Fusarium solani Upregulated Sesquiterpene Synthase Expression, Sesquiterpene Production and Allelopathic Activity in Piper betle

doi:10.1016/j.rsci.2019.08.003

Abstract

Abstract:

Activators of sesquiterpene synthase (STS) gene expression and sesquiterpene production in Piper betle L. were examined using quantitative real time PCR and gas chromatography mass spectrometry methods, and the allelopathic activity of untreated and Fusarium solani-treated betel extracts was tested on seed germination and on the shoot and root growth of Thai rice variety PSL2 (Oryza sativa cv. Phitsanulok 2) and three dominant paddy weeds (Eclipta prostrata, Echinochloa crus-galli and Chloris barbata). The results demonstrated that F. solani dramatically upregulated STS expression and productions of β-cubebene, β-caryophyllene and germacrene D sesquiterpene when compared with the untreated control, and that betel extracts had a greater inhibitory effect on weeds than on rice. The effects were more clearly detected on seed germination and root growth than on shoot growth, and they were found to be dose-dependent. It is also noted that F. solani-treated extract had stronger effects than the untreated extract. The species most sensitive to the allelopathic effects was C. barbata, germination of which was completely inhibited even at a dose of 0.1 mg/mL untreated extract. With regards to rice, although betel extract at 1.0 mg/mL showed no inhibition on germination, it affected the elongation of rice roots, in addition to those of the tested weeds. The obtained data suggested that F. solani has potential as an activator of sesquiterpene allelochemical production via STS expression, the latter leading to the treated betel extract having a stronger phytotoxic effect. These results were beneficial in the promotion of natural herbicide production using biotechnology.

Key words: Piper betle L., paddy weed, Fusarium solani, sesquiterpene synthase, allelopathic activity

Woranoot Kamrai, Buaruang Rattikarn, Aranyakanon Kodchakorn, Ratanasut Kumrop, Kongbangkerd Anupan, Jannoey Panatda, Nangngam Pranee, Choopayak Chonnanit. Fusarium solani Upregulated Sesquiterpene Synthase Expression, Sesquiterpene Production and Allelopathic Activity in Piper betle[J]. Rice Science, 2019, 26(5): 290-299.

Figures/Tables 6

Fig. 1. Expression analysis by quantitative real-time PCR of STS in P. betle tissue culture following Fungi infection. Bars are SE (n = 3), and the lowercase letters indicate significant difference at the 0.05 level.

Fig. 2. Volatile organic compounds (VOCs) obtained from P. betle tissue culture after 3 d of F. solani treatment by gas chromatography-mass spectrometry. A shows VOCs, including phenylpropanoids and sesquiterpenes, while B shows only sesquiterpene peaks for retention time of 11.4 to13.2 min.

Table 1 Volatile organic compounds in untreated and F. solani-treated leaf tissue culture extracts.

Peak	Retention time (min)	Compound	Property	Relative area (%)		Fold change ^a
Peak	Retention time (min)	Compound	Property	Untreatment	Treatment	Fold change ^a
1	9.75	Chavicol	Phenylpropanoid	1.39	0.93	1.5
2	11.39	Isoeugenol	Phenylpropanoid	8.6	15.25	1.8
3	11.53	α-copaene	Sesquiterpene	0.18	0.51	2.8
4	11.7	β-cubebene	Sesquiterpene	0.21	0.79	3.8
5	12.13	β-caryophyllene	Sesquiterpene	0.78	2.41	3.1
6	12.56	α-humulene	Sesquiterpene	0.21	0.58	2.8
7	12.81	α-amorphene	Sesquiterpene	0.89	2.5	2.8
8	12.9	Germacrene D	Sesquiterpene	3.31	9.46	2.9
9	13.09	Bicyclogermacrene	Sesquiterpene	0.9	2.41	2.7
10	13.35	Eugenol acetate	Phenylpropanoid	4.5	7.94	1.8
11	14.73	Allylpyrocatechol-3,4-diacetate	Phenylpropanoid	17.43	14.73	1.2

Fig. 3. Effects of F. solani-treated extracts of P. betle on the germination of O. sativa (A), E. prostrata (B), C. barbata (C) and E. crus-galli (D). T1, 0.5% Tween 80 (control); T2, 0.1 mg/mL ethyl acetate extracts; T3, 0.5 mg/mL ethyl acetate extracts; T4, 1.0 mg/mL ethyl acetate extracts.

Fig. 4. Effects of untreated and F. solani-treated extracts of P. betle on the root length of O. sativa (A), E. prostrata (B), C. barbata (C) and E. crus-galli (D). T1, 0.5% Tween 80 (control); T2, 0.1 mg/mL ethyl acetate extracts; T3, 0.5 mg/mL ethyl acetate extracts; T4, 1.0 mg/mL ethyl acetate extracts.

Fig. 5. Effects of untreated and F. solani-treated extracts of P. betle on the shoot length of O. sativa (A), E. prostrata (B), C. barbata (C) and E. crus-galli (D). T1, 0.5% Tween 80 (control); T2, 0.1 mg/mL ethyl acetate extracts; T3, 0.5 mg/mL ethyl acetate extracts; T4, 1.0 mg/mL ethyl acetate extracts.Bars are SE (n = 3) and the lowercase letters indicate significant difference at the 0.05 level.

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