR, ESIMS, IR and UV methods. The herbicidal activity of dirhamnolipid was evaluated by the inhibition rate on the primary root length and the fresh plant weight of nine test plants, and the synergistic effect when combining with commercial herbicides. Dirhamnolipid exhibited broad herbicidal activity against eight weed species with IC50 values ranging from 28.91 to 217.71 mg/L and no toxicity on Oryza sativa, and the herbicidal activity could be synergistically improved combining dirhamnolipid with commercial herbicides. Thus, dirhamnolipid that originated from C. gloeosporioides BWH-1 displayed the potential to be used as a bioherbicide alone, or as an adjuvant added into commercial herbicides, leading to a decrease in herbicides concentration and increased control efficiency.

rhamnolipid, biosurfactant, secondary metabolite, bioherbicide, phytotoxin, biopesticide, Colletotrichum, synergy

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6	C7	C8	C9	C10
3,3,2	aLRhap	104.32	72.02	72.37	73.96	70.38	18.11
3,3	aLRhap	99.34	80.55	72.02	74.38	70.26	18.16
3	lR3HODco	172.63	41.39	75.55	34.39	?	?	?	?	?	14.52
	lR3HODco	174.50	40.04	72.50	35.14	?	?	?	?	?	14.52


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6	H7	H8	H9	H10
3,3,2	aLRhap	4.90	3.98	3.67-3.76	3.33	3.67-3.76	1.25
3,3	aLRhap	4.94	3.67-3.76	3.67-3.76	3.40	3.67-3.76	1.27
3	lR3HODco	-	2.50-2.59	4.06	1.57	1.33	1.33	1.33	1.33	1.33	0.92
	lR3HODco	-	2.59	5.26	1.64	1.33	1.33	1.33	1.33	1.33	0.92


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6	C7/H7	C8/H8	C9/H9	C10/H10
3,3,2	aLRhap	104.32/4.90	72.02/3.98	72.37/3.67-3.76	73.96/3.33	70.38/3.67-3.76	18.11/1.25
3,3	aLRhap	99.34/4.94	80.55/3.67-3.76	72.02/3.67-3.76	74.38/3.40	70.26/3.67-3.76	18.16/1.27
3	lR3HODco		41.39/2.50-2.59	75.55/4.06	34.39/1.57	?/1.33	?/1.33	?/1.33	?/1.33	?/1.33	14.52/0.92
	lR3HODco		40.04/2.59	72.50/5.26	35.14/1.64	?/1.33	?/1.33	?/1.33	?/1.33	?/1.33	14.52/0.92

The spectrum also has 10 signals at unknown positions (not plotted).

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

CCCCCCC[C@H](CC(=O)O)OC(=O)C[C@@H](CCCCCCC)O[C@@H]1O[C@@H](C)[C@H](O)[C@@H](O)[C@H]1O[C@@H]1O[C@@H](C)[C@H](O)[C@@H](O)[C@H]1O

650.803 g/mol (C₃₂H₅₈O₁₃)